Bovine Viral Diarrhea Virus Infection Research Articles

Seroprevalence and its associated risk factors of Bovine Neosporosis and Bovine Viral Diarrhea in cattle of Tilottama municipality, Rupandehi, Nepal

Background: Bovine Neosporosis (BN) and Bovine Viral Diarrhea (BVD) are economically significant livestock disease that causes reproductive and respiratory problems in cattle and buffalo caused by protozoan parasite Neospora caninum and Bovine Viral Diarrhea Virus (BVDV) respectively. The burden of production and economic loss due to infectious reproductive diseases are still unknown in the various location of Nepal. Objectives: To overview the prevalence of bovine neosporosis and bovine viral diarrhea infection in a commercially important livestock herd from Tilottama Municipality, Rupandehi, Nepal. Methods: A cross-sectional purposive study was conducted through a questionnaire survey to collect demographic and farm characteristics information. In total 92 serum samples from jugular veins were collected from a different herd of Tilottama Municipality of Rupandehi, Nepal. The antibody against BVDV was tested using the competitive Enzyme-Linked Immunosorbent Assay (c-ELISA). Results: The individual animal-wise apparent prevalence was 9.78% with a true prevalence of 9.70% (5.09 - 17.57) and 3.26% with a true prevalence of 3.10%; CI (0.93 - 9.06) for BN and BVD respectively. The risk factor history of abortion was associated significantly with BN and BVD seropositivity (p < 0.05). Animals in the mixed farming system and contact with dogs were associated significantly with the occurrence of bovine viral diarrhea and bovine neosporosis respectively at (p < 0.05). Conclusion and recommendation: The stuy indicated the circulation of Neospora caninum and bovine viral diarrhea in dairy herds of the study area. It is suggested that the stakeholders in the study area should be sensitized to the impact of Neospora caninum and bovine viral diarrhea virus infection in dairy herds and include both diseases in the regional and national surveillance system.

Berbamine hydrochloride inhibits bovine viral diarrhea virus replication via interfering in late-stage autophagy

Bovine viral diarrhea virus (BVDV) is a harmful pathogen that easily causes large-scale infections and huge economic losses to the cattle industry. Berbamine hydrochloride (BBH) is a natural product extracted from berberis and has a wide range of pharmacological effects. However, the antiviral effect of BBH against BVDV needs to be further elucidated. This study aimed to evaluate the antiviral activities of BBH against BVDV infection. We mainly used RT-qPCR, Western blotting, immunofluorescence, and TEM assays to assess the inhibitory activity of BBH against BVDV. The results showed that BBH had an inhibitory effect on BVDV and higher inhibitory activity in the viral attachment and release in MDBK cells. This study found that BVDV could induce and use autophagy to replicate itself. Further results showed that BBH inhibited BVDV infection by inhibiting autophagy integrity in BVDV-infected cells, which was proven by the detection of autophagy-related proteins. Our data show that in BBH-treated BVDV-infected cells, the expression of p62 and LC3 increased over time. After the addition of an autophagy inhibitor, chloroquine (CQ), and an autophagy promoter, rapamycin (Rapa), we found that promoting autophagy was beneficial to the replication of BVDV, while inhibiting autophagy could reduce the number of infections by BVDV, which was evidenced by the expression of the BVDV E2 protein. Furthermore, BBH blocked the normal binding of LC3 and LAMP1 in BVDV-infected cells. In conclusion, BBH inhibited BVDV infection by inhibiting BVDV-induced autophagy in cells, and its inhibitory effect was obvious in the viral attachment and release stages. Therefore, our study provides a new idea for exploring novel anti-BVDV drugs.

A novel competitive ELISA for detection of antibodies against bovine viral diarrhea virus infection.

Diagnosis of bovine viral diarrhea (BVD) relies on the detection of antibodies against its viral causing agent, bovine viral diarrhea virus (BVDV). Here, we designed a novel competitive ELISA (cELISA) using the most immunogenic part of BVDV nonstructural protein 3 (NS3), as a single ELISA recombinant antigen, along with a monoclonal antibody to detect antibodies against BVDV in sera of infected animals. Hence, 197 serum samples were tested by this cELISA and the results were compared to the results obtained from virus neutralization test (VNT) as the gold standard method for diagnosis of BVD. McNemar's test indicated that there was no significant difference between the results of this newly designed cELISA and VNT. Meanwhile, kappa coefficients showed that there was a high correlation between these two assays. The relative sensitivity and specificity of cELISA with respect to VNT were 93.90% and 100%, respectively, suggesting that this newly designed cELISA could be a useful diagnostic tool for detection of BVDV infection. Moreover, as NS3 is highly conserved among Pestiviruses and the developed ELISA is a competitive one, it could potentially be applied to detect BVDV infection in other domestic and wildlife species.

Genetic features of bovine viral diarrhea virus subgenotype 1c in newborn calves at nucleotide and synonymous codon usages.

Bovine viral diarrhea virus (BVDV), serving as an important pathogen for newborn calves, poses threat to reproductive and economic losses in the cattle industry. To survey the infection rate and genetic diversity of BVDV in newborn calves in northern China, a total of 676 sera samples of newborn calves were collected from four provinces between 2021 and 2022. All sera samples were individually detected for BVDV infection by RT-PCR and ELISA. Our results showed that the overall serological rate was 9.76% (66/676) and the average positive rate of BVDV RNA was 8.14% (55/676) in the newborn calves. Eight BVDV strains were successfully isolated from RT-PCR positive sera samples, and four isolates displayed the cytopathic effect (CPE). Based on phylogenetic tree at the genome level, the eight strains were classified into subgenotype 1c. Moreover, the BVDV isolates had a close genetic relationship with the GSTZ strain at either nucleotide or codon usage level. Interestingly, in comparison of synonymous codon usage patterns between the BVDV isolates with CPE and ones without CPE, there were four synonymous codons (UCG, CCC, GCA, and AAC) which displayed the significant differences (p < 0.05) at codon usage pattern, suggesting that synonymous codon usage bias might play a role in BVDV-1c biotypes. In addition, the usage of synonymous codons containing CpG dinucleotides was suppressed by the BVDV-1c isolates, reflecting one of strategies of immune evasion of BVDV to its host. Taken together, our study provided data for monitoring and vaccination strategies of BVDV for newborn calves in northern China.

Forsythiaside A Improves the Inhibitory Efficiency of Recombinant Protein Vaccines against Bovine Viral Diarrhea Virus Infection.

Bovine viral diarrhea virus (BVDV) is a critical animal pathogen that leads to cattle production losses associated with acute disease, immune dysregulation, reproductive failure, and respiratory disease. Due to the monotonous control technique and neglect of BVDV, increasing prevalence of BVDV has caused significant economic losses in the cattle industry worldwide. Therefore, novel anti-BVDV drugs are essential to prevent and control BVDV. Our previous studies have found that Forsythoside A (FTA) could inhibit the replication of BVDV via TRAF2-dependent CD28-4-1BB signaling in bovine peripheral blood mononuclear cells (PBMCs), but whether they can directly inhibit the BVDV remains unclear. Here, we further investigated the effects of FTA on BVDV and its underlying mechanisms of action. We found that FTA significantly inhibited the replication of BVDV in the MDBK cell directly. The results demonstrated that FTA could reduce the functional activation of Caspase-1 to inhibit the inflammatory response caused by BVDV infection and increase the expression of type I interferon (IFN-I) to clear the virus in vitro. The animal experiment was performed to evaluate the antiviral effect of FTA in vivo. Notably, after challenged with BVDV, mice with FTA + Erns-E2 protein displayed alleviated pathological damage and decreased the viral load in the spleen compared with mice inoculated with Erns-E2 protein. Furthermore, treatment with FTA enhanced body defense and delayed infection by the BVDV. Our results reveal that FTA suppresses BVDV replication both in vitro and in vivo and therefore shows promise as an anti-BVDV agent.

Risk-associated factors associated with the bovine viral diarrhea virus in dromedary camels, sheep, and goats in abattoir surveillance and semi-closed herd system.

Background and Aim:Bovine viral diarrhea virus (BVDV) is one of the most important viral pathogens causing high economic losses in cattle of all ages. Despite the active vaccination campaigns against BVDV, many outbreaks are still detected in various populations of cattle worldwide. Other species of animals such as dromedary camels, sheep, and goats may harbor BVDV infection and cause variable clinical syndromes. Thus, they may act as a source of infection to the cattle population around them. However, little is still known about the roles of these animals in the viral transmission and sustainability of BVDV in the environment. This study aimed to explore if the dromedary camels, sheep, and goats may seroconvert against BVDV and to study some associated risk factors for BVDV in these species of animals.Materials and Methods:We tested 1012 serum samples from dromedary camels, 84 from goats, and 21 from sheep for BVDV antibodies using commercial enzyme-linked immunosorbent assay (ELISA) kits. Meanwhile, we selected 211 serum samples from dromedary camels to be tested for the BVDV antigen using the commercial ELISA kits.Results:Our results show that 49/1117 serum samples were positive for the BVDV antibodies in dromedary camels (46/1012), goats (3/84), and none of the tested sheep samples were positive. However, none of the collected serum samples tested positive for the BVDV antigen.Conclusion:Seroconversion of some dromedary camels, sheep, and goats to the BVDV with no history of vaccination against BVDV strongly suggests the potential roles of these species of animals in the virus transmission cycle. The main limitations of the current study are (1) the lack of samples from other species of animals that lived close by these animals, particularly cattle. (2) lack of follow-up samples from the same animal over a long period. We believe the long-term longitudinal study of BVDV in various species of animals, particularly dromedary camels, goats, and sheep, is one of our future research directions. This will provide more information about the dynamics of BVDV antibodies in these species of animals.

Prevalence characteristic of BVDV in some large scale dairy farms in Western China.

The aim of this study was to analyze the prevalence characteristic of Bovine viral diarrhea virus (BVDV) in some large scale dairy farms in Western China. BVDV was detected in 30 samples of bulk tank milk (BTM) collected from 30 large dairy farms in 7 provinces of western China, 93.33% (28/30) of the farms were infected with BVDV, and S/P ratio was over 0.3 in 28 positive farms. The individual status was further estimated in the dairy farm (No. 10) with the highest positive rate (S/P ratio = 1.37) and the dairy farm (No. 17) with the lowest positive rate (S/P ratio = 0.39). Two hundred cows were, respectively, selected from calf, young cows and lactating cows in farm No. 10 and farm No. 17 and the serum sample of each enrolled cow was collected. The individual positive rate of serum antibody (Ab) was 87.17% (523/600) in farm No. 10 and 31.33% (188/600) in farm No. 17. The individual positive ratio of serum antibody in calves, young cows and lactating cows were 41.75 % (167/400), 58.75% (235/400) and 77.25% (309/400), respectively. BTM Ab of farm No. 10 has an S/P ratio more than 1.0, which indicated there were emergent or persistent infection (PI) cases, and further test showed that PI cases were 0.51% in farm No. 10. Pathogens were positive in 42.34% (163/385) of nasal mucus samples collected from cows with respiratory symptom, and BVDV cases were 57 in 163 positive samples. Three strains of NCP BVDV-1, one strain of CP BVDV-1, one strain of NCP BVDV-2 and one strain of CP BVDV-2 were successfully isolated. Phylogenetic analysis revealed that the subtypes of BVDV currently prevalent in western China were BVDV-1a, BVDV-1m, BVDV-1q and BVDV-2. The findings suggested that the BVDV infection is serious in some Large Scale Dairy Farms in Western China.

The recombinant Erns and truncated E2-based indirect enzyme-linked immunosorbent assays to distinguishably test specific antibodies against classical swine fever virus and bovine viral diarrhea virus

BackgroundClassical swine fever (CSF) virus is the causative agent of an economically important, highly contagious disease of pigs. CSFV is genetically and serologically related to bovine viral diarrhea virus (BVDV). BVDV infection in pigs can mimic CSF clinical signs, which cause difficulty in differentiation. Serological test for detection of virus specific antibodies is a valuable tool for diagnosis and surveillance of CSFV and BVDV infections in animals. The aim of this study was to develop the CSFV Erns and BVDV tE2 -based ELISAs to distinguishably test specific antibodies against CSFV and BVDV.MethodsThe CSFV Erns and truncated E2 (tE2, residues 690–865) of BVDV were expressed in E. coli and purified by Ni–NTA affinity chromatography, respectively. Employing Erns or tE2 protein as diagnostic antigen, indirect ELISAs were developed to distinguishably test specific antibodies against CSFV and BVDV. The specificity and sensitivity of ELISAs were evaluated using a panel of virus specific sera of pigs, immunized rabbits and immunized mice. A total 150 clinical serum samples from farm pigs were measured by the developed ELISAs and compared with virus neutralizing test (VNT).ResultsIndirect ELISA was established based on recombinant CSFV Erns or BVDV tE2 protein, respectively. No serological cross-reaction between antibodies against CSFV and BVDV was observed in sera of immunized rabbits, immunized mice or farm pigs by detections of the Erns and tE2 -based ELISAs. Compared to VNT, the CSFV Erns -based ELISA displayed a high sensitivity (93.3%), specificity (92.0%) and agreement rate (92.7%), and the sensitivity, specificity and agreement rate of BVDV tE2 -based ELISA was 92.3%, 95.2% and 94.7%, respectively.ConclusionThe newly developed ELISAs are highly specific and sensitive and would be valuable tools for serological diagnosis for CSFV and BVDV infections.

Coinfection with PEDV and BVDV induces inflammatory bowel disease pathway highly enriched in PK-15 cells

BackgroundFrom the 1078 diarrhea stools tested in our survey from 2017 to 2020 in local area of China, PEDV was the key pathogen that was closely related to the death of piglets with diarrhea. In addition, coinfection of PEDV-positive samples with BVDV reached 17.24%. Although BVDV infection in swine is typically subclinical, the effect of PEDV and BVDV coinfection on disease severity and the potential molecular mechanism of coinfection with these two viruses remain unknown.MethodsIn this study, we developed a model of coinfection with porcine epidemic diarrhea virus (PEDV) and bovine viral diarrhea virus (BVDV) in PK15 cells, and a tandem mass tag (TMT) combined with LC–MS/MS proteomic approach was used to identify differential protein expression profiles. Additionally, we performed drug experiments to explore the inflammatory response induced by PEDV or BVDV mono- or coinfection.ResultsA total of 1094, 1538, and 1482 differentially expressed proteins (DEPs) were identified upon PEDV monoinfection, BVDV monoinfection and PEDV/BVDV coinfection, respectively. KEGG pathway analysis revealed that PEDV and BVDV coinfection led to a highly significantly enrichment of the inflammatory bowel disease (IBD) pathway. In addition, the NF-κB signaling pathway was more intensively activated by PEDV and BVDV coinfection, which induced higher production of inflammatory cytokines, than PEDV or BVDV monoinfection.ConclusionsOur study indicated that cattle pathogens might play synergistic roles in the pathogenesis of porcine diarrhea, which might also improve our understanding of the pathogenesis of multiple infections in diarrhea.

Development of an in-house Indirect ELISA for detection of bovine viral diarrhoea virus antibodies in bovine sera

Bovine viral diarrhoea virus (BVDV) infection is a worldwide distributed animal disease. BVDV is the causal agent of congenital defects, diarrhea, reproductive failure, and contaminating biological products. Virus Neutralization test (VNT) as a gold standard method is used for detection of BVDV. Although this assay is very sensitive and specific, it has disadvantages including requires to an experienced person and cell culture facilities. VNT is time-consuming. It is important to design a method that does not have the mentioned disadvantages. So, in-house indirect enzyme linked immunosorbent assay (i-ELISA) was developed for laboratories where it is not possible to perform VNT. The system was made using NADL strain of BVDV and MDBK cell line. This ELISA system was compared with a commercial ELISA kit using 99 bovine sera. Coefficient of variation (CV) was calculated 3.9% and 4.8% for the positive and negative control, respectively for the designed i-ELISA system. The sensitivity, specificity, and accuracy of i-ELISA system was 88%, 53.6%, and 70.7% respectively. Based on our result correlation between in- house and commercial ELISA kit for detection of antibody against BVDV in bovine sera was significant (Kappa coefficient =0.41, p < 0.05). Results of the present study suggested that an in-house ELISA as an affordable and confident system for primary screening of the sera used for biological product.

Estimating the Effect of a Bovine Viral Diarrhea Virus Control Program: An Empirical Study on the Performance of Dutch Dairy Herds.

More and more European countries have implemented a bovine viral diarrhea virus (BVDV) control program. The economic effects of such programs have been evaluated in simulations, but empirical studies are lacking, especially in the final stage of the program. We investigated the economic (gross margin) and production effects (milk yield, somatic cell count, and calving interval) of the herds obtaining BVDV-free certification based on longitudinal annual accounting and herd performance data from Dutch dairy herds between 2014 and 2019, the final stages of the Dutch national BVDV-free program. This study was designed as a case-control study: two types of case herds were defined for two analyses. The case herds in the first analysis are herds where the BVDV status changed from “BVDV not free” to “BVDV free” during the study period. The not-free status refers to a herd that participated in the BVDV-free program but had not yet obtained the BVDV-free certification. In the second analysis, the case herds started participating in the Dutch BVDV-free program during the study period and obtained the BVDV-free certification. Control herds in both analyses were BVDV-free during the entire study period. Potential bias between the covariates of the two herd groups was reduced by matching case and control herds using the propensity score matching method. To compare the differences between case and control herds before and after BVDV-free certification, we used the time-varying Difference-in-Differences estimation (DID) methodology. The results indicate that there was no significant change in milk yield, somatic cell count, calving interval, and gross margin upon BVDV-free certification. There are several possible explanations for the non-significant effects observed in our study, such as the final stage of the BVDV control program, not knowing the true BVDV infection situation in case herds and not knowing if control measures were implemented in case herds prior to participating in the BVDV-free program. In our study, the effects of BVDV-free certification might have been underestimated, given that the Dutch BVDV control program became mandatory during the study period, and some of the case herds might have never experienced any BVDV infection. The results of this study suggest that in the final stage of the BVDV control program, the program may no longer have a clear benefit to the herd performance of participating dairy herds. When designing national programs to eradicate BVDV, it is therefore important to include incentives for such farms to motivate them to join the program.

Disease surveillance in England and Wales, June 2022.

Disease surveillance in England and Wales, June 2022.

Molecular Diversity and Histopathological Findings of Novel Bovine Viral Diarrhea Virus Strains Isolated from Bull Semen

Bovine viral diarrhea virus (BVDV) is one of the most common viral pathogens affecting the cattle industry worldwide. The present study aimed to molecularly characterize BVDV isolates that are currently circulating in breeding bulls farmed with cattle suffering from reproductive disorders, and also to assess the consequences of BVDV infection on bulls’ semen quality and conception, and its pathological effects on the structure of testicular tissue and spermatozoa. For this purpose, semen, serum, and testicular samples were collected from four breeding bulls in four private dairy farms in the governorates of Kafr-El Sheik, Beni-Suef, Giza, and Assuit, in Egypt from April 2019 to May 2020. An evaluation of sperm abnormalities was carried out by assessing the integrity of the plasma and acrosomal membranes where severe damage and abnormalities were found. Ultrastructure analysis of the spermatozoa by transmission electron microscopy revealed the presence of a swollen plasma membrane with segmented outer acrosomal membrane of spermatozoa and vacuolar degenerated mitochondria. Histopathological examination of testicular and epididymal tissues indicated moderate to severe degenerative effects of virus infection on seminiferous tubules with hypospermatogenesis. By detection of virus antigen in the serum samples using ELISA, bulls were identified as persistently infected with BVDV. Virus isolation revealed four noncytopathic (NCP-BVDV) strains that were confirmed by fluorescent antibody technique (FAT) and amplification of the 5′ untranslated genomic region (5’UTR) and molecularly typed by amplification of the Erns glycoprotein region. Isolates’ Phylogenetic analysis revealed two subgenotypes: BVDV-1b (Genbank accession numbers; LC634512, LC634513, LC634515) and BVDV-1d (LC634516). According to the knowledge of the authors of the present study, the circulation of the BVDV-1d subgenotype is not reported in Egypt. Therefore, it would be of great importance to track circulating strains in specific countries for successful vaccination programs or accurate diagnostic tests, and this necessitates regular updates.

Transcriptomic Analysis of MDBK Cells Infected with Cytopathic and Non-Cytopathic Strains of Bovine Viral Diarrhea Virus (BVDV).

Bovine viral diarrhea virus (BVDV) belongs to the Flaviviridae family and the Pestivirus genus. Infection with BVDV causes a disease with a wide spectrum of clinical symptoms, most often mild, although infections with this virus constitute a serious economic problem all over the world. The virus is characterized by a high genetic variability, while the accumulation of single mutations leads to the formation of its new variants. The aim of this study was to better understand the complicated pathogenesis of this disease at the molecular level via the analysis of the transcriptome of cells infected with this virus. The bovine kidney cell line (MDBK), the cytopathic (cp) reference strain, and two non-cytopathic (ncp) BVD virus field strains were used in transcriptomic studies. The cell transcriptome was tested 24 and 72 h after infection. The results of the microarray analysis revealed changes in the expression levels of numerous genes. Genes with changed expression as a result of infection with the cp strain caused changes in the expression levels of a large number of genes and enriched a number of pathways. Genes with increased expression levels were enriched among other pathways involved in the cell cycle, while genes with reduced expression levels enriched pathways mostly related to metabolism. Genes with increased expression levels as a result of infection with ncp strains enriched a much smaller number of pathways, among them, pathways related to signaling activity 24 h post-infection and serine biosynthetic pathways both 24 and 72 h post-infection. Pathways enriched by genes with reduced expression levels were related to the innate immune response (72 h post-infection) or metabolism (24 and 72 h post-infection). The results of microarray studies can help us to better understand the host’s response to BVDV infection.

Seroprevalence of bovine viral diarrhea virus antibodies and risk factors in dairy cattle from the central desert of Iran

Bovine viral diarrhea virus (BVDV) infects cattle worldwide and causes one of the most important economic diseases of the dairy industry. BVDV infection reduces reproductive efficiency, suppresses the immune system, and causes gastrointestinal and respiratory diseases. A first cross-sectional study was conducted in the central desert of Iran (Yazd and South Khorasan provinces) to estimate the seroprevalence and identify BVDV-related risk factors in dairy cattle. A total of 800 cows were randomly selected of 76 herds, and their serum samples were tested by the indirect enzyme-linked immunosorbent assay (ELISA) method for BVDV antibody detection. Data were analyzed using the logistic regression model. The serum prevalence of BVDV at animal and herd levels was 66.83% and 91.6%, respectively. Traditional housing system (OR = 3.22; 95% CI = 1.20-9.09) and cattle introduction to the herd (OR = 2.12; 95% CI = 1.21-3.70) were the important risk factors for BVDV seropositivity (p < 0.05). Increasing of age per year caused adding in 0.33 log (odds) of BVDV seropositivity (p < 0.05). It is necessary to implement control and eradication programs because of the high seroprevalence at the individual level and at the herd in the central desert of Iran.

Molecular and serological investigation of Bovine Viral Diarrhea Virus in dairy cattle in Konya province

Bovine viral diarrhea virus (BVDV) infection is a viral disease observed in many parts of the world and causing significant economic losses in dairy cattle enterprises. The present study was carried out to determine the seropositivity of BVDV and perform the molecular detection of the virus in cattle in dairy enterprises situated in Konya province and its surroundings in the Central Anatolia Region. In this cross-sectional study performed between March 2017 and April 2019, a total of 393 serum samples were collected from twenty-four dairy cattle enterprises by random sampling. The presence of antibodies in the obtained blood serum samples was investigated by the virus neutralization test using NADL, the BVDV reference strain. Samples were controlled for BVDV specific antibody (Ab) presence and titter values using serum neutralization test. The serum samples were examined in terms of the presence of BVDV-specific antigens and specific RNA using a commercial ELISA kit and the RT-PCR method, respectively. According to the results of the analysis, the animal and herd-level seropositivity was 55.72% (219/393) and 79.16% (19/24), respectively. It was revealed that seropositivity between age groups was statistically significant (χ2:11.81; p=0.002). Moreover, the samples were determined to be 45.13%, 60.53%, and 73.07% seropositive in the 6 months-2 years, 2-5 years, and above 5 years age ranges, respectively. It was revealed that all of the samples tested to detect persistently infected animals were negative for antigen and BVDV-specific RNA. As a result, it indicates the presence of BVDV infection in dairy cattle enterprises in Konya province. Therefore, it is essential for the country's economy to prevent the spread of the infection in question and implement voluntary eradication programs.

The epidemiology of bovine viral diarrhea virus infection on a dairy farm - clinical signs, seroprevalence, virus detection and genotyping

In this study we investigated the presence of Bovine Viral Diarrhoea Virus (BVDV) antibodies and antigens in a dairy cow herd that had reported clinical signs related to BVDV infection. The aim of the study was to confirm BVDV infection on the dairy farm with 1500 Holstein-Friesian cattles, and genetically analyse the BVDV type/s circulating in the herd. Serum samples (n=233) were collected from clinically infected cattles and tested for the presence of BVDV antibodies and antigens. The virus neutralization test showed that 194 out of 233 sera of the tested cows were positive (83.26%), and by Ab-ELISA 203 sera samples of the tested cows were shown to be positive (87.12%). The presence of antigens was proven with the Ag-ELISA test in 2 out of 233 samples (0.86%). After RT-PCR and phylogenetic analysis of two BVD virus isolates, it was found that both isolates belonged to genotype 1, subtype BVDV-1d. This is the first time that the BVDV-1d subtype has been confirmed in Croatia, although the BVDV-1b and BVDV-1f subtypes have been detected previously on Croatian dairy farms.

EXPLORING THE USE OF THE ERYTHROCYTE SEDIMENTATION RATE AS AN INFLAMMATORY MARKER FOR FREE-RANGING WILDLIFE: A CASE STUDY IN AFRICAN BUFFALO (SYNCERUS CAFFER).

Measuring inflammatory markers is critical to evaluating both recent infection status and overall human and animal health; however, there are relatively few techniques that do not require specialized equipment or personnel for detecting inflammation among wildlife. Such techniques are useful in that they help determine individual and population-level inflammatory status without the infrastructure and reagents that many more-specific assays require. One such technique, known as the erythrocyte sedimentation rate (ESR), is a measure of how quickly erythrocytes (red blood cells) settle in serum, with a faster rate indicating a general, underlying inflammatory process is occurring. The technique is simple, inexpensive, and can be performed in the field without specialized equipment. We took advantage of a population of African buffalo (Syncerus caffer), well studied from June 2014 to May 2017, to understand the utility of ESR in an important wildlife species. When ESR was compared with other markers of immunity in African buffalo, it correlated to known measures of inflammation. We found that a faster ESR was significantly positively correlated with increased total globulin levels and significantly negatively correlated with increased red blood cell count and albumin levels. We then evaluated if ESR correlated to the incidence of five respiratory pathogens and infection with two tick-borne pathogens in African buffalo. Our results suggest that elevated ESR is associated with the incidence of bovine viral diarrhea virus infection, parainfluenza virus, and Mannheimia haemolytica infections as well as concurrent Anaplasma marginale and Anaplasma centrale coinfection. These findings suggest that ESR is a useful field test as an inflammatory marker in individuals and herds, helping us better monitor overall health status in wild populations.

First study on genetic variability of bovine viral diarrhea virus isolated from Sapera dairy goats with reproductive disorders in Yogyakarta, Indonesia.

Background and Aim:Bovine viral diarrhea (BVD) virus (BVDV) is an important viral pathogen of cattle that can infect diverse artiodactyl species. The clinical manifestations caused by BVDV in heterologous hosts, as they do in cattle, vary, although respiratory and reproductive failures are commonly reported. BVDV infections commonly result in reproductive failure in goats, with abortion being the primary clinical sign. In central Java, Indonesia, BVDV infection has been reported in two clinically healthy local goat species, and the testing indicated infection by BVDV Type 1. However, the genetic diversity of viruses has not been described in healthy or ill goats. The objectives of the present study were as follows: (1) To investigate the genetic variation of BVDV isolated from Sapera dairy goats with naturally occurring reproductive disorders in Yogyakarta, Indonesia, using the 5’ untranslated region (5’ UTR) and (2) to study the possible correlation between reproductive disorders and the presence of BVDV in the flock.Materials and Methods:Blood samples were collected in October 2021 from 39 goats that had been individually reported to have reproductive disorders. The serum samples were subjected to molecular detection and genetic characterization of BVDV based on the 5’ UTR of the viral genome, followed by sequencing and phylogenetic analyses. Viral isolation was performed on BVDV-positive samples to analyze the viral biotypes.Results:BVDV infection was detected in five out of 39 female goats. The clinical status of the BVDV-infected goats was abortion (n=2), metritis (n=1), and repeated breeding (n=2). All antigen-positive samples were confirmed as BVDV type 1a (BVDV-1a) and noncytopathic (NCP)-BVDV biotype.Conclusion:The BVDV-1a and NCP biotypes are the main subtypes and biotypes present in Sapera dairy goats exhibiting reproductive failure. This result is consistent with previous results in dairy cattle in Yogyakarta. The reported results can facilitate the design of methods for the prevention and control of BVD circulating in Indonesia.

Comparison of the confidence in freedom from infection based on different control programmes between EU member states: STOC free

The STOC free project constructed a generic framework that allows a standardised and harmonised description of different control programmes (CP) for cattle diseases. The STOC free model can be used to determine the confidence of freedom from infection that has been achieved in disease CPs, in support of an ongoing assessment of progress towards output-based standards as outlined in the EU Animal Health Law. With this information, and as required, further CP actions can be taken to mitigate the risks of persistence and (re-)introduction on the probability of freedom from infection. Bovine viral diarrhoea virus (BVDV) was chosen as the case disease because of the diversity in CPs in the six participating countries. A Bayesian hidden Markov model was considered the best modelling method. Detailed BVDV CP information was collected in the participating countries and the key aspects for inclusion in the STOC free model were identified. A first version of STOC free model was developed and tested on simulated data. The risk factors for BVDV infection that needed to be included in the model were defined and default values for these risk factors were quantified. A data collection tool was finalised with which the data for the STOC free model was collected. Subsequently, the developed model was tested and validated using real BVDV CP data from partner countries. Based on the feedback, the model was finalised and the report and corresponding computer code were made publicly available. There were roughly three different BVDV situations that occurred in the partner countries: 1. Endemic situation with a CP operating at herd level, 2. Endemic situation with a CP operating at animal level and 3. BVD free situation. The STOC free model is able to include herd level data only and animal level data has to be aggregated to herd level before the model can be applied. The STOC free model is not applicable for a country that is completely BVDV free given that it needs some infections to estimate its parameters and converge. In the latter situation, a scenario tree model could be a better suited tool, and this was evaluated in the Swedish case study. Further work is needed for generalisation of the method to other diseases and expansion of the method to include socioeconomic aspects of CPs.