Quantitative proteomics reveals significant variation in host responses of cattle with differing buffalo fly susceptibility.

Background: Control of buffalo flies (Haematobia irritans exigua, BFs) relies mainly on chemical methods; however, resistance to insecticides is widespread in BF populations. Breeding for resistance to BFs represents a possible alternative, but direct phenotyping of animals is laborious and often inaccurate. The availability of reliable diagnostic biomarker(s) to identify low BF carrier cattle would facilitate rapid and accurate selection for genetic improvement. However, limited information is available regarding differences amongst cattle in host responses to BF infestation. Methods: This study investigated the variation in Brangus cattle serum proteomic profiles before (naïve) and after peak BF exposure, in low (LF) and high BF burden (HF) cattle. Cattle were phenotyped for susceptibility based on BF counts on multiple dates using visual and photographic techniques. The relative abundance of serum proteins in cattle before and after exposure to BFs was analysed using sequential window acquisition of all theoretical fragment ion mass spectrometry (SWATH-MS). Results: Exposure to BFs elicited similar responses in HF and LF cattle, with 79 and 70 proteins, respectively, showing significantly different abundances post exposure as compared to their relevant naïve groups. The comparison of serum samples from naïve HF and LF cattle identified 44 significantly differentially abundant (DA) proteins, while 37 significantly DA proteins were identified from the comparison between HF and LF cattle post-exposure to BFs. Proteins with higher abundance in naïve LF cattle were enriched in blood coagulation mechanisms that were sustained after exposure to BFs. Strong immune response mechanisms were also identified in naïve LF cattle, whereas these responses developed in HF cattle only after exposure to BF. High BF cattle also showed active anticoagulation mechanisms in response to BF exposure, including downregulation of coagulation factor IX and upregulation of antithrombin-III, which might facilitate BF feeding. Conclusion: Underlying differences in the abundance of proteins related to blood coagulation and immune response pathways could potentially provide indirect indicators of susceptibility to BF infestation and biomarkers for selecting more BF-resistant cattle.

Mixed practice was my plan then cattle health specialisation caught my attention.

Gaining experience in mixed practice was the starting point for Nicola Gladden, then locuming and travel gave her the space to decide which career route to follow next, choosing a residency pathway.

Phenotypic and genotypic characterization of ticks and tick-borne pathogens from cattle in selected villages of Greater Letaba Municipality in Limpopo Province, South Africa.

Ticks are blood ectoparasites that feed on domestic, wild animals and humans. They spread a variety of infections such as protozoa, viruses, and bacteria. Moreover, cattle reared by smallholder farmers are susceptible to ticks and tick-borne pathogens. Therefore, accurate identification of ticks and detection of tick-borne pathogens is crucial. The main aim of this study was to identify and characterize ticks and tick-borne pathogens from selected villages in Greater Letaba Municipality, Limpopo Province, using morphological and molecular techniques. A total of 233 ticks were collected from cattle and identified morphologically using appropriate morphological keys. The following tick species were identified: Amblyomma hebraeum, Hyalomma rufipes, Hyalomma truncatum, Rhipicephalus appendiculatus, Rhipicephalus (Boophilus) decoloratus, Rhipicephalus (Boophilus) microplus, Rhipicephalus evertsi evertsi, and Rhipicephalus sanguineus. Rhipicephalus spp. was the most common species accounting to 73.8% of the identified ticks. The genomic DNA was extracted from the whole tick for tick identification and from midguts of the ticks for the detection of tick-borne pathogens, followed by amplification and sequencing. A total of 27 samples were positive for tick-borne pathogens: 23 samples tested positive for Theileria and four samples tested positive for Ehrlichia. Anaplasma and Rickettsial OmpB could not be detected from any of the samples. There was no obvious grouping of ticks and tick-borne pathogens on the bases of their locality. The findings of this study confirm previous reports that indicated that cattle reared by smallholder farmers harbor various ticks and tick-borne pathogens of veterinary, public health, and economic importance. Regular monitoring of tick infestations in villages around the study areas is recommended to avoid disease outbreaks.

First detection of Cupriavidus gilardii in a bovine neonatal diarrhea outbreak.

BACKGROUND: Cupriavidus gilardii is an aerobic, gram-negative, motile, glucose-nonfermenting bacillus, first described in 1999. Typically, it exhibits low pathogenicity in humans, causing opportunistic infections primarily in individuals with compromised immune systems. This bacterium has been also found in various environmental sources such as plants and contaminated soils. Notably, there have been no documented cases of C. gilardii infections in animals. CASE PRESENTATION: This case report outlines a bovine neonatal diarrhea outbreak that occurred in Northern Greece, during which C. gilardii was isolated. Faecal samples from 5-day-old calves were collected and transported to the laboratory for further examination. Bacterial culture and next generation sequencing techniques were employed to confirm the presence of this bacterium in the samples. Following the isolation and identification of C. gilardii from the samples, an autogenous vaccine was produced and administered to the cows within the farm. Subsequent to vaccination, a progressive reduction in calf diarrhea and deaths was observed, leading to their eventual complete resolution. To the best of our knowledge, this represents the first documentation of C. gilardii isolation from cases of bovine neonatal diarrhea. CONCLUSION: This case report presents the first isolation case of C. gilardii from animal samples and more specifically from calf faecal samples. It represents an important observation, providing evidence that this opportunistic human pathogen could contribute to clinical symptoms in animals.

An evaluation form to aid dairy producers to systematically assess cows prior to culling.

Objective: The objective of this prospective observational research project was to have dairy producers use and assess the utility of a cull cow evaluation form. Animals: Cull dairy cows. Procedure: Veterinarians were recruited to enrol a purposively selected group of dairy producers into a project to evaluate a cull cow evaluation form. Producers were provided with evaluation forms and asked to complete a form for every cow they culled from their herd from January to June 2017, inclusive. Results: A total of 44 producers used the form to record information on 323 cows prior to transport. Conclusion and clinical relevance: Despite the completion of 323 forms, only ~1/3 were completed fully, with compliance highest for body condition score, lameness, and temperature recordings (> 90% of forms). A cull cow evaluation form may improve the thoroughness and consistency of dairy producer assessment of cull dairy cows for fitness for transport.

Un formulaire d'évaluation pour aider les producteurs laitiers à évaluer systématiquement les vaches avant la réforme. Objectif: L'objectif de ce projet de recherche observationnelle prospective était d'amener les producteurs laitiers à utiliser et à évaluer l'utilité d'un formulaire d'évaluation des vaches de réforme. Animaux: Vaches laitières réformées. Procédure: Des vétérinaires ont été recrutés pour inscrire un groupe de producteurs laitiers sélectionnés à dessein dans un projet visant à évaluer un formulaire d'évaluation des vaches réformées. Les producteurs ont reçu des formulaires d'évaluation et ont été invités à remplir un formulaire pour chaque vache qu'ils ont éliminée de leur troupeau de janvier à juin 2017 inclusivement. Résultats: Au total, 44 producteurs ont utilisé le formulaire pour enregistrer des informations sur 323 vaches avant le transport. Conclusion et pertinence clinique: Malgré la complétion de 323 formulaires, seulement environ 1/3 ont été entièrement remplis, avec une conformité plus élevée pour le score d'état corporel, les boiteries et les enregistrements de température (> 90 % des formulaires). Un formulaire d'évaluation des vaches laitières réformées peut améliorer la rigueur et la cohérence de l'évaluation par le producteur laitier des vaches laitières réformées quant à leur aptitude au transport.(Traduit par Dr Serge Messier).

Comparative efficacy of modified-live and inactivated vaccines in boosting responses to bovine respiratory syncytial virus, bovine parainfluenza virus Type 3, and bovine coronavirus following neonatal mucosal priming of beef calves.

Objective: This study compared clinical and immunological responses to coinfection challenge of beef calves mucosally primed and differentially boosted with commercial combination vaccines containing antigens against bovine coronavirus (BCoV), bovine parainfluenza virus Type 3 (BPIV3), and bovine respiratory syncytial virus (BRSV). Animals: Nineteen commercial beef heifers. Procedure: At birth, calves were mucosally (IN) primed with modified-live virus (MLV) vaccines, differentially boosted by injection of either combination MLV (IN-MLV) or inactivated virus (IN-KV) vaccines at a mean age of 44 d, and then challenged by coinfection with BCoV, BPIV3, and BRSV at weaning. Results: Both groups were similarly protected from clinical disease and had anamnestic neutralizing antibody responses to all 3 viruses. The IN-KV group shed more BCoV, and less BPIV3 and BRSV, than the IN-MLV group. Conclusion: These data indicated similar clinical and immunological protection between IN-MLV and IN-KV; however, shed of virus varied. Clinical relevance: Whereas boosting with KV or MLV appeared to have similar efficacy, viral shed differences may affect disease control.

Efficacité comparative des vaccins vivants modifiés et inactivés pour stimuler les réponses au virus respiratoire syncytial bovin, au virus parainfluenza bovin de type 3 et au coronavirus bovin après amorçage via la muqueuse de veaux de boucherie nouveau-nés. Objectif: Cette étude a comparé les réponses cliniques et immunologiques à une co-infection de veaux de boucherie amorcés par voie muqueuse et différentiellement stimulés avec des vaccins combinés commerciaux contenant des antigènes contre le coronavirus bovin (BCoV), le virus parainfluenza bovin de type 3 (BPIV3) et le virus respiratoire syncytial bovin (BRSV). Animaux: Dix-neuf génisses de boucherie commerciales. Procédure: À la naissance, les veaux ont été vaccinés au niveau des muqueuses (IN) avec des vaccins à virus vivants modifiés (MLV), stimulés de manière différentielle par l'injection de vaccins combinés MLV (IN-MLV) ou de virus inactivés (IN-KV) à un âge moyen de 44 jours. puis provoqué par une co-infection avec BCoV, BPIV3 et BRSV au sevrage. Résultats: Les deux groupes étaient protégés de la même manière contre la maladie clinique et présentaient des réponses anamnestiques en anticorps neutralisants contre les 3 virus. Le groupe IN-KV a excrété plus de BCoV et moins de BPIV3 et de BRSV que le groupe IN-MLV. Conclusion: Ces données indiquent une protection clinique et immunologique similaire entre IN-MLV et IN-KV; cependant, l'excrétion du virus variait. Pertinence clinique: Alors que le rappel avec KV ou MLV semble avoir une efficacité similaire, les différences d'excrétion virale peuvent affecter la limitation de la maladie.(Traduit par Dr Serge Messier).

Molecular findings and virological assessment of bladder papillomavirus infection in cattle.

Bovine and ovine papillomaviruses (BPVs - OaPVs) are infectious agents that have an important role in bladder carcinogenesis of cattle. In an attempt to better understand territorial prevalence of papillomavirus genotypes and gain insights into their molecular pathway(s), a virological assessment of papillomavirus infection was performed on 52 bladder tumors in cattle using droplet digital polymerase chain reaction (ddPCR), an improved version of conventional PCR. ddPCR detected and quantified BPV DNA and mRNAs in all tumor samples, showing that these viruses play a determinant role in bovine bladder carcinogenesis. OaPV DNA and mRNA were detected and quantified in 45 bladder tumors. BPV14, BPV13, BPV2, OaPV2, OaPV1, and OaPV3 were the genotypes most closely related to bladder tumors. ddPCR quantified BPV1 and OaPV4 DNA and their transcripts less frequently. Western blot analysis revealed a significant overexpression of the phosphorylated platelet derived growth factor ß receptor (PDGFßR) as well as the transcription factor E2F3, which modulate cell cycle progression in urothelial neoplasia. Furthermore, significant overexpression of calpain1, a Cys protease, was observed in bladder tumors related to BPVs alone and in BPV and OaPV coinfection. Calpain1 has been shown to play a role in producing free transcription factors of the E2F family, and molecular findings suggest that calpain family members work cooperatively to mutually regulate their protease activities in cattle bladder tumors. Altogether, these results showed territorial prevalence of BPV and OaPV genotypes and suggested that PDGFßR and the calpain system appeared to be molecular partners of both BPVs and OaPVs.

Empirical and model-based evidence for a negligible role of cattle in peste des petits ruminants virus transmission and eradication.

Peste des petits ruminants virus (PPRV) is a multi-host pathogen with sheep and goats as main hosts. To investigate the role of cattle in the epidemiology of PPR, we simulated conditions similar to East African zero-grazing husbandry practices in a series of trials with local Zebu cattle (Bos taurus indicus) co-housed with goats (Capra aegagrus hircus). Furthermore, we developed a mathematical model to assess the impact of PPRV-transmission from cattle to goats. Of the 32 cattle intranasally infected with the locally endemic lineage IV strain PPRV/Ethiopia/Habru/2014 none transmitted PPRV to 32 co-housed goats. However, these cattle or cattle co-housed with PPRV-infected goats seroconverted. The results confirm previous studies that cattle currently play a negligible role in PPRV-transmission and small ruminant vaccination is sufficient for eradication. However, the possible emergence of PPRV strains more virulent for cattle may impact eradication. Therefore, continued monitoring of PPRV circulation and evolution is recommended.

Integrating multi-wet laboratory diagnostics to study staphylococci in animals in Uganda.

BACKGROUND: Several diagnostic environments in Uganda lack real-time, robust and high-throughput technologies for comprehensive typing of microbes, which is a setback to infectious disease surveillance. This study combined various wet laboratory diagnostics to understand the epidemiology of pathogenic staphylococci isolated from animals in Uganda and the implications for global health security priorities. METHODS: A retrospective study was conducted employing records and pathogenic staphylococci (from animals) archived at the Central Diagnostic Laboratory (CDL), Makerere University, Uganda, between January 2012 and December 2019. The bacteria were speciated by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and tested for virulence factors [beta lactamases, lecithinase, deoxyribonuclease (DNase), haemolysins] and resistance to ten antimicrobials of clinical and veterinary relevance. Tetracycline and methicillin resistance genes were also tested. RESULTS: The prevalent diseases were mastitis in cattle and skin infections in dogs. Of the 111 staphylococci tested by MALDI-TOF MS, 79 (71.2%) were Staphylococcus aureus, 27 (24.3%) were Staphylococcus pseudintermedius and 5 (4.5%) were Staphylococcus schleiferi. All these strains expressed haemolysins. The prevalence of strains with lecithinase, penicillinase, cephalosporinase and DNase was 35.9% (14/39), 89.7% (35/39), 0.0% (0/39) and 87.2% (34/39), respectively. Staphylococci were primarily resistant to early penicillins (over 80%), tetracycline (57.7%), and chloramphenicol (46.2%). Minimal resistance was noted with cloxacillin (0.0%), ciprofloxacin (9.6%), and cefoxitin (3.8%). The prevalence of multidrug resistance (MDR) was 78.8% for general staphylococci, 82.2% for S. aureus, 73.1% for S. pseudintermedius, and 60.0% for S. schleiferi. Multidrug resistant staphylococci were significantly more prevalent in the cattle isolates than in the dog isolates (P < 0.05). The prevalence of methicillin-resistant staphylococci (MRS) tested by resistance to cefoxitin and mecA carriage was 3.8%. These four strains were all isolated from dog skin infections. The tetK gene was the most predominant (35.4%), followed by tetM (25.0%). CONCLUSION: In resource-constrained settings, the approach of integrated diagnostics promises sustainable disease surveillance and the addressing of current capacity gaps. The emergence of MRS (zoonotic bacteria) in companion animals creates a likelihood of reduced treatment options for related human infections, a threat to global health.

Diversifying selection identified in immune epitopes of bovine coronavirus isolates from Irish cattle.

Bovine betacoronavirus (BoCoV) is a pneumoenteric pathogen of cattle that is closely related to human coronavirus OC43. Vaccines are administered to protect against diseases caused by BoCoV, but knowledge gaps exist with regard to correlates of protection and the effect of immune evasion on driving evolution. In this study, immune epitopes were mapped onto BoCoV structural proteins, including spike and haemagglutinin esterase (HE), and then supported with targeted gene sequencing of Irish clinical isolates and selective pressure analysis. Increased prevalence of diversifying selection and amino acid changes in some mapped immune epitopes suggests that immune escape is selecting for non-synonymous mutations arising in these regions. Selection analysis and sequencing provided increased support for neutralising antibody (nAb) epitopes compared to others, suggesting that nAbs are an important arm of the immune response to BoCoV. Phylogenetic analysis of spike and HE sequences showed that Irish isolates from this study were in the European clade, except for one HE sequence that sat in the Asian/American clade, while the spike gene of this sample was in the European clade. Recombination between a European and an Asian/American isolate would give rise to such a sequence. This study has gathered evidence suggesting that pressure to evade the nAb response is contributing to BoCoV evolution.

Alterations in the diversity and composition of the fecal microbiota of domestic yaks (Bos grunniens) with pasture alteration-induced diarrhea.

Diarrhea is a common issue in domestic yaks (Bos grunniens) that can occur with pasture alterations and significantly impacts growth performance. Previous research has examined the microbiota of diarrhetic yaks; however, the structural changes in gut bacterial community and microbial interactions in yaks with grassland alteration-induced diarrhea remain poorly understood. To explore variations in gut microbiota homeostasis among yaks suffering from diarrhea, fecal microbiota diversity and composition were analyzed using 16 S rRNA amplicon sequencing. Gut fecal microbiota diversity was lower in diarrhetic yaks than in non-diarrhetic yaks. Furthermore, the bacterial community composition (including that of Proteobacteria and Actinobacteria) in the feces of diarrhetic yaks displayed significant alterations. Co-occurrence network analysis further underscored the compromised intestinal flora stability in yaks with diarrhea relative to that in non-diarrhetic yaks. Interestingly, the abundance of beneficial bacteria, such as Lachnospiraceae_AC2044_group and Lachnospiraceae_NK4A136_group, were decreased in yaks with diarrhea, and the reductions were negatively correlated with the fecal water content. Collectively, these findings indicate that diminished microbial stability and increased abundance of certain bacteria in the gut may contribute to diarrhea occurrence in yaks.

A meta-analysis of livestock brucellosis prevalence in Indonesia.

Indonesia has a long history of livestock brucellosis, but the overall pooled prevalence remains unclear. This study aims to determine the pooled estimated prevalence of livestock brucellosis in Indonesia using the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines. Five databases were searched and screened using predefined inclusion and exclusion criteria. Data from included studies were extracted and analyzed using a random effects model in R 4.3.0 for pooled estimated prevalence, subgroup meta-analysis, and meta-regression. Publication bias and sensitivity tests were conducted using Egger's test, funnel plot, trim and fill plot, and leave-one-out. The screening process identified 46 included studies, representing 47,057 samples for brucellosis testing. The pooled estimated prevalence for livestock brucellosis was 3.25% (95% CI, 1.81%-5.78%) with high heterogeneity (Q = 2130.91, p = 0, I2=98%). Subgroup meta-analysis indicated no significant difference in the prevalence of livestock brucellosis across the main islands in Indonesia (p = 0.44) and across provinces in Sulawesi Island (p = 0.83), but significant differences were found among provinces in Java (p < 0.01). The subgroup meta-analysis based on animal type showed no significant difference between cattle, small ruminants, and pig brucellosis estimated prevalence (p = 0.26). Between serological tests, no significant difference was found (p = 0.77). Meta-regression showed no significant difference in brucellosis prevalence from 1988-2023. Egger's test and funnel plot showed publication bias. Trim and fill test indicated 21 studies should be added. As most studies were conducted in Java and Sulawesi Islands, caution should be exercised in interpreting the results, emphasizing the necessity of increasing the study of brucellosis in other regions.

Molecular identification of haemoparasites in animals using blood lysate PCR: a quick and inexpensive alternative to purified whole genomic DNA.

Haemoparasitic diseases constitute a significant constraint to economic livestock farming. Diagnostic techniques that are inexpensive, rapid, reliable, and precise are crucial for the management of diseases. In this context, PCR assays are very valuable yet expensive since the samples must be processed before being included in the PCR reaction. Accordingly, the goal of the current study was to lower the PCR costs without jeopardizing the assay's sensitivity and specificity. For that purpose, the alkaline solution was optimized for low cost and quick DNA extraction (blood lysate), and PCR reagents were modified for optimum reaction. In comparison to purified whole blood genomic DNA, the currently developed and optimized blood lysate method was found to be 95.5% less expensive, as well as being equally sensitive and specific for the molecular detection (PCR) of haemoparasites like Babesia, Theileria, Trypanosoma and rickettsiales in cattle, buffaloes, horses, and dogs. The assay was also demonstrated to be quick, less likely to cross-contaminate, and appropriate for use in laboratories with limited resources. Therefore, the currently developed and optimized blood lysate method could serve as a viable alternative to purified whole blood genomic DNA for molecular detection (PCR) of haemoparasites in animals particularly in resource-limited settings.

Administering the maternal bovine appeasing substance improves overall productivity and health in high-risk cattle during a 60-d feedlot receiving period.

This experiment evaluated health, physiological, and performance responses of high-risk cattle administered the maternal bovine appeasing substance (mBAS) during feedlot receiving. Angus-influenced, newly weaned male calves (n = 120) were purchased from an auction facility. Calves arrived at the research feedyard on day -1 and body weight (BW) was recorded upon arrival (199 ±â€…1 kg). Calves were ranked by arrival BW and received 1 of 2 treatments prior to initial processing (day 0): (1) 10 mL of an mBAS (Ferappease; FERA Diagnostics and Biologicals; College Station, TX) or (2) 10 mL of mineral oil (CON; placebo). Treatments were applied topically to the nuchal skin area (5 mL) and above the muzzle (5 mL). Calves were vaccinated against Clostridium and respiratory pathogens, dewormed, implanted, band-castrated, and received metaphylaxis at initial processing, and then distributed into 10 drylot pens according to arrival BW and treatment (n = 12 calves/pen, 5 pens/treatment). Calves were reapplied treatments (mBAS or CON) concurrently with booster vaccination on d 14. Feed intake and incidence of bovine respiratory disease (BRD) were recorded daily. Blood and hair samples from the tail-switch were collected on days 0, 14, 28, 42, and 60 for analysis of physiological variables. Calves were sampled for nasal microbiota analysis via swab on days 0, 14, and 28. Final shrunk BW was recorded on day 61 after 16 h of feed and water restriction. Calf BW gain and final BW did not differ between treatments (P ≥ 0.40). Incidence of BRD was similar (P = 0.99) between mBAS and CON (56.7% for both treatments; SEM = 6.45). A greater (P = 0.04) proportion of mBAS calves diagnosed with BRD required a single antibiotic treatment to regain health (70.6 vs. 47.0%; SEM = 8.32), and mortality was greater (P = 0.03) in CON calves diagnosed with BRD (17.6 vs. 2.94%; SEM = 5.133). Relative abundance of Mycoplasma in nasal microbiota was reduced (P = 0.04) in mBAS calves compared with CON (34.7 vs. 27.4%; SEM = 2.35). Cortisol concentration in hair from the tail-switch was less (P = 0.01) on day 28 for mBAS calves compared with CON. Administering mBAS improved (P = 0.04) total pen-based liveweight change during the experiment (498 vs. 309 kg/pen; SEM = 65.2) and final pen-based total liveweight (2,676 vs. 2,484 kg/pen; SEM = 65.4). Administration of mBAS to high-risk cattle decreased physiological stress markers, reduced mortality, and increased pen-based productivity during a 60-d receiving period.

Beef cattle are exposed to several stressors during the initial days in the feedlot, which impair their immunity and lead to bovine respiratory disease (BRD). Strategies to mitigate stress upon feedlot arrival are warranted, including administration of the maternal bovine appeasing substance (mBAS). The mBAS is a mixture of fatty acids that replicate the composition of the original bovine appeasing pheromone. Administering mBAS has been shown to alleviate the physiological consequences elicited by stressful management procedures in beef cattle. In this study, male beef calves were purchased from a commercial auction facility soon after weaning, transported to the research feedyard, and assigned initial processing within a 48-h period. Calves were castrated and received a metaphylactic antibiotic treatment during initial processing, which are relevant management practices in US feedyards. In general, mBAS administration at initial processing and 14 d later decreased physiological stress markers, improved immunity parameters, and reduced mortality by 83% during a 60-d feedlot receiving period. Calf growth and BRD incidence were not affected, but the reduced mortality from mBAS administration increased total liveweight production by the end of the experiment. Therefore, this study provides additional evidence of the benefits from administering mBAS to beef cattle upon feedlot arrival.

Lining the small intestine with mycobacteriophages protects from Mycobacterium avium subsp. paratuberculosis and eliminates fecal shedding.

Johne's disease (JD), a chronic, infectious enteritis caused by Mycobacterium avium subsp. paratuberculosis (MAP), affects wild and domestic ruminants. There is no cure or effective prevention, and current vaccines have substantial limitations, leaving this disease widespread in all substantial dairy industries causing economic, and animal welfare implications. Mycobacteriophages (MPs) have been gaining interest in recent years and are proposed as a promising solution to curtailing MAP infection. Using a well-validated infection model, we have demonstrated the preventative potential of MPs to protect dairy calves against MAP infection. Calves were supplemented daily with a phage cocktail from birth till weaning at 2 m of age and inoculated with MAP at 2 wk of age. Infection status was measured for 4.5 mo through blood, fecal, and postmortem tissue samples. Our findings highlight the remarkable efficacy of orally administered MPs. Notably, fecal shedding of MAP was entirely eliminated within 10 wk, in contrast to the infected control group where shedding continued for the entirety of the trial period. Postmortem tissue culture analysis further supported the effectiveness of MPs, with only 1 out of 6 animals in the phage-treated group testing positive for MAP colonized tissues compared to 6 out of 6 animals in the infected control group. Additionally, plaque assay results demonstrated the ability of phages to persist within the intestinal tract. Collectively, these results underscore the potential of orally administered MP cocktails as a highly effective intervention strategy to combat JD in dairy calves and by extension in the dairy industry.

Bluetongue virus serotype 3 in ruminants in the Netherlands: Clinical signs, seroprevalence and pathological findings.

BACKGROUND: The bluetongue virus serotype 3 (BTV-3) outbreak in the Netherlands in 2023 caused severe clinical signs in ruminants. The clinical and pathological signs in ruminants and their spread during the outbreak in 2023 are described. METHODS: Data from the Dutch monitoring and surveillance system were available to describe clinical signs and pathological findings related to BTV-3 in sheep, cattle and goats. During the outbreak, 13 farms (five sheep, five cattle and three dairy goats) were closely monitored. RESULTS: In 2023, BTV-3 infections were confirmed by real-time polymerase chain reaction in sheep flocks (n = 1807), cattle herds (n = 1864), goat herds (n = 62), alpaca and/or llama herds (n = 15) and one dog. Sheep exhibited the most severe clinical signs and had the highest mortality. In other animal species, a large variation in both occurrence and severity of clinical signs was observed. LIMITATION: Only 13 farms were closely monitored. CONCLUSIONS: The clinical signs observed in affected animals during the 2023 BTV-3 outbreak seem to be more severe than those observed during the BTV-8 outbreak between 2006 and 2008. It seems likely that BTV-3 will overwinter, similar to BTV-8. Therefore, the availability of an effective and safe vaccine is crucial to limit the future impact of BTV-3.

The CAMKK/AMPK Pathway Contributes to Besnoitia besnoiti-Induced NETosis in Bovine Polymorphonuclear Neutrophils.

Besnoitia besnoiti is an obligate intracellular apicomplexan parasite and the causal agent of bovine besnoitiosis. Bovine besnoitiosis has a considerable economic impact in Africa and Asia due to reduced milk production, abortions, and bull infertility. In Europe, bovine besnoitiosis is classified as an emerging disease. Polymorphonuclear neutrophils (PMN) are one of the most abundant leukocytes in cattle blood and amongst the first immunological responders toward invading pathogens. In the case of B. besnoiti, bovine PMN produce reactive oxygen species (ROS), release neutrophil extracellular traps (NETs), and show increased autophagic activities upon exposure to tachyzoite stages. In that context, the general processes of NETosis and autophagy were previously reported as associated with AMP-activated protein kinase (AMPK) activation. Here, we study the role of AMPK in B. besnoiti tachyzoite-induced NET formation, thereby expanding the analysis to both upstream proteins, such as the calcium/calmodulin-dependent protein kinase kinase 2 (CAMKK), and downstream signaling and effector molecules, such as the autophagy-related proteins ULK-1 and Beclin-1. Current data revealed early AMPK activation (<30 min) in both B. besnoiti-exposed and AMPK activator (AICAR)-treated bovine PMN. This finding correlated with upstream responses on the level of CAMKK activation. Moreover, these reactions were accompanied by an augmented autophagic activity, as represented by enhanced expression of ULK-1 but not of Beclin-1. Referring to neutrophil effector functions, AICAR treatments induced both AMPK phosphorylation and NET formation, without affecting cell viability. In B. besnoiti tachyzoite-exposed PMN, AICAR treatments failed to affect oxidative responses, but led to enhanced NET formation, thereby indicating that AMPK and autophagic activation synergize with B. besnoiti-driven NETosis.

Seroprevalence and associated risk factors for Neospora caninum infection in dairy cattle in South Africa.

Bovine neosporosis is a widespread parasitic disease associated with significant economic losses. Its effects on the reproductive performance of cows have resulted in losses that run into the hundreds of millions of US dollars in dairy industries in various countries (Reichel et al., Int J Parasitol 43:133-142, 2013). Due to outdated and scant information on the occurrence of Neospora caninum infection in South Africa, the study aimed to determine the seroprevalence and risk factors associated with infection in dairy cattle in South Africa. A total of 1401 blood samples were randomly collected from cattle on 48 dairy farms in seven of the nine provinces in South Africa. A close-ended questionnaire was used in a cross-sectional study to obtain farm-level and animal-level data. Serological testing was done using a commercial IDvet Screen® Neospora caninum Indirect ELISA. An overall seroprevalence, adjusted for test sensitivity and specificity, of 2.3% (95% CI, 1.3-4.1) was detected and 48% (23/48) of sampled farms had at least one animal testing positive. The highest seroprevalence of N. caninum was in the KwaZulu-Natal province with 7.5% (95% CI, 3.8-14.3), and the lowest in Western Cape with 0.1% (95% CI, 0-1.2). The highest within-farm seroprevalence of 25% was detected on a farm in the North West Province. In a multivariable logistic regression model, the odds of N. caninum seropositivity were higher in Holstein-Friesian cattle when compared to other breeds. Good hygiene was identified as a protective factor. Cattle left out on pasture had increased odds of testing positive for N. caninum compared to those that were penned. The odds of testing seropositive for N. caninum was higher on farms that practised segregation of cattle into different age groups. The purchase of replacement animals was a significant risk factor, as open herds had increased odds of N. caninum seropositivity. Cattle on farms that did not have a specific calving location were more likely to be seropositive. This is the first such study in South Africa and shows that N. caninum is widely distributed in the country at a low seroprevalence, but it may be a cause of concern on certain farms.