Soren Alexandersen

Evolutionary analysis of human parechovirus type 3 and clinical outcomes of infection during the 2017–18 Australian epidemic

Publisher: Springer Science and Business Media LLC

Date: 20-06-2019

DOI: 10.1038/S41598-019-45445-Z

Abstract: Human parechovirus type 3 (HPeV3) can cause severe sepsis-like illness in young infants and may be associated with long term neurodevelopmental delay later in childhood. We investigated the molecular epidemiology of HPeV infection in thirty three infants requiring hospitalization before, during and after the peak of the 2017/18 HPeV epidemic wave in Australia. During the peak of the epidemic, all cases were infected with an HPeV3, while before and after the peak, HPeV1 was the predominant type detected. The predominant HPeV3 was the recombinant HPeV3 also detected in the 2013/14 and 2015/16 Australian epidemics. Sepsis-like or meningitis-like symptoms were only reported in cases infected with the recombinant HPeV3. Phylogenetic analysis of the recombinant HPeV3 revealed that the virus continued to evolve, also between the Australian outbreaks, thus indicating continued circulation, despite not being detected and reported in Australia or elsewhere in between epidemic waves. The recombinant HPeV3 continued to show a remarkable stability in its capsid amino acid sequence, further strengthening our previous argument for development of a vaccine or immunotherapeutics to reduce the severity of HPeV3 outbreaks due to this virus.

Experimental Foot-and-Mouth Disease Virus Infection in White Tailed Deer

Publisher: Elsevier BV

Date: 08-2012

DOI: 10.1016/J.JCPA.2012.01.010

Abstract: White tailed deer (Odocoileus virginianus) were inoculated with foot-and-mouth disease virus (FMDV) O UKG 11/2001 and monitored for the development of clinical signs, histopathological changes and levels of virus replication. All FMDV-infected deer developed clinical signs starting at 2 days post inoculation and characterized by an increase in body temperature, increased salivation and lesions in the mouth and on the feet. Virus spread to various tissues was determined by quantifying the amount of FMDV RNA using quantitative reverse transcriptase polymerase chain reaction. Virus or viral antigen was also detected in tissues using traditional isolation techniques, enzyme linked immunosorbent assay and immunohistochemistry. Deer-to-cattle transmission of the virus was observed in this experimental setting however, inoculated deer were not found to become carriers of FMDV.

35. Further studies to quantify the dose of natural aerosols of foot-and-mouth disease virus for pigs

Publisher: Elsevier BV

Date: 04-2002

DOI: 10.1016/S0034-5288(02)90037-8

An outbreak of severe infections among Australian infants caused by a novel recombinant strain of human parechovirus type 3.

Publisher: Springer Science and Business Media LLC

Date: 14-03-2017

DOI: 10.1038/SREP44423

Abstract: Human parechovirus types 1–16 (HPeV1–16) are positive strand RNA viruses in the family Picornaviridae . We investigated a 2015 outbreak of HPeV3 causing illness in infants in Victoria, Australia. Virus genome was extracted from clinical material and isolates and sequenced using a combination of next generation and Sanger sequencing. The HPeV3 outbreak genome was 98.7% similar to the HPeV3 Yamagata 2011 lineage for the region encoding the structural proteins up to nucleotide position 3115, but downstream of that the genome varied from known HPeV sequences with a similarity of 85% or less. Analysis indicated that recombination had occurred, may have involved multiple types of HPeV and that the recombination event/s occurred between March 2012 and November 2013. However the origin of the genome downstream of the recombination site is unknown. Overall, the capsid of this virus is highly conserved, but recombination provided a different non-structural protein coding region that may convey an evolutionary advantage. The indication that the capsid encoding region is highly conserved at the amino acid level may be helpful in directing energy towards the development of a preventive vaccine for expecting mothers or antibody treatment of young infants with severe disease.

Foot-and-mouth disease virus infection in young lambs: Pathogenesis and tissue tropism

Publisher: Elsevier BV

Date: 18-03-2008

DOI: 10.1016/J.VETMIC.2007.08.029

Abstract: Foot-and-mouth disease (FMD) in adult sheep usually causes milder clinical signs than in cattle or pigs, and is often subtle enough to go undiagnosed. In contrast, FMD in lambs has been reported to cause high mortality during field outbreaks. In order to investigate the pathogenesis of FMD in lambs, two groups, aged 10-14 days, were infected with foot-and-mouth disease virus (FMDV) type O UKG. One group of lambs (n=8) was inoculated with FMDV in the coronary band, while the other (n=4) was infected by direct contact with FMDV-inoculated ewes. Daily serum s les and temperature measurements were taken. Lambs were killed sequentially and tissue s les taken for analysis. Using real-time RT-PCR, viral RNA levels in tissue s les and serum were measured, and a novel strand-specific real-time RT-PCR assay was used to quantify viral replication levels in tissues. Tissue sections were examined for histopathological lesions, and in situ hybridisation (ISH) was used to localise viral RNA within histological sections. The contact-infected lambs became infected approximately 24h after the ewes were inoculated. Vesicular lesions developed on the feet of all lambs and on the caudo-lateral part of the tongues of six of the eight inoculated lambs and three of the four contact-infected lambs. Although no lambs developed severe clinical signs, one of the contact-infected lambs died acutely at 5 days post-exposure. Histological examination of the heart from this lamb showed multi-focal areas of lymphocytic-plasmacytic myocarditis similar lesions were also observed in the hearts of three of the inoculated lambs. Using ISH, viral RNA was localised within cardiac and skeletal muscle cells from the lamb which had died, and also from vesicular lesions on the coronary band and tongue of inoculated lambs. These results provide a detailed description of the pathogenesis of the disease in lambs.

Investigation of the pathogenesis of transplacental transmission of Aleutian mink disease parvovirus in experimentally infected mink

Publisher: American Society for Microbiology

Date: 03-1996

DOI: 10.1128/JVI.70.3.1455-1466.1996

Abstract: The transplacental transmission of Aleutian mink disease parvovirus (ADV) was studied in experimental infection of 1-year-old female non-Aleutian mink. The ADV-seronegative female mink were inoculated with ADV prior to mating or after the expected implantation of the embryos during pregnancy. A group of uninfected females served as a control group. Animals from each group were killed prior to or shortly after parturition. The in situ hybridization technique with radiolabeled strand-specific RNA probes was used to determine target cells of virus infection and virus replication. In both infected groups, ADV crossed the endotheliochorial placental barrier, although animals infected before mating already had high antibody titers against ADV at the time of implantation. The percentage of dead and resorbed fetuses was much higher in dams infected before mating. In the placentae of these mink, virus DNA and viral mRNA were detected in cells in the mesenchymal stroma of the placental labyrinth and hematoma but only occasionally in the cytotrophoblast of the placental hematoma. Placentae of animals infected during pregnancy showed in addition very high levels of virus and also viral replication in a large number of cytotrophoblast cells in the placental hematoma, which exhibited distinct inclusion bodies. In both groups, neither virus nor virus replication could be detected in maternal endothelial cells or fetal syncytiotrophoblast of the placental labyrinth. Fetuses were positive for virus and viral replication at high levels in a wide range of tissues. Possible routes of transplacental transmission of ADV and the role of trophoblast cells as targets for viral replication are discussed.

Acute Interstitial Pneumonia in Mink Kits Inoculated with Defined Isolates of Aleutian Mink Disease Parvovirus

Publisher: SAGE Publications

Date: 03-1994

DOI: 10.1177/030098589403100209

Abstract: The present study addressed the causal role of Aleutian mink disease parvovirus (ADV) in acute interstitial pneumonia in mink kits. All the examined isolates of ADV caused interstitial pneumonia in newborn kits, although the severity of disease and the mortality varied. These findings indicate that ADV is the direct causal agent of this disease in mink kits and that cofactors, which could have been present in the original ADV-K isolate, do not play a role. Acute interstitial pneumonia characterized by hypertrophy and hyperplasia of alveolar type II cells, intranuclear viral inclusions, interstitial edema, and hyaline membrane formation was experimentally reproduced in mink kits infected as newborns with five different isolates of ADV. Four hundred forty-nine newborn mink kits were included in the study, of which 247 were necropsied. The lesions caused by the different isolates were indistinguishable by histopathologic examination, but the incidence (50–100%) and severity (mortality of 30–100%. n = 218) of disease among the mink kits varied. Also, the content of ADV antigens in the lungs of infected kits varied among the groups. According to these features, the examined isolates could be placed in groups of high and low virulence. ADV-K, ADV-Utah I, and ADV-DK were in a highly virulent group producing a mortality of 90–100% ( n = 110) in mink inoculated as newborns. ADV-GL and ADV-Pullman belonged to a group of low virulence, with an incidence of clinical disease of 50–70% and a mortality of approximately 30–50% ( n = 118) in kits inoculated as newborns. The mortality in the control group receiving a mock inoculum was around 12% ( n = 34). The period from infection to development of fatal disease varied from approximately 12 days for the highly virulent isolates up to around 20 days for the isolates of low virulence. The 107 mink kits that survived inoculation with ADV as newborns developed lesions typical of classical Aleutian disease irrespective of the ADV isolate used. The lesions consisted of chronic immune complexmediated glomerulonephritis and infiltrations with mononuclear cells, including plasma cells in lung, liver, spleen, kidney, mesenteric lymph node, and intestine. Surviving kits also had hypertrophy of the bronchusassociated lymphoid tissue and focal subpleural, intraalveolar accumulations of large cells with foamy cytoplasm, so-called lipid pneumonia.

Differences in the susceptibility of dromedary and Bactrian camels to foot-and-mouth disease virus

Publisher: Cambridge University Press (CUP)

Date: 08-08-2009

DOI: 10.1017/S0950268808001088

Abstract: In this study, two sheep, eight dromedary camels and two Bactrian camels were inoculated with foot-and-mouth disease virus (FMDV) type A SAU 22/92. Five naive dromedary camels and four sheep were kept in direct or indirect contact with the inoculated camels. The inoculated sheep, which served as positive controls, displayed typical moderate clinical signs of FMD and developed viraemia and high antibody titres. The presence of the virus was also detected in probang and mouth-swab s les for several days after inoculation. In contrast, the inoculated dromedary camels were not susceptible to FMDV type A infection. None of them showed clinical signs of FMD or developed viraemia or specific anti-FMDV antibodies despite the high dose of virus inoculated. All the contact sheep and contact dromedaries that were kept together with the inoculated camels remained virus-negative and did not seroconvert when tested up to 28 days post-inoculation (p.i.). In comparison with the non-susceptible dromedaries, the two inoculated Bactrian camels showed moderate to severe clinical signs of FMD however, the clinical signs of FMD appeared rather late, between 8 and 14 days p.i., compared to the inoculated sheep. Characteristic FMD lesions in the Bactrian camels, accompanied with severe lameness, were only observed on the hind feet. The presence of the virus in the serum s les of both Bactrian camels was detected by real-time RT–PCR in one of the animals on days 3 and 7 p.i. and in the second animal from days 1 to 3 p.i. and subsequently again on day 21 p.i. The Bactrian camels developed high titres of antibodies to the inoculated FMDV which appeared at 7–10 days p.i. and lasted up to 130 days p.i. Only low and transient amounts of FMDV were detected in the mouth-swab and probang s les collected from both Bactrian camels.

Serological and genetic characterisation of bovine respiratory syncytial virus (BRSV) indicates that Danish isolates belong to the intermediate subgroup: No evidence of a selective effect on the variability of G protein nucleotide sequence by prior cell culture adaption and passages in cell culture or calves

Publisher: Elsevier BV

Date: 08-1998

DOI: 10.1016/S0378-1135(98)00226-0

Abstract: Danish isolates of bovine respiratory syncytial virus (BRSV) were characterised by nucleotide sequencing of the G glycoprotein and by their reactivity with a panel of monoclonal antibodies (MAbs). Among the six Danish isolates, the overall sequence ergence ranged between 0 and 3% at the nucleotide level and between 0 and 5% at the amino acid level. Sequence ergences of 7-8%, 8-9% and 2-3% (nucleotide) and 9-11%, 12-16% and 4-6% (amino acid) were obtained in the comparison made between the group of Danish isolates and the previously sequenced 391-2USA, 127UK and 220-69Bel isolates, respectively. Phylogenetic analysis showed that the Danish isolates formed three lineages within a separate branch of the phylogenetic tree. Nevertheless, the Danish isolates were closely related to the 220-69Bel isolate, the prototype of the intermediate antigenic subgroup. The sequencing of the extracellular part of the G gene of additional 11 field BRSV viruses, processed directly from lung s les without prior adaption to cell culture growth, revealed sequence variabilities in the range obtained with the propagated virus. In addition, several passages in cell culture and in calves had no major impact on the nucleotide sequence of the G protein. These findings indicated that the previously established variabilities of the G protein of RS virus isolates were not attributable to mutations induced during the propagation of the virus. The reactivity of the Danish isolates with G protein-specific MAbs were similar to that of the 220-69Bel isolate. Furthermore, the sequence of the immunodominant region was completely conserved among the Danish isolates on one side and the 220-69Bel isolate on the other. When combined, these data strongly suggested that the Danish isolates belong to the intermediate subgroup.

HPeV3 Diversity, Recombination and Clinical Impact Across 7-years: an Australian Story

Publisher: Oxford University Press (OUP)

Date: 22-07-2022

DOI: 10.1093/INFDIS/JIAC311

Abstract: A novel human parechovirus 3 Australian recombinant (HPeV3-AR) strain emerged in 2013 and coincided with biennial outbreaks of sepsis-like illnesses in infants. We evaluated the molecular evolution of the HPeV3-AR strain and its association with severe HPeV infections. HPeV3-positive s les collected from hospitalized infants aged 5–252 days in 2 Australian states (2013–2020) and from a community-based birth cohort (2010–2014) were sequenced. Coding regions were used to conduct phylogenetic and evolutionary analyses. A recombinant-specific polymerase chain reaction was designed and utilized to screen all clinical and community HPeV3-positive s les. Complete coding regions of 54 cases were obtained, which showed the HPeV3-AR strain progressively evolving, particularly in the 3′ end of the nonstructural genes. The HPeV3-AR strain was not detected in the community birth cohort until the initial outbreak in late 2013. High-throughput screening showed that most (& %) hospitalized HPeV3 cases involved the AR strain in the first 3 clinical outbreaks, with declining prevalence in the 2019–2020 season. The AR strain was not statistically associated with increased clinical severity among hospitalized infants. HPeV3-AR was the dominant strain during the study period. Increased hospital admissions may have been from a temporary fitness advantage and/or increased virulence.

Rocket line immunoelectrophoresis: An improved assay for simultaneous quantification of a mink parvovirus (aleutian disease virus) antigen and antibody

Publisher: Elsevier BV

Date: 02-1985

DOI: 10.1016/0166-0934(85)90100-4

Abstract: A rocket line immunoelectrophoretic assay (RLIE) was developed for the simultaneous quantification of viral antigens and antiviral antibodies of the important mink parvovirus, Aleutian disease virus (ADV). The sensitivity of the RLIE assay was found to be 5 log2 higher than that of the counter current immunoelectrophoresis which is the assay routinely used for diagnostic purposes.

123. Quantitation of the expression of antiviral and proinflammatory cytokine mRNA in tissues from FMDV infected pigs

Publisher: Elsevier BV

Date: 2003

DOI: 10.1016/S0034-5288(03)90122-6

Comparative subgenomic mRNA profiles of SARS-CoV-2 Alpha, Delta and Omicron BA.1, BA.2 and BA.5 sub-lineages using Danish COVID-19 genomic surveillance data

Publisher: Elsevier BV

Date: 07-2023

DOI: 10.1016/J.EBIOM.2023.104669

31. Evaluation of automated RT-PCR systems to accelerate FMD diagnosis

Publisher: Elsevier BV

Date: 04-2002

DOI: 10.1016/S0034-5288(02)90033-0

Subgenomic and negative sense RNAs are not markers of active replication of SARS-CoV-2 in nasopharyngeal swabs

Publisher: Cold Spring Harbor Laboratory

Date: 03-07-2021

DOI: 10.1101/2021.06.29.21259511

Abstract: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has spread rapidly in the global population since its emergence in humans in late 2019. Replication of SARS-CoV-2 is characterised by transcription and replication of genomic length RNA and shorter subgenomic RNAs to produce virus proteins and ultimately progeny virions. Here we explore the pattern of both genome-length and subgenomic RNAs and positive and negative strand SARS-CoV-2 RNAs in diagnostic nasopharyngeal swabs using sensitive probe based PCR assays as well as Ampliseq panels designed to target subgenomic RNAs. We successfully developed a multiplex PCR assay to simultaneously measure the relative amount of SARS-CoV-2 full length genomic RNA as well as subgenomic N gene and subgenomic ORF7a RNA. We found that subgenomic RNAs and both positive and negative strand RNA can be readily detected in swab s les taken up to 19 and 17 days post symptom onset respectively, and are strongly correlated with the amount of genomic length RNA present within a s le. Their detection and measurement is therefore unlikely to provide anymore insight into the stage of infection and potential infectivity of an in idual beyond what can already be inferred from the total viral RNA load measured by routine diagnostic SARS-CoV-2 PCRs. Using both an original commercial and two custom SARS-CoV-2 Ampliseq mini-panels, we identified that both ORF7a and N gene subgenomic RNAs were consistently the most abundant subgenomic RNAs. We were also able to identify several non-canonical subgenomic RNAs, including one which could potentially be used to translate the ORF7b protein and others which could be used to translate ORF9b and the ORF N* which has arisen from a new transcription regulatory sequence recently created by mutations after SARS-CoV-2 jumped into people. SARS-CoV-2 genomic length and subgenomic length RNA’s were present in s les even if cellular RNA was degraded, further indicating that these molecules are likely protected from degradation by the membrane structures seen in SARS-CoV-2 infected cells.

Further studies to quantify the dose of natural aerosols of foot-and-mouth disease virus for pigs

Publisher: Cambridge University Press (CUP)

Date: 04-2002

DOI: 10.1017/S0950268801006501

Abstract: Foot-and-mouth disease virus (FMDV) can be spread by a variety of mechanisms, including wind. Simulation models, developed to predict the risk of airborne spread, have played an important part in decision making in some outbreaks. The amount of airborne virus excreted as well as the minimal infectious dose (MID) of FMDV for different species are important determinants of airborne spread. The objective of this study was to obtain data for the O 1 Lausanne, O SKR 2000 and O UKG 2001 strains of FMDV to enhance the capability of such models. Pigs were exposed to naturally generated aerosols of the three strains using an experimental design which delivered high doses of the two strains O 1 Lausanne and O SKR 2000 over a short period, or of the O UKG 2001 strain over an extended period. The average excretion of the O 1 Lausanne strain was 10 6·4 TCID 50 per pig per hour. The excretion of the O SKR 2000 strain averaged 10 5·8 and the O UKG 2001 strain 10 6·1 TCID per pig per 24 h. The results show that the previous estimate of ‘above’ 800 TCID 50 as the MID50 for the O 1 Lausanne strain is a considerable under-estimate and that the real dose may be as high as 6000 TCID 50 . A dose of around 650 TCID 50 of the O SKR 2000 strain failed to infect any pigs. Thus, the aerosol MID 50 for pigs for this isolate is at least 1000 TCID 50 and likely to be as high or higher than the O 1 Lausanne strain. The exposure of pairs of recipient pigs kept physically separated from donor pigs in a series of rooms to aerosol exposure doses of the O UKG 2001 strain of around 50 TCID 50 per min for 24–48 h failed to infect any of eight pigs. Thus, the present experiment confirms our previous findings [1, 2] that pigs, compared to cattle and sheep, are relatively resistant to infection with airborne FMDV.

Appearance of acute PRRS-like symptoms in sow herds after vaccination with a modified live PRRS vaccine.

Publisher: Wiley

Date: 11-1997

DOI: 10.1136/VR.141.19.497

Abstract: Esophagectomy is the gold standard in the surgical therapy of esophageal cancer. It is either performed thoracoabdominal with a intrathoracic anastomosis or in proximal cancers with a three-incision esophagectomy and cervical reconstruction. Delayed gastric conduit emptying (DGCE) is the most common functional postoperative disorder after Ivor-Lewis esophagectomy (IL). Pneumonia is significantly more often in patients with DGCE. It remains unclear if DGCE anastomotic leakage (AL) is associated. Aim of our study is to analyze, if AL is more likely to happen in patients with a DGCE. 816 patients were included. All patients have had an IL due to esophageal/esophagogastric-junction cancer between 2013 and 2018 in our center. Intrathoracic esophagogastric end-to-side anastomosis was performed with a circular stapling device. The collective has been ided in two groups depending on the occurrence of DGCE. The diagnosis DGCE was determined by clinical and radiologic criteria in accordance with current international expert consensus. 27.7% of all patients suffered from DGCE postoperatively. Female patients had a significantly higher chance to suffer from DGCE than male patients (34.4% vs. 26.2% vs., p = 0.040). Pneumonia was more common in patients with DGCE (13.7% vs. 8.5%, p = 0.025), furthermore hospitalization was longer in DGCE patients (median 17 days vs. 14d, p < 0.001). There was no difference in the rate of type II anastomotic leakage, (5.8% in both groups DGCE). All patients with ECCG type II AL (n = 47 5.8%) were treated successfully by endoluminal/endoscopic therapy. The subgroup analysis showed that ASA ≥ III (7.6% vs. 4.4%, p = 0.05) and the histology squamous cell carcinoma (9.8% vs. 4.7%, p = 0.01) were independent risk factors for the occurrence of an AL. Our study confirms that DGCE after IL is a common finding in a standardized collective of patients in a high-volume center. This functional disorder is associated with a higher rate of pneumonia and a prolonged hospital stay. Still, there is no association between DGCE and the occurrence of an AL after esophagectomy. The hypothesis, that an DGCE results in a higher pressure on the anastomosis and therefore to an AL in consequence, can be refuted. DGCE is not a pathogenetic factor for an AL.

Predicting the spread of foot and mouth disease by airborne virus

Publisher: O.I.E (World Organisation for Animal Health)

Date: 12-2002

DOI: 10.20506/RST.21.3.1362

Abstract: Foot and mouth disease (FMD) can spread by a variety of mechanisms which, under certain climatic and epidemiological conditions, includes the windborne spread of disease. Recent advances in knowledge of the aerobiological features of FMD are described. The strain of virus and species of infected animal are major determinants of airborne virus emission. Pigs emit most virus, cattle and sheep lesser but similar amounts to each other. Peak excretion of airborne virus by sheep occurs before the clinical phase of disease, whereas with cattle and pigs, it coincides with the development of early clinical disease. The probability of aerogenous infection differs greatly between livestock species. Cattle are the most susceptible, followed by sheep, whereas pigs are very resistant. Computer-based simulation models have been developed to analyse and predict the risk of airborne spread of FMD and have been used successfully during outbreaks to support decision-making. Further research is required to refine and extend the models for operational use.

Investigation of airborne foot-and-mouth disease virus transmission during low-wind conditions in the early phase of the UK 2001 epidemic

Publisher: Copernicus GmbH

Date: 28-11-2003

DOI: 10.5194/ACP-3-2101-2003

Abstract: Abstract. Foot-and-mouth disease (FMD) is a highly contagious viral disease of cloven-hoofed domesticated and wild animals. The highly contagious nature of FMD is a reflection of the wide range of host species, the enormous quantities of virus liberated by infected animals, the range of excretions and secretions which can be infectious, the stability of the virus in the environment, the multiplicity of routes of infection and the very small doses of the virus that can initiate infection. One of the mechanisms of spread is the carriage of droplets and droplet nuclei exhaled in the breath of infected animals. Such spread can be rapid and extensive, and it is known in certain circumstances to have transmitted disease over a distance of several hundred kilometres. During the 2001 FMD epidemic in the United Kingdom (UK), atmospheric dispersion models were applied in real time in order to assess the potential for atmospheric dispersion of the disease. The operational value of such modelling is primarily to identify premises which may have been exposed so that the human resources for surveillance and disease control purposes are employed most effectively. The paper describes the combined modelling techniques and presents the results obtained of detailed analyses performed during the early stages of the UK 2001 epidemic. This paper investigates the potential for disease spread in relation to two outbreaks (Burnside Farm, Heddon-on-the-Wall and Prestwick Hall Farm, Ponteland, Northumberland). A separate paper (Gloster et al., 2002) provides a more detailed analysis of the airborne disease transmission in the vicinity of Burnside Farm. The combined results are consistent with airborne transmission of disease to livestock in the Heddon-on-the-Wall area. Local topography may have played a significant role in influencing the pattern of disease spread.

Investigation into the Role of Potentially Contaminated Feed as a Source of the First-Detected Outbreaks of Porcine Epidemic Diarrhea in Canada

Publisher: Hindawi Limited

Date: 07-08-2014

DOI: 10.1111/TBED.12269

Molecular cloning and phylogenetic analysis of integrins αvβ1 and αvβ6 of one-humped camel (Camelus dromedarius)

Publisher: Elsevier BV

Date: 05-2010

DOI: 10.1016/J.VETIMM.2009.11.008

Abstract: Bactrian camels can relatively easily be infected with FMDV, but dromedary camels remain resistant even to high doses of the virus. To understand the different susceptibility between the two camel species from the standpoint of viral receptors, this work reports the sequences of the dromedary camel integrin cDNAs encoding alphavbeta1 and alphavbeta6 and compare them to those of other species, especially to Bactrian camels. The complete coding sequences for the dromedary camel alphav, beta1 and beta6 subunits were found to be 3147, 2397, and 2364 nucleotides in length, encoding 1048, 798, and 787 amino acids, respectively. The dromedary camel integrin alphav, beta1, and beta6 subunit shares common structural and functional elements with their counterparts from the other species. Phylogenetic trees showed that the dromedary camel alphav, beta1, and beta6 were clustered into the Artiodactyla group, together with those of Bactrian camel, pig, sheep, and cattle that are susceptible to FMDV infection. Compared with the Bactrian camel integrins, 4, 10, and 8 amino acid changes were found in the dromedary camel alphav, beta1, and beta6 subunits, respectively. This study will be of importance in understanding the differences of integrins as FMDV receptors among dromedary camel and other species.

Importance of arginine 20 of the swine vesicular disease virus 2A protease for activity and virulence

Publisher: American Society for Microbiology

Date: 2005

DOI: 10.1128/JVI.79.1.428-440.2005

Abstract: A major virulence determinant of swine vesicular disease virus (SVDV), an Enterovirus that causes an acute vesicular disease, has been mapped to residue 20 of the 2A protease. The SVDV 2A protease cleaves the 1D-2A junction in the viral polyprotein, induces cleavage of translation initiation factor eIF4GI, and stimulates the activity of enterovirus internal ribosome entry sites (IRESs). The 2A protease from an attenuated strain of SVDV (Ile at residue 20) is significantly defective at inducing cleavage of eIF4GI and the activation of IRES-dependent translation compared to the 2A protease from a pathogenic strain (J1/73, Arg at residue 20), but the two proteases have similar 1D-2A cleavage activities (Y. Sakoda, N. Ross-Smith, T. Inoue, and G. J. Belsham, J. Virol. 75:10643-10650, 2001). Residue 20 has now been modified to every possible amino acid, and the activities of each mutant 2A protease has been analyzed. Selected mutants were reconstructed into full-length SVDV cDNA, and viruses were rescued. The rate of virus growth in cultured swine kidney cells reflected the efficiency of 2A protease activity. In experimentally infected pigs, all four of the mutant viruses tested displayed much-reduced virulence compared to the J1/73 virus but a significant, albeit reduced, level of viral replication and excretion was detected. Direct sequencing of cDNA derived from s les taken early and late in infection indicated that a gradual selection-reversion to a more efficient protease occurred. The data indicated that extensive sequence change and selection may introduce a severe bottleneck in virus replication, leading to a decreased viral load and reduced or no clinical disease.

Effect of the initial dose of foot-and-mouth disease virus on the early viral dynamics within pigs

Publisher: The Royal Society

Date: 19-11-2009

DOI: 10.1098/RSIF.2008.0434

Abstract: This paper investigates the early viral dynamics of foot-and-mouth disease (FMD) within infected pigs. Using an existing within-host model, we investigate whether in idual variation can be explained by the effect of the initial dose of FMD virus. To do this, we consider the experimental data on the concentration of FMD virus genomes in the blood (viral load). In this experiment, 12 pigs were inoculated with one of three different doses of FMD virus: low medium or high. Measurements of the viral load were recorded over a time course of approximately 11 days for every 8 hours. The model is a set of deterministic differential equations with the following variables: viral load virus in the interstitial space and the proportion of epithelial cells available for infection, infected and uninfected. The model was fitted to the data for each animal in idually and also simultaneously over all animals varying only the initial dose. We show that the general trend in the data can be explained by varying only the initial dose. The higher the initial dose the earlier the development of a detectable viral load.

In situ hybridization for the detection of chicken anaemia virus in experimentally-induced infection and field outbreaks.

Publisher: Informa UK Limited

Date: 03-1995

DOI: 10.1080/03079459508419055

Abstract: A double stranded DNA probe, biotinylated by random priming, was used for in situ hybridization of chicken anaemia virus (CAV) in thymus tissue. Hybridization was visualized with an alkaline phosphatase conjugated streptavidin biotin complex and the substrate New Fuchsin. The test was performed on both uninfected and CAV-, adenovirus- or infectious bursal disease virus-infected specified pathogen free White Leghorn chickens. In addition, both healthy commercial broilers and broilers with blue wing disease or with colisepticemia, were tested. Genomic CAV was detected in all CAV-infected and blue wing-diseased chickens. With the exception of one chicken with colisepticemia, which showed a unexpectedly positive reaction, all control chickens tested negative in the test developed.

Patterns, risk factors and characteristics of reported and perceived foot-and-mouth disease (FMD) in Uganda

Publisher: Springer Science and Business Media LLC

Date: 06-06-2010

DOI: 10.1007/S11250-010-9605-3

Abstract: Patterns of outbreaks of foot-and-mouth disease (FMD) in Uganda were elucidated from spatial and temporal retrospective data retrieved from monthly reports from District Veterinary Officers (DVOs) to the central administration for the years spanning 2001-2008. An assessment of perceived FMD occurrence, risk factors and the associated characteristics was made based on semi-structured questionnaires administered to the DVOs. During this period, a total of 311 FMD outbreaks were reported in 56 (70%) out of Uganda's 80 districts. The number of reported FMD outbreaks changed over time and by geographical regions. Occurrence of FMD was significantly associated with the dry season months (p = 0.0346), the time when animals movements are more frequent. The average number of FMD outbreaks was higher for some sub-counties adjacent to national parks than for other sub-counties, whilst proximity to international border only seemed to play a role at the southern border. DVOs believed that the major risk factor for FMD outbreaks was animal movements (odds ratio OR 50.8, confidence interval CI 17.8-144.6) and that most outbreaks were caused by introduction of sick animals.

Comparative subgenomic mRNA profiles of SARS-CoV-2 Alpha, Delta and Omicron BA.1, BA.2 and BA.5 sub-lineages using Danish COVID-19 genomic surveillance data

Publisher: Research Square Platform LLC

Date: 03-02-2023

DOI: 10.21203/RS.3.RS-2542939/V1

Abstract: Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has rapidly spread worldwide in the population since it was first detected in late 2019. The transcription and replication of coronaviruses, although not fully understood, is characterised by the production of genomic length RNA and shorter subgenomic RNAs to make viral proteins and ultimately progeny virions. Observed levels of subgenomic RNAs differ between sub-lineages and open reading frames but their biological significance is presently unclear. Using a large and erse panel of virus sequencing data produced as part of the Danish COVID-19 routine surveillance together with information in electronic health registries, we assessed the association of subgenomic RNA levels with demographic and clinical variables of the infected in iduals. Our findings suggest no causative relationships between levels of subgenomic RNAs and host-related factors. Differences between lineages and subgenomic ORFs may be related to differences in target cell tropism, early virus replication/transcription kinetics or sequence features.

Detection of all seven serotypes of foot-and-mouth disease virus by real-time, fluorogenic reverse transcription polymerase chain reaction assay

Publisher: Elsevier BV

Date: 08-2002

DOI: 10.1016/S0166-0934(02)00081-2

Abstract: A fluorogenic RT-PCR (5'-nuclease probe-based) assay using a primer robe set designed from the internal ribosomal entry site region of the virus genome was developed for the specific detection of all seven serotypes of foot-and-mouth disease (FMD) virus in epithelial suspensions and cell culture virus preparations. The reverse transcription polymerase chain reaction (RT-PCR) specifically detected FMD virus in s le submissions from the UK 2001 FMD outbreak with greater sensitivity than our conventional RT-PCR procedure and our routine diagnostic procedures of ELISA and virus isolation in cell culture. The fluorogenic RT-PCR provides relatively fast results, enables a quantitative assessment to be made of virus amounts and can handle more s les and/or replicates of s les in a single assay than the conventional RT-PCR procedure. Therefore it is seen as a valuable tool to complement the routine diagnostic procedures for FMD virus diagnosis.

Thin‐layer counter current line absorption immuno‐electrophoretic analysis of antigens and antibodies to aleutian disease virus ‐ a mink parvovirus

Publisher: Wiley

Date: 1985

DOI: 10.1002/ELPS.1150061104

Occurrence and significance of Omicron BA.1 infection followed by BA.2 reinfection

Publisher: Cold Spring Harbor Laboratory

Date: 22-02-2022

DOI: 10.1101/2022.02.19.22271112

Abstract: The newly found Omicron SARS-CoV-2 variant of concern has rapidly spread worldwide. Omicron carries numerous mutations in key regions and is associated with increased transmissibility and immune escape. The variant has recently been ided into four subvariants with substantial genomic differences, in particular between Omicron BA.1 and BA.2. With the surge of Omicron subvariants BA.1 and BA.2, a large number of reinfections from earlier cases has been observed, raising the question of whether BA.2 specifically can escape the natural immunity acquired shortly after a BA.1 infection. To investigate this, we selected a subset of s les from more than 1,8 million cases of infections in the period from November 22, 2021, until February 11, 2022. Here, in iduals with two positive s les, more than 20 and less than 60 days apart, were selected. From a total of 187 reinfection cases, we identified 47 instances of BA.2 reinfections shortly after a BA.1 infection, mostly in young unvaccinated in iduals with mild disease not resulting in hospitalization or death. In conclusion, we provide evidence that Omicron BA.2 reinfections do occur shortly after BA.1 infections but are rare.

Increased pulmonary secretion of tumor necrosis factor-α in calves experimentally infected with bovine respiratory syncytial virus

Publisher: Elsevier BV

Date: 10-2000

DOI: 10.1016/S0165-2427(00)00214-2

ACUTE INTERSTITIAL PNEUMONITIS CAUSED BY ALEUTIAN DISEASE VIRUS IN MINK KITS

Publisher: Wiley

Date: 03-1984

DOI: 10.1111/J.1699-0463.1984.TB04419.X

Abstract: In four Danish mink ranches acute interstitial pneumonitis caused excessive mortality among kits within the first 2 1/2 months after parturition. The disease was found to be due to an Aleutian disease virus (ADV) and could be reproduced experimentally in neonatal kits by inoculation with material from spontaneous cases, as well as with other strains of ADV. Experimental reproduction was only possible in kits from dams free of Aleutian disease (AD) whereas kits from dams experimentally or naturally infected with ADV developed no lung changes. Presently available evidence indicates that the initial lung lesions result from primary viral injury to type II alveolar cells, and that immune mechanisms, essential for the development of traditional AD, are not involved in the pathogenesis.

Transcription and localization of growth factor mRNA in the bovine oviduct

Publisher: Wiley

Date: 09-1995

DOI: 10.1002/MRD.1080420105

Abstract: It has become evident that certain growth factors are involved in the regulation of the initial bovine embryogenesis. In the present study, we examined by means of Northern blot and in situ hybridization, the expression and localization in the bovine oviduct of mRNAs encoding for platelet-derived growth factor (PDGF-B), basic fibroblast growth factor (bFGF), and insulin-like growth factor (IGF-I). Northern blot analysis on oviduct tissue demonstrated transcripts for PDGF-B and bFGF, but not IGF-I mRNAs. Two bands with estimated sizes of 5.0 and 1.5 kb were detected for PDGF-B and two bands with sizes of 7.5 and 4.9 kb for bFGF. In situ hybridization analysis demonstrated localization of PDGF-B mRNA in the lamina epithelialis and tunica muscularis of the oviduct whereas bFGF mRNA was detected in the lamina propria. It is concluded that the lamina epithelialis and lamina propria of the oviduct represent sites of synthesis of PDGF-B and bFGF mRNA, respectively.

30. Measurement of cytokine mRNA in pigs infected with foot-and-mouth-disease virus using quantitative real-time RT-PCR

Publisher: Elsevier BV

Date: 04-2002

DOI: 10.1016/S0034-5288(02)90032-9

Nucleotide sequence analysis of aleutian mink disease parvovirus shows that multiple virus types are present in infected mink

Publisher: American Society for Microbiology

Date: 08-1991

DOI: 10.1128/JVI.65.8.4378-4386.1991

Abstract: Different isolates of Aleutian mink disease parvovirus (ADV) were cloned and nucleotide sequenced. Analysis of in idual clones from two in vivo-derived isolates of high virulence indicated that more than one type of ADV DNA were present in each of these isolates. Analysis of several clones from two preparations of a cell culture-adapted isolate of low virulence showed the presence of only one type of ADV DNA. We also describe the nucleotide sequence from map units 44 to 88 of a new type of ADV DNA. The new type of ADV DNA is compared with the previously published ADV sequences, to which it shows 95% homology. These findings indicate that ADV, a single-stranded DNA virus, has a considerable degree of variability and that several virus types can be present simultaneously in an infected animal.

Evaluation of real-time reverse transcription polymerase chain reaction assays for the detection of swine vesicular disease virus

Publisher: Elsevier BV

Date: 03-2004

DOI: 10.1016/J.JVIROMET.2003.11.007

Abstract: Differential detection of swine vesicular disease virus (SVDV) from the other vesicular disease viruses of foot-and-mouth disease (FMD), vesicular stomatitis (VS) and vesivirus is important as the vesicular lesions produced by these viruses are indistinguishable in pigs. Two independent sets of primers and probe, designed from nucleotide sequences within the 5' untranslated region (UTR) of the SVDV genome, were evaluated in a real-time (5' nuclease probe-based or fluorogenic) PCR format. Although both primers robe sets failed to detect one isolate, the assays successfully lified RNA extracted from epithelial suspensions (ES) and cell culture grown virus preparations from clinical s les representing all currently designated phylogenetic groups of SVDV. Furthermore, no cross-reactivity was demonstrated when these primer robe sets were tested with RNA prepared from all seven serotypes of FMD virus (FMDV) and from selected isolates of VS virus (VSV), vesivirus and teschoviruses. These assays provide sensitive and rapid alternatives to supplement the routine procedures of ELISA and virus isolation for SVDV diagnosis. The two independent sets of primers robe can be used routinely while only one of the primers robe sets would typically be used in SVDV diagnosis during an outbreak.

Ability to detect antibodies to beak and feather disease virus in blood on filter paper decreases with duration of storage

Publisher: PeerJ

Date: 20-12-2021

DOI: 10.7717/PEERJ.12642

Abstract: Beak and feather disease virus (BFDV) is a circovirus that infects captive and wild psittacine birds, and is of conservation concern. The haemagglutination inhibition (HI) assay is used to determine antibody titres against BFDV, and the use of dried blood spots (DBS) on filter paper stored at room temperature has been suggested to be an equally valid technique to the use of frozen serum. However, research on other pathogens has found variable results when investigating the longevity of antibodies stored on DBS at room temperature. Consequently, we aimed to test the temporal stability of antibodies to BFDV in DBS s les stored long-term at room temperature. A further goal was to add to the current knowledge of antibody response to naturally acquired BFDV infection in crimson rosellas ( Platycercus elegans ). Blood was collected from wild P. elegans in Victoria, Australia, that had been live-trapped ( n = 9) or necropsied ( n = 11). BFDV virus load data were obtained from blood stored in ethanol by real-time quantitative PCR (qPCR) antibody titres were obtained by HI assay from either DBS or serum s les, which had been collected concurrently. All HI assays were performed commercially by the Veterinary Diagnostic Laboratory (VDL) in Charles Sturt University, Australia, who were blind to BFDV blood status. HI titres from DBS stored at room temperature declined significantly over time (~80 weeks). By contrast, frozen serum s les assayed after 80 weeks in storage all had high HI titres, only varying up to one dilution step from the initial HI titres obtained from DBS at 3–6 weeks after s ling. Weak HI titres from DBS s les all came back negative when the test was repeated only nine weeks later. Novel high HI titres were reported in P. elegans , and while most birds with high antibody titres had corresponding negative qPCR results, a single subadult presented with high HI titres and virus load simultaneously. Detection of antibodies on filter paper stored at room temperature decreases over time, increasing the chances of false negatives in these s les, and in repeated testing of s les with weak HI titres. Consequently, serum should be the preferred s le type to use for seroepidemiological studies on BFDV in parrots and other bird species. When not possible, it may help to store DBS on filter paper at −20 °C or lower. However, prompt testing of DBS s les ( e.g. , weeks in storage) is recommended pending further research on antibody temporal stability. We also show that P. elegans , especially adults, can produce high antibody titres against BFDV, which may help them resist infection.

SARS-CoV-2 genomic and subgenomic RNAs in diagnostic samples are not an indicator of active replication

Publisher: Springer Science and Business Media LLC

Date: 27-11-2020

DOI: 10.1038/S41467-020-19883-7

Abstract: Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) was first detected in late December 2019 and has spread worldwide. Coronaviruses are enveloped, positive sense, single-stranded RNA viruses and employ a complicated pattern of virus genome length RNA replication as well as transcription of genome length and leader containing subgenomic RNAs. Although not fully understood, both replication and transcription are thought to take place in so-called double-membrane vesicles in the cytoplasm of infected cells. Here we show detection of SARS-CoV-2 subgenomic RNAs in diagnostic s les up to 17 days after initial detection of infection and provide evidence for their nuclease resistance and protection by cellular membranes suggesting that detection of subgenomic RNAs in such s les may not be a suitable indicator of active coronavirus replication/infection.

The VetMet Veterinary Decision Support System for Airborne Animal Diseases

Publisher: Springer Netherlands

Date: 2008

DOI: 10.1007/978-1-4020-8453-9_22

Evaluation of automated RT-PCR to accelerate the laboratory diagnosis of foot-and-mouth disease virus

Publisher: Elsevier BV

Date: 02-2003

DOI: 10.1016/S0166-0934(02)00210-0

Abstract: Automated fluorogenic (5' nuclease probe-based) reverse transcription polymerase chain reaction (RT-PCR) procedures were evaluated for the diagnosis of foot-and-mouth disease (FMD) using suspensions of vesicular epithelium, heparinised or clotted blood, milk and oesophageal-pharyngeal fluid ('probang') s les from the United Kingdom (UK) 2001 epidemic and on sera from animals experimentally infected with the outbreak serotype O FMD virus strain. A MagNA Pure LC was initially programmed to automate the nucleic acid extraction and RT procedures with the PCR lification carried out manually by fluorogenic assay in a GeneAmp 5700 Sequence Detection System. This allowed 32 s les to be tested by one person in a typical working day or 64 s les by two people within 10-12 h. The PCR lification was later automated and a protocol developed for one person to complete a single test incorporating 96 RT-PCR results within 2 working days or for two people to do the same thing in around 12 h. The RT-PCR results were directly compared with those obtained by the routine diagnostic tests of ELISA and virus isolation in cell culture. The results on blood, probang and milk s les were in broad agreement between the three procedures but specific RT-PCR protocols for such material have to be fully optimised as perhaps have the positive-negative acceptance criteria. However, the automated RT-PCR achieved definitive diagnostic results (positive or negative) on supernatant fluids from first passage inoculated cell cultures and its sensitivity was greater than ELISA on suspensions of vesicular epithelium (ES) and at least equivalent to that of virus isolation in cell culture. The combined tests of ELISA, virus isolation in cell culture and RT-PCR might, therefore, only be required for confirmation of a first outbreak of FMD in a previously FMD-free country. Should a prolonged outbreak subsequently occur, then either ELISA plus RT-PCR or else RT-PCR alone could be used as the laboratory diagnostic tool(s). Either approach would eliminate the requirement for s le passage in cell culture and considerably advance the issue of laboratory diagnostic test results.

Diverse Bacterial Resistance Genes Detected in Fecal Samples From Clinically Healthy Women and Infants in Australia—A Descriptive Pilot Study

Publisher: Frontiers Media SA

Date: 17-09-2021

DOI: 10.3389/FMICB.2021.596984

Abstract: The gut microbiota is an immense reservoir of antimicrobial resistance genes (ARGs), the so-called “resistome.” In Australia, where antibiotic use is high and resistance rates in some common pathogens are increasing, very little is known about the human resistome. To assess the presence and ersity of ARGs in the gut of Australians from south-eastern Victoria, we investigated fecal s les from clinically healthy infants and pregnant women using non-targeted (shotgun metagenomics sequencing or SMS) and targeted sequencing (two Ion Ampliseq TM panels). All methods detected ARGs in all s les, with the detection overall of 64 unique genes conferring resistance to 12 classes of antibiotics. Predominant ARGs belonged to three classes of antibiotics that are the most frequently prescribed in Australia: tetracycline, β-lactams and MLS B (macrolide, lincosamide, streptogramin B). The three bacterial Orders commonly identified as carrying ARGs were Clostridiales, Bacteroidales, and Enterobacteriales. Our preliminary results indicate that ARGs are ubiquitously present and erse among the gut microbiota of clinically healthy humans from south-eastern Victoria, Australia. The observed resistance pattern partly overlaps with antimicrobial usage in human medicine in Australia, but ARGs to tetracycline are more common than could be expected. Our current s le is small and limited to south-eastern Victoria, and more data on healthy in iduals will be needed to better depict resistance patterns at the population level, which could guide population and/or environmental monitoring and surveillance of antibiotic resistance on various spatio-temporal scales in Australia. For future studies, we recommend using the Ion Ampliseq TM Antimicrobial Resistance Research panel, which is sensitive and user-friendly, or combining several methods to increase the detected ersity.

Early protection in sheep against intratypic heterologous challenge with serotype O foot-and-mouth disease virus using high-potency, emergency vaccine

Publisher: Elsevier BV

Date: 2015

DOI: 10.1016/J.VACCINE.2014.11.043

Abstract: In 2009-2011, spread of a serotype O foot-and-mouth disease virus (FMDV) belonging to the South East Asia topotype led to the culling of over 3.5 million cattle and pigs in Japan and Korea. The O1 Manisa vaccine (belonging to the Middle East-South Asian topotype) was used at high potency in Korea to limit the expansion of the outbreak. However, no data are available on the spread of this virus or the efficacy of the O1 Manisa vaccine against this virus in sheep. In this study, the early protection afforded with a high potency (>6 PD50) FMD O1 Manisa vaccine against challenge with the O/SKR/2010 virus was tested in sheep. Sheep (n=8) were vaccinated 4 days prior to continuous direct-contact challenge with donor sheep. Donor sheep were infected with FMDV O/SKR/2010 by coronary band inoculation 24h prior to contact with the vaccinated animals, or unvaccinated controls (n=4). Three of the four control sheep became infected, two clinically. All eight O1 Manisa vaccinated sheep were protected from clinical disease. None had detectable antibodies to FMDV non-structural proteins (3ABC), no virus was isolated from nasal swabs, saliva or oro-pharyngeal fluid and none became carriers. Using this model of challenge, sheep were protected against infection as early as 4 days post vaccination.

Evolutionary and network analysis of virus sequences from infants infected with an Australian recombinant strain of human parechovirus type 3

Publisher: Springer Science and Business Media LLC

Date: 20-06-2017

DOI: 10.1038/S41598-017-04145-2

Abstract: We present the near complete virus genome sequences with phylogenetic and network analyses of potential transmission networks of a total of 18 Australian cases of human parechovirus type 3 (HPeV3) infection in infants in the period from 2012–2015. Overall the results support our previous finding that the Australian outbreak strain/lineage is a result of a major recombination event that took place between March 2012 and November 2013 followed by further virus evolution and possibly recombination. While the nonstructural coding region of unknown provenance appears to evolve significantly both at the nucleotide and amino acid level, the capsid encoding region derived from the Yamagata 2011 lineage of HPeV3 appears to be very stable, particularly at the amino acid level. The phylogenetic and network analyses performed support a temporal evolution from the first Australian recombinant virus sequence from November 2013 to March/April 2014, onto the 2015 outbreak. The 2015 outbreak s les fall into two separate clusters with a possible common ancestor between March/April 2014 and September 2015, with each cluster further evolving in the period from September to November/December 2015.

Breeding season and pregnancy-associated increase in plasma levels of fibrin/fibrinogen in the mink

Publisher: Elsevier BV

Date: 04-1988

DOI: 10.1016/0378-4320(88)90033-4

Prevalence estimates of antibodies towards foot-and-mouth disease virus in small ruminants in uganda

Publisher: Hindawi Limited

Date: 09-11-2009

DOI: 10.1111/J.1865-1682.2009.01094.X

Abstract: Foot-and-mouth disease (FMD) is endemic in Uganda with control strategies focusing on vaccination of cattle, while small ruminants are largely ignored. In order for Uganda to establish effective control strategies, it is crucial that the epidemiology of the disease is fully understood. This study summarizes results of serological investigations of sheep and goats for antibodies to FMDV from four districts in 2006 following an FMD outbreak in the region and from an attempted comprehensive random s ling in two districts in 2007. Antibodies were quantified and serotyped using competitive ELISA for antibodies towards non-structural proteins (NSP) and structural proteins towards serotype O, and blocking ELISA for antibodies towards the seven serotypes of FMD virus (FMDV). In 2006, sheep and goats in Bushenyi and Isingiro districts were free from antibodies towards FMDV, while herds in Kasese and Mbarara districts excluding Kahendero village were all positive for antibodies towards NSP and SP-O. In 2007, mean prevalence estimates of antibodies towards FMDV NSP was 14% in goats and 22% in sheep in Kasese district, while Bushenyi was still free. The difference between these two districts probably reflects different levels of FMDV challenge attributed to the variation in exposure rates which again in part may be as a result of the differing husbandry practices. Contrary to 2006, with clear antibodies towards serotype O, the serotype-specificity of the antibodies was less clear in 2007, as antibodies towards both serotype O and SAT serotypes were identified. Our results show that goats and sheep are infected during FMD outbreaks, and that they may be useful for determining the serotype of FMD outbreaks in Uganda, if they are s led shortly after an outbreak.

Identification of alternatively spliced mRNAs encoding potential new regulatory proteins in cattle infected with bovine leukemia virus

Publisher: American Society for Microbiology

Date: 1993

DOI: 10.1128/JVI.67.1.39-52.1993

Abstract: The polymerase chain reaction was used to detect and characterize low-abundance bovine leukemia virus (BLV) mRNAs. In infected cattle we could detect spliced mRNA with a splice pattern consistent with a Tax/Rex mRNA, as well as at least four alternatively spliced RNAs. Two of the alternatively spliced mRNAs encoded hitherto unrecognized BLV proteins, designated RIII and GIV. The Tax/Rex and alternatively spliced mRNAs could be detected at their highest levels in BLV-infected cell cultures the next highest levels were found in s les from calves experimentally infected at 6 weeks postinoculation. Alternatively spliced mRNAs were also expressed, albeit at lower levels, in naturally infected animals they were detected by a nested polymerase chain reaction. Interestingly, the GIV mRNA was specifically detected in naturally infected cows with persistent lymphocytosis and in two of five calves at 6 months after experimental infection with BLV. Furthermore, the calf with the strongest signal for GIV had the highest lymphocyte counts. These data may suggest a correlation between expression of the GIV product and development of persistent lymphocytosis. Some of the donor and acceptor sites in the alternatively spliced mRNAs were highly unusual. The biological mechanisms and significance of such a choice of unexpected splice sites are currently unknown.

Effective animal health disease surveillance using a network-enabled approach

Publisher: Hindawi Limited

Date: 15-09-2010

DOI: 10.1111/J.1865-1682.2010.01166.X

Abstract: There are many benefits that derive from real-time knowledge of the health status of the national livestock population. Effective animal disease surveillance is a requirement for countries that trade in live animals and their products in order to comply with the World Organization for Animal Health (OIE) guidelines. Rapid identification of introduced and emerging disease allows rapid response and mitigation of the economic consequences. Connections between animal and human disease caused by a common pathogen can be recognized and control measures implemented, thereby protecting public health and maintaining public confidence in the food supply. Production-limiting diseases can be monitored, and control programmes be evaluated with benefits accruing from decreased economic losses associated with disease as well as reducing the welfare concerns associated with diseased animals. Establishing a surveillance programme across a wide area with erse ecosystems and political administrations as Canada is a complex challenge. When funding became available from a government programme to enable early detection of a bio-terrorist attack on livestock, the Canadian Animal Health Surveillance Network (CAHSN) became officially established. An existing web-based information platform that supports intelligence exchange, surveillance and response for public health issues in Canada was adapted to link the network animal health laboratories. A minimum data set was developed that facilitated sharing of results between participating laboratories and jurisdictions as the first step in creating the capacity for national disease trend analysis. In each of the network laboratories, similar quality assurance and bio-containment systems have been funded and supported, and diagnostic staff have been trained and certified on a suite of diagnostic tests for foreign animal diseases. This ensures that national standards are maintained throughout all of the diagnostic laboratories. This paper describes the genesis of CAHSN, its current capability and governance, and potential for future development.

Re-assessing the likelihood of airborne spread of foot-and-mouth disease at the start of the 1967-1968 UK foot-and-mouth disease epidemic

Publisher: Cambridge University Press (CUP)

Date: 12-04-2005

DOI: 10.1017/S0950268805004073

Abstract: The likelihood of airborne spread of foot-and-mouth disease at the start of the 1967–1968 epidemic is re-assessed in the light of current understanding of airborne disease spread. The findings strongly confirm those made at the time that airborne virus was the most likely cause of the rapid early development of the disease out to 60 km from the source. This conclusion is reached following a detailed epidemiological, meteorological and modelling study using original records and current modelling techniques. The role played by ‘lee waves’ as the mechanism for the spread is investigated. It is thought that they played little part in influencing the development of the epidemic. A number of lessons learned from the work are drawn, identifying the need for further research on the quantity and characteristics of airborne virus. The results are also used to illustrate what advice would have been available to disease controllers if the outbreak had occurred in 2004.

Epithelial Distribution and Replication of Foot-and-Mouth Disease Virus RNA in Infected Pigs

Publisher: Elsevier BV

Date: 08-2008

DOI: 10.1016/J.JCPA.2008.05.004

Abstract: Although the pathogenesis of foot-and-mouth disease (FMD) has been extensively investigated, relatively few studies have addressed the localization of FMD virus (FMDV) and in particular its replication in relation to the typical in-vivo sites of FMD lesions. In the present study, pigs were infected experimentally with FMDV serotype O UKG 34/2001 and tissue s les were collected from 1 to 4 days post-infection. S les were stored at -70 degrees C and frozen sections were prepared for in-situ hybridization (ISH). A digoxigenin-labelled RNA probe complementary to a coding part of the RNA-dependent RNA polymerase (3D) genomic region was prepared. The FMDV positive strand RNA was prominent in the basal layers of the epithelium. A diffuse positive signal was also noted in the cytoplasm of cells of the stratum spinosum of lesional epithelium, but there was no signal in the stratum corneum. Detection of FMDV negative strand RNA was observed in basal cells above the basement membrane and along the dermal papillae. The basal cells therefore demonstrate the highest signal for detection of the FMDV positive and negative strand RNAs in both tongue and foot epithelium. These novel results suggest that the epithelial basal cells could be an early replication site of FMDV in vivo.

Characterization of H1N1 swine influenza viruses circulating in Canadian pigs in 2009

Publisher: American Society for Microbiology

Date: 09-2011

DOI: 10.1128/JVI.00801-11

Abstract: The 2009 pandemic H1N1 (pH1N1), of apparent swine origin, may have evolved in pigs unnoticed because of insufficient surveillance. Consequently, the need for surveillance of influenza viruses circulating in pigs has received added attention. In this study we characterized H1N1 viruses isolated from Canadian pigs in 2009. Isolates from May 2009 were comprised of hemagglutinin and neuraminidase (NA) genes of classical SIV origin in combination with the North American triple-reassortant internal gene (TRIG) cassette, here termed contemporary SIV (conSIV) H1N1. These conSIV H1N1 viruses were contiguous with the North American αH1 cluster, which was distinct from the pH1N1 isolates that were antigenically more related to the γH1 cluster. After the initial isolation of pH1N1 from an Alberta pig farm in early May 2009, pH1N1 was found several times in Canadian pigs. These pH1N1 isolates were genetically and antigenically homogeneous. In addition, H1N1 viruses bearing seasonal human H1 and N1 genes together with the TRIG cassette and an NA encoding an oseltamivir-resistance marker were isolated from pigs. The NS gene of one of these seasonal human-like SIV (shSIV) H1N1 isolates was homologous to pH1N1 NS, implicating reassortment between the two strains. Antigenic cross-reactivity was observed between pH1N1 and conSIV but not with shSIV H1N1. In summary, although there was cocirculation of pH1N1 with conSIV and shSIV H1N1 in Canadian pigs after May 2009, there was no evidence supporting the presence of pH1N1 in pigs prior to May 2009. The possibility for further reassortants being generated exists and should be closely monitored.

Infection Dynamics of Swine Influenza Virus in a Danish Pig Herd Reveals Recurrent Infections with Different Variants of the H1N2 Swine Influenza A Virus Subtype

Publisher: MDPI AG

Date: 10-09-2020

DOI: 10.3390/V12091013

Abstract: Influenza A virus (IAV) in swine, so-called swine influenza A virus (swIAV), causes respiratory illness in pigs around the globe. In Danish pig herds, a H1N2 subtype named H1N2dk is one of the main circulating swIAV. In this cohort study, the infection dynamic of swIAV was evaluated in a Danish pig herd by s ling and PCR testing of pigs from two weeks of age until slaughter at 22 weeks of age. In addition, next generation sequencing (NGS) was used to identify and characterize the complete genome of swIAV circulating in the herd, and to examine the antigenic variability in the antigenic sites of the virus hemagglutinin (HA) and neuraminidase (NA) proteins. Overall, 76.6% of the pigs became PCR positive for swIAV during the study, with the highest prevalence at four weeks of age. Detailed analysis of the virus sequences obtained showed that the majority of mutations occurred at antigenic sites in the HA and NA proteins of the virus. At least two different H1N2 variants were found to be circulating in the herd one H1N2 variant was circulating at the sow and nursery sites, while another H1N2 variant was circulating at the finisher site. Furthermore, it was demonstrated that in idual pigs had recurrent swIAV infections with the two different H1N2 variants, but re-infection with the same H1N2 variant was also observed. Better understandings of the epidemiology, genetic and antigenic ersity of swIAV may help to design better health interventions for the prevention and control of swIAV infections in the herds.

Development of a novel real-time RT-PCR assay for quantitation of foot-and-mouth disease virus in diverse porcine tissues

Publisher: Elsevier BV

Date: 03-2001

DOI: 10.1016/S0166-0934(00)00265-2

Abstract: Pigs are more difficult to immunise and more variable in their response to foot-and-mouth disease (FMD) than other livestock species. This has important consequences for FMD control during both prophylactic vaccination programmes in endemic situations and when emergency vaccination may be used as an adjunct to st ing out during outbreaks in countries normally free from the disease. The rapid and effective control of FMD in pigs is especially important in regions of high pig density since infected pigs have the potential to generate plumes of airborne virus and spread infection beyond the immediate control area. Increased knowledge of the kinetics of FMDV replication in pigs, especially in their respiratory tracts, could create opportunities for strategies to improve FMD vaccines for pigs. A fluorogenic TaqMan RT-PCR assay specific for the IRES (internal ribosome entry site) sequence of the O(1) Kaufbeuren/Lausanne strain of FMDV has been developed for this purpose. The assay had a sensitivity of 0.1 TCID(50)/ml, and a dynamic range of at least eight orders of magnitude. It was found that an established method of quantitation, relative to a housekeeping gene, exhibited two significant shortcomings when applied to a set of 16 anatomically highly erse solid tissues: (i) tissue differences in housekeeping gene expression caused errors of up to 60-fold in estimated FMDV concentrations and (ii) variability in total RNA yields caused unpredictable saturation of RT reactions, which in turn caused errors of up to 250-fold in the estimated FMDV concentration. A novel quantitation strategy, designated the C(t)(min) method, was developed to overcome these problems. The C(t)(min) method was based on the the RT-PCR examination of a dilution series of spectrophotometrically quantitated total RNA, spanning the optimum for the RT-PCR system used. The new method was influenced minimally by any tissue-specific RT-PCR inhibitors and was used to determine FMDV concentrations in tissues from four experimentally infected pigs. The results suggest that the lungs play a less important role in the replication of FMDV in pigs than was thought previously.

Foot-and-Mouth Disease in Red Deer - Experimental Infection and Test Methods Performance

Publisher: Hindawi Limited

Date: 23-04-2015

DOI: 10.1111/TBED.12363

Abstract: The aim of this study was to evaluate a number of foot-and-mouth disease (FMD) test methods for use in red deer. Ten animals were intranasally inoculated with the FMD virus (FMDV) O UKG 11/2001, monitored for clinical signs, and s les taken regularly (blood, serum, oral swabs, nasal swabs, probang s les and lesion swabs, if present) over a 4-week period. Only one animal, deer 1103, developed clinical signs (lesions under the tongue and at the coronary band of the right hind hoof). It tested positive by 3D and IRES real-time reverse transcription polymerase chain reaction (rRT-PCR) in various swabs, lesion materials and serum. In a non-structural protein (NSP) in-house ELISA (NSP-ELISA-IH), one commercial ELISA (NSP-ELISA-PR) and a commercial antibody NSP pen side test, only deer 1103 showed positive results from day post-inoculation (dpi) 14 onwards. Two other NSP-ELISAs detected anti-NSP serum antibodies with lower sensitivity. It also showed rising antibody levels in the virus neutralization test (VNT), the in-house SPO-ELISA-IH and the commercial SPO-ELISA-PR at dpi 9, and in another two commercial SPO-ELISAs at dpi 12 (SPO-ELISA-IV) and dpi 19 (SPO-ELISA-IZ), respectively. Six of the red deer that had been rRT-PCR and antibody negative were re-inoculated intramuscularly with the same O-serotype FMDV at dpi 14. None of these animals became rRT-PCR or NSP-ELISA positive, but all six animals became positive in the VNT, the in-house SPO-ELISA-IH and the commercial SPO-ELISA-PR. Two other commercial SPO-ELISAs were less sensitive or failed to detect animals as positive. The rRT-PCRs and the four most sensitive commercial ELISAs that had been used for the experimentally inoculated deer were further evaluated for diagnostic specificity (DSP) using 950 serum s les and 200 nasal swabs from non-infected animals. DSPs were 100% for the rRT-PCRs and between 99.8 and 100% for the ELISAs.

Detection of a Reassortant H9N2 Avian Influenza Virus with Intercontinental Gene Segments in a Resident Australian Chestnut Teal

Publisher: MDPI AG

Date: 13-01-2020

DOI: 10.3390/V12010088

Abstract: The present study reports the genetic characterization of a low-pathogenicity H9N2 avian influenza virus, initially from a pool and subsequently from in idual faecal s les collected from Chestnut teals (Anas castanea) in southeastern Australia. Phylogenetic analyses of six full gene segments and two partial gene segments obtained from next-generation sequencing showed that this avian influenza virus, A/Chestnut teal/Australia/CT08.18/12952/2018 (H9N2), was a typical, low-pathogenicity, Eurasian aquatic bird lineage H9N2 virus, albeit containing the North American lineage nucleoprotein (NP) gene segment detected previously in Australian wild birds. This is the first report of a H9N2 avian influenza virus in resident wild birds in Australia, and although not in itself a cause of concern, is a clear indication of spillover and likely reassortment of influenza viruses between migratory and resident birds, and an indication that any lineage could potentially be introduced in this way.

Use of automated real-time Reverse Transcription-Polymerase Chain Reaction (RT-PCR) to monitor experimental swine vesicular disease virus infection in pigs

Publisher: Elsevier BV

Date: 11-2004

DOI: 10.1016/J.JCPA.2004.05.003

Abstract: Automated real-time RT-PCR was evaluated as a diagnostic tool for swine vesicular disease virus (SVDV) infection on a range of s les (vesicular epithelium, serum, nasal swabs, faeces) from four inoculated and three in-contact pigs over a period of 28 days. Traditional diagnostic procedures (virus isolation, and ELISAs for antigen and antibody) were used in parallel. Each inoculated pig developed a significant viraemia and clinical disease, and excreted virus, which was transmitted to the in-contact animals. The latter, however, developed only a short-lived, low-level viraemia and no clinical disease. The RT-PCR and virus isolation were generally comparable in detecting SVDV in the serum and nasal swabs from inoculated and in-contact pigs up to day 6 after infection it was possible, however, to isolate virus for a longer period from the faeces of a few pigs. This suggested that further optimization of the template extraction method was required to counteract the effects of RT-PCR inhibitors in faeces. It was concluded that the automated real-time RT-PCR is a useful diagnostic method for SVD in clinically or subclinically affected pigs and contributed to the study of the pathogenesis of SVD in the pigs.

Aspects of the persistence of foot-and-mouth disease virus in animals—the carrier problem

Publisher: Elsevier BV

Date: 08-2002

DOI: 10.1016/S1286-4579(02)01634-9

Abstract: Foot-and-mouth disease virus (FMDV) is a member of the Aphthovirus genus in the Picornaviridae family. Seven distinct serotypes, each including a wide range of variants, have been defined. FMD, affects wild and domesticated ruminants and pigs, is difficult to control and is the major constraint to international trade in livestock and animal products. After the acute stage of infection, FMDV may cause a prolonged, asymptomatic but persistent infection in ruminants. Also, vaccinated or naturally immune animals subsequently exposed to live virus may become persistently infected (the so-called carriers), a situation which can result in export embargoes if vaccination is included in a country's control policy.

Vaccination with aleutian mink disease parvovirus (AMDV) capsid proteins enhances disease, while vaccination with the major non-structural AMDV protein causes partial protection from disease

Publisher: Elsevier BV

Date: 07-1998

DOI: 10.1016/S0264-410X(98)80114-X

Abstract: Vaccination studies were performed with partially purified recombinant AMDV VP1/2 capsids as well as with the major AMDV non-structural protein (NS1). All vaccine constructs induced an antibody response, but did not prevent infection upon challenge with AMDV. The severity of Aleutian disease (AD) was judged by the serum gammaglobulin level, the quantity of peripheral blood CD8 lymphocytes, antibody titers to VP1/2 and NS1 proteins and mink death rates. The VP1/2 vaccine constructs enhanced the disease process with drastic death rates for the vaccinated mink. On the contrary, the NS1 vaccine constructs resulted in milder AD than seen in the non-vaccinated mink.

Determinants of Early Foot-and-Mouth Disease Virus Dynamics in Pigs

Publisher: Elsevier BV

Date: 11-2004

DOI: 10.1016/J.JCPA.2004.05.002

Abstract: This paper provides a quantitative description of the early infectious process in pigs experimentally infected with foot-and-mouth disease virus (FMDV), obtained by dose-dependent, time course studies of viral load in serum. Pigs were inoculated by the intravenous or intradermal/subcutaneous route with FMDV and housed together in groups or in idually. The effects of dose, inoculation route and exposure intensity on the replication of FMDV in vivo and the development of disease were studied. It was shown that the higher the dose, the shorter was the time to the start of active viraemia and to the onset of clinical signs. Exposure intensity and housing conditions influenced the viral dynamics of FMDV. Increasing the exposure intensity, by increasing the number of infected pigs housed together, had the effect of synchronizing the infection and reducing the variance in the start of active viraemia. Increasing the number of pigs housed together also increased the interaction between the pigs and the activity of in idual pigs, which had the effect of shortening the time to the onset of clinical signs such as vesicle formation. Intradermal inoculation was more effective than intravenous inoculation for transmitting FMDV to pigs, resulting in shorter times to the start of active viraemia and in higher clinical scores.

Subcellular localization of aleutian mink disease parvovirus proteins and DNA during permissive infection of crandell feline kidney cells

Publisher: American Society for Microbiology

Date: 05-1996

DOI: 10.1128/JVI.70.5.3242-3247.1996

Abstract: Confocal microscopy allowed us to localize viral nonstructural (NS) and capsid (VP) proteins and DNA simultaneously in cells permissively infected with Aleutian mink disease parvovirus (ADV). Early after infection, NS proteins colocalized with viral DNA to form intranuclear inclusions, whereas VP proteins formed hollow intranuclear shells around the inclusions. Later, nuclei had irregular outlines and were virtually free of ADV products. In these cells, inclusions of viral DNA with or without associated NS protein were embedded in cytoplasmic VP protein. These findings implied that ADV replication within an infected cell is regulated spatially as well as temporally.

Reassortant Highly Pathogenic Influenza A H5N2 Virus Containing Gene Segments Related to Eurasian H5N8 in British Columbia, Canada, 2014

Publisher: Springer Science and Business Media LLC

Date: 25-03-2015

DOI: 10.1038/SREP09484

Abstract: In late November 2014 higher than normal death losses in a meat turkey and chicken broiler breeder farm in the Fraser Valley of British Columbia initiated a diagnostic investigation that led to the discovery of a novel reassortant highly pathogenic avian influenza (HPAI) H5N2 virus. This virus, composed of 5 gene segments (PB2, PA, HA, M and NS) related to Eurasian HPAI H5N8 and the remaining gene segments (PB1, NP and NA) related to North American lineage waterfowl viruses, represents the first HPAI outbreak in North American poultry due to a virus with Eurasian lineage genes. Since its first appearance in Korea in January 2014, HPAI H5N8 spread to Western Europe in November 2014. These European outbreaks happened to temporally coincide with migratory waterfowl movements. The fact that the British Columbia outbreaks also occurred at a time associated with increased migratory waterfowl activity along with reports by the USA of a wholly Eurasian H5N8 virus detected in wild birds in Washington State, strongly suggest that migratory waterfowl were responsible for bringing Eurasian H5N8 to North America where it subsequently reassorted with indigenous viruses.

Antibodies against foot-and-mouth disease (FMD) virus in African Buffalos syncerus caffer in selected national parks in Uganda (2001-2003)

Publisher: Hindawi Limited

Date: 18-06-2010

DOI: 10.1111/J.1865-1682.2010.01147.X

Abstract: In East Africa, the foot-and-mouth disease (FMD) virus (FMDV) isolates have over time included serotypes O, A, C, Southern African Territories (SAT) 1 and SAT 2, mainly from livestock. SAT 3 has only been isolated in a few cases and only in African buffalos (Syncerus caffer). To investigate the presence of antibodies against FMDV serotypes in wildlife in Uganda, serological studies were performed on buffalo serum s les collected between 2001 and 2003. Thirty-eight s les from African buffalos collected from Lake Mburo, Kidepo Valley, Murchison Falls and Queen Elizabeth National Parks were screened using Ceditest FMDV NS to detect antibodies against FMDV non-structural proteins (NSP). The seroprevalence of antibodies against non-structural proteins was 74%. To characterize FMDV antibodies, s les were selected and titrated using serotype-specific solid phase blocking enzyme linked immunosorbent assay (ELISAs). High titres of antibodies (> or =1 : 160) against FMDV serotypes SAT 1, SAT 2 and SAT 3 were identified. This study suggests that African buffalos in the different national parks in Uganda may play an important role in the epidemiology of SAT serotypes of FMDV.

Neonatal parechovirus infection: Possibility of in‐utero transmission

Publisher: Wiley

Date: 04-10-2021

DOI: 10.1111/JPC.15763

118. Detection and quantification of swine vesicular disease virus by TaqMan® RT-PCR assay

Publisher: Elsevier BV

Date: 2003

DOI: 10.1016/S0034-5288(03)90117-2

37. Evaluation of the Cepheid SmartCycler real-time PCR machine for rapid and portable diagnosis of foot-and-mouth disease

Publisher: Elsevier BV

Date: 04-2002

DOI: 10.1016/S0034-5288(02)90039-1

Epidemiological and Evolutionary Inference of the Transmission Network of the 2014 Highly Pathogenic Avian Influenza H5N2 Outbreak in British Columbia, Canada

Publisher: Springer Science and Business Media LLC

Date: 04-08-2016

DOI: 10.1038/SREP30858

Abstract: The first North American outbreak of highly pathogenic avian influenza (HPAI) involving a virus of Eurasian A/goose/Guangdong/1/1996 (H5N1) lineage began in the Fraser Valley of British Columbia, Canada in late November 2014. A total of 11 commercial and 1 non-commercial (backyard) operations were infected before the outbreak was terminated. Control measures included movement restrictions that were placed on a total of 404 in idual premises, 150 of which were located within a 3 km radius of an infected premise(s) (IP). A complete epidemiological investigation revealed that the source of this HPAI H5N2 virus for 4 of the commercial IPs and the single non-commercial IP likely involved indirect contact with wild birds. Three IPs were associated with the movement of birds or service providers and localized/environmental spread was suspected as the source of infection for the remaining 4 IPs. Viral phylogenies, as determined by Bayesian Inference and Maximum Likelihood methods, were used to validate the epidemiologically inferred transmission network. The phylogenetic clustering of concatenated viral genomes and the median-joining phylogenetic network of the viruses supported, for the most part, the transmission network that was inferred by the epidemiologic analysis.

Foot-and-mouth disease virus infection of sheep: implications for diagnosis and control.

Publisher: Wiley

Date: 06-2002

DOI: 10.1136/VR.150.23.724

Analysis of parvovirus infections using strand-specific hybridization probes

Publisher: Elsevier BV

Date: 09-1989

DOI: 10.1016/0168-1702(89)90066-X

Abstract: The autonomous parvoviruses cause a broad spectrum of acute and chronic infections of animals and man. The discrimination of sites of viral replication from sites of viral sequestration is an important goal in elucidating the pathogenesis of these diseases. It is possible to employ strand-specific RNA hybridization probes in such analyses because a 'plus' sense probe will react with single stranded virion DNA and duplex replicative form DNA, but a 'minus' sense probe will react preferentially with obligate replicative intermediates (duplex replicative form DNA and mRNA). Strand-specific RNA hybridization probes were developed for the Aleutian mink disease parvovirus (ADV) and were used to study acute and chronic infections of mink. Such probes were capable of differentiating replicative intermediates (duplex replicative form DNA and mRNA) from single-stranded virion DNA in Southern blot analysis and in strand-specific in situ hybridization. ADV infection of seronegative newborn mink kits causes an acute, cytopathic infection of type II alveolar cells. Replication in these cells is highly permissive and is characterized by high levels of replicative intermediates and virion DNA. A fatal respiratory distress syndrome and hyaline membrane formation result from impaired surfactant production by the infected type II cells. On the other hand, ADV infection of adult mink is associated with a persistent infection and a disorder of the immune regulation. The target cells for viral replication in adult mink are confined to the lymphoid system and the bone marrow. Replication in these cells, which are probably lymphocytes, is restricted, and characterized by greatly reduced levels of replicative intermediates and virion DNA. It, therefore, seems that disease in the infected adult mink results from a restricted infection by ADV. Large amounts of virion DNA can also be demonstrated in locations where replication cannot be detected and apparently represents sequestration of virion particles by elements of the reticuloendothelial system. Thus, replication and sequestration can, in fact, be distinguished by the strand-specific in situ hybridization. These studies indicate that strand-specific in situ hybridization is a potentially valuable method for studying the pathogenesis of parvovirus infections.

The αvβ6 integrin receptor for Foot-and-mouth disease virus is expressed constitutively on the epithelial cells targeted in cattle

Publisher: Microbiology Society

Date: 10-2005

DOI: 10.1099/VIR.0.81172-0

Abstract: Field strains of Foot-and-mouth disease virus (FMDV) use a number of α v-integrins as receptors to initiate infection on cultured cells, and integrins are believed to be the receptors used to target epithelial cells in animals. In this study, immunofluorescence confocal microscopy and real-time RT-PCR were used to investigate expression of two of the integrin receptors of FMDV, α v β 6 and α v β 3, within various epithelia targeted by this virus in cattle. These studies show that α v β 6 is expressed constitutively on the surfaces of epithelial cells at sites where infectious lesions occur during a natural infection, but not at sites where lesions are not normally formed. Expression of α v β 6 protein at these sites showed a good correlation with the relative abundance of β 6 mRNA. In contrast, α v β 3 protein was only detected at low levels on the vasculature and not on the epithelial cells of any of the tissues investigated. Together, these data suggest that in cattle, α v β 6, rather than α v β 3, serves as the major receptor that determines the tropism of FMDV for the epithelia normally targeted by this virus.

Metagenomics detection and characterisation of viruses in faecal samples from Australian wild birds

Publisher: Springer Science and Business Media LLC

Date: 06-06-2018

DOI: 10.1038/S41598-018-26851-1

Abstract: We present an optimised metagenomics method for detection and characterisation of all virus types including single and double stranded DNA/RNA and enveloped and non-enveloped viruses. Initial evaluation included both spiked and non-spiked bird faecal s les as well as non-spiked human faecal s les. From the non-spiked bird s les (Australian Muscovy duck and Pacific black ducks) we detected 21 viruses, and we also present a summary of a few viruses detected in human faecal s les. We then present a detailed analysis of selected virus sequences in the avian s les that were somewhat similar to known viruses, and had good quality (Q20 or higher) and quantity of next-generation sequencing reads, and was of interest from a virological point of view, for ex le, avian coronavirus and avian paramyxovirus 6. Some of these viruses were closely related to known viruses while others were more distantly related with 70% or less identity to currently known/sequenced viruses. Besides detecting viruses, the technique also allowed the characterisation of host mitochondrial DNA present and thus identifying host species, while ribosomal RNA sequences provided insight into the “ribosomal activity microbiome” of gut parasites and of food eaten such as plants or insects, which we correlated to non-avian host associated viruses.

Relative risks of the uncontrollable (airborne) spread of FMD by different species.

Publisher: Wiley

Date: 05-2001

DOI: 10.1136/VR.148.19.602

Detection and characterisation of canine astrovirus, canine parvovirus and canine papillomavirus in puppies using next generation sequencing

Publisher: Springer Science and Business Media LLC

Date: 14-03-2019

DOI: 10.1038/S41598-019-41045-Z

Abstract: Gastroenteritis in young animals is a clinical presentation with many infectious and non- infectious aetiologies. We used next generation sequencing (NGS) to investigate the possible infectious causes of gastroenteritis in puppies from a dog kennel in Victoria, Australia. The near complete genome of a canine astrovirus was obtained from pooled faecal s les, and was found to be 94.7% identical with a canine astrovirus detected in the United Kingdom in 2012. The phylogenetic analysis of the capsid gene found similarities to those of canine astroviruses identified in Italy in 2005 and in UK and Hungary in 2012, but distant from that of a canine astrovirus previously identified in Australia in 2012. Thus, different serotypes of canine astrovirus are likely circulating in Australia. The close relationship to European astroviruses also suggested that there had been recent movements of ancestor canine astroviruses between Australia and Europe. NGS also detected other infections in the puppies including several canine papillomaviruses and a canine parvovirus (vaccine strain) as well as a very low level of c ylobacter. Canine astrovirus was the probable cause of diarrhoea in these puppies, with the possible involvement of c ylobacter bacteria. NGS was effective as a non-targeted method to determine the likely infectious cause of gastroenteritis.

Exploring the Cause of Diarrhoea and Poor Growth in 8–11-Week-Old Pigs from an Australian Pig Herd Using Metagenomic Sequencing

Publisher: MDPI AG

Date: 13-08-2021

DOI: 10.3390/V13081608

Abstract: Diarrhoea and poor growth among growing pigs is responsible for significant economic losses in pig herds globally and can have a wide range of possible aetiologies. Next generation sequencing (NGS) technologies are useful for the detection and characterisation of erse groups of viruses and bacteria and can thereby provide a better understanding of complex interactions among microorganisms potentially causing clinical disease. Here, we used a metagenomics approach to identify and characterise the possible pathogens in colon and lung s les from pigs with diarrhoea and poor growth in an Australian pig herd. We identified and characterized a wide ersity of porcine viruses including RNA viruses, in particular several picornaviruses—porcine sapelovirus (PSV), enterovirus G (EV-G), and porcine teschovirus (PTV), and a porcine astrovirus (PAstV). Single stranded DNA viruses were also detected and included parvoviruses like porcine bocavirus (PBoV) and porcine parvovirus 2 (PPV2), porcine parvovirus 7 (PPV7), porcine bufa virus (PBuV), and porcine adeno-associated virus (AAV). We also detected single stranded circular DNA viruses such as porcine circovirus type 2 (PCV2) at very low abundance and torque teno sus viruses (TTSuVk2a and TTSuVk2b). Some of the viruses detected here may have had an evolutionary past including recombination events, which may be of importance and potential involvement in clinical disease in the pigs. In addition, our metagenomics data found evidence of the presence of the bacteria Lawsonia intracellularis, Brachyspira spp., and C ylobacter spp. that may, together with these viruses, have contributed to the development of clinical disease and poor growth.

RESTRICTED HETEROGENEITY OF THE EARLY ANTIBODY RESPONSE TO ALEUTIAN DISEASE VIRUS IN MINK KITS

Publisher: Wiley

Date: 11-1986

DOI: 10.1111/J.1699-0463.1986.TB02103.X

Abstract: Mink kits born of Aleutian disease (AD) negative dams were infected neonatally with different isolates of Aleutian disease virus (ADV). They were then sacrified at different days after infection (10 — 45 days) and their sera were analysed by different techniques for concentrations of immunoglobulins and quality of the produced antibodies to ADV. The infected mink kits produced significantly higher concentrations of IgM and IgG immunoglobulins than non‐infected‐age matched control mink (p .001). Eight percent of the sera from the infected mink kits exhibited a restricted heterogeneous gammaglobulin profile as shown by serum electrophoresis. These findings were further investigated by other techniques such as SDS‐PAGE and crossed serum line immunoelectrophoresis. It is concluded that mink kits when infected neonatally with ADV start to develop hypergammaglobulinemia soon after infection, and that a small percentage of the mink react with a gammaglobulin response of restricted heterogeneity.

Middle east respiratory syndrome coronavirus antibody reactors among camels in Dubai, United Arab Emirates, in 2005

Publisher: Hindawi Limited

Date: 24-01-2014

DOI: 10.1111/TBED.12212

Lack of susceptibility of the dromedary camel (Camelus dromedarius) to foot‐and‐mouth disease virus serotype O

Publisher: Wiley

Date: 02-2006

DOI: 10.1136/VR.158.6.201

Diagnosis of foot‐and‐mouth disease by real‐time fluorogenic PCR assay

Publisher: Wiley

Date: 11-2001

DOI: 10.1136/VR.149.20.621

Foot‐and‐mouth disease viral loads in pigs in the early, acute stage of disease

Publisher: Wiley

Date: 2010

DOI: 10.1136/VR.B5583

Abstract: The progress and pathogenesis of foot-and-mouth disease virus (FMDV) was studied in infected pigs by observing the development of clinical signs in two separate experiments. Viral loads were determined by real-time quantitative RT-PCR in the liver, spleen, cervical lymph node, mandibular lymph node, retropharyngeal lymph node, soft palate, pharynx, tonsil, tongue and skin (coronary band area). Tissue s les were collected from both inoculated and contact-infected pigs at several time points during infection, and blood s les were collected to assess viraemia and its relationship to tissue viral load. Virus first appeared in the lymph nodes, followed by viraemia and then clinical signs. The results suggested that FMDV accumulated in lymphoid tissue up to six hours after infection, in the tissues drained by the mandibular lymph node and tonsil and then disseminated throughout the body where epithelial cells were the favoured sites of replication.

New directions

Publisher: Elsevier BV

Date: 2004

DOI: 10.1016/J.ATMOSENV.2003.10.014

Validation of a foot-and-mouth disease antibody screening solid-phase competition ELISA (SPCE)

Publisher: Elsevier BV

Date: 02-2004

DOI: 10.1016/J.JVIROMET.2003.09.016

Abstract: This paper describes the validation of a solid-phase competition enzyme-linked immunosorbent assay (SPCE) for the serological detection of antibody to serotype O foot-and-mouth disease (FMD) in sheep, cattle and pigs. The specificity of the SPCE was calculated from the results of testing known negative sera from sheep, cattle and pigs (n=3030, 1418 and 1495, respectively). The mean percentage inhibition (PI) for known negative sheep, cattle and pig sera were 19.3, 24.1 and 20.8%, respectively. The specificity of the SPCE at a cut-off point (COP) of 60 PI was 99.50% for sheep sera, 99.44% for cattle sera and 100% for pig sera. The analytical sensitivity of the SPCE was examined by testing sera from sheep, cattle and pigs. Based on the testing of serial bleeds from experimentally infected animals, seroconversion at the 60 PI COP occurred between 4 and 9 days post-infection or -exposure, similar to that observed using the virus neutralisation test (VNT) with a COP of 1/45. When applied to 267 sheep and 143 pig s les, that were obtained in Great Britain (GB) during the 2001 FMD UK outbreak, the SPCE identified more positive s les than did the VNT. Estimates of the accuracy, repeatability and reproducibility of the SPCE were verified during the large-scale serosurveillance necessitated by the 2001 outbreak. Results from field and experimental sera showed that when compared against the VNT, the sensitivity of the SPCE was less affected by the choice of virus strain used in the test. Using the O(1) UKG 2001 FMD virus in the VNT with s les representative of the uninfected GB sheep population, the test specificity was 100% at a COP of 1/45.

Influence of Exposure Intensity on the Efficiency and Speed of Transmission of Foot-and-Mouth Disease

Publisher: Elsevier BV

Date: 05-2009

DOI: 10.1016/J.JCPA.2008.12.002

Abstract: Foot-and-mouth disease virus (FMDV) can be spread by direct animal-to-animal contact, indirect contact facilitated by contaminated materials or by airborne spread. The rate of spread and the incubation period, as well as the severity of disease, depends on many variables including the dose received, the route of introduction, the virus strain, the animal species and the conditions under which the animals are kept. Quantitative data related to these variables are needed if model predictions are to be used in practical disease control. This experimental study quantifies the risk of transmission of FMDV in pigs exposed by contact, sheep exposed by indirect contact with pigs and sheep exposed to airborne FMDV. Groups of pigs were inoculated with the FMDV O UKG 34/2001 strain and susceptible pigs were then exposed to the inoculated animals at different stages of the infection cycle. The mean incubation period in the susceptible pigs ranged from 1 to 10 days. The length of the incubation period, severity of clinical disease and efficiency of spread were related to dose (i.e. infectiousness of source and intensity of contact). Low intensity transmission increased the proportion of subclinical or abortive infections. Local conditions are important in the efficiency and speed of transmission of FMDV. The results of the experiments described above suggest that transmission is frequency dependent rather than density dependent. The sheep experiments provided further evidence that development of infection and clinical disease is dependent upon local conditions. Dose, infectiousness, intensity of contact and local factors are thus important determinants for the outcome of an initial outbreak and must be truthfully accounted for in mathematical models of epidemiological spread.

Cytokine and Toll-like Receptor mRNAs in the Nasal-associated Lymphoid Tissues of Cattle During Foot-and-mouth Disease Virus Infection

Publisher: Elsevier BV

Date: 2006

DOI: 10.1016/J.JCPA.2005.06.011

Abstract: The pharyngeal region is known to play an important role in foot-and-mouth disease virus (FMDV) infection in relation to acute disease and viral persistence. In this study, the local mucosal immune response in nasal-associated lymphoid tissue (NALT) of cattle infected with FMDV (strain O UKG 34/2001) was examined. Quantitative "real-time" RT-PCR assays were used to measure mRNA expression of cytokines (IFN-alpha, beta and gamma, IL-2, IL-1alpha and TNF-alpha) and Toll-like receptors (TLR)-3 and -4. NALTs from dorsal soft palate were collected from cattle at 7 days post-infection (dpi) and from carriers and non-carriers at 64 dpi. Expression of IFN-alpha mRNA was significantly greater in NALT during acute disease than in uninfected animals. Increased expression of IFN-gamma and IL-1alpha mRNA was also observed but was much lower than IFN-alpha expression. There was a slight increase in mRNA expression of TNF-alpha and IL-2. During persistence, TNF-alpha mRNA expression in carrier cattle was much higher than in non-carrier cattle. Expression of TLR-4 in NALT during the acute stage of infection was greater than in uninfected animals. Carrier and non-carrier cattle did not differ in respect of expression of TLR-3 and -4 mRNA in NALT.

Evaluation of the portable Cepheid SmartCycler real‐time PCR machine for the rapid diagnosis of foot‐and‐mouth disease

Publisher: Wiley

Date: 05-2002

DOI: 10.1136/VR.150.20.625

Abstract: The ability of the portable Cepheid SmartCycler real-time PCR machine to detect foot-and-mouth disease (FMD) virus sensitively and accurately was evaluated by comparing the results of the analyses of nasal swab and serum s les from experimentally infected animals with those obtained from the real-time PCR assay currently in use in the laboratory. The results indicated that the ability of the machine to detect viral RNA is greatly affected by the PCR reagents used for the assay. When it was used with PCR beads it was unable to detect weakly positive s les, but when TaqMan core reagents were used for the assay, its sensitivity was significantly increased. The machine could be used for the laboratory-based detection of FMD however, as with all assays, significant optimisation of assay conditions as well as solid validation of the technique is required.

Clinical and laboratory investigations of five outbreaks of foot-and-mouth disease during the 2001 epidemic in the United Kingdom.

Publisher: Wiley

Date: 04-2003

DOI: 10.1136/VR.152.16.489

Abstract: Clinical and laboratory investigations of five outbreaks of foot-and-mouth disease (FMD) were made during the early stages of the 2001 epidemic in the UK. The first outbreak, confirmed on February 20, was at an abattoir in Essex which specialised in the processing of culled sows and boars. On February 23, the disease was confirmed at a pig farm in Northumberland which held cull sows and boars fed on waste food the findings indicated that it was the first of the five premises to be infected. The disease had probably been present since early February, and it was the most likely origin of the epidemic. The other premises investigated were a waste food-fed cull sow/boar pig unit in Essex, approximately 30 km from the abattoir, which was probably infected at the same time or before the abattoir, a sheep and cattle farm approximately 6 km from the Northumberland pig farm, which was probably infected by airborne virus from it in the period immediately before February 13, and a sheep and cattle farm in Devon which had clinical disease from February 20 and was probably infected by sheep transported from Northumberland on February 13 which arrived on February 15.

Epidemic and Inter-epidemic Burden of Pediatric Human Parechovirus Infection in New South Wales, Australia, 2017–2018

Publisher: Ovid Technologies (Wolters Kluwer Health)

Date: 06-2020

DOI: 10.1097/INF.0000000000002615

Sequence comparison of the non-structural genes of four different types of Aleutian mink disease parvovirus indicates an unusual degree of variability

Publisher: Springer Science and Business Media LLC

Date: 09-1994

DOI: 10.1007/BF01379127

Detection of carrier cattle and sheep persistently infected with foot-and-mouth disease virus by a rapid real-time RT-PCR assay

Publisher: Elsevier BV

Date: 08-2003

DOI: 10.1016/S0166-0934(03)00165-4

Abstract: The detection of foot-and-mouth disease virus (FMDV) in persistently infected carriers among exposed ruminants is of great importance in disease control. For this purpose, a real time, fluorogenic reverse transcription polymerase chain reaction (real-time RT-PCR) assay was evaluated for the identification of FMDV carrier animals. The results indicate that this real time RT-PCR assay may be suitable for detection of FMDV carrier animals.

Picornaviruses

Publisher: Wiley

Date: 29-03-2019

DOI: 10.1002/9781119350927.CH40

Two parvoviruses that cause different diseases in mink have different transcription patterns

Publisher: American Society for Microbiology

Date: 07-1997

DOI: 10.1128/JVI.71.7.4990-4996.1997

Abstract: The two parvoviruses of mink cause very different diseases. Mink enteritis virus (MEV) is associated with rapid, high-level viral replication and acute disease. In contrast, infection with Aleutian mink disease parvovirus (ADV) is associated with persistent, low-level viral replication and chronic severe immune dysregulation. In the present report, we have compared viral transcription in synchronized CRFK cells infected with either MEV or ADV using a nonradioactive RNase protection assay. The overall level of viral transcription was 20-fold higher in MEV- than in ADV-infected cells. Furthermore, MEV mRNA encoding structural proteins (MEV mRNA R3) was dominant throughout the infectious cycle, comprising approximately 80% of the total viral transcription products. In marked contrast, in ADV-infected cells, transcripts encoding nonstructural proteins (ADV mRNA R1 and R2) comprised more than 84% of the total transcripts at all times after infection, whereas ADV mRNA R3 comprised less than 16%. Thus, the ADV mRNA coding for structural proteins (ADV mRNA R3) was present at a level at least 100-fold lower than the corresponding MEV mRNA R3. These findings paralleled previous biochemical studies analyzing in vitro activities of the ADV and MEV promoters (J. Christensen, T. Storgaard, B. Viuff, B. Aasted, and S. Alexandersen, J. Virol. 67:1877-1886, 1993). The overall low levels of ADV mRNA and the paucity of the mRNA coding for ADV structural proteins may reflect an adaptation of the virus for low-level restricted infection.

Acute interstitial pneumonia in mink kits

Publisher: SAGE Publications

Date: 09-1986

DOI: 10.1177/030098588602300506

Abstract: Organ homogenates from kits that died of interstitial pneumonia were inoculated into adult Aleutian disease virus (ADV)-negative mink and shown to contain infectious ADV. Acute interstitial pneumonia was experimentally reproduced with the organ homogenate but only by inoculation of newborn kits born from ADV-negative dams. Older kits and kits from ADV-positive dams did not develop interstitial pneumonia, but later developed the classic form of Aleutian disease. Electron microscopic examination was done on purified suspensions of defined ADV isolates and on purified organ homogenates from kits with spontaneous or experimental interstitial pneumonia. In kits from both groups a virus, morphologically resembling the defined ADV isolates, was demonstrated. Findings of intranuclear inclusion bodies and intranuclear ADV antigen in alveolar type-II cells in affected lungs and the lack of immunologically mediated lesions suggest that lung lesions result from primary viral injury to alveolar type-II cells. Experiments also showed that infection of dams with ADV before pregnancy decreased the number of kits per mated dam and infection with ADV in mid-pregnancy caused fetal death, fetal resorption, or abortion.

Use of confocal immunofluorescence microscopy to localize viral nonstructural proteins and potential sites of replication in pigs experimentally infected with foot-and-mouth disease virus

Publisher: American Society for Microbiology

Date: 15-05-2005

DOI: 10.1128/JVI.79.10.6410-6418.2005

Abstract: Replication of foot-and-mouth disease virus in infected pig epithelium has been studied by immunofluorescence labeling of the viral nonstructural protein 3ABC and confocal microscopy. The results were correlated with viral RNA copy numbers in tissue s les from adjacent sites determined by reverse transcription-PCR (RT-PCR). Lesion formation was seen in the tongues and coronary band epithelia of infected pigs 2 days after infection. Viral replication was observed in cells of the epithelium of the tongue and coronary band but not in the associated stromal cells. Infected epithelial cells were present in the stratum spinosum, away from the lesion, with small lesions formed above the basement membrane. Viral replication was markedly reduced in tongue epithelium by day 3 postinfection but remained apparent in the coronary band tissue up to 5 days postinfection. These results were confirmed by the RNA copy number determined by RT-PCR.

Demonstration of non-degraded aleutian disease virus (ADV) proteins in lung tissue from experimentally infected mink kits

Publisher: Springer Science and Business Media LLC

Date: 03-1986

DOI: 10.1007/BF01310549

Abstract: Previous studies suggest a link between gut microbiota and the development of ulcerative colitis (UC) and irritable bowel syndrome (IBS). Our aim was to investigate any quantitative differences in faecal bacterial compositions in UC and IBS patients compared to healthy controls, and to identify in idual bacterial species that contribute to these differences. Faecal microbiota of 13 UC patients, 11 IBS patients and 22 healthy volunteers were analysed by PCR-Denaturing Gradient Gel Electrophoresis (DGGE) using universal and Bacteroides specific primers. The data obtained were normalized using in-house developed statistical method and interrogated by multivariate approaches. The differentiated bands were excised and identified by sequencing the V3 region of the 16S rRNA genes. Band profiles revealed that number of predominant faecal bacteria were significantly different between UC, IBS and control group (p < 10-4). By assessing the mean band numbers in UC (37 ± 5) and IBS (39 ± 6), compared to the controls (45 ± 3), a significant decrease in bacterial species is suggested (p = 0.01). There were no significant differences between IBS and UC. Bio ersity of the bacterial species was significantly lower in UC (μ = 2.94, σ = 0.29) and IBS patients (μ = 2.90, σ = 0.38) than controls (μ = 3.25, σ = 0.16 p = 0.01). Moreover, similarity indices revealed greater biological variability of predominant bacteria in UC and IBS compared to the controls (median Dice coefficients 76.1% (IQR 70.9 - 83.1), 73.8% (IQR 67.0 - 77.5) and 82.9% (IQR 79.1 - 86.7) respectively). DNA sequencing of discriminating bands suggest that the presence of Bacteroides vulgatus, B. ovatus, B. uniformis, and Parabacteroides sp. in healthy volunteers distinguishes them from IBS and UC patients. DGGE profiles of Bacteroides species revealed a decrease of Bacteroides community in UC relative to IBS and controls. Molecular profiling of faecal bacteria revealed abnormalities of intestinal microbiota in UC and IBS patients, while different patterns of Bacteroides species loss in particular, were associated with UC and IBS.

Genetic and pathobiologic characterization of pandemic H1N1 2009 influenza viruses from a naturally infected swine herd

Publisher: American Society for Microbiology

Date: 03-2010

DOI: 10.1128/JVI.02118-09

Abstract: Since its initial identification in Mexico and the United States, concerns have been raised that the novel H1N1 influenza virus might cause a pandemic of severity comparable to that of the 1918 pandemic. In late April 2009, viruses phylogenetically related to pandemic H1N1 influenza virus were isolated from an outbreak on a Canadian pig farm. This outbreak also had epidemiological links to a suspected human case. Experimental infections carried out in pigs using one of the swine isolates from this outbreak and the human isolate A/Mexico/InDRE4487/2009 showed differences in virus recovery from the lower respiratory tract. Virus was consistently isolated from the lungs of pigs infected with A/Mexico/InDRE4487/2009, while only one pig infected with A/swine/Alberta/OTH-33-8/2008 yielded live virus from the lung, despite comparable amounts of viral RNA and antigen in both groups of pigs. Clinical disease resembled other influenza virus infections in swine, albeit with somewhat prolonged virus antigen detection and delayed viral-RNA clearance from the lungs. There was also a noteworthy amount of genotypic variability among the viruses isolated from the pigs on the farm. This, along with the somewhat irregular pathobiological characteristics observed in experimentally infected animals, suggests that although the virus may be of swine origin, significant viral evolution may still be ongoing.

Sequence analysis of travel-related SARS-CoV-2 cases in the Greater Geelong region, Australia

Publisher: Cold Spring Harbor Laboratory

Date: 09-06-2020

DOI: 10.1101/2020.06.08.20125898

Abstract: This study reports the sequence analysis of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) from infected in iduals within the Greater Geelong region, Victoria, Australia. All but one in idual had recently returned from travelling abroad, and all had clinical signs consistent with SARS-CoV-2 infection. SARS-CoV-2 belonging to three lineages were detected and represent separate introductions of the virus into the region. Sequence data were consistent with the recent travel history of each case. Full virus genome sequencing can play an important role in supporting local epidemiological tracing and monitoring for community transmission. Quality of the SARS-CoV-2 sequences obtained was highly dependent on appropriate s le collection and handling.

Purification and characterization of the major nonstructural protein (NS- 1) of Aleutian mink disease parvovirus

Publisher: American Society for Microbiology

Date: 03-1995

DOI: 10.1128/JVI.69.3.1802-1809.1995

Abstract: We have previously described the expression of the major nonstructural protein (NS-1) of Aleutian mink disease parvovirus (ADV) in insect cells by using a baculovirus vector (J. Christensen, T. Storgaard, B. Bloch, S. Alexandersen, and B. Aasted, J. Virol. 67:229-238, 1993). To study its biochemical properties, ADV NS-1 was expressed in Sf9 insect cells and purified to apparent homogeneity with a combination of nuclear extraction, Zn2+ ion chromatography, and immunoaffinity chromatography on monoclonal antibodies. The purified protein showed ATP binding and ATPase- and ATP- or dATP-dependent helicase activity requiring either Mg2+ or Mn2+ as a cofactor. The ATPase activity of NS-1 was efficiently stimulated by single-stranded DNA and, to a lesser extent, double-stranded DNA. We also describe the expression, purification, and characterization of a mutant NS-1 protein, in which a lysine in the putative nucleotide binding consensus sequence of the molecule was replaced with serine. The mutated NS-1 was expressed at 10-fold higher levels than wild-type NS-1, but it exhibited no ATP binding. ATPase, or helicase activity. The availability of large amounts of purified functional NS-1 protein will facilitate studies of the biochemistry of ADV replication and gene regulation leading to disease in mink.

Nucleotide sequence of the 5'-terminal palindrome of Aleutian mink disease parvovirus and construction of an infectious molecular clone

Publisher: American Society for Microbiology

Date: 07-1990

DOI: 10.1128/JVI.64.7.3551-3556.1990

Abstract: The 5'-terminal palindrome of the ADV-G strain of Aleutian mink disease parvovirus (ADV) was molecularly cloned and sequenced. A full-length molecular clone of ADV-G, denoted pXVB, was then constructed. When this clone was transfected into cell cultures, infectious ADV could be rescued. Virus derived from pXVB was nonpathogenic for adult mink, as is the parent ADV-G strain.

Replication and Clearance of Respiratory Syncytial Virus

Publisher: Elsevier BV

Date: 12-2002

DOI: 10.1016/S0002-9440(10)64496-3

Relative resistance of pigs to infection by natural aerosols of FMD virus.

Publisher: Wiley

Date: 05-2001

DOI: 10.1136/VR.148.19.600

Severe Human Case of Zoonotic Infection with Swine-Origin Influenza A Virus, Denmark, 2021

Publisher: Centers for Disease Control and Prevention (CDC)

Date: 12-2022

DOI: 10.3201/EID2812.220935

Abstract: During routine surveillance at the National Influenza Center, Denmark, we detected a zoonotic swine influenza A virus in a patient who became severely ill. We describe the clinical picture and the genetic characterization of this variant virus, which is distinct from another variant found previously in Denmark.

Novel reverse transcription loop-mediated isothermal amplification for rapid detection of foot-and-mouth disease virus

Publisher: Springer Science and Business Media LLC

Date: 02-02-2006

DOI: 10.1007/S00705-005-0708-5

Abstract: Speed is paramount in the diagnosis of foot-and-mouth disease (FMD) and simplicity is required if a test is to be deployed in the field. The development of a one-step, reverse transcription loop-mediated lification (RT-LAMP) assay enables FMD virus (FMDV) to be detected in under an hour in a single tube without thermal cycling. A fragment of the 3D RNA polymerase gene of the virus is lified at 65 degrees C in the presence of a primer mixture and both reverse transcriptase and Bst DNA polymerase. Compared with real-time RT-PCR, RT-LAMP was consistently faster, and ten copies of FMDV transcript were detected in twenty-two minutes. Amplification products were detected by visual inspection, agarose gel electrophoresis, or in real-time by the addition of a fluorescent dye. The specificity of the reaction was demonstrated by the absence of lification of RNA from other viruses that cause vesicular diseases and from that of genetically related picornaviruses. Diagnostic sensitivity was validated by the lification of reference FMDV strains and archival material from field cases of FMD. In comparison with the performance of the established diagnostic TaqMan assay, RT-LAMP appears to be sensitive, rapid, specific, and cost-effective, with the potential for field deployment and use by developing countries for FMDV surveillance.

41. Detection of swine vesicular disease virus genome by fluorogenic RT-PCR assay

Publisher: Elsevier BV

Date: 04-2002

DOI: 10.1016/S0034-5288(02)90043-3

Genetic characterisation of the recent foot-and-mouth disease virus subtype A/IRN/2005

Publisher: Springer Science and Business Media LLC

Date: 15-11-2007

DOI: 10.1186/1743-422X-4-122

Abstract: According to the World Reference Laboratory for FMD, a new subtype of FMDV serotype A was detected in Iran in 2005. This subtype was designated A/IRN/2005, and rapidly spread throughout Iran and moved westwards into Saudi Arabia and Turkey where it was initially detected from August 2005 and subsequently caused major disease problems in the spring of 2006. The same subtype reached Jordan in 2007. As part of an ongoing project we have also detected this subtype in Pakistan with the first positive s les detected in April 2006. To characterise this subtype in detail, we have sequenced and analysed the complete coding sequence of three subtype A/IRN/2005 isolates collected in Pakistan in 2006, the complete coding sequence of one subtype A/IRN/2005 isolate collected during the first outbreak in Turkey in 2005 and, in addition, the partial 1D coding sequence derived from 4 epithelium s les and 34 swab-s les from Asian buffaloes or cattle subsequently found to be infected with the A/IRN/2005 subtype. The phylogenies of the genome regions encoding for the structural proteins, displayed, with the exception of 1A, distinct, serotype-specific clustering and an evolutionary relationship of the A/IRN/2005 sublineage with the A22 sublineage. Potential recombination events have been detected in parts of the genome region coding for the non-structural proteins of FMDV. In addition, amino acid substitutions have been detected in the deduced VP1 protein sequence, potentially related to clinical or subclinical outcome of FMD. Indications of differential susceptibility for developing a subclinical course of disease between Asian buffaloes and cattle have been detected. Furthermore, hitherto unknown insertions of 2 amino acids before the second start codon, as well as sublineage specific amino acids have been detected in the genome region encoding for the leader proteinase of A/IRN/2005 sublineage. Our findings indicate that the A/IRN/2005 sublineage has undergone two different paths of evolution for the structural and non-structural genome regions. The structural genome regions have had their evolutionary starting point in the A22 sublineage. It can be assumed that, due to the quasispecies structure of FMDV populations and the error-prone replication process, advantageous mutations in a changed environment have been fixed and lead to the occurrence of the new A/IRN/2005 sublineage. Together with this mechanism, recombination within the non-structural genome regions, potentially modifying the virulence of the virus, may be involved in the success of this new sublineage. The possible origin of this recombinant virus may be a co-infection with Asia1 and a serotype A precursor of the A/IRN/2005 sublineage potentially within Asian Buffaloes, as these appears to relatively easy become infected, but usually without developing clinical disease and consequently showing not a strong acute inflammatory immune response against a second FMDV infection.

The phosphoprotein gene of a dolphin morbillivirus isolate exhibits genomic variation at the editing site

Publisher: Microbiology Society

Date: 12-1995

DOI: 10.1099/0022-1317-76-12-3051

Abstract: The nucleotide sequence of the phosphoprotein (P) gene of a dolphin morbillivirus (DMV) isolate was determined. Like those of other morbilliviruses the DMV P gene encoded P and C proteins in overlapping open reading frames and V protein by editing the P gene transcript. Among P mRNA based clones the editing site variants GGGC, GGGG, GAGC and GGGGGGC predicting a P protein, and the variants GGGGC and GGGGGG predicting a V protein, were found. Surprisingly, the three variants GGGC, GGGG and GAGC were also found among clones generated from genomic RNA of the DMV isolate. Thus, more than one viral genome type appeared to be present in cells infected with the DMV isolate. By a similar analysis of the virus genomes in the tissue from which the DMV isolate was obtained, only the GGGC type was found, indicating that the GGGG and GAGC types arose during adaptation of the virus to growth in cell cultures. No editing site variants likely to have arisen by editing the GAGC type were encountered, and it remains ot be determined whether mRNA encoding V protein can be transcribed from genomes with this editing site. Using antisera raised against the common N terminus and unique C termini of the predicted P and V proteins, the in vivo expression of these proteins was demonstrated.

Molecular characterization of pandemic H1N1 influenza viruses isolated from Turkeys and pathogenicity of a human pH1N1 isolate in Turkeys

Publisher: American Association of Avian Pathologists (AAAP)

Date: 12-2010

DOI: 10.1637/9422-061410-REG.1

Passive transfer of antiviral antibodies restricts replication of Aleutian mink disease parvovirus in vivo.

Publisher: American Society for Microbiology

Date: 1989

DOI: 10.1128/JVI.63.1.9-17.1989

Abstract: When mink kits were infected neonatally with a highly virulent strain of Aleutian disease virus (ADV), 100% of both Aleutian and non-Aleutian genotype mink died of interstitial pneumonia characterized by permissive ADV infection of alveolar type II cells. Treatment of infected kits with either mink anti-ADV gamma globulin or mouse monoclonal antibodies against ADV structural proteins reduced mortality by 50 to 75% and drastically reduced the severity of clinical signs. Interestingly, mink kits that survived the acute pulmonary disease all developed the chronic form of immune complex-mediated Aleutian disease. Thus, the antibodies directed against ADV structural proteins were capable of modulating the in vivo pathogenicity from an acute fulminant disease to a chronic immune complex-mediated disorder. The mechanism of this modulation was examined by strand-specific in situ hybridization. We found that the number of ADV-infected type II cells was the same in both untreated and antibody-treated kits. However, in the treated kits, viral replication and transcription were restricted at the cellular level. These data suggested that antibodies prevented acute viral pneumonia by restricting the intracellular level of viral replication and that the relevant antigenic determinants were contained within the viral structural proteins. The restricted levels of viral replication and transcription seen in antibody-treated mink kits resembled the levels observed in infected adult mink and suggested a role of antiviral antibodies in development of persistent infection and chronic immune complex disease.

Epidemiology of foot-and-mouth disease in Landhi Dairy Colony, Pakistan, the world largest Buffalo colony

Publisher: Springer Science and Business Media LLC

Date: 29-04-2008

DOI: 10.1186/1743-422X-5-53

Abstract: Foot-and-mouth disease (FMD) is endemic in Pakistan and causes huge economic losses. This work focus on the Landhi Dairy Colony (LDC), located in the suburbs of Karachi. LDC is the largest Buffalo colony in the world, with more than 300,000 animals (around 95% buffaloes and 5% cattle, as well as an unknown number of sheep and goats). Each month from April 2006 to April 2007 we collected mouth-swabs from apparently healthy buffaloes and cattle, applying a convenient s ling based on a two-stage random s ling scheme, in conjunction with participatory information from each selected farm. Furthermore, we also collected epithelium s les from animals with clinical disease, as well as mouth-swabs s les from those farms. In addition, we analysed a total of 180 serum s les randomly collecting 30 s les each month at the local slaughterhouse, from October 2006 to March 2007. S les have been screened for FMDV by real-time RT-PCR and the partial or full 1D coding region of selected isolates has been sequenced. Serum s les have been analysed by applying serotype-specific antibody ELISA and non-structural proteins (NSP) antibody ELISA. FMDV infection prevalence at aggregate level shows an endemic occurrence of FMDV in the colony, with peaks in August 2006, December 2006 and February 2007 to March 2007. A significant association of prevalence peaks to the rainy seasons, which includes the coldest time of the year and the muslimic Eid-festival, has been demonstrated. Participatory information indicated that 88% of all questioned farmers vaccinate their animals. Analysis of the serum s les showed high levels of antibodies for serotypes O, A, Asia 1 and C. The median endpoint-titre for all tested serotypes, except serotype C, in VNT titration is at a serum dilution of equal or above 1/100. All 180 serum s les collected have been tested for antibodies against the non-structural proteins and all but four have been found positive. Out of the 106 swab-s les from apparently healthy and affected animals positive in real-time RT-PCR, we sequenced the partial or full 1D coding region from 58 s les. In addition we sequenced the full 1D coding region of 17 epithelium s les from animals with clinical signs of FMD. From all sequenced s les, swabs and epithelium, 19 belong to the regional PanAsia II lineage of serotype O and 56 to the A/Iran/2005 lineage of serotype A. For an effective and realisable FMD control program in LDC, we suggest to introduce a twice annually mass vaccination of all buffaloes and cattle in the colony. These mass vaccinations should optimally take place shortly before the beginning of the two rainy periods, e.g. in June and September. Those vaccinations should, in our opinion, be in addition to the already in idually performed vaccinations of single animals, as the latter usually targets only newly introduced animals. This suggested combination of mass vaccination of all large ruminants with the already performed in idually vaccination should provide a continuous high level of herd immunity in the entire colony. Vaccines used for this purpose should contain the matching vaccine strains, i.e. as our results indicate antigens for A/Iran/2005 and the regional type of serotype O (PanAsia II), but also antigens of the, in this world region endemic, Asia 1 lineage should be included. In the long term it will be important to control the vaccine use, so that subclinical FMD will be avoided.

Foot-and-Mouth Disease: Host Range and Pathogenesis

Publisher: Springer-Verlag

Date: 2005

DOI: 10.1007/3-540-27109-0_2

Abstract: In this chapter the host range of foot-and-mouth disease (FMD) under natural and experimental conditions is reviewed. The routes and sites of infection, incubation periods and clinical and pathological findings are described and highlighted in relation to progress in understanding the pathogenesis of FMD.

Application of the ceditest® FMDV type O and FMDV-NS enzyme-linked immunosorbent assays for detection of antibodies against Foot-and-mouth disease virus in selected livestock and wildlife species in Uganda

Publisher: SAGE Publications

Date: 29-02-2012

DOI: 10.1177/1040638711435807

Abstract: Diagnosis and control of Foot-and-mouth disease virus (FMDV) requires rapid and sensitive diagnostic tests. Two antibody enzyme-linked immunosorbent assay (ELISA) kits, Ceditest® FMDV-NS for the detection of antibodies against the nonstructural proteins of all FMDV serotypes and Ceditest® FMDV type O for the detection of antibodies against serotype O, were evaluated under African endemic conditions where the presence of multiple serotypes and the use of nonpurified vaccines complicate serological diagnosis. Serum s les from 218 African buffalo, 758 cattle, 304 goats, and 88 sheep were tested using both kits, and selected s les were tested not only in serotype-specific ELISAs for antibodies against primarily FMDV serotype O, but also against other serotypes. The FMDV-NS assay detected far more positive s les (93%) than the FMDV type O assay (30%) in buffalo ( P 0.05), with predominant antibodies against the South African Territories (SAT) serotypes, while the seroprevalence was generally comparable in cattle with antibodies against serotype O elicited by infection and/or vaccination. However, some districts had higher seroprevalence using the FMDV type O assay indicating vaccination without infection, while 1 cattle herd with antibodies against the SAT serotypes had far more positive s les (85%) using the FMDV-NS versus the FMDV type O (10%), consistent with the latter test’s lower sensitivity for antibodies against SAT serotypes. Based on the current investigation, the FMDV type O ELISA may be limited by the presence of SAT serotypes. The FMD NS assay worked well as a screening test for antibodies against all FMDV serotypes present in Uganda however, as long as nonpurified vaccines are applied in the region, this test cannot be used to differentiate between vaccinated and infected animals.

The early pathogenesis of foot-and-mouth disease in pigs infected by contact: a quantitative time-course study using TaqMan RT–PCR

Publisher: Microbiology Society

Date: 04-2001

DOI: 10.1099/0022-1317-82-4-747

Abstract: Foot-and-mouth disease (FMD) is a highly contagious, economically important virus disease of cloven-hoofed animals. The objective of the present study was to examine the early pathogenesis of FMD in pigs by a quantitative time-course study. Under experimental conditions, recipient pigs were infected by contact with donor pigs affected by FMD. Every 24 h from day 1 to day 4 after exposure, two recipient pigs were selected randomly, killed and necropsied. A range of tissues were analysed by a quantitative TaqMan RT–PCR method and by titration of FMD virus on primary bovine thyroid cells. The titres of virus determined by assay in cell culture and calculated from the quantitative TaqMan data correlated strongly ( r ·9), thereby establishing the validity of the TaqMan calculations. The data indicated that the replication of virus in the lungs contributes only in small part to airborne virus excretion. Sites in the pharynx, trachea and nasal mucosa are probably more important in that regard. The sites of earliest virus infection and possibly replication in recipient pigs appeared to be in the pharynx (soft palate, tonsil and floor of pharynx). The data indicated that FMD virus replication in pigs is rapid and that the majority of virus lification occurs in the skin. A model for the progression of infection is proposed, indicating initial spread from the pharyngeal region, through regional lymph nodes and via the blood to epithelial cells, resulting in several cycles of virus lification and spread.

The protective capacity of high payload FMDV A22 IRQ vaccine in sheep against direct-contact challenge with a heterologous, contemporary FMDV A strain from South East Asia

Publisher: Public Library of Science (PLoS)

Date: 18-06-2018

DOI: 10.1371/JOURNAL.PONE.0195302

EXAMINATION OF ALEUTIAN DISEASE VIRUS IN CHARGE‐SHIFT CROSSED IMMUNOELECTROPHORESIS

Publisher: Wiley

Date: 09-1984

DOI: 10.1111/J.1699-0463.1984.TB02842.X

Abstract: The surface properties of Aleutian disease virus were studied by charge‐shift crossed immunoelectrophoresis. When different strains of Aleutian disease virus were treated with non‐charged detergent followed by charged detergents, they showed bi‐directional migration velocity shifts in electrophoresis, indicating hiphilic surface properties of the virus.

59. Quantitative and modelling aspects of foot-and-mouth disease virus infection in pigs

Publisher: Elsevier BV

Date: 2003

DOI: 10.1016/S0034-5288(03)90058-0

Studies of Quantitative Parameters of Virus Excretion and Transmission in Pigs and Cattle Experimentally Infected with Foot-and-Mouth Disease Virus

Publisher: Elsevier BV

Date: 11-2003

DOI: 10.1016/S0021-9975(03)00045-8

Abstract: Foot-and-mouth disease virus (FMDV) can be spread by a variety of mechanisms and the rate of spread, the incubation period and the severity of disease depend on a multitude of parameters, including the strain of virus, the dose received, the route of introduction, the animal species and the husbandry conditions. More knowledge with regard to these parameters is urgently needed to improve resource-efficient disease control. This report describes detailed studies of FMDV load, excretion and transmission in pigs infected with FMDV O UKG 2001, O TAW 1997 and C Noville virus and in cattle infected with the O UKG 2001 virus to facilitate use of a "FMDV load framework" for the assessment of transmission risks. Virus replicated rapidly in pigs and cattle exposed by direct contact. The mean incubation period was around 3-4 days for cattle-to-cattle and 1-3 days for pig-to-pig transmission, depending on the intensity of contact. The results confirmed that a strong relation exists between dose and length of incubation period. Clinical disease was severe in pigs but relatively mild in inoculated cattle contact infection of cattle appeared to increase the severity of lesions. FMDV RNA was recovered in nasal and mouth swabs from inoculated animals soon after they developed a viraemia and probably reflected the early production and excretion of virus. FMDV RNA in nasal and mouth swabs from contact animals could be detected several days before they showed other signs of infection, indicating the possibility of detecting exposed animals during the incubation period. FMDV RNA could also be detected in swab s les after the viraemic phase. This may have represented background environmental virus that had been trapped in the respiratory tract and mouth. Alternatively, it may have indicated a somewhat slower clearance or half-life of viral RNA or an extended low level of FMDV replication at these sites. The pattern of FMDV RNA concentrations in pigs was closely similar to that in cattle, but the amounts of FMDV RNA were higher.

Extent of reduction of foot-and-mouth disease virus RNA load in oesophageal–pharyngeal fluid after peak levels may be a critical determinant of virus persistence in infected cattle

Publisher: Microbiology Society

Date: 02-2004

DOI: 10.1099/VIR.0.19538-0

Abstract: To investigate whether foot-and-mouth disease virus (FMDV) RNA loads in oesophageal–pharyngeal fluid (OP-fluid) in the early course of infection is related to the outcome of virus persistence, viral RNA in OP-fluid s les from cattle experimentally infected with FMDV type O was quantitatively analysed by using a quantitative real-time RT-PCR. Viral RNA was detected within 24 h post-infection (p.i.) in all infected animals. Rapid virus replication led to peak levels of viral RNA load by 30–53 h p.i., and then the load declined at various rates. In some animals ( n =12, so-called non-carriers) viral RNA became undetectable between 7 and 18 days p.i. In contrast, in persistently infected animals ( n =12, so-called carriers) viral RNA persisted in OP-fluid s les at detectable levels beyond 28 days p.i. Analysis of early viral decay/clearance and virus clearance half-life in OP-fluid s les showed that the extent of reduction of viral RNA in OP-fluid s les immediately following peak levels is a critical determinant of the outcome of FMDV persistence.

Expression of Aleutian mink disease parvovirus proteins in a baculovirus vector system

Publisher: American Society for Microbiology

Date: 1993

DOI: 10.1128/JVI.67.1.229-238.1993

Abstract: We have previously published a detailed transcription map of Aleutian mink disease parvovirus (ADV) and proposed a model for the translation of the two virion structural proteins (VP1 and VP2) and three nonstructural proteins (NS-1, NS-2, and NS-3) (S. Alexandersen, M. E. Bloom, and S. Perryman, J. Virol. 62:3684-3994, 1988). To verify and further characterize this model, we cloned the predicted open reading frames for NS-1, NS-2, NS-3, VP1-VP2, and VP2 alone into a recombinant baculovirus and expressed them in Sf9 insect cells. Expression of VP1-VP2 or VP2 alone in cDNA and in the genomic form was achieved. The expressed proteins had molecular weights similar to those of the corresponding proteins of wild-type ADV-G, although the ratio of VP1 to VP2 was altered. The recombinant baculovirus-expressed ADV VP1 and VP2 showed nuclear localization in Sf9 cells and were able to form particles indistinguishable, by electron microscopy, from wild-type virus. The large nonstructural protein, NS-1, showed predominantly nuclear localization in Sf9 cells when analyzed by immunofluorescence and had a molecular weight similar to that of wild-type ADV NS-1. Moreover, expression of NS-1 in Sf9 cells caused a change in morphology of the cells and resulted in 10-times-lower titers of recombinant baculovirus during infection, suggesting a cytostatic or cytotoxic action of this protein. The smaller NS-2 gene product seems to be located in the cytoplasm. When analyzed by Western immunoblotting, NS-2 comigrated with an approximately 16-kDa band seen in lysates of ADV-infected feline kidney cells. The putative NS-3 gene product exhibited a diffuse distribution in Sf9 cells and had a molecular weight of approximately 10,000. All of the expressed ADV-encoded proteins were recognized by sera from ADV-infected mink. Thus, expression of ADV cDNAs allowed assignment of the different mRNAs to the viral proteins observed during ADV infection in cell culture and supported our previously proposed ADV transcriptional and translational scheme. Moreover, the production of structural proteins from a full-length NS-2 mRNA may add to the repertoire of parvovirus gene expression.

Detection and characterisation of coronaviruses in migratory and non-migratory Australian wild birds

Publisher: Springer Science and Business Media LLC

Date: 13-04-2018

DOI: 10.1038/S41598-018-24407-X

Abstract: We evaluated the presence of coronaviruses by PCR in 918 Australian wild bird s les collected during 2016–17. Coronaviruses were detected in 141 s les (15.3%) from species of ducks, shorebirds and herons and from multiple s ling locations. Sequencing of selected positive s les found mainly gammacoronaviruses, but also some deltacoronaviruses. The detection rate of coronaviruses was improved by using multiple PCR assays, as no single assay could detect all coronavirus positive s les. Sequencing of the relatively conserved Orf1 PCR licons found that Australian duck gammacoronaviruses were similar to duck gammacoronaviruses around the world. Some sequenced shorebird gammacoronaviruses belonged to Charadriiformes lineages, but others were more closely related to duck gammacoronaviruses. Australian duck and heron deltacoronaviruses belonged to lineages with other duck and heron deltacoronaviruses, but were almost 20% different in nucleotide sequence to other deltacoronavirus sequences available. Deltacoronavirus sequences from shorebirds formed a lineage with a deltacoronavirus from a ruddy turnstone detected in the United States. Given that Australian duck gammacoronaviruses are highly similar to those found in other regions, and Australian ducks rarely come into contact with migratory Palearctic duck species, we hypothesise that migratory shorebirds are the important vector for moving wild bird coronaviruses into and out of Australia.

Quantitative analysis of foot-and-mouth disease virus RNA loads in bovine tissues: implications for the site of viral persistence

Publisher: Microbiology Society

Date: 09-2004

DOI: 10.1099/VIR.0.80011-0

Abstract: To understand better the pathogenesis of foot-and-mouth disease (FMD), the levels of viral RNA in various bovine tissues during the acute and persistent stages of FMD virus (FMDV) infection were investigated by using quantitative RT-PCR. The viral RNA levels in the tissues examined had peaked by day 1 post-infection (p.i.) and were markedly different among the tissues examined. The epithelium collected from sites of lesion development, i.e. the interdigital area and coronary band on the feet, and the tongue, contained the highest level of viral RNA, indicating the predominant tissue sites of viral infection and lification during the acute stage of infection. Clearance of viral RNA from most of the tissues occurred relatively rapidly and the rate of clearance was largely independent of the level of viral RNA. The viral RNA load in most of the tissues declined slower than in serum, in which viral clearance is rapid. Beyond 28 days p.i., a proportion of pharyngeal region tissues (soft palate, pharynx, tonsil and mandibular lymph node) from infected animals still contained a detectable level of viral RNA, while viral RNA in non-pharyngeal region tissues was generally only detectable for variable periods ranging from 4 to 14 days p.i. The presence of viral RNA in dorsal soft palate tissue had a good correlation with the presence of infectious virus in oesophageal-pharyngeal fluid (OP-fluid) s les, a finding indicative of the specific tissue sites of FMDV persistence.

Serotype specificity of antibodies against foot-and-mouth disease virus in cattle in selected districts in Uganda

Publisher: Hindawi Limited

Date: 02-08-2010

DOI: 10.1111/J.1865-1682.2010.01157.X

Abstract: Uganda had an unusually large number of foot-and-mouth disease (FMD) outbreaks in 2006, and all clinical reports were in cattle. A serological investigation was carried out to confirm circulating antibodies against foot-and-mouth disease virus (FMDV) by ELISA for antibodies against non-structural proteins and structural proteins. Three hundred and forty-nine cattle sera were collected from seven districts in Uganda, and 65% of these were found positive for antibodies against the non-structural proteins of FMDV. A subset of these s les were analysed for serotype specificity of the identified antibodies. High prevalences of antibodies against non-structural proteins and structural proteins of FMDV serotype O were demonstrated in herds with typical visible clinical signs of FMD, while prevalences were low in herds without clinical signs of FMD. Antibody titres were higher against serotype O than against serotypes SAT 1, SAT 2 and SAT 3 in the sera investigated for serotype-specific antibodies. Only FMDV serotype O virus was isolated from one probang s le. This study shows that the majority of the FMD outbreaks in 2006 in the region studied were caused by FMDV serotype O however, there was also evidence of antibodies to both SAT 1 and SAT 3 in one outbreak in a herd inside Queen Elizabeth national park area.

The role of African buffalos (syncerus caffer) in the maintenance of foot-and-mouth disease in Uganda

Publisher: Springer Science and Business Media LLC

Date: 12-2010

DOI: 10.1186/1746-6148-6-54

Abstract: To study the role of African buffalos ( Syncerus caffer ) in the maintenance of foot-and-mouth disease in Uganda, serum s les were collected from 207 African buffalos, 21 impalas ( Aepyceros mel us ), 1 giraffe ( Giraffa camelopardalis ), 1 common eland ( Taurotragus oryx ), 7 hartebeests ( Alcelaphus buselaphus ) and 5 waterbucks ( Kobus ellipsiprymnus ) from four major National Parks in Uganda between 2005 and 2008. Serum s les were screened to detect antibodies against foot-and-mouth disease virus (FMDV) non-structural proteins (NSP) using the Ceditest ® FMDV NS ELISA. Solid Phase Blocking ELISAs (SPBE) were used to determine the serotype-specificity of antibodies against the seven serotypes of FMDV among the positive s les. Virus isolation and sequencing were undertaken to identify circulating viruses and determine relatedness between them. Among the buffalo s les tested, 85% (95% CI = 80-90%) were positive for antibodies against FMDV non-structural proteins while one hartebeest s le out of seven (14.3% 95% CI = -11.6-40.2%) was the only positive from 35 other wildlife s les from a variety of different species. In the buffalo, high serotype-specific antibody titres (≥ 80) were found against serotypes O (7/27 s les), SAT 1 (23/29 s les), SAT 2 (18/32 s les) and SAT 3 (16/30 s les). Among the s les titrated for antibodies against the four serotypes O, SAT 1, SAT 2 and SAT 3, 17/22 (77% CI = 59.4-94.6%) had high titres against at least two serotypes. FMDV isolates of serotypes SAT 1 (1 s le) and SAT 2 (2 s les) were obtained from buffalo probang s les collected in Queen Elizabeth National Park (QENP) in 2007. Sequence analysis and comparison of VP1 coding sequences showed that the SAT 1 isolate belonged to topotype IV while the SAT 2 isolates belonged to different lineages within the East African topotype X. Consistent detection of high antibody titres in buffalos supports the view that African buffalos play an important role in the maintenance of FMDV infection within National Parks in Uganda. Both SAT 1 and SAT 2 viruses were isolated, and serological data indicate that it is also likely that FMDV serotypes O and SAT 3 may be present in the buffalo population. Detailed studies should be undertaken to define further the role of wildlife in the epidemiology of FMDV in East Africa.

Dromedaries (Camelus dromedarius) are of Low Susceptibility to Inoculation with Foot-and-Mouth Disease Virus Serotype O

Publisher: Elsevier BV

Date: 11-2008

DOI: 10.1016/J.JCPA.2008.06.006

Abstract: Two sheep and five dromedaries were inoculated with a highdose of a cattle-passaged type O strain of foot-and-mouth disease virus (FMDV). The sheep developed typical FMD. The inoculated camels, which were placed in contact with five further dromedaries and four sheep, showed no visible signs of illness or vesicular lesions. However, one of them had a raised body temperature at 3 days post-inoculation (pi) and a viraemia from days 2 to 10 probang s les from this animal were negative for infectious virus, but a low level of FMDV RNA was detected in a s le taken on day 6 pi, five other s les taken from days 3 to 28 being negative. Examination of mouth swabs indicated a low level of FMDV RNA at days 1-5 pi in four of the five inoculated camels, but no infectious FMDV or FMDV RNA was detected in serum, probang or mouth swab s les from contact-exposed animals (camels and sheep). All the contact-exposed camels and sheep and two of the inoculated camels were serologically negative for FMD when tested up to day 28. In contrast, the camel with viraemia became serologically positive from day 14, and the other two inoculated camels (which had been exposed to FMDV in an earlier experiment) became serologically positive from day 10. The experiment suggested that dromedaries (1) are of low susceptibility to FMDV serotype O, (2) do not transmit infection, even by close contact, and (3) are unlikely to play a significant epidemiological role in FMD.

Localization of growth factor mRNA in the bovine oviduct

Publisher: Wiley

Date: 10-1993

DOI: 10.1111/J.1439-0531.1993.TB01011.X

The Pathogenesis and Diagnosis of Foot-and-Mouth Disease

Publisher: Elsevier BV

Date: 07-2003

DOI: 10.1016/S0021-9975(03)00041-0

Abstract: The pathogenesis of foot-and-mouth disease (FMD) is reviewed, taking account of knowledge gained from field and experimental studies and embracing investigations at the level of the virus, the cell, the organ, the whole animal and the herd or flock. The review also addresses the immune response and the carrier state in FMD. Progress made in understanding the pathogenesis of the disease is highlighted in relation to developments in diagnosis and methods of control.

Experimental studies with foot-and-mouth disease virus, strain O, responsible for the 2001 epidemic in the United Kingdom

Publisher: Elsevier BV

Date: 06-2002

DOI: 10.1016/S0264-410X(02)00178-0

Abstract: In 2001, the United Kingdom experienced its worst epidemic of foot-and-mouth disease (FMD). To date approximately 3.9 million animals have been culled and direct and indirect revenue losses are probably in excess of pound 12 billion. This study was carried out to investigate the biological characteristics of the FMD virus strain O/UKG/2001 responsible for the epidemic. Animal transmission experiments indicated that this strain is not host restricted and will infect the three main susceptible livestock species (cattle, sheep and pigs). Immunisation with high potency emergency vaccine derived from O(1) Manisa strain of FMD virus protected all three species against clinical disease when challenged with FMD virus strain O/UKG/2001.

An emerging human parechovirus type 5 causing sepsis-like illness in infants in Australia

Publisher: MDPI AG

Date: 03-10-2019

DOI: 10.3390/V11100913

Abstract: Human parechovirus (HPeV), particularly type 3 (HPeV3), is an important cause of sepsis-/meningitis-like illness in young infants. Laboratory records identified a total of ten HPeV-positive cases in Southeastern Australia between January and July 2019. The HPeV present in these cases were typed by Sanger sequencing of the partial viral capsid protein 1 (VP1) region and selected cases were further characterised by additional Sanger or Ion Torrent near-full length virus sequencing. In seven of the ten cases, an HPeV type 5 (HPeV5) was identified, and in the remaining three cases, an HPeV type 1 was identified. The HPeV5-positive cases were infants under the age of 3 months admitted to hospital with fever, rash, lethargy and/or sepsis-like clinical signs. Near full-length virus sequencing revealed that the HPeV5 was most likely a recombinant virus, with structural genes most similar to an HPeV5 from Belarus in 2018, and a polymerase gene most similar to an HPeV3 from Australia in 2013/14. While HPeV5 is not typically associated with severe clinical signs, the HPeV5 identified here may have been able to cause more severe disease in young infants through the acquisition of genes from a more virulent HPeV.

Airborne transmission of foot-and-mouth disease virus from Burnside Farm, Heddon-on-the-Wall, Northumberland, during the 2001 epidemic in the United Kingdom.

Publisher: Wiley

Date: 04-2003

DOI: 10.1136/VR.152.17.525

Abstract: The results of a detailed assessment of the atmospheric conditions when foot-and-mouth disease (FMD) virus was released from Burnside Farm, Heddon-on-the-Wall, Northumberland at the start of the 2001 epidemic in the UK are consistent with the hypothesis that the disease was spread to seven of the 12 farms in the immediate vicinity of the source by airborne virus, and airborne infection could not be ruled out for three other premises the remaining two premises were unlikely to have been infected by airborne virus. The distances involved ranged from less than 1 km up to 9 km. One of the farms which was most probably infected by airborne virus from Burnside Farm was Prestwick Hall Farm, which is believed to have been key to the rapid spread of the disease throughout the country. In contrast, the results of detailed atmospheric modelling, based on a combination of clinical evidence from the field and laboratory experiments have shown that by assuming a relationship between the 24-hour average virus concentrations and subsequent infection, threshold infection levels were seldom reached at the farms close to Burnside Farm. However, significant short-term fluctuations in the concentration of virus can occur, and short-lived high concentrations may have increased the probability of infection and explain this discrepancy.

The effects of gamma interferon on replication of foot-and-mouth disease virus in persistently infected bovine cells

Publisher: Springer Science and Business Media LLC

Date: 10-2002

DOI: 10.1007/S00705-002-0867-6

Abstract: Foot-and-mouth disease virus (FMDV) causes a highly contagious viral disease of cloven-hoofed animals, which has a considerable socio-economic impact on the countries affected. In addition, persistent infection can occur following clinical or sub-clinical disease in either vaccinated or non-vaccinated cattle. The mechanism(s) by which FMDV persistence is established and maintained is not fully understood. To better understand the basic mechanisms controlling the virus infection in cattle, the effects of interferon gamma (IFN-gamma) on the replication of FMDV was evaluated in vitro in persistently infected-epithelial cells isolated from FMDV infected cattle. Initially primary bovine thyroid (BTY) cells were treated with varying doses of bovine recombinant IFN-gamma. The cytokine activity was measured by detection of viral antigen in cell supernatants and viral RNA expression compared with cells without INF-gamma treatment. Pretreatment with IFN-gamma profoundly affected FMDV growth in BTY cells. The replication of FMDV was affected in the presence of more than 2.5 u/ml of IFN-gamma and the effect was both dose-dependent and related to the time of exposure. Analysis of the mechanism of inhibition suggests that IFN-gamma did not inhibit the viral replication through induction of nitric oxide. More interesting is the finding that continuous treatment with IFN-gamma severely restricts FMDV replication or even cures persistently infected bovine epithelial cells, indicating that a cytokine-mediated pathway may be involved in the in vivo clearance of persistent FMDV.

Synovial fluid proteins in degenerative joint disease in dogs

Publisher: Elsevier BV

Date: 09-1989

DOI: 10.1016/0165-2427(89)90060-3

Abstract: Concentrations of three immunoglobulins, albumin, ceruloplasmin, alpha-2 macroglobulin and pregnancy zone protein were estimated by immunoelectrophoresis in paired s les of synovial fluid and serum from 12 dogs with degenerative joint disease (DJD) and six normal dogs. The ratios of synovial fluid to serum concentrations (SF/S) of the four non-immunoglobulins showed an almost inverse linear relationship with their molecular weight in both groups. The SF/S were higher in the DJD synovial fluid than in normal synovial fluid. The difference increased with increasing molecular weight and was highly significant for the largest molecules, reflecting an increased permeability and inflammation in the synovial membrane of DJD joints. The SF/S ratios of the three immunoglobulins studied were compared to the diffusion curves of the four non-immunoglobulins. The SF/S ratios of IgM from dogs with DJD exceeded those calculated from the molecular weights. The present observations support the concept that DJD should be considered an inflammatory disease and suggest that immunologic processes may initiate and/or sustain the inflammation.

Rinderpest virus lineage differentiation using RT-PCR and SNAP-ELISA

Publisher: Elsevier BV

Date: 2003

DOI: 10.1016/S0166-0934(02)00186-6

Abstract: An RT-PCR/ELISA system has been developed that detects and differentiates Rinderpest virus (RPV) from the other closely related morbillivirus of ruminants, Peste des petits Ruminants virus (PPRV). In addition, using lineage specific probes, it is possible to determine whether the virus s le is wild-type or vaccine, and the likely origin of the outbreak if it is wild-type. It involves carrying out a RT-PCR with one digoxygenin (Dig)-labelled primer followed by a hybridisation step with a virus-specific, biotin-labelled, probe. The hybridisation step is carried out in an ELISA format on a streptavidin-coated plate. The DIG-labelled products are detected using a specific anti-DIG monoclonal antibody and an anti-mouse horseradish peroxidase conjugate. The hybridisation step replaces nucleotide sequencing or nested PCR for confirmation of the identity of DNA product. The assay is fast and easy to carry out and can give semi-quantitative estimates of the virus content of s les.

Metagenomic characterisation of additional and novel avian viruses from Australian wild ducks

Publisher: Springer Science and Business Media LLC

Date: 17-12-2020

DOI: 10.1038/S41598-020-79413-9

Abstract: Birds, notably wild ducks, are reservoirs of pathogenic and zoonotic viruses such as influenza viruses and coronaviruses. In the current study, we used metagenomics to detect and characterise avian DNA and RNA viruses from wild Pacific black ducks, Chestnut teals and Grey teals collected at different time points from a single location. We characterised a likely new species of duck aviadenovirus and a novel duck gyrovirus. We also report what, to the best of our knowledge, is the first finding of an avian orthoreovirus from Pacific black ducks and a rotavirus F from Chestnut teals. Other viruses characterised from the s les from these wild ducks belong to the virus families Astroviridae, Caliciviridae and Coronaviridae. Some of the viruses may have potential cross-species transmissibility, while others indicated a wide genetic ersity of duck viruses within a genus. The study also showed evidence of potential transmission of viruses along the East Asian—Australasian Flyway potentially facilitated by migrating shorebirds. The detection and characterisation of several avian viruses not previously described, and causing asymptomatic but potentially also symptomatic infections suggest the need for more virus surveillance studies for pathogenic and potential zoonotic viruses in wildlife reservoirs.

Experimental Transmission of Aleutian Disease virus (ADV) to Different Animal Species

Publisher: Wiley

Date: 09-1985

DOI: 10.1111/J.1699-0463.1985.TB02876.X

Abstract: Two animals from each of 8 different species (mink, Finn raccoon, cat, dog, ferret, blue fox, mouse and rabbit) were inoculated with the highly virulent Utah I strain of ADV. Only the mink developed hypergammaglobulinemia. All animals produced antibodies to ADV antigens, but with different litres. Mink sera had much higher antibody titres than the other animal sera. Antibodies to the ADV‐coded, non‐structural polypeptide (p71) were found in mink, Finn raccoons and dogs only. Presence of this kind of antibodies was taken as evidence of ADV replication, because p71 was not present in the ADV inoculum. The animals were killed 4 weeks after virus inoculation. Homogenates of different organs from mink, Finn raccoons, ferrets, dogs, mice and the cat were shown to infect ADV‐negative mink, which developed antibodies to ADV following inoculation. We conclude from the present studies that mink and Finn raccoons are potential threats as ADV transmitters. Cats, ferrets, dogs and mice may be considered potential ADV reservoirs, because they possibly harbour ADV for 4 weeks or longer. Blue foxes and rabbits did not seem to be a risk factor for ADV transmission.

Sites of replication of bovine respiratory syncytial virus in naturally infected calves as determined by in situ hybridization

Publisher: SAGE Publications

Date: 07-1996

DOI: 10.1177/030098589603300403

Abstract: Replication of bovine respiratory syncytial virus (BRSV) was studied in three naturally infected calves by in situ hybridization using strand-specific RNA probes. One of the calves was a 5-month-old Friesian, the other two calves were a 3-month-old and a 3-week-old Jersey. Two Jersey calves, 3 months and 3 weeks of age, served as controls. Replication of BRSV took place in the luminal lining of the respiratory tract. In one of the BRSV infected animals (calf No. 1), replication was especially seen in the bronchi, whereas in the two other animals (calf Nos. 2 and 3) replication of BRSV was demonstrated in the bronchiolar epithelial cells and in alveolar cells. Syncytia were often observed in the bronchiolar walls and in alveoli and such syncytia were always replicating BRSV. By immunohistochemistry it was possible to demonstrate BRSV antigen at the same location as replication of BRSV was detected. In tissue outside the respiratory tract neither BRSV antigen nor replication of BRSV could be demonstrated.

Metagenomic characterisation of avian parvoviruses and picornaviruses from Australian wild ducks

Publisher: Springer Science and Business Media LLC

Date: 30-07-2020

DOI: 10.1038/S41598-020-69557-Z

Abstract: Ducks can shed and disseminate viruses and thus play a role in cross-species transmission. In the current study, we detected and characterised various avian parvoviruses and picornaviruses from wild Pacific black ducks, Chestnut teals, Grey teals and Wood ducks s led at multiple time points from a single location using metagenomics. We characterised 46 different avian parvoviruses belonging to three different genera Dependoparvovirus, Aveparvovirus and Chaphamaparvovirus, and 11 different avian picornaviruses tentatively belonging to four different genera Sicinivirus , Anativirus , Megrivirus and Aalivirus . Most of these viruses were genetically different from other currently known viruses from the NCBI dataset. The study showed that the abundance and number of avian picornaviruses and parvoviruses varied considerably throughout the year, with the high number of virus reads in some of the duck s les highly suggestive of an active infection at the time of s ling. The detection and characterisation of several parvoviruses and picornaviruses from the in idual duck s les also suggests co-infection, which may lead to the emergence of novel viruses through possible recombination. Therefore, as new and emerging diseases evolve, it is relevant to explore and monitor potential animal reservoirs in their natural habitat.

Temporal assessment of seroconversion in response to inactivated foot-and-mouth disease vaccine in Arabian oryx (Oryx leucoryx).

Publisher: Wiley

Date: 12-2008

DOI: 10.1136/VR.163.24.717

Abstract: Ten male Arabian oryx (Oryx leucoryx) were vaccinated with a commercially available standard aqueous foot-and-mouth-disease vaccine containing aluminium hydroxide as an adjuvant, and their antibody titres against serotypes O and A were measured using solid-phase blocking elisa and the virus neutralisation test. Mean elisa antibody titres greater than 1.45 log(10) were recorded for serotype A, but low elisa titres were recorded for serotype 0 low titres were recorded by VNT for both serotypes.

SARS-CoV-2 genomic and subgenomic RNAs in diagnostic samples are not an indicator of active replication

Publisher: Cold Spring Harbor Laboratory

Date: 05-06-2020

DOI: 10.1101/2020.06.01.20119750

Abstract: Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) emerged in China in late December 2019 and has spread worldwide. Coronaviruses are enveloped, positive sense, single-stranded RNA viruses and employ a complicated pattern of virus genome length RNA replication as well as transcription of genome length and leader containing subgenomic RNAs. Although not fully understood, both replication and transcription are thought to take place in so-called double-membrane vesicles in the cytoplasm of infected cells. We here describe detection of SARS-CoV-2 subgenomic RNAs in diagnostic s les up to 17 days after initial detection of infection, provide evidence for their nuclease resistance and likely protection by cellular membranes consistent with being part of virus-induced replication organelles. Furthermore, we show that the ratios of genomic to subgenomic RNA as well as the ratios of plus to negative strand RNA of genomic and subgenomic RNA are consistent with what have been detected for other coronaviruses in cell culture albeit with the caveat that in vivo diagnostic s les, even in relatively early infection, the ratios of these RNAs are most reminiscent of late culture, semi-purified virus preparations shown to have a relatively constant ratio of genomic to subgenomic RNAs of around 5-10 or higher, while the ratios of positive to negative strands are more than 100 for the genomic RNA and around 20 for the subgenomic RNAs. Overall, our results may help explain the extended PCR positivity of some s les, and may also, at least in part, help explain discrepancies in results of different diagnostic PCR methods described by others in particular for s les with a low virus load or of poor quality. Overall, we present evidence that subgenomic RNAs may not be an indicator of active coronavirus replication/infection, but that these RNAs, similar to the virus genome RNA, may be rather stable, and thus detectable for an extended period, most likely due to their close association with cellular membranes.

Options for decentralized testing of suspected secondary outbreaks of foot-and-mouth disease

Publisher: Hindawi Limited

Date: 05-2010

DOI: 10.1111/J.1865-1682.2010.01141.X

Abstract: This article reviews the options for use of virus detection techniques for decentralized testing of s les from suspected secondary outbreaks of foot-and-mouth disease (FMD). These options have been expanded by the advent of new tests including disposable lateral flow devices (LFDs) that detect viral proteins and portable RT-PCR equipment that detects viral RNA. LFDs have been developed with similar sensitivity to antigen detection ELISA but with the ability to provide a result 1-30 min after the addition of epithelium or vesicular fluid. Portable RT-PCR platforms are being developed that can detect FMD viral RNA in blood, epithelium or other materials with minimal s le processing and with high sensitivity, in as little as 60 min in some cases. These devices may be used on infected farms as pen-side tests, in regional, local or mobile laboratories, or in National Reference Laboratories (NRL). Advantages and disadvantages of different testing options are considered to inform decisions on the optimal strategies for different national circumstances. Issues include validation and quality control, containment needs, availability of test devices and reagents, the decision tree for declaring an outbreak, training issues and provision of s les for subsequent viral characterization. Tests to confirm the diagnosis of the index case of an outbreak of FMD should continue to be carried out in the NRL.

Deakin University

Location: Australia

View Organisation

Statens Serum Institut

Location: Denmark

View Organisation

Aarhus University

Location: Denmark

View Organisation

Geelong Centre For Emerging Infectious Diseases

Location: Australia

View Organisation

Barwon Health

Location: Australia

View Organisation

Discovery Projects - Grant ID: DP180103494

Start Date: 06-2018

End Date: 12-2024

Amount: $309,762.00

Funder: Australian Research Council