Serological Epidemiological Survey Of Bovine Diarrhea In Large-Scale Cattle Farms In Inner Mongolia, China from 2019 to 2022 (preprint)

Bovine diarrhea is a multi-factorial disease, and remains one of the most serious health problems in cattle husbandry. However, the endemic trends of the main pathogens responsible for bovine diarrhea have not been analyzed before, therefore, the purpose of this study was to estimate the prevalence of bovine diarrhea pathogens in the large-scale cattle farms of Inner Mongolia in China, we assessed the differences in the prevalence of infection based on age and area, as well as determined local prevalence rates and the rates of mixed infections. Using stratified random sampling strategy, cattle were divided into four age stages (0-2 months, 3-6 months, 7-18 months and ＞18 months), 2,216 serum samples were collected from 38 cattle herds in 12 leagues and cities in Inner Mongolia of China. Positive rates of Bovine Viral Diarrhoea Virus (BVDV), Bovine Rotavirus (BRV), Bovine Coronavirus (BCoV), Escherichia coli K99 (E.coli K99), and M. paratuberculosis (Mptb) antibodies were detected in the samples by ELISA to determine the epidemic trends and differences of the five pathogens in Inner Mongolia. The positive rate of BVDV vaccine higher than 90% in each age category of cattle, and the prevalence rates of BRV and Mptb in other ages significantly higher than ＞18 months (P <0.05). Except BVDV, the prevalence of BCoV was the highest in 0-2 months old (74.88%), the highest positive rate in ＞18 months old is BRV (63.31%), the infection of viral pathogens were serious in Inner Mongolia. With the highest mixed infection (33.74%) being BRV and BCoV in the category of 0-2 months, and there was a mixed infection of BRV and BCoV at four age stages, Only double and triple infections were detected in cattle aged 2-6 months and 6-18 months, quadruple infections were detected in cattle aged 0-2 months and ＞18 months. Furthermore, obvious regional epidemiological differences among the five pathogens were observed, 5 pathogens were detected in 11 regions, Hulun Buir city was seriously infected with bacterial pathogens, only antibodies to viral pathogens were detected in Ulanqab city. All the above detected pathogens can trigger high mortality and poor prognosis in cattle. This study is the first to report the prevalence and characteristics of the main pathogens of bovine diarrhea in large-scale cattle farms in Inner Mongolia, China, which has practical significance for the future prevention and control of bovine diarrhea and reduce economic losses in cattle farming.

[Genetic diversity of Siberian bovine coronavirus isolates (Coronaviridae: Coronavirinae: Betacoronavirus-1: Bovine-Like coronaviruses)].

INTRODUCTION: Bovine coronaviruses (BCoVs) are causative agents of diarrhea, respiratory diseases in calves and winter cow dysentery. The study of genetic diversity of these viruses is topical issue. The purpose of the research is studying the genetic diversity of BCoV isolates circulating among dairy cattle in Siberia. MATERIALS AND METHODS: Specimens used in this study were collected from animals that died or was forcedly slaughtered before the start of the study. The target for amplification were nucleotide sequences of S and N gene regions. RESULTS: Based on the results of RT-PCR testing, virus genome was present in 16.3% of samples from calves with diarrheal syndrome and in 9.9% with respiratory syndrome. The nucleotide sequences of S gene region were determined for 18 isolates, and N gene sequences - for 12 isolates. Based on S gene, isolates were divided into two clades each containing two subclades. First subclade of first clade (European line) included 11 isolates. Second one included classic strains Quebec and Mebus, strains from Europe, USA and Korea, but none of sequences from this study belonged to this subclade. 6 isolates belonged to first subclade of second clade (American-Asian line). Second subclade (mixed line) included one isolate. N gene sequences formed two clades, one of them included two subclades. First subclade included 3 isolates (American-Asian line), and second subclade (mixed) included one isolate. Second clade (mixed) included 8 sequences. No differences in phylogenetic grouping between intestinal and respiratory isolates, as well as according to their geographic origin were identified. CONCLUSION: The studied population of BCoV isolates is heterogeneous. Nucleotide sequence analysis is a useful tool for studying molecular epidemiology of BCoV. It can be beneficial for choice of vaccines to be used in a particular geographic region.

Detection of Influenza D-Specific Antibodies in Bulk Tank Milk from Swedish Dairy Farms.

Influenza D virus (IDV) has been detected in bovine respiratory disease (BRD) outbreaks, and experimental studies demonstrated this virus's capacity to cause lesions in the respiratory tract. In addition, IDV-specific antibodies were detected in human sera, which indicated that this virus plays a potential zoonotic role. The present study aimed to extend our knowledge about the epidemiologic situation of IDV in Swedish dairy farms, using bulk tank milk (BTM) samples for the detection of IDV antibodies. A total of 461 and 338 BTM samples collected during 2019 and 2020, respectively, were analyzed with an in-house indirect ELISA. In total, 147 (32%) and 135 (40%) samples were IDV-antibody-positive in 2019 and 2020, respectively. Overall, 2/125 (2%), 11/157 (7%) and 269/517 (52%) of the samples were IDV-antibody-positive in the northern, middle and southern regions of Sweden. The highest proportion of positive samples was repeatedly detected in the south, in the county of Halland, which is one of the counties with the highest cattle density in the country. In order to understand the epidemiology of IDV, further research in different cattle populations and in humans is required.

Evolutionary adaptation of bovine coronavirus (BCoV): Screening of natural recombinations across the complete genomes.

Bovine coronavirus (BCoV) is a member of pathogenic Betacoronaviruses that has been circulating for several decades in multiple host species. Given the similarity between BCoV and human coronaviruses, the current study aimed to review the complete genomes of 107 BCoV strains available on the GenBank database, collected between 1983 and 2017 from different countries. The maximum-likelihood based phylogenetic analysis revealed three main BCoV genogroups: GI, GII, and GIII. GI is further divided into nine subgenogroups: GI-a to GI-i. The GI-a to GI-d are restricted to Japan, and GI-e to GI-i to the USA. The evolutionary relationships were also inferred using phylogenetic network analysis, revealing two major distinct networks dominated by viruses identified in the USA and Japan, respectively. The USA strains-dominated Network Cluster includes two sub-branches: France/Germany and Japan/China in addition to the United States, while Japan strains-dominated Network Cluster is limited to Japan. Twelve recombination events were determined, including 11 intragenogroup (GI) and one intergenogroup (GII vs. GI-g). The breakpoints of the recombination events were mainly located in ORF1ab and the spike glycoprotein ORF. Interestingly, 10 of 12 recombination events occurred between Japan strains, one between the USA strains, and one from intercontinental recombination (Japan vs. USA). These findings suggest that geographical characteristics, and population density with closer contact, might significantly impact the BCoV infection and co-infection and boost the emergence of more complex virus lineages.

Association of ovine gammaherpesvirus 2 with an outbreak of acute respiratory disease in dairy cattle.

This study investigated the cause of an outbreak of an acute respiratory disease syndrome followed by episodes of diarrhea in a dairy cattle herd from Southern Brazil. Deep nasal swabs (DNS) from asymptomatic calves, calves with pulmonary discomfort, and diarrheic calves after episodes of respiratory distress were used in molecular assays designed to detect the principal pathogens associated with bovine respiratory disease (BRD). Fecal samples were used for the molecular detection of bovine enteric disease agents. Pulmonary tissues from three calves and a cow that died were evaluated by molecular assays to identify 11 agents associated with the development of BRD. The intestinal and pulmonary fragments of one calf and the cow revealed atrophic enteritis and interstitial pneumonia by histopathology, respectively. Immunohistochemistry (IHC) identified intralesional antigens of a malignant catarrhal fever virus, genus Macavirus, within epithelial cells of the lungs and intestines. Molecular assays amplified ovine gammaherpesvirus 2 (OvGHV2) from most of the DNS, and the pulmonary and intestinal fragments from the animals that died, confirming that the Macavirus identified by IHC was OvGHV2. Concomitant pulmonary infections of OvGHV2 with bovine gammaherpesvirus 6 and bovine coronavirus were identified. Additionally, bovine viral diarrhea virus 1b and Aichivirus B were detected in the fecal samples. These findings demonstrated that OvGHV2, a Macavirus, was the disease agent most frequently (81.2%; 13/16) associated with singular pulmonary infections during this outbreak of BRD, suggesting that this virus may be another potential agent of respiratory disease of cattle.

Bronchopneumonia with interstitial pneumonia in beef feedlot cattle: Characterization and laboratory investigation.

Bronchopneumonia with interstitial pneumonia (BIP) has been considered a variant of acute interstitial pneumonia (AIP) rather than a distinct disease. This study compared 18 BIP, 24 bronchopneumonia (BP), and 13 AIP cases in feedlot beef cattle. Grossly, BIP cases typically had cranioventral lung lesions of similar morphology and extent as BP cases, but the caudodorsal lung appeared overinflated, bulged on section, and had interlobular edema and emphysema. Gross diagnosis of BIP had 83% sensitivity and 73% specificity relative to histopathology. Histologic lesions of BIP in cranioventral areas were of chronic BP, while caudodorsal lesions included alveolar and bronchiolar damage and inflammation, interstitial hypercellularity, and multifocal hemorrhages. In BIP cases, cranioventral lung lesions were more chronic than caudodorsal lesions. Histologic scores and microbiology data were comparable in cranioventral lung of BIP versus BP cases and caudodorsal lung of BIP versus AIP cases, with differences reflecting a more chronic disease involving less virulent bacteria in BIP versus BP. Mycoplasma bovis infection was similarly frequent among groups, and a viral cause of BIP was not identified. Lesion morphology and similar blood cytokine concentrations among groups argued against sepsis as a cause of lung injury. Surfactant dysfunction was identified in BIP and BP, and was only partially the result of protein exudation. These and other findings establish BIP as a distinct condition in which chronic cranioventral BP precedes acute caudodorsal interstitial lung disease, supporting a role of chronic inflammation in heightened sensitivity to 3-methylindole or another lung toxicant.

Multiplex RT-qPCR Application in Early Detection of Bovine Respiratory Disease in Healthy Calves.

Bovine respiratory diseases (BRD) are associated with various predisposing factors, such as physical and physiological stress factors, and bacterial and viral pathogens. These stressors and viruses suppress immune defenses, leading to bacterial growth in the upper respiratory tract and invasion of pathogens into the lower respiratory tract. Therefore, continuous monitoring of the causative pathogens would contribute to the early detection of BRD. Nasal swabs and sera from 63 clinically healthy calves were continuously collected from seven farms in Iwate prefecture from 2019 to 2021. We attempted to monitor dynamics of BRD-associated pathogens by multiplex real-time RT-PCR (RT-qPCR) using their nasal swab samples. In addition, we attempted to monitor fluctuation of antibody titers against each BRD-associated pathogen by virus neutralization test (VNT) using their sera. In contrast, nasal swabs from 89 calves infected with BRD were collected from 28 farms in Iwate prefecture from 2019 to 2021. We attempted to analyze their nasal swab samples by multiplex RT-qPCR aim to detect BRD-associated pathogens that are dominant in this region. As a result, our analyses using samples from clinically healthy calves showed that positive results by multiplex RT-qPCR were closely related to a significant increase of antibody titers by VNT in bovine coronavirus (BCoV), bovine torovirus (BToV), and bovine respiratory syncytial virus (BRSV). In addition, our data exhibited that BCoV, BToV, BRSV, bovine parainfluenza virus 3, and Mycoplasma bovis have been more frequently detected in calves infected with BRD compared to those detected in clinically healthy calves. Moreover, the data presented herein revealed co-infections by combination multiple viral pathogens with bacterial pathogens are closely involved in the onset of BRD. Taken together, our study demonstrates multiplex RT-qPCR which can simultaneously analyze multiple pathogens, including viruses and bacteria, and is useful for the early detection of BRD.

Epidemic of lumpy skin disease in Pakistan.

Lumpy skin disease (LSD) is a viral disease that affects farm animals including water buffalo. It is caused by the contagious LSD virus, a member of the Poxiviridae family's Capripox genus. Skin sores are thought to be the most common site of infection since the virus may live for lengthy periods in lesions or scabs. The first clinical indications of LSD were described in Zambia, in 1929. Pakistan has also been afflicted by LSD, with a high number of animals infected at many cattle ranches in Karachi, 190,000 cases of LSD have been reported nationwide, with greater than 7500 deaths attributable to the illness. LSD has a huge influence on Pakistan's economic status, resulting in the loss of cattle and a decrease in milk output. The Ministry of Research and National Food Safety in Pakistan has formed a working group to create a framework for controlling the spread of LSD in cattle and buffalo. Official and private veterinarians, both field and slaughterhouse, veterinary students, farmers, cattle merchants, cattle truck drivers, and artificial inseminators should all participate in awareness efforts.

SARS coronavirus 2-reactive antibodies in bovine colostrum.

Objective: To determine if bovine colostrum and sera have antibodies that react with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Animals: Dairy and beef cattle from North America and Europe, sampled before and after the SARS-CoV-2 pandemic. Procedures: Indirect ELISAs using whole bovine coronavirus (BCoV) and SARS-CoV-2; whole SARS-CoV-2 Spike 1, Spike 2, and nucleocapsid proteins; and SARS-CoV-2-specific nucleocapsid peptide as antigens. Virus neutralization assay for BCoV. Surrogate virus neutralization assay for SARS-CoV-2. Results: Antibodies reactive to BCoV were highly prevalent in samples collected from cattle before and after the SARS-CoV-2 pandemic. Antibodies reactive with SARS-CoV-2 were present in the same samples, and apparently increased in prevalence after the SARS-CoV-2 pandemic. These antibodies had variable reactivity with the spike and nucleocapsid proteins of SARS-CoV-2 but were apparently not specific for SARS-CoV-2. Conclusions: Bovine coronavirus continues to be endemic in cattle populations, as indicated by the high prevalence of antibodies to the virus in colostrum and serum samples. Also, the prevalent antibodies to SARS-CoV-2 in bovine samples, before and after the pandemic, are likely the result of responses to epitopes on the spike and nucleocapsid proteins that are shared between the 2 betacoronaviruses. Cross-reactive antibodies in bovine colostrum could be examined for prophylactic or therapeutic effects on SARS-CoV-2 infections in humans.

Anticorps réactifs au coronavirus du SRAS 2 dans le colostrum bovin. Objectif: Déterminer si le colostrum et des échantillons de sérum bovins contiennent des anticorps qui réagissent avec le coronavirus 2 du syndrome respiratoire aigu sévère (SRAS-CoV-2). Animaux: Bovins laitiers et bovins de boucherie d'Amérique du Nord et d'Europe, échantillonnés avant et après la pandémie de SARS-CoV-2. Procédures: Épreuves ELISA indirectes utilisant le coronavirus bovin entier (BCoV) et le SARS-CoV-2; ensemble des protéines SARS-CoV-2 Spicule 1, Spicule 2 et nucléocapside; et le peptide de nucléocapside spécifique du SARS-CoV-2 comme antigènes. Test de neutralisation du virus pour le BCoV. Virus de substitution pour le test de neutralisation du SRAS-CoV-2. Résultats: Les anticorps réactifs au BCoV étaient très répandus dans les échantillons prélevés sur les bovins avant et après la pandémie de SRAS-CoV-2. Des anticorps réactifs au SRAS-CoV-2 étaient présents dans les mêmes échantillons et leur prévalence a apparemment augmenté après la pandémie de SRAS-CoV-2. Ces anticorps avaient une réactivité variable avec les protéines de spicule et de nucléocapside du SARS-CoV-2 mais n'étaient apparemment pas spécifiques du SARS-CoV-2. Conclusion: Le coronavirus bovin continue d'être endémique dans les populations bovines, comme l'indique la forte prévalence d'anticorps dirigés contre le virus dans les échantillons de colostrum et de sérum. De plus, les anticorps prévalents contre le SRAS-CoV-2 dans les échantillons de bovins, avant et après la pandémie, sont probablement le résultat de réponses à des épitopes sur les protéines de spicule et de nucléocapside qui sont partagées entre les 2 bêtacoronavirus. Les anticorps à réaction croisée dans le colostrum bovin pourraient être examinés pour leurs effets prophylactiques ou thérapeutiques sur les infections par le SRAS-CoV-2 chez l'humain.(Traduit par Dr Serge Messier).

Evaluation of practices used to reduce the incidence of bovine respiratory disease in Australian feedlots (to November 2021).

Bovine respiratory disease (BRD) has been identified as the most significant infectious disease of feedlot cattle in eastern Australia.1 Bovine respiratory disease causes economic loss due to medication costs, mortalities, excessive feed inputs associated with increased time on feed, reduced sale prices and associated labour costs. Bovine respiratory disease is a complex multifactorial condition with multiple animal, environmental and management risk factors predisposing cattle to illness. A range of microorganisms are implicated in BRD with at least four viral and five bacterial species commonly involved individually or in combination. The viruses most commonly associated with BRD in Australia are bovine herpesvirus 1 (BHV1), bovine viral diarrhoea virus (BVDV or bovine pestivirus), bovine parainfluenza 3 virus (PI3) and bovine respiratory syncytial virus (BRSV). More recently, bovine coronavirus has been identified as a potential viral contributor to BRD in Australia.2 A number of bacterial species have also been recognised as important to the BRD complex; these include Mannheimia haemolytica, Pasteurella multocida, Histophilus somni, Trueperella pyogenes and Mycoplasma bovis. Although one or more of the pathogens listed above can be isolated from clinical cases of BRD, there is no evidence that infection alone causes serious illness. This indicates that, in addition to specific infectious agents, other factors are crucial for the development of BRD under field conditions. These can be categorised as environmental, animal and management risk factors. These risk factors are likely to exert their effects through multiple pathways including reductions in systemic and possibly local immunity. For example, stressors such as weaning, handling at saleyards, transport, dehydration, weather conditions, dietary changes, comingling and pen competition might reduce the effectiveness of the immune system. Reduced immunocompetence can allow opportunistic infection of the lower airways with potential pathogens leading to the development of BRD. The objective of this paper is to critically review the evidence for management practices aimed at reducing the incidence of BRD in Australian feedlot cattle. Predisposing factors (Table 1) largely beyond the control of most feedlots, such as weather and exposure to respiratory viruses, are discussed separately, but these factors can generate indirect prevention responses that are discussed under the preventative practices categories. The current practices are classified as either animal preparation practices (Table 2) or feedlot management practices (Table 3).

Bovine Enteroids as an In Vitro Model for Infection with Bovine Coronavirus.

Bovine coronavirus (BCoV) is one of the major viral pathogens of cattle, responsible for economic losses and causing a substantial impact on animal welfare. Several in vitro 2D models have been used to investigate BCoV infection and its pathogenesis. However, 3D enteroids are likely to be a better model with which to investigate host-pathogen interactions. This study established bovine enteroids as an in vitro replication system for BCoV, and we compared the expression of selected genes during the BCoV infection of the enteroids with the expression previously described in HCT-8 cells. The enteroids were successfully established from bovine ileum and permissive to BCoV, as shown by a seven-fold increase in viral RNA after 72 h. Immunostaining of differentiation markers showed a mixed population of differentiated cells. Gene expression ratios at 72 h showed that pro-inflammatory responses such as IL-8 and IL-1A remained unchanged in response to BCoV infection. Expression of other immune genes, including CXCL-3, MMP13, and TNF-α, was significantly downregulated. This study shows that the bovine enteroids had a differentiated cell population and were permissive to BCoV. Further studies are necessary for a comparative analysis to determine whether enteroids are suitable in vitro models to study host responses during BCoV infection.

Rabies surveillance in the United States during 2021.

OBJECTIVE: To provide epidemiological information on the occurrence of animal and human rabies in the US during 2021 and summaries of 2021 rabies surveillance for Canada and Mexico. PROCEDURES: State and territorial public health departments and USDA Wildlife Services provided data on animals submitted for rabies testing in 2021. Data were analyzed temporally and geographically to assess trends in domestic animal and wildlife rabies cases. RESULTS: During 2021, 54 US jurisdictions reported 3,663 rabid animals, representing an 18.2% decrease from the 4,479 cases reported in 2020. Texas (n = 456 [12.4%]), Virginia (297 [8.1%]), Pennsylvania (287 [7.8%]), North Carolina (248 [6.8%]), New York (237 [6.5%]), California (220 [6.0%]), and New Jersey (201 [5.5%]) together accounted for > 50% of all animal rabies cases reported in 2021. Of the total reported rabid animals, 3,352 (91.5%) involved wildlife, with bats (n = 1,241 [33.9%]), raccoons (1,030 [28.1%]), skunks (691 [18.9%]), and foxes (314 [8.6%]) representing the primary hosts confirmed with rabies. Rabid cats (216 [5.9%]), cattle (40 [1.1%]), and dogs (36 [1.0%]) accounted for 94% of rabies cases involving domestic animals in 2021. Five human rabies deaths were reported in 2021. CLINICAL RELEVANCE: The number of animal rabies cases reported in the US decreased significantly during 2021; this is thought to be due to factors related to the COVID-19 pandemic.

Development of an IgY-Based Treatment to Control Bovine Coronavirus Diarrhea in Dairy Calves.

Bovine Coronavirus (BCoV) is a major pathogen associated with neonatal calf diarrhea. Standard practice dictates that to prevent BCoV diarrhea, dams should be immunized in the last stage of pregnancy to increase BCoV-specific antibody (Ab) titers in serum and colostrum. For the prevention to be effective, calves need to suck maternal colostrum within the first six to twelve hours of life before gut closure to ensure a good level of passive immunity. The high rate of maternal Ab transfer failure resulting from this process posed the need to develop alternative local passive immunity strategies to strengthen the prevention and treatment of BCoV diarrhea. Immunoglobulin Y technology represents a promising tool to address this gap. In this study, 200 laying hens were immunized with BCoV to obtain spray-dried egg powder enriched in specific IgY Abs to BCoV on a large production scale. To ensure batch-to-batch product consistency, a potency assay was statistically validated. With a sample size of 241, the BCoV-specific IgY ELISA showed a sensitivity and specificity of 97.7% and 98.2%, respectively. ELISA IgY Abs to BCoV correlated with virus-neutralizing Ab titers (Pearson correlation, R2 = 0.92, p < 0.001). Most importantly, a pilot efficacy study in newborn calves showed a significant delay and shorter duration of BCoV-associated diarrhea and shedding in IgY-treated colostrum-deprived calves. Calves were treated with milk supplemented with egg powder (final IgY Ab titer to BCoV ELISA = 512; VN = 32) for 14 days as a passive treatment before a challenge with BCoV and were compared to calves fed milk with no supplementation. This is the first study with proof of efficacy of a product based on egg powder manufactured at a scale that successfully prevents BCoV-associated neonatal calf diarrhea.

Effect of degalactosylated bovine glycoprotein formulations MAF and M сapsules on lymphopenia and clinical outcomes in hospitalized Covid-19 patients: a randomized clinical trial (preprint)

BACKGROUND Targeting mucosal immunity of the gut, which is known to provide antigen processing, while avoiding excessive or unnecessary inflammation, was tested as a way to modulate Covid-19 severity. METHODS Randomized open-label trial in 204 adults hospitalized with non-critical Covid-19 who received for 14 days in addition to standard of care (SOC) degalactosylated bovine glycoproteins formulations of either MAF Capsules (MAF group) or M Capsules (M group) or SOC only (control group). RESULTS Median recovery time when patients did not require supplemental oxygen was 6 days in both study groups compared to 9 days in the control (MAF vs. control; P=0.020 and M vs. control; P=0.004). A greater reduction in mortality was seen in the MAF group compared to the control by day 14 (8.3% vs. 1.6%; P=0.121) and by day 29 (15.3% vs. 3.2%; P=0.020), and similarly in the M group by day 14 (8.3% vs. 2.9%; P=0.276) and by day 29 (15.3% vs. 2.9%; P=0.017). The proportion of those who had baseline absolute lymphocyte count (ALC) lower than 0.8x10^9/L was 13/63 (20.6%), 17/69 (27.0%), and 18/72 (25.0%) of patients in MAF, M, and control group respectively. Day 29 mortality among these lymphopenic patients was three times higher than for the intent-to-treat population (21% vs. 7%) and consisted in above subgroups: 2/13 (15%), 2/17 (12%), and 6/18 (33%) of patients. There showed decreased mortality in both study subgroups correlated with greater ALC restoration above 0.8x109/L level seen on day 14 in 77% and 59% of patients in MAF and M subgroups respectively compared to 22% of patients in control subgroup. Incidences of any ALC decrease below the baseline level on day 14 occurred in 25.4% of patients in the MAF group and 29.0% of patients in the M group compared to 45.8% in control and ALC depletion by ≥50% from the baseline level consisted of 7.9%, 5.8%, and 15.3% of cases in these groups respectively. CONCLUSION This study showed that both study agents prevented ALC depletion and accelerated its restoration, which is believed one of the mechanisms of improved crucial clinical outcomes in hospitalized Covid-19 patients. Trial registration: ClinicalTrials.gov NCT04762628, https://www.clinicaltrials.gov/ct2/show/NCT04762628.

The Prevalence and Evolutionary Characteristics of Bovine Coronavirus in China (preprint)

Four viral intestinal pathogens such as bovine coronavirus (BCoV), bovine rotavirus (BRV), bovine viral diarrhea virus (BVDV) and bovine astrovirus (BoAstV) are the most common causes of adult cattle diarrhea. In this study, 797 samples were collected from January 2020 to February 2023. Among them, BCoV is major pathogen we detected and the positive rate was 48.18% (471/794). Farther more, among 307 diarrhea samples and 487 asymptomatic samples, 159 and 312 were BCoV positive, respectively. The results reveal that BCoV is associated with bovine diarrhea (p=0.001). In addition, we also amplified and analyzed Hemagglutinin-esterase (HE), Spike (S) and whole genome of partially positive sample. Getting 8 HE complete sequence, 8 S complete sequence and 5 whole genomes. The molecular characterization provides reveal that 5 strains were branched Chinese strains, Japan and part of American strains in GⅡb，and Korea strains branched in GⅡa. BCoV-Neimeng S in this study was mutated 143H/A to 143R, which is uncommon in S gene. As a residue on the side chain of the core binding site of S1-NTD, this mutation changed the tertiary structure and may be possible to affect the glycoside binding spectrum. The noteworthy that BCoV-SJZ2 and BCoV-YZ1 have four amino acid insertions on HE, which is the same as SWUN/A10/2018. We also analysis 4.8kDa and 4.9kDa non-structural protein large-scale truncation and membrane protein mutation. This article will discuss epidemiology and genome analysis.

Cluster analysis of bovine respiratory disease (BRD)-associated pathogens shows the existence of two epidemiological patterns in BRD outbreaks.

A hierarchical cluster analysis was used to classify outbreaks of bovine respiratory disease (BRD; n = 156) in natural groups according to the detection of nine pathogens (parainfluenza 3 virus (PI-3), bovine respiratory syncytial virus (BRSV), bovine coronavirus (BCV), bovine viral diarrhea virus (BVDV), and bovine herpesvirus 1 (BHV-1), Mannheimia haemolytica, Pasteurella multocida, Histophilus somni, and Mycoplasma bovis. Pathogens were detected by individual q-PCRs. Two clusters were identified. Cluster 1 was characterized by a relatively high frequency (40-72%) of four BRD-associated viruses, supporting their primary involvement in BRD. Cluster 2 was characterized by frequencies of PI-3, BRSV, or BVDV below 10% each. P. multocida and M. haemolytica were detected with high frequencies in both clusters (P > 0.05), while M. bovis and H. somni showed a significantly higher frequency in cluster 1and 2, respectively. Outbreaks in cluster 1 were associated with preweaning calves younger than 5 months (OR 2.2; 95% CI 1.1-4.5) and with cold months, whereas cluster 2 was associated with fattening calves older than 5 months after arrival to feedlots and without any seasonality. Thus, in addition to the classic epidemiological BRD pattern characterized by the primary involvement of viruses occurring preferably during winter and affecting young calves, there is a second pattern in which viruses would be less relevant, affecting mainly calves older than 5 months at any time of the year. This study allows a better understanding of the BRD epidemiology, which can be useful when implementing management and prophylaxis measures for a better control of this disease.

Coping with drug resistant tuberculosis alongside COVID-19 and other stressors in Zimbabwe: a qualitative study. (preprint)

Background Households in low-resource settings are more vulnerable to events which adversely affect their livelihoods, including shocks such as the death of a family member, inflation, droughts and more recently COVID-19. Drug Resistant Tuberculosis (DR-TB) is also another shock that inflicts physical, psychological and socioeconomic burden on individuals and households. We describe experiences and coping strategies among people affected by DR-TB and their households in Zimbabwe during the COVID-19 pandemic, 2020 to 2021. Methods We conducted 16 in-depth interviews with adults who had just completed or were completing DR-TB treatment. Interview themes included health seeking behaviour, impact of DR-TB on livelihoods and coping strategies adopted during treatment. We analysed data using thematic analysis. Results Health seeking from providers outside the public sector, extra-pulmonary TB and health system factors resulted in delayed DR-TB diagnosis and treatment and increased financial drain on households. DR-TB reduced productive capacity and narrowed job opportunities leading to income loss that continued even after completion of treatment. Household livelihood was further adversely affected by lockdowns due to COVID-19, outbreaks of bird flu and cattle disease. Stockouts of DR-TB medicines, common during COVID-19, exacerbated loss of productive time and transport costs as medication had to be accessed from other clinics that were further away. Reversible coping strategies included: reducing number of meals; relocating in search of caregivers and/or family support; spending savings; negotiating with school authorities to keep children in school. Some households had to adopt irreversible coping strategies such as selling productive assets and withdrawing children from school. Conclusion DR-TB combined with COVID-19 and other stressors pushed households into deeper poverty, and vulnerability. Multi-sectoral approaches that combine health systems, psychosocial and economic interventions are crucial to mitigate diagnostic delays and suffering, and meaningfully support people with DR-TB and their households to compensate the loss of livelihoods during and post DR-TB treatment.

Histophilus somni disease conditions with simultaneous infections by ovine gammaherpesvirus 2 in cattle herds from Southern Brazil.

This report investigated the cause of cattle mortality in two farms in Southern Brazil. The tissues of one animal from each farm (animals #1 and #2) respectively were used in pathological and molecular investigations to determine the possible cause of death. The principal pathological findings observed in animal #1 were pulmonary, myocardial, and encephalitic hemorrhages with vasculitis, and lymphoplasmacytic interstitial pneumonia with proliferative vascular lesions (PVL). The main pathological findings observed in animal #2 were purulent bronchopneumonia, hemorrhagic myocarditis, and lymphoplasmacytic interstitial pneumonia with PVL. An immunohistochemical assay detected intralesional antigens of a malignant catarrhal fever virus (MCFV) from multiple tissues of animal #2 while PCR confirmed that the MCFV amplified was ovine gammaherpesvirus 2 (OvGHV2), genus Macavirus, subfamily Gammaherpesvirinae; OvGHV2 was also amplified from multiple tissues of animal #1. Furthermore, PCR assays amplified Histophilus somni DNA from multiple fragments of both animals. However, the nucleic acids of Mannheimia haemolytica, Pasteurella multocida, Mycoplasma bovis, bovine respiratory syncytial virus, bovine alphaherpesvirus virus 1 and 5, bovine coronavirus, and bovine parainfluenza virus 3 were not amplified from any of the tissues analyzed, suggesting that these pathogens did not participate in the development of the lesions herein described. These findings demonstrated that both animals were concomitantly infected by H. somni and OvGHV2 and developed the septicemic and encephalitic manifestations of H. somni. Furthermore, the interstitial pneumonia observed in cow #2 was more likely associated with infection by OvGHV2.

Characterisation of the Upper Respiratory Tract Virome of Feedlot Cattle and Its Association with Bovine Respiratory Disease.

Bovine respiratory disease (BRD) is a major health problem within the global cattle industry. This disease has a complex aetiology, with viruses playing an integral role. In this study, metagenomics was used to sequence viral nucleic acids in the nasal swabs of BRD-affected cattle. The viruses detected included those that are well known for their association with BRD in Australia (bovine viral diarrhoea virus 1), as well as viruses known to be present but not fully characterised (bovine coronavirus) and viruses that have not been reported in BRD-affected cattle in Australia (bovine rhinitis, bovine influenza D, and bovine nidovirus). The nasal swabs from a case-control study were subsequently tested for 10 viruses, and the presence of at least one virus was found to be significantly associated with BRD. Some of the more recently detected viruses had inconsistent associations with BRD. Full genome sequences for bovine coronavirus, a virus increasingly associated with BRD, and bovine nidovirus were completed. Both viruses belong to the Coronaviridae family, which are frequently associated with disease in mammals. This study has provided greater insights into the viral pathogens associated with BRD and highlighted the need for further studies to more precisely elucidate the roles viruses play in BRD.

Prevalence of bovine coronavirus in cattle in China: A systematic review and meta-analysis.

Bovine coronavirus (BCoV) is one of the important pathogens that cause calf diarrhea (CD), winter dysentery (WD), and the bovine respiratory disease complex (BRDC), and spreads worldwide. An infection of BCoV in cattle can lead to death of young animals, stunted growth, reduced milk production, and milk quality, thus bringing serious economic losses to the bovine industry. Therefore, it is necessary to prevent and control the spread of BCoV. Here, a systematic review and meta-analysis was conducted to assess the prevalence of BCoV in cattle in China before 2022. A total of 57 articles regarding the prevalence of BCoV in cattle in China were collected from five databases (PubMed, ScienceDirect, CNKI, VIP, and Wan Fang). Based on the inclusion criteria, a total of 15,838 samples were included, and 6,136 were positive cases. The overall prevalence of BCoV was 30.8%, with the highest prevalence rate (60.5%) identified in South China and the lowest prevalence (15.6%) identified in Central China. We also analyzed other subgroup information, included sampling years, sample sources, detection methods, breeding methods, age, type of cattle, presence of diarrhea, and geographic and climatic factors. The results indicated that BCoV was widely prevalent in China. Among all subgroups, the sample sources, detection methods, breeding methods, and presence or absence of diarrheal might be potential risk factors responsible for BCoV prevalence. It is recommended to strengthen the detection of BCoV in cattle, in order to effectively control the spread of BCoV.