Identifying environmental risk factors for louping Ill virus seroprevalence in sheep and the potential to inform wildlife management policy

Identifying the risk factors for disease is crucial for developing policy and strategies for controlling exposure to pathogens. However, this is often challenging, especially in complex disease systems, such as vector-borne diseases with multiple hosts and other environmental drivers. Here we combine seroprevalence data with GIS-based environmental variables to identify the environmental risk factors associated with an endemic tick-borne pathogen—louping ill virus—in sheep in Scotland. Higher seroprevalences were associated with (i) upland/moorland habitats, in accordance with what we predicted from the habitat preferences of alternative LIV transmission hosts (such as red grouse), (ii) areas of higher deer density, which supports predictions from previous theoretical models, since deer are the key Ixodes ricinus tick reproduction host in this system, and (iii) a warmer climate, concurring with our current knowledge of how temperature affects tick activity and development rates. The implications for policy include adopting increased disease management and awareness in high risk habitats and in the presence of alternative LIV hosts (e.g., grouse) and tick hosts (especially deer). These results can also inform deer management policy, especially where there may be conflict between contrasting upland management objectives, for example, revenue from deer hunting vs. sheep farmers

Bovine viral diarrhoea virus loses quasispecies diversity rapidly in culture

Bovine viral diarrhoea (BVD) is an important disease of cattle, with significant impacts on animal health and welfare. The wide host range of the causative pestiviruses may lead to formation of virus reservoirs in other ruminant or wildlife species, presenting a concern for the long-term success of BVD eradication campaigns. It is likely that the quasispecies nature of these RNA viruses contributes to their interspecies transmission by providing genetic plasticity. Understanding the spectrum of sequence variants present in persistently infected (PI) animals is, therefore, essential for studies of virus transmission. To analyse quasispecies diversity without amplification bias, we extracted viral RNA from the serum of a PI cow, and from cell culture fluid after three passages of the same virus in culture, to produce cDNA without amplification. Sequencing of this material using Illumina 250 bp paired-read technology produced full-length virus consensus sequences from both sources and demonstrated the quasispecies diversity of this pestivirus A genotype 1a field strain within serum and after culture. We report the distribution and diversity of over 800 SNPs and provide evidence for a loss of diversity after only three passages in cell culture, implying that cultured viruses cannot be used to understand quasispecies diversity and may not provide reliable molecular markers for source tracing or transmission studies. Additionally, both serum and cultured viruses could be sequenced as a set of 25 overlapping PCR amplicons that demonstrated the same consensus sequences and the presence of many of the same quasispecies variants. The observation that aspects of the quasispecies structure revealed by massively parallel sequencing are also detected after PCR and Sanger sequencing suggests that this approach may be useful for small or difficult to analyse samples

Demonstration of early functional compromise of bone marrow derived hematopoietic progenitor cells during bovine neonatal pancytopenia through in vitro culture of bone marrow biopsies

<p>Abstract</p> <p>Background</p> <p>Bovine neonatal pancytopenia (BNP) is a syndrome characterised by thrombocytopenia associated with marked bone marrow destruction in calves, widely reported since 2007 in several European countries and since 2011 in New Zealand. The disease is epidemiologically associated with the use of an inactivated bovine virus diarrhoea (BVD) vaccine and is currently considered to be caused by absorption of colostral antibody produced by some vaccinated cows (“BNP dams”). Alloantibodies capable of binding to the leukocyte surface have been detected in BNP dams and antibodies recognising bovine MHC class I and β-2-microglobulin have been detected in vaccinated cattle. In this study, calves were challenged with pooled colostrum collected from BNP dams or from non-BNP dams and their bone marrow hematopoietic progenitor cells (HPC) cultured <it>in vitro</it> from sternal biopsies taken at 24 hours and 6 days post-challenge.</p> <p>Results</p> <p>Clonogenic assay demonstrated that CFU-GEMM (colony forming unit-granulocyte/erythroid/macrophage/megakaryocyte; pluripotential progenitor cell) colony development was compromised from HPCs harvested as early as 24 hour post-challenge. By 6 days post challenge, HPCs harvested from challenged calves failed to develop CFU-E (erythroid) colonies and the development of both CFU-GEMM and CFU-GM (granulocyte/macrophage) was markedly reduced.</p> <p>Conclusion</p> <p>This study suggests that the bone marrow pathology and clinical signs associated with BNP are related to an insult which compromises the pluripotential progenitor cell within the first 24 hours of life but that this does not initially include all cell types.</p

Assessing Needs and Outcomes of Children and Youth Receiving Intensive Services

This study investigated whether children/youth in Ontario triaged to residential services showed a higher intensity of need than those referred to outpatient services, and whether residential treatment gains were sufficient for transition to community services. Participants included 2053 children/youth assessed at 23 diverse mental health agencies across Ontario using the interRAI™ Child and Youth Mental Health (ChYMH) instrument. Various presenting problems were examined utilizing scales including: Disruptive/Aggressive Behavior, Hyperactive/Distraction, Social Disengagement, Anxiety, and Sleep Difficulties. Analyses were conducted separately for boys and girls. Notable differences were found in the initial assessment, with residential boys scoring higher on all scales than outpatient boys, and residential girls scoring higher on the externalizing scales (Disruptive/Aggressive Behavior, Hyperactive/Distraction) than outpatient girls. Treatment gains at residential discharge included improvements in Anxiety, Social Disengagement, Hyperactive/Distraction and Sleep Difficulties for boys and girls to levels at or below the initial scores of outpatient peers. Disruptive/Aggressive Behavior is still a high need following residential services. The results highlight differences in severity of mental health presentation between children/youth receiving residential and outpatient services, and how multiple agencies in Ontario are providing services that successfully reduce the severity of mental health needs

One-step multiplex real time RT-PCR for the detection of bovine respiratory syncytial virus, bovine herpesvirus 1 and bovine parainfluenza virus 3

<p>Abstract</p> <p>Background</p> <p>Detection of respiratory viruses in veterinary species has traditionally relied on virus detection by isolation or immunofluorescence and/or detection of circulating antibody using ELISA or serum neutralising antibody tests. Multiplex real time PCR is increasingly used to diagnose respiratory viruses in humans and has proved to be superior to traditional methods. Bovine respiratory disease (BRD) is one of the most common causes of morbidity and mortality in housed cattle and virus infections can play a major role. We describe here a one step multiplex reverse transcriptase quantitative polymerase chain reaction (mRT-qPCR) to detect the viruses commonly implicated in BRD.</p> <p>Results</p> <p>A mRT-qPCR assay was developed and optimised for the simultaneous detection of bovine respiratory syncytial virus (BRSV), bovine herpes virus type 1 (BoHV-1) and bovine parainfluenza virus type 3 (BPI3 i & ii) nucleic acids in clinical samples from cattle. The assay targets the highly conserved glycoprotein B gene of BoHV-1, nucleocapsid gene of BRSV and nucleoprotein gene of BPI3. This mRT-qPCR assay was assessed for sensitivity, specificity and repeatability using <it>in vitro </it>transcribed RNA and recent field isolates. For clinical validation, 541 samples from clinically affected animals were tested and mRT-qPCR result compared to those obtained by conventional testing using virus isolation (VI) and/or indirect fluorescent antibody test (IFAT).</p> <p>Conclusions</p> <p>The mRT-qPCR assay was rapid, highly repeatable, specific and had a sensitivity of 97% in detecting 10²copies of BRSV, BoHV-1 and BPI3 i & ii. This is the first mRT-qPCR developed to detect the three primary viral agents of BRD and the first multiplex designed using locked nucleic acid (LNA), minor groove binding (MGB) and TaqMan probes in one reaction mix. This test was more sensitive than both VI and IFAT and can replace the aforesaid methods for virus detection during outbreaks of BRD.</p

Pancreatitis associated with azathioprine and 6-mercaptopurine use in Crohn’s disease: A systematic review

Background Thiopurines are proven agents in the treatment of Crohn’s disease. While pancreatitis is recognised as an adverse event associated with therapy, the effect-size and morbidity of thiopurine-induced pancreatitis is not known. The aim of this systematic review and meta-analysis was to quantify the risk of pancreatitis with azathioprine and 6-mercaptopurine within Crohn’s disease. Methods We searched six electronic databases from inception to 29th October 2019. The primary outcomes measures were the occurrence of pancreatitis. We calculated pooled odds ratio with corresponding 95% confidence intervals for risk of pancreatitis. A number needed to harm analysis was performed. Results The search identified 4,418 studies, of which 25 RCTs met the criteria for inclusion. The number of patients treated with azathioprine to cause an episode of pancreatitis was 36 (induction of remission) and 31 (maintenance of remission).The risk of pancreatitis in patients receiving azathioprine across all contexts was 3.80%, compared to a control risk of 0.2% (placebo) and 0.5% (5-ASA agents). There was no difference seen between 6-MP and placebo, although this was a low certainty result due to imprecision from very low event numbers and patient numbers. Conclusion There is a probably increased occurrence of pancreatitis when azathioprine is used in Crohn’s disease (moderate certainty), with incidence overall approximately 3.8%. Most cases are mild and resolve on cessation of therapy and no mortality was reported. There was no increased occurrence seen when using 6-MP, although this is a low certainty finding

BSE situation and establishment of Food Safety Commission in Japan

Eight major policies were implemented by Japanese Government since Oct. 2001, to deal with bovine spongiform encephalopathy (BSE). These are; 1) Surveillance in farm by veterinarian, 2) Prion test at healthy 1.3mi cows/yr, by veterinarian, 3) Elimination of specified risk material (SRM), 4) Ban of MBM for production, sale use, 5) Prion test for fallen stocks, 6) Transparent information and traceability, 7) New Measures such as Food Safety Basic Law, and 8) Establish of Food Safety Commission in the Cabinet Office. At this moment, the extent of SRM risk has only been indicated by several reports employing tests with a limited sensitivity. There is still a possibility that the items in the SRM list will increase in the future, and this indiscriminately applies to Japanese cattle as well. Although current practices of SRM elimination partially guarantee total food safety, additional latent problems and imminent issues remain as potential headaches to be addressed. If the index of SRM elimination cannot guarantee reliable food safety, we have but to resort to total elimination of tissues from high risk-bearing and BSE-infected animals. However, current BSE tests have their limitations and can not yet completely detect high-risk and/or infected animals. Under such circumstances, tissues/wastes and remains of diseased, affected fallen stocks and cohort animals have to be eliminated to prevent BSE invading the human food chain systems. The failure to detect any cohort should never be allowed to occur, and with regular and persistent updating of available stringent records, we are at least adopting the correct and useful approach as a reawakening strategy to securing food safety. In this perspective, traceability based on a National Identification System is required