Bovine Disease Research Articles

The prevailing infection of Schistosoma japonicum and other zoonotic parasites in bubaline reservoir hosts in the ricefield of lake ecosystem: the case of Lake Mainit, Philippines.

Bovines are important reservoir hosts of schistosomiasis, placing humans and animals in rice fields areas at risk of infection. This study reported the prevailing infection of zoonotic parasites from bovine feces in the rice fields adjacent to Lake Mainit, Philippines. Formalin Ethyl Acetate Sedimentation was performed on 124 bovine fecal samples from rice fields and documented eggs and cysts from seven parasites: Schistosoma japonicum, Fasciola gigantica, Ascaris sp., Strongyloides sp., Balantidium coli, coccidian oocyst and a hookworm species. Among these parasites, F. gigantica harboured the highest infection with a 100% prevalence rate, followed by hookworms (51.61%), B. coli (30.64%) and S. japonicum (12.09%), respectively. The intensity of infection of S. japonicum eggs per gram (MPEG = 4.19) among bovines is categorized as ‘light.’ Bovine contamination index (BCI) calculations revealed that, on average, infected bovines in rice fields excrete 104 750 S. japonicum eggs daily. However, across all ricefield stations, bovines were heavily infected with fascioliasis with BCI at 162 700 F. gigantica eggs per day. The study reports that apart from the persistent cases of schistosomiasis in the area, bovines in these rice fields are also heavily infected with fascioliasis. The study confirms the critical role of bovines as a reservoir host for continued infection of schistosomiasis, fascioliasis and other diseases in the rice fields of Lake Mainit. Immediate intervention to manage the spread of these diseases in bovines is recommended.

Occurrence and surveillance of Taiwanese bovine arboviruses using hematophagous insects in dairy farms during 2012-2019.

Culicoides-borne viruses are an important arbovirus group causing bovine diseases. During 2012-2019, 2,525 pools consisting of 108,937 specimens of vectors were subjected to PCR detection of bovine arbovirus belonging to Orthobunyavirus, Orbivirus, and Ephemerovirus. Twelve virus RNAs, of which 6, that is, Shuni virus, Shamonda virus, and Sathuperi virus in Orthobunyavirus and Sathuvachari virus and epizootic hemorrhagic disease virus serotypes 4 and 7 in Orbivirus were detected for the first time in the area. Potential vector species were evaluated by the minimum infection rate, and the population abundance of Culicoides oxystoma, Culex tritaeniorhynchus, and Anopheles sinensis indicated that they were the main potential vector species in dairy farms in Taiwan.

Effects of DNA Methylation on Gene Expression and Phenotypic Traits in Cattle: A Review.

Gene expression in cells is determined by the epigenetic state of chromatin. Therefore, the study of epigenetic changes is very important to understand the regulatory mechanism of genes at the molecular, cellular, tissue and organ levels. DNA methylation is one of the most studied epigenetic modifications, which plays an important role in maintaining genome stability and ensuring normal growth and development. Studies have shown that methylation levels in bovine primordial germ cells, the rearrangement of methylation during embryonic development and abnormal methylation during placental development are all closely related to their reproductive processes. In addition, the application of bovine male sterility and assisted reproductive technology is also related to DNA methylation. This review introduces the principle, development of detection methods and application conditions of DNA methylation, with emphasis on the relationship between DNA methylation dynamics and bovine spermatogenesis, embryonic development, disease resistance and muscle and fat development, in order to provide theoretical basis for the application of DNA methylation in cattle breeding in the future.

A novel triplex real-time PCR assay for the differentiation of lumpy skin disease virus, goatpox virus, and sheeppox virus.

Three members of Capripoxvirus (CaPV) genus, including lumpy skin disease virus (LSDV), goatpox virus (GTPV), and sheeppox virus (SPPV), are mentioned as notifiable forms by World Organization for Animal Health. These viruses have negatively impacted ruminant farming industry worldwide, causing great economic losses. Although SPPV and GTPV cause more severe clinical disease in only one animal species, they can transfer between sheep and goats. Both homologous and heterologous immunization strategies are used to protect animals against CaPVs. However, development of accurate and rapid methods to distinguish these three viruses is helpful for the early detection, disease surveillance, and control of CaPV infection. Therefore, we developed a novel triplex real-time PCR (qPCR) for the differentiation of LSDV, GTPV, and SPPV. Universal primers were designed to detect pan-CaPV sequences. Species-specific minor groove binder (MGB)-based probes were designed, which were labeled with FAM for LSDV, HEX for GTPV, and ROX for SPPV. The sensitivity, specificity, reproducibility, and ability of detecting mixed infections were evaluated for the triplex qPCR. Further, 226 clinical samples of the infection and negative controls were subjected to the triplex qPCR, and the results were verified using PCR-restriction fragment length polymorphism (PCR-RFLP) and sequencing methods for PRO30 gene. The triplex qPCR could successfully distinguish LSDV, GTPV, and SPPV in one reaction, and the assay sensitivity was 5.41, 27.70, and 17.28 copies/μL, respectively. No cross-reactivity was observed with other viruses causing common ruminant diseases, including des petits ruminants virus, foot-and-mouth disease virus, bluetongue virus, ovine contagious pustular dermatitis virus, infectious bovine rhinotracheitis virus, and bovine viral diarrhea-mucosal disease virus. Inter-and intra-assay variabilities were < 2.5%. The results indicated that the triplex qPCR was highly specific, sensitive, and reproducible. Simulation experiments revealed that this assay could successfully distinguish two or three viruses in case of mixed infections without any cross-reaction. For clinical samples, the results were completely consistent with the results of PCR-RFLP and sequencing. This demonstrated that the assay was reliable for clinical application. The triplex qPCR is a robust, rapid, and simple tool for identifying various types of CaPV as it can successfully distinguish LSDV, GTPV, and SPPV in one reaction. Furthermore, the assay can facilitate more accurate disease diagnosis and surveillance for better control of CaPV infection.

Global Risk Assessment of the Occurrence of Bovine Lumpy Skin Disease: Based on an Ecological Niche Model

Lumpy skin disease (LSD) is a highly contagious disease in bovine animals. An outbreak of LSD can cause devastating economic losses to the cattle industry. To investigate the distribution characteristics of historical LSD epidemics, LSD was divided into four phases for directional distribution analysis based on trends in epidemic prevalence. Ecological niche models were developed for LSD as well as for two vectors (Stomoxys calcitrans and Aedes aegypti), and global predictive maps were generated for the probability of LSD occurrence and the potential distribution of the two LSD vectors. The models had good predictive performance (the AUC values were 0.894 for the LSD model, 0.911 for the S. calcitrans model, and 0.950 for the A. aegypti model). The LSD combined vector prediction map was generated by combining the distribution maps of Stomoxys calcitrans and Aedes aegyptiwith fuzzy overlay tool in ArcGIS. The LSD combined vector prediction map was combined with the LSD prediction map to generate the LSD vector transmission risk map. The eastern and northwestern regions of North America, the eastern and northern regions of South America, the central and southern regions of Africa, the southern region of Europe, the northwestern and southeastern regions of Asia, and the eastern region of Australia were predicted to provide suitable environmental conditions for the occurrence of LSD. Cattle density, buffalo density, and bio2 (mean diurnal range) were identified as key variables for the occurrence of LSD. The findings of this study can be useful to policymakers in developing and implementing preventive measures of LSD for the health of cattle and the cattle industry.

Application of Flow Cytometry in the Diagnosis of Bovine Epidemic Disease.

As science and technology continue to advance, the use of flow cytometry is becoming more widespread. It can provide important information about cells in the body by detecting and analysing them, thereby providing a reliable basis for disease diagnosis. In the diagnosis of bovine epidemic diseases, flow cytometry can be used to detect bovine viral diarrhoea, bovine leukaemia, bovine brucellosis, bovine tuberculosis, and other diseases. This paper describes the structure of a flow cytometer (liquid flow system, optical detection system, data storage and analysis system) and its working principles for rapid quantitative analysis and sorting of single cells or biological particles. Additionally, the research progress of flow cytometry in the diagnosis of bovine epidemic diseases was reviewed in order to provide a reference for future research and application of flow cytometry in the diagnosis of bovine epidemic diseases.

New Insights into the Role and Therapeutic Potential of Heat Shock Protein 70 in Bovine Viral Diarrhea Virus Infection.

Bovine viral diarrhea virus (BVDV), a positive-strand RNA virus of the genus Pestivirus in the Flaviviridae family, is the causative agent of bovine viral diarrhea-mucosal disease (BVD-MD). BVDV's unique virion structure, genome, and replication mechanism in the Flaviviridae family render it a useful alternative model for evaluating the effectiveness of antiviral drugs used against the hepatitis C virus (HCV). As one of the most abundant and typical heat shock proteins, HSP70 plays an important role in viral infection caused by the family Flaviviridae and is considered a logical target of viral regulation in the context of immune escape. However, the mechanism of HSP70 in BVDV infection and the latest insights have not been reported in sufficient detail. In this review, we focus on the role and mechanisms of HSP70 in BVDV-infected animals/cells to further explore the possibility of targeting this protein for antiviral therapy during viral infection.

An outbreak of mucosal disease in cattle caused by &lt;i&gt;Pestivirus H&lt;/i&gt;

An outbreak of infection caused by Pestivirus H (virus of bovine viral diarrhea – mucosal disease of the third kind, BVDV-3) in a dairy farm with high morbidity and mortality in animals of different ages is described. In some sick animals a full complex of pronounced symptoms characteristic of "classical" bovine mucosal disease was registered: erosions and ulcers on the nasal mirror and tongue, foaming from the mouth, serous discharge from the nose, hemorrhagic inflammation and pronounced longitudinal erosions on the mucosa of the esophagus, rennet stomach and intestine. Cows miscarried at different stages of pregnancy. The coefficient of effective insemination decreased to 20%. The course of the disease was complicated by the involvement of the bovine herpes virus type 4, bacteria of the family Pasteurellaceae and Clostridium spp. in the infectious process. The BVDV-3 genome was found in a wide range of internal organs of aborted fetuses, calves, and adult animals. According to sequencing data, the pathogen was classified as subtype 3a. Phylogenetic analysis of the 5'-untranslated region of the virus genome (5'-UTR) showed its close relationship to the strains isolated in Italy and Brazil, most of which were previously identified as contaminants of fetal bovine serum and live vaccines against viral infections of cattle. No specific prophylaxis against BVDV-3 infection has been developed at this time, therefore, it is necessary to update and improve diagnostic methods, optimize control measures to prevent the spread of virulent strains of the pathogen, and control the safety of the vaccines used.

Integration of Network Pharmacology and Molecular Docking to Analyse the Mechanism of Action of Oregano Essential Oil in the Treatment of Bovine Mastitis.

The active components, potential targets, and mechanisms of action of oregano essential oil in the treatment of bovine mastitis disease were investigated using network pharmacology and molecular docking approaches. The TCMSP and literature databases were examined for the main compounds in oregano essential oil. Afterward, the physical, chemical, and bioavailability characteristics of the components were evaluated. The PubChem, BATMAN, PharmMapper, and Uniprot databases were utilized to predict the target genes of the major components of oregano essential oil. Via the databases of DrugBank, OMIM, GeneCards, TTD, and DisGenet, the disease targets of bovine mastitis were discovered. We analyzed common targets and built protein-protein interaction (PPI) networks using the STRING database. Key genes were analyzed, obtained, and compound-target-pathway-disease visualization networks were created using Cytoscape. For the GO function and KEGG pathway enrichment analysis, the DAVID database was utilized. Molecular docking via Autodock Tools was utilized to evaluate the reliability of the interactions between oregano essential oil and hub targets. Thymol, carvacrol, and p-cymene are the three major components found in oregano essential oil. The potential targets (TNF, TLR4, ALB, IL-1β, TLR2, IL-6, IFNG, and MyD88) were screened according to the visual network. The enrichment analysis suggested that the major signaling pathways in network pharmacology may include PI3K-Akt, MAPK, IL-17, and NF-κ B. Molecular docking analysis shows that thymol had good docking activity with TNF, IL-6, and MyD88, carvacrol had good docking activity with TNF, and p-cymene had good docking activity with ALB. This study clarified the mechanism of action of oregano essential oil in the treatment of bovine mastitis, thus providing data supporting the potential for the use of oregano essential oil in the development of new therapeutics for bovine mastitis.

Seroprevalence of Infectious Bovine Rhinotracheitis (IBR) in Indian Buffaloes

Background: Infectious bovine rhinotracheitis (IBR) is an infectious disease of bovines causing respiratory problem, abortions and reduced milk yield leading to huge economic losses. IBR is endemic in India. India ranks first in buffalo population. Reports on seroprevalence of IBR in cattle in India are available but not in buffaloes. The study was aimed at conducting a seroprevalence of IBR in Indian buffaloes. Methods: A total of 1000 buffalo serum samples collected randomly from top 10 milk producing states of India and were tested for IBR antibodies using Avidin-Biotin (AB) ELISA. Result: Cumulative seropositivity was found to be 43.30%. Punjab and Andhra Pradesh states showed highest (58%) and lowest (27%) seropositivity respectively and were part of north and south zone of the country. India has highest number of buffalo population. Presently, India do not vaccinate bovines against IBR and should take call on vaccination of dairy buffalo after having witnessed a high seroprevalence of IBR. This is the first report of such kind of number of buffalo samples tested for IBR antibodies.

Bacteriophages and their Endolysin: An Alternative Therapeutic Approach for Bovine Mastitis

Mastitis is one of the most prevalent and serious disease in bovines and led to high economic loss in the dairy industry. Mastitis dramatically reduces the reproduction abilities of cows. Despite significant progress in controlling and treating this disease, it is still frequent. Mastitis risks public health if milk from treated animals is taken by humans. Multiple factors are responsible for this disease, but bacterial mastitis is the most prevalent and threatening. The emergence of drug-resistant bacterial strains makes mastitis untreatable. Misuse of antibiotics in animal therapy is responsible for this issue, which resulted in the creation of strains that are multidrug resistant. This antibiotic resistance among bacteria is alarming. There is a need for an alternative treatment to cure mastitis. Bacteriophages are viruses, which kill bacteria. The effectiveness of bacteriophages and their endolysin against different bacterial infections causing mastitis has been approved from the results of various studies. Phage therapy is used as both a treatment and preventive measure.

An In Silico Functional Analysis of Non-Synonymous Single-Nucleotide Polymorphisms of Bovine CMAH Gene and Potential Implication in Pathogenesis.

The sugar molecule N-glycolylneuraminic acid (Neu5Gc) is one of the most common sialic acids discovered in mammals. Cytidine monophospho-N-acetylneuraminic acid hydroxylase (CMAH) catalyses the conversion of N-acetylneuraminic acid (Neu5Ac) to Neu5Gc, and it is encoded by the CMAH gene. On the one hand, food metabolic incorporation of Neu5Gc has been linked to specific human diseases. On the other hand, Neu5Gc has been shown to be highly preferred by some pathogens linked to certain bovine diseases. We used various computational techniques to perform an in silico functional analysis of five non-synonymous single-nucleotide polymorphisms (nsSNPs) of the bovine CMAH (bCMAH) gene identified from the 1000 Bull Genomes sequence data. The c.1271C>T (P424L) nsSNP was predicted to be pathogenic based on the consensus result from different computational tools. The nsSNP was also predicted to be critical based on sequence conservation, stability, and post-translational modification site analysis. According to the molecular dynamic simulation and stability analysis, all variations promoted stability of the bCMAH protein, but mutation A210S significantly promoted CMAH stability. In conclusion, c.1271C>T (P424L) is expected to be the most harmful nsSNP among the five detected nsSNPs based on the overall studies. This research could pave the way for more research associating pathogenic nsSNPs in the bCMAH gene with diseases.

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.

Genomic and Molecular Epidemiological Analyses and Antimicrobial Susceptibility of Bovine Mycoplasmas in Japan

Mycoplasma bovis and M. californicum cause a variety of bovine diseases. Spread of these mycoplasmas is difficult to control because it occurs predominantly through asymptomatically infected individuals according to the results of molecular epidemiological analyses. After the 2000s, M. bovis strains having low susceptibility to 16-membered macrolides and tetracyclines have spread in Japan. With the spread of these low-susceptibility M. bovis strains seeming to be involved in the refractoriness to antimicrobial therapy often found in cases of M. bovis infection, it is speculated that these M. bovis strains entered Japan through the importation of bovine biological resources, such as breeding dairy cows, semen, and fertilized eggs. Additionally, appearance of M. bovis strains having low susceptibility to fluoroquinolone is suggested to have relevance to the increase of the amount of fluoroquinolone purchased, being as it is a frequent characteristic of beef cattle-derived strains. The identification of mutations involved in antimicrobial susceptibility and the development of methods to directly detect such mutations have significantly reduced the time required to understand a causal strain’s antimicrobial susceptibility. Selection of antimicrobial agents by this method is expected not only to improve the cure rate but also to greatly contribute to the reduction of antibiotic usage.

First Molecular Investigation of lumpy Skin Disease in Buffaloes Using the Qualitative and Quantitative Polymerase Chain Reaction Assays, Iraq.

Lumpy skin disease (LSD) is one of the most important infectious bovine diseases in Iraq in the last 10 years; however, the current study represents the first investigation to confirm the disease in buffaloes as well as ticks with estimation the association of positivity to clinical vital signs and risk factors. A total of 150 buffaloes were subjected for blood sampling, skin lesions and ticks. All the collected samples; 150 blood, 13 skin lesions, and 29 tick samples, were examined molecularly using the conventional and real-time PCR assays. The total positive results of blood, skin and ticks by conventional PCR were 5.33, 7.69 and 0%, respectively; while for real-time PCR, it was 15.33, 7.69 and, 0%, respectively. Insignificant differences were showed between values of temperature, pulse and respiratory rates of LSD positive and negative buffaloes by the conventional and real-time PCR assays. The association of positive conventional PCR results to risk factors (age, sex and region) was revealed a significant increase in prevalence and risk of LSD in buffaloes aged < 1 year; but for gender, insignificant variation in prevalence but not risk was seen between females and males. In case of different geographical region, significant higher prevalence was reported in Wasit; while, buffaloes of Maysan and Wasit were appeared at higher risk than those of Dhi-Qar. Regarding real-time PCR, insignificant differences were found between values of < 1, 1-4 and > 4-8 years age old, but not in group of >8 that showed a significant decline in positivity (0%). For sex, insignificant variation in prevalence, but not risk, was seen between females and males. Concerning region, buffaloes of Wasit province were recorded a significant higher values of prevalence and risk than other regions. LSD in buffaloes is mainly sub-acute, and PCR appeared to be a suitable diagnostic method in detection of infection; however, furthermore studies are necessary.

Development of a dual vaccine against East Coast fever and lumpy skin disease.

East Coast fever is an acute bovine disease caused by the apicomplexan parasite Theileria parva and is regarded as one of the most important tick-vectored diseases in Africa. The current vaccination procedure has many drawbacks, as it involves the use of live T. parva sporozoites. As a novel vaccination strategy, we have constructed the recombinant lumpy skin disease virus (LSDV) named LSDV-SODis-p67HA-BLV-Gag, encoding a modified form of the T. parva p67 surface antigen (p67HA), as well as the bovine leukemia virus (BLV) gag gene for the formation of virus-like particles (VLPs) to potentially enhance p67 immunogenicity. In place of the native sequence, the chimeric p67HA antigen has the human tissue plasminogen activator signal sequence and the influenza hemagglutinin A2 transmembrane domain and cytoplasmic tail. p67HA was detected on the surface of infected cells, and VLPs comprising BLV Gag and p67HA were produced. We also show that higher multiple bands observed in western blot analysis are due to glycosylation of p67. The two vaccines, pMExT-p67HA (DNA) and LSDV-SODis-p67HA-BLV-Gag, were tested for immunogenicity in mice. p67-binding antibodies were produced by vaccinated animals, with higher titers detected in mice vaccinated with the recombinant LSDV. This candidate dual vaccine warrants further testing in cattle.

Bioactive Feed Additive for the Prevention of Clostridial Disease in High-Yielding Dairy Cattle

The purpose of this research is to develop and test a new approach to prevent clostridial disease in cattle, based on the use of a new compound biologically active feed additive (BFA). Some properties of the separate components of BFA are characterized. The research showed that a strain of the bacterium Bacillus amyloliquefaciens159 has an expressed antagonism to toxin-producing strains of C. perfringens. When using the test strains of C. perfringens from the ATCC collection (13,124 as type A, 10,543 as type C, 12,916 as type F), the anticlostridial activity of the tested strains varied, with size range of 14.0 ± 0.95–15.0 ± 1.28 mm of delayed growth zones. The bactericidal properties of lauric acid and the sorption properties of diatomaceous earth, included in BFA, were confirmed. The experiment was conducted on Holstein cows at the beginning of lactation (control, C (n = 15) vs. experimental E48 (n = 15), E80 (n = 15) and E112 (n = 15), 48, 80 and 112 g/head/day BFA, respectively. All cows were vaccinated with “Coglavax” (vaccine against bovine and sheep clostridial disease, Ceva-Phylaxia VeterinaryBiologicals, Hungary), reinjected two weeks before the experiment. At the end of the experiment (3.5 months after the vaccination and 3 months after the start of BFA feeding according to the scheme of the experiment), the immune response in the control and Group E48 to C. perfringens β-toxin remained at the initial level, while the response in Group E80 and Group E112 became higher under the influence of BFA feeding. Cows fed BFA saw a guaranteed improvement in non-specific resistance. The increase in serum lysozyme concentration in cows of Groups E was 1.01–2.91 mkg/mL vs. control (p &lt; 0.001). TP, GLB, ALB/GLB vs. Groups C and E48 (p &lt; 0.001); this stabilized and normalized while feeding Group E80 and E112 animals with BFA. They also had improved nitrogen, fat, mineral metabolism, as indicated by significant increase in ALB (p &lt; 0.05), UREA (p &lt; 0.01), CHOL (p &lt; 0.01), and CHL (p &lt; 0.01) vs. Groups C and E48. Consumption of BFA increased the amount of anti-oxidants in the blood (highest TAWSA values in Group E80 14.45 mg/g, p = 0.002). Serum TBA–AP/ CP ratio was directly related to TBA–AP (r = 0.87, p &lt; 0.001), and decreased in Group E80. The milk productivity increased under the action of BFA; the average daily milk yield of the cows from the experimental groups for the period of the experiment (d0–d98) was 1.24–1.66 kg higher than that of the control. At the same time, Group E112 cows had a significant increase in milk yield (by 5.1%, p = 0.03 vs. Control). Thus, feeding BFA to dairy cows was found to improve resistance, prevent toxicoses and increase milk production of cattle, which can serve as an additional strategy for bioprotection of cattle against infection.

Validation of stable reference genes in peripheral blood mononuclear cells for expression studies involving vector-borne haemoparasitic diseases in bovines

Normalization of gene expression data using appropriate reference genes is critical to diminish any technical bias in an experiment involving quantitative real-time PCR (qPCR). To the best of our knowledge, this is the first report offering a systematic assessment of 14 potential reference genes (RPLP0, ACTB, RPS28, YWHAZ, SDHA, PPIA, RPS9, RPS15, UXT, GAPDH, B2M, BACH1, HMBS, and PPIB) for the identification of the most stable normalizers for qPCR of target genes in peripheral blood mononuclear cells (PBMCs) of bovines for vector-borne haemoparasitic diseases such as anaplasmosis, babesiosis, theileriosis, and trypanosomiasis. A total of 38 blood samples were collected from healthy as well as diseased cattle and buffaloes representing different haemoparasitic diseases. RNA isolated from the PBMCs was subjected to qPCR for the 14 prospective internal control genes. The comprehensive ranking of the genes was accomplished by the RefFinder tool that integrates the results of three algorithms (geNorm, NormFinder, and BestKeeper) and the comparative CT method. RPS15, B2M, and GAPDH were ranked to be the most stable genes, whereas, PPIA and HMBS emerged to be the least suitable genes. Validation of the selected reference genes by the qPCR analysis of two immunity genes, ISG15 and GPX7 was congruent with the observations of this study. We recommend that a panel of three reference genes including RPS15, B2M, and GAPDH could prove useful in delineating the transcriptional landscape of PBMCs for vector-borne haemoparasitic diseases in bovines.

In Vivo and In Vitro Expression of iC1, a Methylation-Controlled J Protein (MCJ) in Bovine Liver, and Response to In Vitro Bovine Fatty Liver Disease Model.

Mitochondrial complex I inhibitor (iC1) is a methylation-controlled J protein (MCJ) that decreases cellular respiration by inhibiting oxidative phosphorylation. Recent rodent studies showed that loss or inhibition of iC1 was associated with preventing lipid accumulation. A common metabolic disorder of dairy cattle is a fatty liver disease (FLD), which often occurs during the periparturient period. In humans and rodents, iC1 is expressed in the liver and acts as a mitochondrial "brake". However, iC1 expression in bovine liver and its possible role in FLD development have not yet been characterized. We hypothesized that iC1 is expressed in the bovine liver and that the expression of iC1 is correlated with FLD in periparturient dairy cattle. To test this hypothesis, we collected bovine liver tissue samples from an abattoir and isolated primary hepatic cells immediately following harvest. Utilizing an in vitro model of bovine FLD developed in our laboratory, we cultured primary hepatic cells in low-glucose DMEM supplemented with 10% FBS. The basal media was made to induce lipid accumulation and cytotoxicity in the primary liver cells with three treatments. To the basal media (control) we added 0.4 mM palmitate (treatment 1) or 20 ng/mL TNFα (treatment 2), or both 0.4 mM palmitate and 20 ng/mL TNFα (treatment 3). Consistent with our hypothesis, we present the novel characterization of iC1 expression in primary bovine liver cells cultured with or without the addition of lipotoxic factors made to emulate bovine FLD. We demonstrate both in situ and in vitro expression of iC1 in bovine liver and mRNA expression in hepatic cells and in the precipitates of conditioned media. The results of RT-qPCR, IHC, and western blot all demonstrated the expression of iC1 in bovine liver. In addition, we isolated precipitates of conditioned media further demonstrated iC1 expression by RT-qPCR. The transcript of iC1 tended to be more concentrated (4-fold; p > 0.05) in TNFα-treated conditioned media when compared with the control. Taken together, we present the novel finding that iC1 transcript and protein are expressed in liver tissue from dairy cattle, primary hepatic cells isolated from that liver tissue, and, finally, in the conditioned media derived from those cells. These novel findings and the prior findings on the role of iC1 in rodents and humans indicate that further investigation of the role of iC1 in the etiology and pathology of FLD in periparturient dairy cows is warranted.

Overview on epidemiological studies of bovine and equine diseases in India

India's most valuable resource is its livestock population. Farmers that raise livestock are almost entirely reliant on animals for a variety of reasons. India is one of the countries that has the greatest population of livestock in the world. In a similar vein, India has been cited as the source of numerous reports of diseases affecting livestock over the course of many years. Farmers are required to have an understanding of the various diseases that can affect bovine and equine animals. This will allow them to be aware of the etiology of the diseases as well as the pattern of disease transmission, allowing them to make the necessary preparations to prevent and control the diseases. Many diseases, including bovine rotaviral diarrhoea, bovine mastitis, bovine brucellosis, bovine herpes viral infection, and equine piroplasmosis and equine herpes viral infection, among others, are regularly reported from India. The purpose of this review is to provide an overview of the bovine and equine illnesses that are widespread in India from an epidemiological standpoint and to do so in a concise manner.