Bovine Viral Diarrhea Research Articles

An end-point multiplex PCR/reverse transcription-PCR for detection of five agents of bovine neonatal diarrhea

Neonatal calf diarrhea (NCD) is frequently associated with single or mixed viral, bacterial and/or protozoal infections. Consequently, laboratory diagnostic of NCD usually requires specific tests for each potential agent; a time-consuming, laborious and expensive process. Herein, we describe an end-point multiplex PCR/reverse transcription-PCR (RT-PCR) for detection of five major NCD agents: bovine rotavirus (BRV), bovine coronavirus (BCoV), Escherichia coli K99 (E. coli K99), Salmonella enterica (S. enterica) and Cryptosporidium parvum (C. parvum). Initially, we selected and/or designed high-coverage primers. Subsequently, we optimized multiplex PCR/RT-PCR conditions. Next, we evaluated the analytical sensitivity of the assay and assessed the performance of the reaction by testing 95 samples of diarrheic calf feces. The analytical specificity was evaluated against bovine viral diarrhea virus (BVDV), E. coli heat-stable enterotoxin (STa) and Eimeria spp. The detection limit of our assay was about 10 infectious units of BRV, 10−2 dilution of a BCoV positive sample pool, about 5 × 10−4 CFU for S. enterica, 5 × 10−6 CFU for E. coli K99 and 50 oocysts for C. parvum. No non-specific amplification of other bovine diarrhea agents was detected. Out of 95 samples analyzed, 50 were positive for at least one target, being 35 single and 15 mixed infections. BRV was the most frequent agent detected in single infections (16/35), followed by Cryptosporidium spp. (11/35), which was the most frequent in mixed infections (11/15). Positive and negative multiplex results were confirmed in individual reactions. In conclusion, we described an end-point multiplex PCR/RT-PCR for faster and easier NCD diagnosis, which may be useful for routine diagnosis and surveillance studies.

First gene-edited calf with reduced susceptibility to a major viral pathogen.

Bovine viral diarrhea virus (BVDV) is one of the most important viruses affecting the health and well-being of bovine species throughout the world. Here, we used CRISPR-mediated homology-directed repair and somatic cell nuclear transfer to produce a live calf with a six amino acid substitution in the BVDV binding domain of bovine CD46. The result was a gene-edited calf with dramatically reduced susceptibility to infection as measured by reduced clinical signs and the lack of viral infection in white blood cells. The edited calf has no off-target edits and appears normal and healthy at 20 months of age without obvious adverse effects from the on-target edit. This precision bred, proof-of-concept animal provides the first evidence that intentional genome alterations in the CD46 gene may reduce the burden of BVDV-associated diseases in cattle and is consistent with our stepwise, in vitro and ex vivo experiments with cell lines and matched fetal clones.

The effect of neonatal vaccination for bovine respiratory disease in the face of a dual challenge with bovine viral diarrhea virus and Mannheimia hemolytica.

Bovine respiratory disease is the greatest threat to calf health. In this study, colostrum-fed dairyXbeef calves were vaccinated at ∼30days of age with an adjuvanted parenteral vaccine containing modified live bovine viral diarrhea virus (BVDV) type 1 and type 2, bovine herpesvirus 1 (BHV-1), bovine parainfluenza type 3 virus (PI3V) and bovine respiratory syncytial virus (BRSV) andM. haemolyticatoxoid (Group 1), or intranasal temperature-sensitive BHV-1, BRSV and PI3V concurrently witha parenteral vaccine containing modified live BVDV type 1 and type 2 andM. haemolyticatoxoid (Group 2) or a placebo (Group 3). The calves were challenged ∼150days post vaccination intranasally with BVDV 1b and then 7days later intratracheally withM. haemolytica. The calves wereeuthanized 6days after theM. haemolyticachallenge. Clinical signs following BVDV infection were similar in all groups. There was increased rectal temperatures in the Groups 2 and 3 on day 3 and in Group 3 on days 8-13. Group 1 animals had a slight leukopenia following BVDV infection while Groups 2 and 3 had greater leukopenia. BVDV type 1 and 2 serum titers increased in Group 1 following vaccination while these titers waned in Groups 2 and 3. There were higher levels of BVDV in the buffy coats and nasal samples in Group 2 and Group 3 versus Group 1 (p<0.01). Interferon-gamma response was higher (p<0.01) in Group 1 animals than Groups 2 and 3. Group 1 had the lowest percent pneumonic tissue (1.6%) while Group 2 vaccinates had 3.7% and the control Group 3 was 5.3%. Vaccination in the face of maternal antibody with a parenteral adjuvanted vaccine resulted in better protection than the regimen of an intranasal vaccine anda parenteral adjuvanted BVDV andM haemolyticacombination vaccine in a BVDV-M. haemolyticadual challenge.

160 Cross-neutralisation between Bovine Viral Diarrhea Virus (BVDV) type 1 and 2 after vaccination with a BVDV-1a modified-live-vaccine

160 Cross-neutralisation between Bovine Viral Diarrhea Virus (BVDV) type 1 and 2 after vaccination with a BVDV-1a modified-live-vaccine

Evaluation of the stress‐reducing effect of trace mineral injection in beef calves

Little is known about the effects of trace mineral supplementation on the stress response in beef calves. To investigate the effect of injectable trace mineral supplementation (ITM) on the stress response in beef calves exposed to different types of stress. Thirty weaned Angus and Angus crossbred calves. The enrolled calves were randomly assigned to 2 groups: ITM, 15 calves received modified-live virus vaccine (MLV) and ITM SC and 15 calves received MLV and saline SC (CONT). The calves were exposed to 3 types of stress: the stress of MLV vaccination (d0), nasal aerosol with bovine viral diarrhea virus-2 (BVDV-2) challenge (d5), and liver biopsy (d26). The calves' body weights and health status were monitored. Leukocyte counts, serum cortisol concentration ([cort]), BVDV-2 serum neutralizing antibodies (SNA), and percentages of CD4+ , CD8+ , WC1+ , and CD25+ T-lymphocytes were measured. Serum cortisol concentration ([cort]) showed strong associations with the percentage of CD8+ (rs = .50), BVDV2-SNA (rs = -.43), and WC1CD25+ (rs = .41) cells, and rectal temperature (rs = .40). The highest [cort] was reported 3 days after aerosol BVDV-2 challenge. Serum [cort] was decreased in ITM-treated calves 3 days post-BVDV-2 challenge, compared with CONT calves, with an average decrease of 18.5 ng/μL (95% confidence interval [CI], -6.07 to -31.3). The ITM-treated calves were heavier and healthier (P < .01)than the CONT calves. Trace mineral supplementation appears to have stress mitigation effects in beef cattle that may reflect positively on growth and health performance. Viral exposure is associated with a high degree of stress, which is considered a major welfare concern.

Hypersphere and Antiviral Activity of Three Alkyl Chain Iminocyclitols with D and L Ribitol Stereochemistry

<I>N</I>-Alkyl-C₁-dialkyl chains iminocyclitols with D or L-ribitol stereochemistry are synthesized with high diastereoselectivity after Grignard reagents addition to <I>N</I>-quaternary pyrrolines salts, and tested for antiviral activity in bovine viral diarrhea virus (BVDV), surrogate for hepatitis C virus (HCV). Dihedral angles are calculated from carbon chemical shift (δ_Cn[ppm]) with 3-sphere method without building units. 3-Sphere, a hypersphere in 4D, under Hopf fibration and Lie algebra mathematics theories enable calculation of the dihedral angles from the NMR data (vicinal coupling constant ³<I>J</I>_HnHn+1[Hz], chemical shift δ_Cn[ppm]). Instead of 3D manifold equations on seven sets unit or six sets units are proposed equations between 4D – 2D, in function of the curvature. The relationship between the antiviral activity and the iminocyclitol structure reveals that monoalkyl chain, <I>N</I>-n-C₁-dodecyl β-L-ribitol trifloroacetate salt 30 (IC₅₀ 1.5 uM) has higher antiviral activity in tangential space, relative to three alkyl chain, <I>N</I>-Methyl-C₁-butil, nonyl-L-ribitol. HCl 26 (IC₅₀ < 2 uM) with torus and Dupin cyclide coordinate, both with coordinates in 2D. Three alkyl chain isopropylidene protected pyrrolidine 25 has in 4D with all equations for calculation of the dihedral angles, and in protected pyrroline 19b double bond moves the coordinates in 2D.

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.

Serological Detection of SRMV, Bovine Viral Diarrhea Virus, BVDV and BEFV in Camels (Camelus dromedarius) in Southwest Iran

Background: Small ruminant morbillivirus (SRMV), bovine viral diarrhea virus (BVDV), bovine herpes virus-1 (BHV-1), and bovine ephemeral fever virus (BEFV) are among the most important viruses of farm animals. Objectives: This study aimed at the serological detection of SRMV, BVDV, BHV-1, and BEFV in the camel population of Khuzestan Province, located in southwest Iran. Methods: A total of 150 camel blood samples were randomly collected from free-ranging, seemingly healthy camels of both sexes and various ages in 8 different regions of Khuzestan Province, Iran. The sera were tested for SRMV, BVDV, BHV-1, and BEFV with serum neutralization test. Results: The seropositive samples for SRMV, BVDV, and BHV-1 were 1(0.67%), 7(4.67%), and 11(7.33%), respectively. There was no seroconversion against BEFV in the serum samples. Conclusion: The camel population of Khuzestan Province is subclinically infected with SRMV, BVDV, and BHV-1 and could play a significant role in the epizootiology of these viral diseases in this region, which is very important for the control and eradication programs.

Causes of fetal death in the Flemish cattle herd in Brazil.

Flemish cattle in Brazil are on the brink of extinction and are found only in one herd in Lages, Santa Catarina State. This study aimed to uncover the reasons for the recurring abortions in the Flemish cattle herd. Seventeen Flemish fetuses underwent postmortem examinations, with samples collected for histopathology and microbiology culture tests, polymerase chain reaction (PCR) test for Neospora caninum, and reverse transcription-PCR (RT-PCR) test for bovine viral diarrhea virus (BVDV) from 2015 to 2020. Of the 17 fetuses, N. caninum was the most common diagnosis and was found in 88% (15/17). One fetus (5.8%) had a coinfection with N. caninum and Citrobacter amalonaticus, leading to fibrinonecrotic pericarditis. All fetuses tested negative for BVDV by RT-PCR. Of the 107 dams tested by indirect immunofluorescence assay, 26 (25.2%) were anti-N. caninum seropositive, with 17 (65.4%) aborting and 5 (19.2%) having estrus repetition. Reverse transcription-PCR results showed that 9 (8.4%) of the serum samples collected from dams tested positive, which tested follow-up test 3 months later, indicating a BVDV transient infection. The factors that contributed to neosporosis included dogs' access to pastures and improper disposal of fetal remains, which made it easier for dogs to consume them. This study warns the occurrence of N. caninum as a cause of reproductive disorders that can lead to abortion in the studied Flemish cattle herd.

Hemorrhagic disease caused by bovine viral diarrhea virus-2a in Korean Indigenous Cattle: case reports

Bovine viral diarrhea virus (BVDV), which is prevalent worldwide, is one of the most important viral pathogens and causes substantial economic loss to the livestock industry. Despite its importance, BVDV is largely unnoticed in the Republic of Korea (ROK). In this study, we report the case of a steer with BVDV that died suddenly due to severe enteritis. Two 1-year-old Korean indigenous cattle in the same herd presented severe hemorrhagic diarrhea. Case 1 had severe dehydration and died after 3 days, whereas case 2 had anorexia, depression, and severe diarrhea with mucus and blood. Only case 2 was necropsied, and BVDV2a was detected in the tissues of its alimentary tract. Gross lesions, including erosion, ulceration, and extensive hemorrhage, were observed in the digestive tract mucosa. Immunohistochemistry revealed marked positive staining for BVDV2a antigen in the large intestine. This report describes the first case of hemorrhagic disease caused by acute BVDV2a infection, which is characterized by high mortality in Korean indigenous cattle. This study will help establish vaccination and control strategies for BVDV in the ROK.

Circulation of classical swine fever virus (CSFV) strains of bovine origin in China and India.

The classical swine fever virus (CSFV) is a species member of the family Flaviviridae. CSFV is widely distributed in the world causing a severe impact on pig industry. This pathogen is considered restricted to domestic and wild suids. However, some reports from 2014 to 2018 showed the presence of the CFSV antigen in the bovine species. The virus was found in commercialized batches of fetal bovine serum (FBS) of Chinese origin and in bovine herds in in the provinces of Henan and Jiangsu, China, and in Tamil Nadu and Meghalaya, southern and north‑eastern states of India, respectively. Detection was done using antigen capture ELISA and RT‑PCR tests. In certain cases, animals with natural infection showed clinical signs and reproduction was also affected. Genetic characterization was performed considering the 5'‑UTR sequences of the bovine strains. In addition, the entire CSFV E2 genomic region could be amplified from two positive animals. The bovine strains were genetically related to the Chinese CSFV live attenuated hog cholera lapinized vaccine (HCLV) strain used in pigs, sharing sequence characteristics. The vaccine strain HCLV was widely used in China to protect bovines and yaks from bovine viral diarrhea, and, as a possible consequence, inducing an adaptation in cattle and a further natural diffusion. Furthermore, a contaminant strain from China was genetically distant from all other previously described genotypes of the CSFV. This suggests also the occurrence of micro evolutive step in the species related to geographical segregation. These observations deserve attention and further investigations, especially relevant in countries where CSFV control and eradication strategies are applied.

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).

Sensitive Detection of BVDV Using Gold Nanoparticle-Modified Few-Layer Black Phosphorus with Affinity Peptide-Based Electrochemical Sensor

The lethality of the bovine viral diarrhea virus (BVDV) in cattle involves inapparent infection and various, typically subclinical, syndromes. Cattle of all ages are vulnerable to infection with the virus. It also causes considerable economic losses, primarily due to reduced reproductive performance. In the absence of treatment that can completely cure infected animals, detection of BVDV relies on highly sensitive and selective diagnosis methods. In this study, an electrochemical detection system was developed as a useful and sensitive system for the detection of BVDV to suggest the direction of diagnostic technology through the development of conductive nanoparticle synthesis. As a countermeasure, a more sensitive and rapid BVDV detection system was developed using the synthesis of electroconductive nanomaterials black phosphorus (BP) and gold nanoparticle (AuNP). To increase the conductivity effect, AuNP was synthesized on the BP surface, and the stability of BP was improved by using dopamine self-polymerization. Moreover, its characterizations, electrical conductivity, selectivity, and sensitivity toward BVDV also have been investigated. The BP@AuNP-peptide-based BVDV electrochemical sensor exhibited a low detection limit of 0.59 copies mL-1 with high selectivity and long-term stability (retaining 95% of its initial performance over 30 days).

Effects of Spatial Expression of Activating Transcription Factor 4 on the Pathogenicity of Two Phenotypes of Bovine Viral Diarrhea Virus by Regulating the Endoplasmic Reticulum-Mediated Autophagy Process.

The endoplasmic reticulum (ER) stress response is a highly conserved stress-defense mechanism and activates the adaptive unfolded protein response (UPR) to mitigate imbalance. The ER stress-activated signaling pathways can also trigger autophagy to facilitate cellular repair. Bovine viral diarrhea virus (BVDV) utilizes the host cellular ER as the primary site of the life cycle. However, the interplay between cellular ER stress and BVDV replication remains unclear. This report reveals that cytopathic (cp) and noncytopathic (ncp) BVDV have distinct strategies to regulate UPR mechanisms and ER stress-mediated autophagy for their own benefit. Immunoblot analysis revealed that cp and ncp BVDV differentially regulated the abundance of ER chaperone GRP78 for viral replication, while the protein kinase RNA-like ER kinase (PERK)-eukaryotic translation initiation factor 2 subunit α (eIF2α)-activating transcription factor 4 (ATF4) pathway of the UPR was switched on at different stages of infection. Pretreatment with ER stress inducer promoted virion replication, but RNA interference (RNAi) knockdown of ATF4 in BVDV-infected cells significantly attenuated BVDV infectivity titers. More importantly, the effector ATF4 activated by cp BVDV infection translocated into the nucleus to mediate autophagy, but ATF4 was retained in the cytoplasm during ncp BVDV infection. In addition, we found that cp BVDV core protein was localized in the ER to induce ER stress-mediated autophagy. Overall, the potential therapeutic target ATF4 may contribute to the global eradication campaign of BVDV. IMPORTANCE The ER-tropic viruses hijack the host cellular ER as the replication platform of the life cycle, which can lead to strong ER stress. The UPR and related transcriptional cascades triggered by ER stress play a crucial role in viral replication and pathogenesis, but little is known about these underlying mechanisms. Here, we report that cytopathic and noncytopathic BVDV use different strategies to reprogram the cellular UPR and ER stress-mediated autophagy for their own advantage. The cytopathic BVDV unconventionally downregulated the expression level of GRP78, creating perfect conditions for self-replication via the UPR, and the noncytopathic BVDV retained ATF4 in the cytoplasm to provide an advantage for its persistent infection. Our findings provide new insights into exploring how BVDV and other ER-tropic viruses reprogram the UPR signaling pathway in the host cells for replication and reveal the attractive host target ATF4 for new antiviral agents.

Serum and Milk Levels of Antibodies to Bovine Viral Diarrhoea Virus, Bovine Herpesvirus-1 and -4, and Circulation of Different Bovine Herpesvirus-4 Genotypes in Dairy Cattle with Clinical Mastitis.

Bovine viral diarrhoea virus (BVDV) and bovine herpesvirus (BoHV)-1 and -4 are important causes of respiratory diseases and reproductive disorders of dairy cattle worldwide. Investigation of BVDV and BoHV-1 and -4 antibody levels in the serum and milk of dairy cattle in a group with clinical mastitis and a healthy group was undertaken using an indirect ELISA, and identification of the BoHV-4 genotypes in clinical mastitis cases was attempted by PCR and sequencing. Antibodies specific to BVDV, BoHV-1 and BoHV-4 were detected in the serum and milk of all dairy cattle with clinical mastitis. The cut-off values for BVDV and BoHV-1 in the sera and milk were extremely high in both healthy and mastitic animals. However, BoHV-4 antibodies were detected only in the clinically mastitic cattle, and BoHV-4 levels were higher in milk than in sera among these animals. Genotypes I and II of BoHV-4 were detected in the milk samples of four seropositive cows with clinical mastitis from the same herd. The results of this investigation demonstrate that clinical mastitis cases in the same herd may have aetiology in different BoHV-4 genotypes.

Characterization of a Highly Virulent Noncytopathic Bovine Viral Diarrhea Virus 2b Isolate Detected in Cattle in Inner Mongolia, China

Bovine viral diarrhea virus (BVDV) infection causes subclinical to severe acute disease in cattle all over the world. Two genotypes of BVDV are recognized, BVDV1 and BVDV2. Currently, the subtypes of BVDV1b and BVDV2a are mainly circulating in China. Here, a newly noncytopathic (ncp) BVDV strain named BVDVNM21 was isolated and identified from cattle. We determined the complete genome sequence of BVDVNM21, phylogenetic analysis of 5′ untranslated region (5′UTR), Npro, and E2 gene, and complete genome showed the virus belongs to BVDV2b subtype and formed an independent clade within BVDV2b. Genome-wide sequence analysis showed that BVDVNM21 had high homology with SD1301 (98.10%) isolated from China. In the experimental infection study of guinea pigs and calves, they all developed similar clinical signs, including a pronounced and prolonged febrile response lasting more than 3 days and pronounced reduction in white blood cells (WBC) and platelets of more than 40%. Moreover, guinea pigs showed more acute infection characteristics after infection, and WBC decreased by more than 50% at 1 day postinfection (dpi), and they recovered in less than 3 days. The results indicated that the BVDVNM21 strain showed high virulence in calves. It was the first confirmed isolation of a highly pathogenic ncpBVDV2b in cattle, as well as the establishment of the BVDV-guinea pig model. This study may provide a basis for further research and control of the prevalence of BVDV2b in China.

Recent advances in antigen targeting to antigen-presenting cells in veterinary medicine.

Advances in antigen targeting in veterinary medicine have gained traction over the years as an alternative approach for diseases that remain a challenge for traditional vaccines. In addition to the nature of the immunogen, antigen-targeting success relies heavily on the chosen receptor for its direct influence on the elicited response that will ensue after antigen uptake. Different approaches using antibodies, natural or synthetic ligands, fused proteins, and DNA vaccines have been explored in various veterinary species, with pigs, cattle, sheep, and poultry as the most frequent models. Antigen-presenting cells can be targeted using a generic approach, such as broadly expressed receptors such as MHC-II, CD80/86, CD40, CD83, etc., or focused on specific cell populations such as dendritic cells or macrophages (Langerin, DC-SIGN, XCR1, DC peptides, sialoadhesin, mannose receptors, etc.) with contrasting results. Interestingly, DC peptides show high specificity to DCs, boosting activation, stimulating cellular and humoral responses, and a higher rate of clinical protection. Likewise, MHC-II targeting shows consistent results in enhancing both immune responses; an example of this strategy of targeting is the approved vaccine against the bovine viral diarrhea virus in South America. This significant milestone opens the door to continuing efforts toward antigen-targeting vaccines to benefit animal health. This review discusses the recent advances in antigen targeting to antigen-presenting cells in veterinary medicine, with a special interest in pigs, sheep, cattle, poultry, and dogs.

The effect of fibroblast growth factor 21 on a mouse model of bovine viral diarrhea.

Previously, we researched that bovine viral diarrhea virus (BVDV) induced a very significant increase in fibroblast growth factor 21 (FGF21) expression in mouse liver and that FGF21 was increased in the peripheral blood of BVD cattle and BVD mice. To determine the role of FGF21 in relieving clinical symptoms and inhibiting the intestinal damage caused by BVDV in BVD development in mice, BALB/c mice were intraperitoneally injected with cytopathic biotype (cp) BVDV-LS01 (isolated and identified by our group) to establish a BVD mouse model. The role of FGF21 in the BVD mouse model was investigated by injecting the mice with FGF21. The animals were divided into control, BVDV challenge, BVDV + FGF21, BVDV + FGF21Ab (anti-FGF21 antibody), and BVDV + IgG (immunoglobulin G) groups. The stool consistency, the degree of bloody diarrhea, histopathological changes, inflammatory cell infiltration, weight loss percentage, and detection of BVDV in the feces of the mice were examined, and the pathological changes and inflammatory cytokine expression were analyzed. The results showed that after BVDV challenge, the average BVD mouse model score of the BVDV mice was 11.6 points. In addition to mild diarrhea and tissue damage, BVDV was detected in the stools of 13 BVDV mice. Only two mice in the control group had scores (both, 1 point each). The comprehensive scoring results demonstrated the successful establishment of the BVD mouse model. FGF21 alleviated the clinical symptoms in the BVD mice and significantly improved weight loss. Furthermore, FGF21 inhibited the BVDV-induced leukocyte, platelet, and lymphocyte reduction while inhibiting the expression of BVDV-induced inflammatory factors. In the BVD mice, FGF21 promoted duodenal epithelial cell proliferation, thereby significantly improving the damage to the cells. In conclusion, FGF21 exerted a good therapeutic effect on the BVD mouse model.

Response to Bovine Viral Diarrhea Virus in Heifers Vaccinated with a Combination of Multivalent Modified Live and Inactivated Viral Vaccines

Bovine viral vaccines contain both live or inactivated/killed formulations, but few studies have evaluated the impact of vaccinating with either live or killed antigens and re-vaccinating with the reciprocal. Commercial dairy heifers were utilized for the study and randomly assigned to three treatment groups. Treatment groups received a commercially available modified-live viral (MLV) vaccine containing BVDV and were revaccinated with a commercially available killed viral (KV) vaccine containing BVDV, another group received the same KV vaccine and was revaccinated with the same MLV vaccine, and yet another group served as negative controls and did not receive any viral vaccines. Heifers in KV/MLV had higher virus neutralizing titers (VNT) at the end of the vaccination period than heifers in MLV/KV and control groups. The frequency of IFN-γ mRNA positive CD4+, CD8+, and CD335+ populations, as well as increased mean fluorescent intensity of CD25+ cells was increased for the MLV/KV heifers as compared to KV/MLV and controls. The data from this study would suggest that differences in initial antigen presentation such as live versus killed could augment CMI and humoral responses and could be useful in determining vaccination programs for optimizing protective responses, which is critical for promoting lifetime immunity.

A One-Year Retrospective Analysis of Viral and Parasitological Agents in Wildlife Animals Admitted to a First Aid Hospital.

This study aimed to provide information on the presence and frequency of viral and parasitic agents in wildlife presented to a Veterinary Teaching Hospital in 2020-2021. Serum and faecal samples were collected from 50 rescued animals (roe deer, fallow deer, foxes, badgers, pine martens, and porcupines) and examined by serological, molecular, and parasitological techniques. Transtracheal wash (TTW) was also collected post-mortem from roe deer. Overall, the results of the different techniques showed infections with the following viral and parasitic agents: Bovine Viral Diarrhea Virus, Small Ruminant Lentiviruses, Kobuvirus, Astrovirus, Canine Adenovirus 1, Bopivirus, gastrointestinal strongyles, Capillaria, Ancylostomatidae, Toxocara canis, Trichuris vulpis, Hymenolepis, Strongyloides, Eimeria, Isospora, Dictyocaulus, Angiostrongylus vasorum, Crenosoma, Dirofilaria immitis, Neospora caninum, Giardia duodenalis, and Cryptosporidium. Sequencing (Tpi locus) identified G. duodenalis sub-assemblages AI and BIV in one roe deer and one porcupine, respectively. Adult lungworms collected from the TTW were identified as Dictyocaulus capreolus (COX1 gene). This is the first molecular identification of G. duodenalis sub-assemblage AI and D. capreolus in roe deer in Italy. These results show a wide presence of pathogens in wild populations and provide an overview of environmental health surveillance.