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Bovine respiratory disease (BRD) is the leading cause of mortality and antimicrobial drug (AMD) use in weaned dairy heifers. Limited information is available regarding antimicrobial resistance (AMR) in respiratory bacteria in this population. This study determined AMR gene presence in 326 respiratory isolates (Pasteurella multocida, Mannheimia haemolytica, and Histophilus somni) from weaned dairy heifers using whole genome sequencing. Concordance between AMR genotype and phenotype was determined. Twenty-six AMR genes for 8 broad classes of AMD were identified. The most prevalent, medically important AMD classes used in calf rearing, to which these genes predict AMR among study isolates were tetracycline (95%), aminoglycoside (94%), sulfonamide (94%), beta-lactam (77%), phenicol (50%), and macrolide (44%). The co-occurrence of AMR genes within an isolate was common; the largest cluster of gene co-occurrence encodes AMR to phenicol, macrolide, elfamycin, β-lactam (cephalosporin, penam cephamycin), aminoglycoside, tetracycline, and sulfonamide class AMD. Concordance between genotype and phenotype varied (Matthews Correlation Coefficient ranged from -0.57 to 1) by bacterial species, gene, and AMD tested, and was particularly poor for fluoroquinolones (no AMR genes detected) and ceftiofur (no phenotypic AMR classified while AMR genes present). These findings suggest a high genetic potential for AMR in weaned dairy heifers; preventing BRD and decreasing AMD reliance may be important in this population.

Ovarian follicular GCs are strongly implicated in the growth, development, and atresia of ovarian follicles. The Wnt/β-catenin and Notch signaling pathways participate in GC proliferation, differentiation, apoptosis, and steroid hormone production during follicular development. However, the crosstalk between Wnt and Notch signaling in GCs remains unclear. This study investigated this crosstalk and the roles of these pathways in apoptosis, cell cycle progression, cell proliferation, and steroid hormone secretion in bovine follicular GCs. The interaction between β-catenin and Notch2 in GCs was assessed by overexpressing CTNNB1, which encodes β-catenin. The results showed that inhibiting the Notch pathway by Notch2 silencing in GCs arrested the cell cycle, promoted apoptosis, reduced progesterone (P4) production, and inhibited the Wnt2-mediated Wnt/β-catenin pathway in GCs. IWR-1 inhibited Wnt2/β-catenin and Notch signaling, reduced GC proliferation, stimulated apoptosis, induced G1 cell cycle arrest, and reduced P4 production. CTNNB1 overexpression had the opposite effect and increased 17β-estradiol (E2) production and Notch2 protein expression. Co-immunoprecipitation assays revealed that Notch2 interacted with β-catenin. These results elucidate the crosstalk between the Wnt/β-catenin and Notch pathways and the role of these pathways in bovine follicular GC development.

Gene therapy is a promising method for treating inherited diseases by directly delivering the correct genetic material into patient cells. However, the limited packaging capacity of vectors poses a challenge. Minimizing promoter size is a viable strategy among various approaches to address this issue. This study aims to optimize the bovine myostatin (MSTN) promoter, enhancing its utility in gene therapy applications. We identified the primary driver of activity as the proximal regulatory region. Isolated from the native promoter and termed M243, this 243-bp sequence was assessed for its potential as a ubiquitous promoter. In a variety of cell types, including bovine embryos and embryonic stem cells, the M243 promoter showed consistent expression, highlighting its suitability for applications requiring compact promoters. The isolation of a highly conserved, compact 243-bp sequence serving as a promoter suggests a solution for overcoming the size constraints of AAV vectors, suggesting potential contributions to the field of gene therapy. [ABSTRACT FROM AUTHOR]

Background: Embryonic genome activation (EGA) is a critical step in early embryonic development, as it marks the transition from relying on maternal factors to the initiation of transcription from embryo's own genome. The factors associated with EGA are not well understood and need further investigation. PRD-like (PRDL) homeodomain transcription factors (TFs) are considered to play crucial roles in this early event during development but these TFs have evolved differently, even within mammalian lineages. Different numbers of PRDL TFs have been predicted in bovine (Bos taurus); however, their divergent evolution requires species-specific confirmation and functional investigations. Results: In this study, we conducted molecular cloning of mRNAs for the PRDL TFs ARGFX, DUXA, LEUTX, NOBOX, TPRX1, TPRX2, and TPRX3 in bovine oocytes or in vitro fertilized (IVF) preimplantation embryos. Our results confirmed the expression of PRDL TF genes in early bovine development at the cDNA level and uncovered their structures. For each investigated PRDL TF gene, we isolated at least one homeodomain-encoding cDNA fragment, indicative of DNA binding and thus potential role in transcriptional regulation in developing bovine embryos. Additionally, our cDNA cloning approach allowed us to reveal breed-related differences in bovine, as evidenced by the identification of a high number of single nucleotide variants (SNVs) across the PRDL class homeobox genes. Subsequently, we observed the prediction of the 9aa transactivation domain (9aaTAD) motif in the putative protein sequence of TPRX3 leading us to conduct functional analysis of this gene. We demonstrated that the TPRX3 overexpression in bovine fibroblast induces not only protein-coding genes but also short noncoding RNAs involved in splicing and RNA editing. We supported this finding by identifying a shared set of genes between our and published bovine early embryo development datasets. Conclusions: Providing full-length cDNA evidence for previously predicted homeobox genes that belong to PRDL class improves the annotation of the bovine genome. Updating the annotation with seven developmentally-important genes will enhance the accuracy of RNAseq analysis with datasets derived from bovine preimplantation embryos. In addition, the absence of TPRX3 in humans highlights the species-specific and TF-specific regulation of biological processes during early embryo development. [ABSTRACT FROM AUTHOR]

Background and Aims: The objective of this case report was to describe an ultrasound‐guided, minimally invasive method for longitudinal mammary gland tissue collection from the bovine species. Materials and Methods: Biopsies were performed on 14 8‐week‐old calves and 113 10‐week‐old calves. A subset of 36 animals had repeated mammary gland biopsies through the first lactation. Mammary gland biopsies were performed using a disposable biopsy punch. The technique was also performed on multiparous cows on other independent research trials. Results: One‐hundred and thirteen animals healed from the 10‐week biopsies with no complications. Of the 36 animals that received repeated biopsies, one developed mastitis due to premature suture removal and one had recurring mastitis in all quarters. Thirty‐three animals underwent all biopsies during gestation. Thirty of the original 36 are currently in lactation and still undergoing repeated biopsies. The method has also been successfully replicated on multiparous cows in separate studies. Discussion and Conclusion: The described technique is a safe, reliable method for cattle mammary gland biopsies beginning at eight weeks of age and can be utilized to obtain repeated tissue collection from individual animals. The technique is also straightforward to perform and utilizes simple tools while providing acceptable amounts of tissue for most applications, with low risk for infection and long‐term tissue damage. [ABSTRACT FROM AUTHOR]

Background: Bovine fascioliasis is a parasitic disease that affects cattle. It leads to direct and indirect great economic loss due to animal mortalities, growth retardation and expenditure on anthelmintics, reduction livestock productivity and essentially condemnation of infected liver by inspection service. The study was carried out to determine the seasonal prevalence and estimated financial losses of fascioliasis in cattle in the Western Highlands of Cameroon. Methodology: A total of 2167 cattle were selected. Later, the cattle, have being the carcasses, were dissected, and the livers were dissected and carefully examined for adult liver flukes. Faeces were collected immediately after the cattle were killed and examined using the formol‐ether concentration technique. Condemned livers were weighted to estimate the financial losses using the average price of a kilogram of liver. Results: A total 428 of cattle were found with infected livers or egg in the faeces giving a total prevalence of 19.75%. Among the infected carcass, 18.64% had both the Fasciola eggs in the faeces and flukes in the liver while 1.10% had only flukes in the liver. Results revealed that prevalence of Fasciola spp. was significantly higher during the rainy season (23.48%) than 16% in the dry season (p = 0.00). A significantly (p = 0.000) higher prevalence was also observed in females (33.9%) than in males (13.1%). Age range showed significant (p = 0.000) influence with the animals of 8–10 years old recorded the highest prevalence of 39.3%. Furthermore, 433.1 kg of liver was condemned giving a direct financial loss of 1221,550 FCFA (2049.64 USD) and an annual financial loss due to liver condemnation estimated to 1814,775 (3045.01 USD). Conclusion: Findings of this study indicated that bovine fascioliasis is prevalent in cattle in the Western Highlands of Cameroon, and it leads to an important financial loss particularly in the rainy season. [ABSTRACT FROM AUTHOR]

The timing of puberty significantly influences subsequent reproductive performance in cattle. N6-methyladenosine (m6A) is a key epigenetic modification involved in the regulation of pubertal onset. However, limited research has investigated alterations in m6A methylation within the hypothalamic–pituitary–ovarian (HPO) axis during the onset of puberty. In this study, combined analysis of methylated RNA immunoprecipitation sequencing (MeRIP-Seq) and RNA sequencing (RNA-seq) is used to describe the overall modification pattern of m6A in the HPO axis, while GSEA, KEGG, and GO analyses are used to describe the enrichment pathways of differentially expressed genes and differentially methylated genes. The m6A modifications of the differential genes KL, IGSF10, PAPPA2, and BMP15 and the pathways of cell adhesion molecules (CAMs), TGF-β, cell cycle, and steroid hormone synthesis may play roles in regulating the function of the HPO axis tissue during pubertal transition. Notably, BMP15′s m6A modification depends on the action of the demethylase ALKBH5, which is recognized by the reader protein YTHDF2, promoting bovine granulosa cell proliferation, steroid production, and estrogen secretion. This study reveals for the first time the modification mechanism of BMP15 m6A during the initiation of bovine puberty, which will provide useful information for improving the reproductive efficiency of Chinese beef cattle. [ABSTRACT FROM AUTHOR]

Background: Due to global warming seasonal heat stress is an increasing problem in temperate zones. Heat stress not only decreases fertility in females, but can also be detrimental to male fertility. Objectives: We studied the effects of natural summer heat stress during spermatogenesis in Holstein bulls on semen quality parameters and on fertilization performance in vitro and possible intergenerational transmission of effects on the next male generation. Materials and Methods: Semen samples from young Holstein breeding bulls, referred to as F0 founders during this study, were collected during summer (F0 "summer" semen) and the following winter (F0 "winter" semen). Parameters such as ejaculate volume, sperm density, motility, thermoresistance, and in vitro blastocyst rates from these F0 semen samples were determined. In addition, after generation of offspring by artificial insemination, semen samples from F1 male offspring were collected and tested for the same quality and performance parameters to capture intergenerational effects. F1 bulls were raised together under identical conditions and semen was collected at about 1 year after birth. Results: The data showed that in vitro blastocyst rates of F0 "summer" semen samples were lower compared with "winter" semen, whereas blastocyst rates of F1 semen samples did not show significant differences. However, whereas F0 semen samples did not indicate significantly different quality parameters we found that motility of F1 semen samples showed significant differences with higher values when collected from bulls generated with F0 "winter" semen. Discussion and Conclusion: From our data, we conclude that (i) natural summer heat stress during spermatogenesis can affect in vitro fertility parameters and (ii) the observed effects on sperm motility of F1 semen samples suggest intergenerational paternal transmission. [ABSTRACT FROM AUTHOR]

The beef cattle industry has experienced a shift driven by a market demand for healthier meat, cost efficiency and environmental sustainability in recent years. Consequently, there has been a growing focus on the fatty acids content and functions of meat in cattle breeding programmes. Besides, a deeper understanding of the biological mechanisms influencing the expression of different phenotypes related to fatty acid profiles is crucial. In this study, we aimed to identify Single‐Nucleotide Variants (SNV) and Insertion/Deletion (InDels) DNA variants in candidate genes related to fatty acid profiles described in genomic, transcriptomic and proteomic studies conducted in beef cattle breeds. Utilizing whole‐genome re‐sequencing data from Brazilian locally adapted bovine breeds, namely Caracu and Pantaneiro, we identified SNVs and InDels associated with 23,947 genes. From these, we identified 318 candidate genes related to fatty acid profiles that contain variants. Subsequently, we select only genes with SNVs and InDels in their promoter, 5′ UTR and coding region. Through the gene–biological process network, approximately 19 genes were highlighted. Furthermore, considering the studied trait and a literature review, we selected the main transcription factors (TF). Functional analysis via gene–TF network allowed us to identify the 30 most likely candidate genes for meat fatty acid profile in cattle. LIPE, MFSD2A and SREBF1 genes were highlighted in networks due to their biological importance. Further dissection of these genes revealed 15 new variants found in promoter regions of Caracu and Pantaneiro sequences. The gene networks facilitated a better functional understanding of genes and TF, enabling the identification of variants potentially related to the expression of candidate genes for meat fatty acid profiles in cattle. [ABSTRACT FROM AUTHOR]

After ejaculation, mammalian sperm undergo a series of molecular events conducive to the acquisition of fertilizing competence. These events are collectively known as capacitation and involve acrosomal responsiveness and a vigorous sperm motility called hyperactivation. When mimicked in the laboratory, capacitating bovine sperm medium contains bicarbonate, calcium, albumin and heparin, among other components. In this study, we aimed at establishing a new capacitation protocol for bovine sperm, using calcium ionophore. Similar to our findings using mouse sperm, bovine sperm treated with Ca2+ ionophore A23187 were quickly immobilized. However, these sperm initiated capacitation after ionophore removal in fresh medium without heparin, and independent of the Protein Kinase A. When A23187-treated sperm were used on in vitro fertilization (IVF) procedures without heparin, eggs showed cleavage rates similar to standardized IVF protocols using heparin containg synthetic oviduct fluid (IVF-SOF). However, when A23187 pre-treated sperm were further used for inseminating eggs in complete IVF-SOF-heparin, a significantly higher percentage of embryo development was observed, suggesting a synergism between two different signaling pathways during bovine sperm capacitation. These results have the potential to improve current protocols for bovine IVF that could also be applied in other species of commercial interest. • Exposure calcium ionophore improves bovine sperm hyperactivation after washing. • Sperm capacitation is achieved by transient exposure to calcium ionophore. • Exposure calcium ionophore trigger capacitation events independent of PKA. • Calcium ionophore capacitated sperm improve embryo production when used in IVF. [ABSTRACT FROM AUTHOR]

Maternal effects in cattle genetics are defined as the causal influence of the phenotype or maternal genotype on the offspring’s phenotype by effects occurring when the genetic and environmental characteristics of the mother influence the phenotype of the offspring beyond the direct inheritance of genes. Its relevance has been strongly described in genetic models focused on the genetic improvement of preweaning traits in cow-calf beef cattle production systems. Here, basic concepts and the importance of maternal effects when using linear and animal model procedures for genetic evaluations of growth and live-weight traits in beef cattle are reviewed and discussed. A brief history of estimation methods from classical studies to recent studies used for the development of animal models for studying maternal effects is also provided. Some important biometric concepts for maternal effect estimation are described, and the antagonism between direct genetic effects and maternal effects, its biological basis, and sources of error in the estimation of direct genetic and maternal covariance are discussed. Finally, some genomic perspectives are presented. [ABSTRACT FROM AUTHOR]

Slaughterhouse inspections play a crucial role in the sanitary control of zoonoses and foodborne diseases. This study aimed to identify and analyze the frequencies of lymph node diseases in cattle slaughtered for human consumption, using the samples sent to the anatomic pathology service of the Federal Laboratory for Agricultural Defense (Laboratório Federal de Defesa Agropecuária), Minas Gerais, Brazil, from January 2015 to September 2022. In total, 2000 lymph node samples were analyzed, and additional information was individually retrieved. Lesions were most frequently identified in thoracic lymph nodes. Bacterial isolation and quantitative polymerase chain reaction (qPCR) were performed using samples suspected of tuberculosis. Tuberculosis cases accounted for 89.3% of the samples. Histopathology was more sensitive than other ancillary tests for diagnosing tuberculosis. Paraffin-embedded tissues from lymphoma cases were subjected to immunophenotyping using anti-CD3 and anti-CD79a immunohistochemistry. Frozen and/or paraffin-embedded tissues from lymphoma cases were used to identify the enzootic bovine leukosis (EBL) retrovirus through qPCR. Other diagnoses included primary (T- and B-cell lymphoma) and metastatic neoplasms (squamous cell carcinoma, pulmonary adenocarcinoma, undifferentiated carcinoma, undifferentiated adenocarcinoma, undifferentiated sarcoma, undifferentiated round cell tumor, mesothelioma, hepatic carcinoid, meningioma, and seminoma), actinogranulomas (pyogranulomatous lymphadenitis [actinobacillosis and actinomycosis]), idiopathic lymphadenitis (neutrophilic and/or histiocytic, granulomatous, and suppurative), and miscellaneous nonspecific lymphadenopathies (depletion/lymphoid atrophy, lymphangiectasia, erythrocyte drainage, parasitic eosinophilic lymphadenitis, follicular hyperplasia, and toxic granulomatous lymphadenitis). The combination of histopathology with complementary techniques is important for successful diagnosis, especially in complex cases of high epidemiological, economic, and zoosanitary importance, such as tuberculosis and EBL. [ABSTRACT FROM AUTHOR]

Background Hypothesis/Objectives Animals Methods Results Conclusions and Clinical Importance Skeletal dysplasia encompasses a heterogeneous group of genetic disorders characterized by an abnormal development of bones, joints, and cartilage. Two Chianina half‐sibling calves from consanguineous mating with congenital skeletal malformations and cardiac abnormalities were identified.To characterize the disease phenotype, to evaluate its genetic cause, and to determine the prevalence of the deleterious alleles in the Chianina population.Two affected calves, their parents and 332 Chianina bulls.The affected animals underwent clinicopathological investigation. Whole‐genome sequencing trio‐approach and PCR‐based assessment of the frequency of TDP‐glucose 4,6‐dehydratase (TGDS) and laminin subunit alpha 4 (LAMA4) alleles were performed.The cases presented with retarded growth, poor nutritional status associated with muscular atrophy and angular deformities of the hindlimbs. Radiologic examination identified generalized osteopenia and shortening of the limb long bones. Necropsy showed osteochondrodysplastic limbs and dilatation of the heart right ventricle. On histological examination, the physeal cartilages were characterized by multifocal mild to moderate loss of the normal columnar arrangement of chondrocytes. Osteopenia also was observed. Genetic analysis identified a missense variant in TGDS and a splice‐site variant in LAMA4, both of which were homozygous in the 2 cases. Parents were heterozygous and allele frequency in the Chianina population for the TGDS variant was 5% and for the LAMA4 variant was 2%.Genetic findings identified 2 potentially pathogenic alleles in TGDS and LAMA4, but no clear mode of inheritance could be determined. [ABSTRACT FROM AUTHOR]

The first lineage differentiation in mammals gives rise to the inner cell mass and the trophectoderm (TE). In mice, TEAD4 is a master regulator of TE commitment, as it regulates the expression of other TE-specific genes and its ablation prevents blastocyst formation, but its role in other mammals remains unclear. Herein, we have observed that TEAD4 ablation in two phylogenetically distant species (bovine and rabbit) does not impede TE differentiation, blastocyst formation and the expression of TE markers, such as GATA3 and CDX2, although a reduced number of cells in the inner cell mass was observed in bovine TEAD4 knockout (KO) blastocysts. Transcriptional analysis in bovine blastocysts revealed no major transcriptional effect of the ablation, although the expression of hypoblast and Hippo signalling-related genes tended to be decreased in KO embryos. Experiments were conducted in the bovine model to determine whether TEAD4 was required for post-hatching development. TEAD4 KO spherical conceptuses showed normal development of the embryonic disc and TE, but hypoblast migration rate was reduced. At later stages of development (tubular conceptuses), no differences were observed between KO and wild-type conceptuses. [ABSTRACT FROM AUTHOR]

Endoplasmic reticulum (ER) stress interferes with developmental processes in oocyte maturation and embryo development. In vitro growth (IVG) is associated with low developmental competence, and ER stress during IVG culture may play a role. Therefore, this study aimed to examine the effect of tauroursodeoxycholic acid (TUDCA), an ER stress inhibitor, on the IVG of bovine oocytes to understand the role of ER stress. Oocyte-granulosa cell complexes (OGCs) were collected from early antral follicles (1.5–1.8 mm) and allowed to grow in vitro for 5 days at 38.5 °C in a humidified atmosphere containing 5 % CO 2. Basic growth culture medium was supplemented with TUDCA at various concentrations (0, 50, 100, 250, and 500 μM). After IVG, oocyte diameters were similar among groups, but the antrum formation rate tended to be higher in the TUDCA 100 μM group. The mRNA expression levels of ER stress-associated genes (PERK , ATF6 , ATF4 , CHOP , BAX , IRE1 , and XBP1) in OGCs were downregulated in the TUDCA 100 μM group than those in the control group. Moreover, the TUDCA 100 μM group exhibited reduced ROS production with higher GSH levels and improved in vitro -grown oocyte maturation compared with those in the control group. In contrast, no difference in the developmental competence of embryos following in vitro fertilization was observed between the control and TUDCA 100 μM groups. These results indicate that ER stress could impair IVG and subsequent maturation rate of bovine oocytes, and TUDCA could alleviate these detrimental effects. These outcomes might improve the quality of oocytes in IVG culture in assisted reproductive technology. • ER stress could impair oocyte development during in vitro (IVG) growth. • ER stress attenuation by TUDCA promotes antrum formation during in vitro growth. • TUDCA downregulates ER stress-associated gene expression in IVG. • TUDCA suppresses oxidative stress and improves subsequent maturation of IVG-grown oocytes. [ABSTRACT FROM AUTHOR]

Repeat breeder (RB) cows are clinically healthy animals with regular oestrous cycles that do not become pregnant after three or more services. This syndrome has an incidence ranging between 10.1% and 24%. Repeat breeder syndrome (RBS) in dairy cows leads to economic losses to dairy farmers by increasing the calving interval and consequently reducing milk and calf production. RBS has a complex oetiopathogenesis as many factors are involved in its onset. The causes can be grouped into two categories: causes leading to fertilisation failure and factors leading to early embryonic death. Accurate identification of the cause and early diagnosis of RBS is essential to minimise the problems that this issue brings to the farm, but is not always possible. Hypothesising the underlying aetiology of the syndrome is also crucial for targeted therapy, whether pharmacological or managerial. The aim of this review is to report the different therapies, proposed in the literature, for the treatment of RBS in cattle, based on the knowledge of possible aetiological causes. [ABSTRACT FROM AUTHOR]

Background Methicillin-resistant Staphylococcus haemolyticus (MRSH) is an important pathogenic agent of bovine mastitis. Among the prominent clone lineages in dairy cows are MRSH sequence types ST3 and ST42. Little information is available on the complete characterization of SCC mec elements in MRSH. Objective In this study, two clinical isolates of MRSH ST3 and ST42 from bovine mastitis milk were selected, and their nontypable SCC mec structures were compared. Methods Two MRSH strains, MRSH-ST3 strain M62.3 and MRSH-ST42 strain M81.1, were identified from bovine mastitis milk in Thailand in 2022. Minimum inhibitory concentration was used to screen for antimicrobial resistance susceptibility. Oxford Nanopore Technologies and Illumina sequencing were performed in combination to complete the genome. Their gene organization and structure of SCC mec types were analysed and compared with the whole sequences of other strains in the same sequence types. Results Both MRSH-ST3 strain M62.3 and MRSH-ST42 strain M81.1 possessed the class C1 mec complex but lacked the ccr gene complex. Notably, MRSH-ST42 strain M81.1 contained a novel variant of C1 mec complex, which consisted of IS 431 - mecA -IS Sha1 - paaZ - upgQ -IS 431 , with IS 431 organized in the same orientation. Apart from class C1 mec and the heavy metal-resistant cluster, the gene composition and order of the SCC mec element varied. In ST3, variations in the SCC mec type, gene content and organization were observed. Conclusions The distinct evolution of the MRSH lineage was indicated by the various SCC mec elements. The insertion of IS Sha1 resulted in a unique variant of class C1 mec complex that demonstrated the important role of the insertion sequence in SCC mec diversification. [ABSTRACT FROM AUTHOR]

Simple Summary: Newborn calves rely on consuming the first milk, colostrum, to transfer important proteins and cells for protection from pathogens. Lymphocytes compose a fraction of the cells transferred through the colostrum to the calf, known as T and B cells. In cattle, a subset of B cells possess a unique receptor with an ultralong protein insert, known as ultralong complementarity determining region 3 (CDR3), which may be crucial in neutralizing viruses. This study found that colostrum had a significantly higher percentage of B cells with ultralong CDR3s than B cells from the blood. This finding may indicate an important role and selective transfer of B cells in colostrum with an ultralong CDR3. In cattle, colostral maternal immunoglobulins and lymphocytes transfer across the neonate's intestinal epithelium to provide protection against pathogens. This study aimed to compare repertoires of B cell populations in blood and colostrum in cows for the first time, with an emphasis on ultralong complementarity determining region 3 (CDR3, ≥40 amino acids). Blood mononuclear cells (BMCs, n= 7) and colostral cells (n = 7) were isolated from Holstein-Friesian dairy cows. Magnetic-activated cell sorting was used to capture IgM and IgG B cells from BMCs. Colostral cells were harvested by centrifugation. RNA was extracted and cDNA was produced; IgM and IgG transcripts were amplified using polymerase chain reactions. Amplicons were sequenced using the Nanopore Native barcoding kit 24 V14 and MinION with R10.4 flow cells. In colostrum, there was a significantly greater percentage of IgM B cells with ultralong CDR3s (8.09% ± 1.73 standard error of the mean) compared to blood (4.22% ± 0.70, p = 0.05). There was a significantly greater percentage of IgG B cells in colostrum with ultralong CDR3s (12.98% ± 1.98) compared to blood (6.61% ± 1.11, p = 0.05). A higher percentage of IgM and IgG B cells with ultralong CDR3s in colostrum may be indicative of a potential role in protecting the neonate. [ABSTRACT FROM AUTHOR]

Nuclear factor (erythroid-derived 2)-like 2 (NRF2) is a transcription factor protein-coding gene, considered a master regulator of the cellular stress response. The genetic variations of the NRF2 could influence its transcriptional profile and, subsequently, the stress resilience in all cell types, including sperm cells. Therefore, the sperm-borne antioxidants abundance in association with the genetic variation of a GCC microsatellite located at the 5′ upstream region of the NRF2 gene was investigated in young (n = 8) and old (n = 8) Holstein bulls' sperm cells at different seasons. The sperm DNA was sequenced using Sanger sequencing, while- the sperm-borne mRNA analysis was carried out using the synthesized cDNA and qPCR. The data were statistically analyzed using GraphPad Prism 10.0.2 software. The results showed that two bulls had a heterozygous genotype of eight and nine GCC repeats, while biallelic of eight, nine, and fifteen repeats were identified in two, ten, and two bulls, respectively. The computational in silico analysis revealed that the NRF2 upstream sequence with 15, 9, and 8 GCC repeats bound with 725, 709, and 707 DNA-binding transcription factor proteins, respectively. Lower quality of sperm DNA was detected in the spring season compared to other seasons and in young bulls compared to old ones, particularly in the summer and autumn seasons. The mRNA expression analysis revealed that the PRDX1 gene was the abundant transcript among the studied sperm-borne antioxidants and was significantly determined in old bulls' spermatozoa. Moreover, two transcripts of the NRF2 gene and antioxidant (SOD1, CAT, GPX1, TXN1, NQO1) genes displayed differential expression patterns between the age groups across seasons in an antioxidant-dependent manner. The bulls with a heterozygous GCC sequence exhibited elevated sperm-borne mRNA levels of NRF2 and PRDX1 transcripts. Taken together, the findings suggest that the NRF2-GCC microsatellite may contribute to the transcription regulation of NRF2 transcripts and their subsequent downstream antioxidants in bovine sperm cells. [ABSTRACT FROM AUTHOR]

Beef and dairy cattle commonly ingest soil when consuming forage-based diets in paddock feeding situations. However, the extent of this soil intake is poorly understood in the Australian environment and under Australian grazing systems. Therefore, the aim of the current literature review was to examine soil ingestion in cattle and the factors that affect ingestion. We found 11 studies containing soil-ingestion data, based in England, France, New Zealand and USA but none from Australia. A wide range in soil ingestion rates was reported and intake varied considerably with season, forage-pasture type, pasture-on-offer, stocking rate and grazing conditions. Generally, soil ingestion was lower in beef cattle than in dairy cattle. We considered the differences in reported levels, variables around those differences, and reliability of methodology used, and compared results with existing international guidelines. For Australian applications, we deduced and recommended that 0.5 kg/head.day is used until field-based research is conducted in Australia which might provide a more specific value for Australian conditions. Our review will inform future livestock management, particularly on contaminated agricultural land. In Australia, there is a poor understanding of how much soil cattle ingest while grazing. We conducted a literature review and found 11 papers to inform our views, but there were no published papers from Australia. Ingestion levels varied among countries, and between dairy and beef enterprises. We estimated that cattle ingest about 0.5 kg/head.day; however, this varies according to circumstance. Research is required in Australian environment to better inform managers. [ABSTRACT FROM AUTHOR]

Background: The genetic test for Expected Progeny Differences (EPDs) is one of the newest technologies to reduce uncertainty in calf production. It provides information about the genetic composition of a bull, which relates to its propensity to produce offspring with specific growth and quality characteristics. Objective: To evaluate the influence of EPDs reports on the variation of Nellore bull semen prices for artificial insemination in the Brazilian beef cattle industry. Methods: Ordinary Least Squares (OLS) regressions were estimated. The study utilized genetic data provided by GENEPLUS as well as market prices of semen and physical measurements of the bulls collected from the Alta Genetics Insemination Agency. Results: Genetic results, except for age at first calving, weaning weight, and weight at 120 days EPDs, have little influence on semen prices. For each additional day expected for the first calving of the progeny average price of semen decreases by 0.66%. For each additional kg expected in weaning weight, the semen price was 1.73% higher. Furthermore, for each additional kg expected in the progeny at 120 days, semen price increased by 2.46%. Conclusion: Joint analyses of genetic reports and physical characteristics of bulls may provide a better explanatory power. [ABSTRACT FROM AUTHOR]

The oviduct epithelium is the initial maternal contact site for embryos after fertilization, offering the microenvironment before implantation. This early gestation period is particularly sensitive to stress, which can cause reduced fertility and reproductive disorders in mammals. Nevertheless, the local impact of elevated stress hormones on the oviduct epithelium has received limited attention to date, except for a few reports on polyovulatory species like mice and pigs. In this study, we focused on the effects of chronic maternal stress on cattle, given its association with infertility issues in this monoovulatory species. Bovine oviduct epithelial cells (BOEC) differentiated at the air–liquid interface (ALI) were stimulated with 250 nmol/L cortisol for 1 or 3 weeks. Subsequently, they were assessed for morphology, bioelectrical properties, and gene expression related to oviduct function, glucocorticoid pathway, cortisol metabolism, inflammation, and apoptosis. Results revealed adverse effects of cortisol on epithelium structure, featured by deciliation, vacuole formation, and multilayering. Additionally, cortisol exposure led to an increase in transepithelial potential difference, downregulated mRNA expression of the major glucocorticoid receptor (NR3C1), upregulated the expression of cortisol-responsive genes (FKBP5, TSC22D3), and significant downregulation of oviductal glycoprotein 1 (OVGP1) and steroid receptors PGR and ESR1. The systematic comparison to a similar experiment previously performed by us in porcine oviduct epithelial cells, indicated that bovine cultures were more susceptible to elevated cortisol levels than porcine. The distinct responses between both species are likely linked to their divergence in the cortisol-induced expression changes of HSD11B2, an enzyme controlling the cellular capacity to metabolise cortisol. These findings provide insights into the species-specific reactions and reproductive consequences triggered by maternal stress. [ABSTRACT FROM AUTHOR]

Background/Objectives: Vaccines may improve the control and eradication of bovine tuberculosis. However, the evaluation of experimental candidates requires the assessment of the protection, excretion, transmission and biosafety. A natural transmission trial among likely infected animals was conducted. Methods: Seventy-four male heifers were randomly distributed (five groups) and vaccinated subcutaneously with attenuated strains (M. bovis Δmce2 or M. bovis Δmce2-phoP), a recombinant M. bovis BCG Pasteur (BCGr) or M. bovis BCG Pasteur. Then, they cohoused with a naturally infected bTB cohort under field conditions exposed to the infection. Results: A 23% of transmission of wild-type strains was confirmed (non-vaccinated group). Strikingly, first vaccination did not induce immune response (caudal fold test and IFN-gamma release assay). However, after 74 days of exposure to bTB, animals were re-vaccinated. Although their sensitization increased throughout the trial, the vaccines did not confer significant protection, when compared to the non-vaccinated group, as demonstrated by pathology progression of lesions and confirmatory tools. Besides, the likelihood of acquiring the infection was similar in all groups compared to the non-vaccinated group (p > 0.076). Respiratory and digestive excretion of viable vaccine candidates was undetectable. To note, the group vaccinated with M. bovis Δmce2-phoP exhibited the highest proportion of animals without macroscopic lesions, compared to the one vaccinated with BCG, although this was not statistically supported. Conclusions: This highlights that further evaluation of these vaccines would not guarantee better protection. The limitations detected during the trial are discussed regarding the transmission rate of M. bovis wild-type, the imperfect test for studying sensitization, the need for a DIVA diagnosis and management conditions of the trials performed under routine husbandry conditions. Re-vaccination of likely infected bovines did not highlight a conclusive result, even suggesting a detrimental effect on those vaccinated with M. bovis BCG. [ABSTRACT FROM AUTHOR]

This study was conducted to investigate the effects and mechanisms of glutamine (Gln) on the repair of acid-induced injury in dairy cow ruminal epithelial cells (RECs) in vitro. Dairy cow RECs were cultured in a medium with pH of 5.5 for 3 h and subsequently treated with various concentrations of Gln (4, 8, 12, 32 mmol/l) for 12 h. The messenger ribonucleic acid (mRNA) expression levels of occludin (OCLN), claudin 1 (CLDN1), toll-like receptor 2 (TLR2), toll-like receptor 4 (TLR4) and genes for inflammatory factors were measured using reverse transcription-quantitative real-time PCR (RT-qPCR). The results showed that cellular activity and OCLN expression were significantly highest at 8 mmol/l Gln (P < 0.05). CLDN1 expression was significantly higher at 4 mmol/l Gln compared to the other groups (P < 0.05). The relative expression levels of tumour necrosis factor (TNF), interleukin 1B (IL1B), C-X-C motif chemokine ligand 8 (CXCL8), TLR2 and TLR4 in the acid treatment group were significantly higher than those in the control group (P < 0.05), but they were lower in the Gln-treated groups than in the acid treatment group (P < 0.05). These findings demonstrate that Gln promotes the proliferation of RECs, enhances the expression of epithelial cell junction proteins, and inhibits the expression of inflammatory factors and surface receptors. In conclusion, Gln shows a potential for repairing acid-induced injury in RECs. [ABSTRACT FROM AUTHOR]

The recent expansion of HPAIV H5N1 infections in terrestrial mammals in the Americas, most recently including the outbreak in dairy cattle, emphasizes the critical need for better epidemiological monitoring of zoonotic diseases. In this work, we detected, isolated, and characterized the HPAIV H5N1 from environmental swab samples collected from a dairy farm in the state of Kansas, USA. Genomic sequencing of these samples uncovered two distinctive substitutions in the PB2 (E249G) and NS1 (R21Q) genes which are rare and absent in recent 2024 isolates of H5N1 circulating in the mammalian and avian species. Additionally, approximately 1.7% of the sequence reads indicated a PB2 (E627K) substitution, commonly associated with virus adaptation to mammalian hosts. Phylogenetic analyses of the PB2 and NS genes demonstrated more genetic identity between this environmental isolate and the 2024 human isolate (A/Texas/37/2024) of H5N1. Conversely, HA and NA gene analyses revealed a closer relationship between our isolate and those found in other dairy cattle with almost 100% identity, sharing a common phylogenetic subtree. These findings underscore the rapid evolutionary progression of HPAIV H5N1 among dairy cattle and reinforces the need for more epidemiological monitoring which can be done using environmental sampling. [ABSTRACT FROM AUTHOR]

Helcococcus ovis (H. ovis) is an opportunistic bacterial pathogen of a wide range of animal hosts including domestic ruminants, swine, avians, and humans. In this study, we sequenced the genomes of 35 Helcococcus sp. clinical isolates from the uterus of dairy cows and explored their antimicrobial resistance and biochemical phenotypes in vitro. Phylogenetic and average nucleotide identity analyses classified four Helcococcus isolates within a cryptic clade representing an undescribed species, for which we propose the name Helcococcus bovis sp. nov. By establishing this new species clade, we also resolve the longstanding question of the classification of the Tongji strain responsible for a confirmed human conjunctival infection. This strain did not neatly fit into H. ovis and is instead a member of H. bovis. We applied whole genome comparative analyses to explore the pangenome, resistome, virulome, and taxonomic diversity of the remaining 31 H. ovis isolates. An overwhelming 97% of H. ovis strains (30 out of 31) harbor mobile tetracycline resistance genes and displayed significantly increased minimum inhibitory concentrations of tetracyclines in vitro. The high prevalence of mobile tetracycline resistance genes makes H. ovis a significant antimicrobial resistance gene reservoir in our food chain. Finally, the phylogenetic distribution of co-occurring high-virulence determinant genes of H. ovis across unlinked and distant loci highlights an instance of convergent gene loss in the species. In summary, this study showed that mobile genetic element-mediated tetracycline resistance is widespread in H. ovis, and that there is evidence of cooccurring virulence factors across clades suggesting convergent gene loss in the species. Finally, we introduced a novel Helcococcus species closely related to H. ovis, called H. bovis sp. nov., which has been reported to cause infection in humans. [ABSTRACT FROM AUTHOR]

Bovine mastitis is one of the main inflammatory diseases that can affect the udder during lactation. Somatic cell counts and sometimes microbiological tests are routinely adopted during monitoring diagnostics in dairy herds. However, subclinical mastitis is challenging to identify, reducing the possibility of early treatments. The main aim of this study was to investigate the miRNome profile of extracellular vesicles isolated from milk as potential biomarkers of subclinical mastitis. Milk samples were collected from a total of 60 dairy cows during routine monitoring tests. Small RNA sequencing technology was applied to extracellular vesicles of milk samples collected from cows classified according to the somatic cell count to identify differences in the miRNome between mastitic and healthy cows. A total of 1997 miRNAs were differentially expressed between both groups. Among them, 68 miRNAs whose FDRs were < 0.05 were mostly downregulated, with only one upregulated miRNA (i.e., miR-361). Functional analysis revealed that miR-455-3p, miR-503-3p, miR-1301-3p and miR-361-5p are involved in the regulation of several biological processes related to mastitis, including immune system-related processes. This study suggests the involvement of extracellular vesicle-derived miRNAs in the regulation of mastitis. Moreover, these findings provide evidence that miRNAs from milk extracellular vesicles can be used to identify biomarkers of mastitis. However, further studies must be conducted to validate these miRNAs, especially for subclinical diagnosis. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate the superestimulatory and superovulatory responses of cattle treated with corifollitropin-alpha, a long-acting human recombinant FSH (rhFSH). In the first and second experiments, we used Nelore (Bos indicus) heifers previously submitted to follicular wave suppression by active immunization against GnRH. In Experiment 1 (a dose-response study), heifers (n = 20) were randomly allocated into five groups, which received placebo (saline) or a single sc dose of 7.5, 15.0, 22.5 or 30.0 μg rhFSH. The heifers were subjected to daily ovarian scan and blood sampling during 11 days. We observed group, time, and group x time effects (P< 0.0001) for both average follicle size and circulating FSH concentrations, with a strong correlation (R = 0.82, P< 0.0001) between the area under curve (AUC) for both parameters. The peak concentration of FSH 24h after treatment and average follicle size at all timepoints, however, were similar (P> 0.05) between groups 22.5 and 30.0 μg. In Experiment 2, heifers (n = 18) were allocated into three groups, which received (0h) either placebo (control), 25 μg rhFSH or 130 mg pFSH (Folltropin). There was no difference (P> 0.05) in average follicle size at any moment, as well as in intrafollicular E2 at 120h or in plasma P4 seven days later between groups rhFSH and pFSH. In Experiment 3, cycling Nelore heifers (n = 20) were subjected to a wave synchronization protocol and superovulated (day 0) using a standard pFSH protocol (120 mg split in eight decreasing im doses) or with a single sc injection of 20 μg rhFSH. The number of follicles >7 mm on day 4 did not differ (P= 0.4370). Heifers receiving rhFSH had greater average follicle size on day 4 (P= 0.0005), ovulation rate (P< 0.0001), and number of CL (P= 0.0155), as well as a trend towards a greater number of ova (P= 0.07) and viable embryos (P= 0.0590). In Experiment 4, superovulation was induced with a single sc injection of 25 μg rhFSH in Girolando and Nelore cows and heifers (n = 20). None of the embryo yield endpoints differed between the two breeds (P> 0.05). In conclusion, cattle superstimulation and superovulation can be successfully induced with a single dose of a long-acting rhFSH (corifollitropin-alpha). • Cattle respond to superstimulation with corifollitropin-alpha in a dose-dependent manner. • Ovarian superovulation can be induced with 20–25 mcg of corifollitropin-alpha. • Corifollitropin-alpha is detected in circulating serum up to 11 days after treatment. [ABSTRACT FROM AUTHOR]

Follicular fluid (FF) is rich in extracellular vesicles (EVs). EVs carries a variety of miRNA involved in regulating follicular development, the function of cells in follicles, primordial follicular formation, follicular recruitment and selection, follicular atresia, oocyte communication, granulosa cells (GCs) function and luteinization and other biological processes of follicular development. Previous studies in our laboratory have shown that bovine follicular fluid (bFF) high density-small extracellular vesicles (HD-sEVs)-miRNA was enriched in autophagy-related pathways. However, the mechanism of bFF EVs carrying miRNA regulating GCs autophagy is not clear. Thus, this study carried out a series of studies on the previous HD-sEVs sequencing data and miR-128–3p contained in bFF HD-sEVs. A total of 38 differentially expressed genes were detected by RNA-Seq after overexpression of miR-128–3p in bovine GCs (bGCs). Through cell transfection, Western blot (WB) and Immunofluorescence (IF), it was proved that overexpression of miR-128–3p could promote the expression of LC3 (microtubule-associated protein I light chain 3), inhibit p62, promote the number of autophagosome, promote the formation of autophagy lysosome and autophagy flow, and activate bGCs autophagy. MiR-128–3p inhibitor significantly inhibited the expression of LC3 and monodansylcadaverine (MDC) in bGCs, and promoted the expression of autophagy substrate p62, indicating that HD-sEVs-miR-128–3p could activate bGCs autophagy. In addition, through double luciferase assay, bioinformatics analysis, WB and RT-qPCR, it was concluded that bFF HD-sEVs-miR-128–3p could target TFEB (transcription factor EB) and FoxO4 (Forkhead box O4) and activate GCs autophagy. • High density-small extracellular vesicles (HD-sEVs) was from bovine follicular fluid and carried miR-128–3p. • MiR-128–3p mimics/inhibitors promoted/inhibited bovine granulosa cells (bGCs) autophagy. • HD-sEVs-miR-128–3p inhibits bGCs apoptosis. HD-sEVs-miR-128–3p promotes bGCs autophagy. • HD-sEVs-miR-128–3p activated bGCs autophagy by targeting FoxO4 and TFEB. [ABSTRACT FROM AUTHOR]

This work explored whether a well-characterized recombinant human interleukin-6 (hIL6) protein will influence in vitro produced (IVP) bovine embryo development and survival after cryopreservation. Cumulus oocyte complexes were collected from abattoir derived ovaries, matured for 24 h, and fertilized using pooled semen from Holstein bulls. Embryos were treated with 0, 25, 50, or 100 ng/mL hIL6 on day 5 post-fertilization. An increase in ICM cell numbers was observed in each hIL6 treatment, with the lowest hIL6 treatment having the same magnitude of response as the middle and highest hIL6 concentration. No effects on TE cell numbers were observed. The second study involved cryopreserving (via slow freezing) of hIL6-treated blastocysts, then examining post-thaw blastocyst survival by incubating for 24 h in the absence of hIL6 treatments. Blastocyst re-expansion and hatching rates were unaffected by any of the IL6 treatments, however, increases in both ICM and TE cell numbers were detected at 24 h post-thawing in blastocysts exposed to 100 ng/mL hIL6 but not lower concentrations before freezing. A reduction in the percentage of TUNEL-positive TE cells was observed after thawing in blastocysts exposed to 25, 50 and 100 ng/mL hIL6 before cryopreservation. No treatment-dependent changes in TUNEL-positive ICM cells were observed. In summary, hIL6 supplementation improves ICM cell numbers in bovine blastocysts to a degree that is commensurate with what has been observed when using bovine recombinant IL6. This positive effect of hIL6 on ICM cell numbers is maintained after freezing and thawing, and a novel improvement in post-thaw TE cell numbers occur in hIL6 treated embryos. This positive effect on TE cell numbers is attributed, at least in part, to an hIL6-dependent reduction in TE cell apoptosis. [Display omitted] • Increases in inner cell mass cell numbers are achieved by supplementing recombinant human interleukin-6. • Positive effects of human interleukin-6 supplementation on both the inner cell mass and trophectoderm were detected after embryo cryopreservation. • Human interleukin-6 may is effective in bovine embryos. [ABSTRACT FROM AUTHOR]

A plethora of infectious and non-infectious causes of bovine abortions and perinatal mortalities (APM) have been reported in literature. However, due to financial limitations or a potential zoonotic impact, many laboratories only offer a standard analytical panel, limited to a preestablished number of pathogens. To improve the cost-efficiency of laboratory diagnostics, it could be beneficial to design a targeted analytical approach for APM cases, based on maternal and environmental characteristics associated with the prevalence of specific abortifacient pathogens. The objective of this retrospective observational study was to implement a machine learning pipeline (MLP) to predict maternal and environmental factors associated with infectious APM. Our MLP based on a greedy ensemble approach incorporated a standard tuning grid of four models, applied on a dataset of 1590 APM cases with a positive diagnosis that was achieved by analyzing an extensive set of abortifacient pathogens. Production type (dairy/beef), gestation length, and season were successfully predicted by the greedy ensemble, with a modest prediction capacity which ranged between 63 and 73 %. Besides the predictive accuracy of individual variables, our MLP hierarchically identified predictor importance causes of associated environmental/maternal characteristics of APM. For instance, in APM cases that happened in beef cows, season at APM (spring/summer) was the most important predictor with a relative importance of 24 %. Furthermore, at the last trimester of gestation Trueperella pyogenes and Neospora caninum were the most important predictors of APM with a relative importance of 22 and 17 %, respectively. Interestingly, herd size came out as the most relevant predictor for APM in multiparous dams, with a relative importance of 12 %. Based on these and other mix of predicted environmental/maternal and pathogenic potential causes, it could be concluded that implementing our MLP may be beneficial to design a more cost-effective, case-specific diagnostic approach for bovine APM cases at the diagnostic laboratory level. • We implement a MLP to predict maternal and environmental factors associated with infectious bovine APM. • Our greedy ensemble approach incorporates a standard tuning grid of four machine learning models. • The MLP correctly predicted production type (beef vs. dairy), gestation length, and season at a range of 63 to 73 %. • Implementation of a MLP allows a cost-effective, case-specific diagnostic approach for bovine APM cases. [ABSTRACT FROM AUTHOR]

Introduction: Bovine subclinical mastitis (SCM) caused by Gram-positive bacteria is a major cause of economic loss in the dairy industry, exacerbated in situations where antimicrobial resistance is present. Pure platelet-rich plasma (P-PRP) may be a therapeutic alternative for SCM, when used alone or with antibiotics, such as sodium cloxacillin (SC). This study aimed 1) to evaluate the therapeutic efficacy of allogeneic P-PRP, SC, and their combination (P-PRP+SC) in cows with SCM caused by Staphylococcus aureus and by streptococci (Staphylococcus aureus and S. dysgalactiae); 2) to determine the concentrations of somatic cells (SCC), interleukin 1 beta (IL-1ß), tumor necrosis factor-alpha (TNF-a) and TGF-ß1 in milk samples of the cows. Methods: 130 cows from 4 dairy herds completed the study, of which 40 cows were treated with P-PRP (10 mL), 28 cows with SC (5g), 36 with P-PRP+SC (10mL/5g), and 26 did not receive no treatment (negative control group, NCG). Results: The overall bacteriological cure was observed in 10/40 (25%) cows in the P-PRP group, 9/28 (32.14%) animals in the SC group, 26/36 (72.22%) cows in the P-PRP+SC group, and 10/26 (38.46%) animals in the NCG. SCM caused by S. aureus (82/130, 63.08%), was cured in 6/24 (25%) cows treated with P-PRP, 7/24 (29.2%) cows treated with SC, 8/16 (50%) animals treated with P-PRP+SC, and in 8/18 (44.4%) cows in NCG. For SCM caused by the streptococci (48/130, 36.91%), the cure was achieved in 4/12 (33.3%) cows treated with P-PRP, 2/4 (50%) cows treated with SC, 18/20 (90%) cows treated with P-PRP+SC, and in 2/8 (25%) cows of the NCG. SCC was significantly (p < 0.001) affected by the treatment, herd, cure, bacteria group, and number of calvings factors. IL-1ß milk concentrations were significantly (p < 0.001) influenced by treatment and farm factors, and the interaction between these factors. TNF-a milk concentrations were significantly (p < 0.001) influenced by time factor. TGF-ß1 milk concentrations were significantly affected by the time and cure factors. Conclusion: The combination of P-PRP and SC showed the best therapeutic response (90%) against bovine SCM caused by streptococci. However, none of the treatments showed an effective therapeutic response against S. aureus. [ABSTRACT FROM AUTHOR]

Background: This study aimed to identify the roles of L-tryptophan (Trp) and its rate-limiting enzymes on the receptivity of bovine endometrial epithelial cells. Real-time PCR was conducted to analyze the differential expression of genes between different groups of bovine endometrial epithelial cells. Western blot was performed to detect Cyclooxygenase-2 (COX2) expression after treatment with Trp or kynurenine (the main metabolites of Trp). The kynurenine assay was used to examine if Trp or prostaglandin E2 (PGE2) can increase the production of kynurenine in the bovine endometrial epithelial cells. Results: Trp significantly stimulates insulin growth factor binding protein 1 (IGFBP1) expression, a common endometrial marker of conceptus elongation and uterus receptivity for ruminants. When bovine endometrial epithelial cells are treated with Trp, tryptophan hydroxylase-1 remains unchanged, but tryptophan 2,3-dioxygenase 2 (TDO2) is significantly increased, suggesting tryptophan is mainly metabolized through the kynurenine pathway. Kynurenine significantly stimulates IGFBP1 expression. Furthermore, Trp and kynurenine significantly increase the expression of aryl hydrocarbon receptor (AHR). CH223191, an AHR inhibitor, abrogates the induction of Trp and kynurenine on IGFBP1. PGE2 significantly induces the expression of TDO2, AHR, and IGFBP1. Conclusions: The regulation between Trp / kynurenine and PGE2 may be crucial for the receptivity of the bovine uterus. [ABSTRACT FROM AUTHOR]

Introduction: Nocardia farcinica is an opportunistic bacterium that causes bovine mastitis and pulmonary, cutaneous, and central nervous system infections in humans. Bovine abortion caused by N. farcinica has been sporadically reported. The purpose of this study was to analyze the pathological findings of bovine abortions caused by N. farcinica in the Republic of Korea and determine the antimicrobial resistance and genotypical characteristics of N. farcinica isolates. Case presentation: Three cases of bovine abortions were submitted to the Animal and Plant Quarantine Agency for differential diagnosis. Grossly, one fetus showed severe lung consolidation following palpation of the entire lobes. Histologically, necrotizing granulomatous interstitial pneumonia was observed in all fetuses; a fetus with a gross lesion demonstrated necrotizing lymphadenitis in the mesenteric lymph nodes and necrotizing dermatitis in the ear. N. farcinica isolates were isolated from the abomasal contents and lungs of all fetuses. Finally, two cases were diagnosed as abortions due to N. farcinica, and one was diagnosed as an N. farcinica abortion coinfected with bovine viral diarrhea virus. According to the multilocus sequence analysis, all isolates were identified as N. farcinica and were determined to be genetically related to isolates from humans. Two N. farcinica isolates were resistant to trimethoprimsulfamethoxazole, which is recommended as the first treatment for human nocardial infections. Conclusion: This is the first pathological report of bovine abortion caused by N. farcinica in the Republic of Korea. Further studies are needed to phenotypically and genotypically characterize N. farcinica isolates with various sources and continuously monitor antimicrobial resistance patterns. [ABSTRACT FROM AUTHOR]

Bovine hemolytic anemia has a negative impact on animal welfare and productivity due to its associated clinical symptoms. Hemolysis is generally known to cause reticulocytosis, increased indirect bilirubin, decreased concentration of haptoglobin, and increased lactate dehydrogenase. Additionally, tissue hypoperfusion due to concomitant anemia increases blood lactate concentration. However, few studies have reported the correlation between these indicators and hemolytic anemia in cattle. We expected that alterations in hematological and biochemical parameters could identify cattle with hemolytic anemia. Therefore, in addition to reporting differences in indicators according to hemolytic anemia, this study aimed to derive indicators and set criteria for identification of bovine hemolytic anemia. In cattle with hemolytic anemia, reticulocytosis, increased indirect bilirubin, and increased L-lactate were observed, and the correlation of these indicators with hematocrit (HCT) was confirmed. And since HCT alone has limitations in identifying hemolytic anemia, we suggest additional criteria to identify hemolytic anemia in cattle. [ABSTRACT FROM AUTHOR]

Embryo cultivation is the longest step during the in vitro embryo production process. In this study, the effect of adding 5% FCS or 3 mg/ml BSA to SOF and KSOM in vitro culture media was investigated. When adding FCS in SOF medium, a better effect on separation was observed (P < 0.05), and the percentage of blastocysts was higher - 58.6%. When BSA was added to SOF medium, the percentage of blastocysts was 40.1% - significantly lower. When cultured in KSOM medium with the addition of FCS, the division rate was lower than when cultured in SOF+FCS and the proportion of blastocysts was also slightly lower. When BSA was added to the KSOM medium, the percentage of blastocysts was almost as in the SOF+BSA group. Аt the conclusion of the conducted experiment, the addition of FCS in KSOM and SOF media is beneficial for in vitro cultivation, resulting in a higher percentage of blastocysts. [ABSTRACT FROM AUTHOR]

Spermatogonial stem cells (SSCs) possess the characteristics of self-renewal and differentiation, as well as the ability to generate functional sperm. Their unique stemness has broad applications in male infertility treatment and species preservation. In rodents, research on SSCs has been widely reported, but progress is slow in large livestock such as cattle and pigs due to long growth cycles, difficult proliferation in vitro, and significant species differences. Previously, we showed that histone 3 (H3) lysine 9 (K9) trimethylation (H3K9me3) is associated with the proliferation of bovine SSCs. Here, we isolated and purified SSCs from calf testicular tissues and investigated the impact of different H3K9me3 levels on the in vitro proliferation of bovine SSCs. The enriched SSCs eventually formed classical stem cell clones in vitro in our feeder-free culture system. These clones expressed glial cell-derived neurotrophic factor family receptor alpha-1 (GFRα1, specific marker for SSCs), NANOG (pluripotency protein), C-KIT (germ cell marker), and strong alkaline phosphatase (AKP) positivity. qRT-PCR analysis further showed that these clones expressed the pluripotency genes NANOG and SOX2, and the SSC-specific marker gene GFRα1. To investigate the dynamic relationship between H3K9me3 levels and SSC proliferation, H3K9me3 levels in bovine SSCs were first downregulated using the methyltransferase inhibitor, chaetocin, or transfection with the siRNA of H3K9 methyltransferase suppressor of variegation 3-9 homologue 1 (SUV39H1). The EDU (5-Ethynyl-2′-deoxyuridine) assay revealed that SSC proliferation was inhibited. Conversely, when H3K9me3 levels in bovine SSCs were upregulated by transfecting lysine demethylase 4D (KDM4D) siRNA, the EDU assay showed a promotion of cell proliferation. In summary, this study established a feeder-free culture system to obtain bovine SSCs and explored its effects on the proliferation of bovine SSCs by regulating H3K9me3 levels, laying the foundation for elucidating the regulatory mechanism underlying histone methylation modification in the proliferation of bovine SSCs. [ABSTRACT FROM AUTHOR]

Inflammation models are widely used in the in vitro investigation of new therapeutic approaches for osteoarthritis. TNFα (tumor necrosis factor alpha) plays an important role in the inflammatory process. Current inflammation models lack uniformity and make comparisons difficult. Therefore, this study aimed to systematically investigate whether the effects of TNFα are concentration-dependent and whether chondrocyte expansion has an effect on the inflammatory model. Bovine chondrocytes were enzymatically isolated, expanded to passages 1–3, and transferred into a 3D pellet culture. Chondrocyte pellets were stimulated with recombinant bovine TNFα at different concentrations for 48 h to induce inflammation. Gene expression of anabolic (collagen 2, aggrecan, cartilage oligomeric protein (COMP)), catabolic (matrix metalloproteinases (MMP3, MMP13)), dedifferentiation (collagen 1) markers, inflammation markers (interleukin-6 (IL-6), nuclear factor kappa B (NFkB), cyclooxygenase-2 (COX), prostaglandin-E-synthase-2 (PTGES2)), and the apoptosis marker caspase 3 was determined. At the protein level, concentrations of IL-6, nitric oxide (NO), and sulfated glycosaminoglycans (GAG) were evaluated. Statistical analysis was performed using the independent t-test, and significance was defined as p < 0.05. In general, TNFα caused a decrease in anabolic markers and an increase in the expression of catabolic and inflammatory markers. There was a concentration-dependent threshold of 10 ng/mL to induce significant inflammatory effects. Most of the markers analyzed showed TNFα concentration-dependent effects (COMP, PRG4, AGN, Col1, MMP3, and NFkB). There was a statistical influence of selected gene expression markers from different passages on the TNFα chondrocyte inflammation model, including Col2, MMP13, IL-6, NFkB, COX2, and PTGES2. Considering the expression of collagen 2 and MMP3, passage 3 chondrocytes showed a higher sensitivity to TNFα stimulation compared to passages 1 and 2. On the other hand, MMP13, IL-6, NFkB, and caspase 3 gene expression were lower in P3 chondrocytes compared to the other passages. On the protein level, inflammatory effects showed a similar pattern, with cytokine effects starting at 10 ng/mL and differences between the passages. TNFα had a detrimental effect on cartilage, with a clear threshold observed at 10 ng/mL. Although TNFα effects showed concentration-dependent patterns, this was not consistent for all markers. The selected passage showed a clear influence, especially on inflammation markers. Further experiments were warranted to explore the effects of TNFα concentration and passage in long-term stimulation. [ABSTRACT FROM AUTHOR]

Ayurvedic medicine utilizes metal-based preparations, known as bhasmas, to treat various health conditions. Yasad bhasma (YB), a zinc-based ayurvedic preparation, shows promise as a potential candidate for developing zinc-based nanomedicines with anti-inflammatory and antioxidant properties. In this study, we synthesized a formulation combining YB and hydroxychloroquine (HC) as a zinc ionophore (YBHC) and investigated its biocompatibility and antiviral effects against buffalo calf coronavirus (BCoV) in Vero cells. Our results demonstrated that the formulation exhibited good conformity and enhanced cell proliferation compared to untreated cells. Additionally, no cytopathic effects were observed in BCoV-infected Vero cells treated with YBHC and YB, while infected control cells exhibited cytopathic effects. YB showed cytoprotection by promoting epithelial tissue turnover. We further explored whether YB/YBHC exerted a lysosomotropic effect to produce antiviral effects on coronavirus-adapted Vero cells, but no cell internalization was observed. In addition to the synergistic antiviral effect of YB and HC, YB may play a vital role in rejuvenating affected tissues. [ABSTRACT FROM AUTHOR]

Simple Summary: Guanidinoacetic acid (GAA) is a natural derivative of amino acids. They are acted upon by enzymes in the liver and transformed into essential components of the energy metabolism of muscle and nervous tissue. Although the animal synthesizes creatine, the amount is less than necessary for high growth rates, according to researchers. GAA consumed by cattle reduces fat between muscle fibers and serum cholesterol. It has a mild but positive effect on the fatty acid profile of meat—i.e., it increases myristoleic, linoleic, and arachidonic acids. We also concluded that GAA consumption increases the amount of volatile fatty acids in the rumen, especially acetic, propionic, and butyric acid. We draw attention to the acetic acid related to lipid synthesis. With the increase in population, it is increasingly necessary to produce food more efficiently. This has expanded the market for additives, which are products that directly (nutritional effect) or indirectly (effect on animal health) favor productivity. Guanidinoacetic acid (GAA) is a natural precursor of creatine. It acts as an energy reserve in skeletal muscle. In addition to being a compound with more significant bioavailability, it is more thermally stable and less expensive than creatine. Therefore, this study aimed to determine whether adding GAA to the cattle diet would alter the meat's composition and fatty acid profile. We used 24 Holstein cattle males (409 ± 5.6 kg), approximately 15 months old, and separated them into four homogeneous groups, one being the control group and three groups with various dosages of GAA in the diets (3.3; 6.6, and 9.9 g/animal/day), for an experimental period of 60 days. Blood, rumen fluid, and animal weighing were performed at three points (days 1, 30, and 60), and daily feed consumption was measured. Steers fed with GAA (9.9 g/d) showed a 16.9% increase in average daily gain (ADG) compared to the control group. These same animals (T-9.9 group) fed with GAA showed a 20% increase in fed efficiency compared to the control group. Lower leukocyte, lymphocyte, and granulocyte counts and lower cholesterol levels were observed in animals that consumed 6.6 g and 9.9 g/d GAA compared to the control group. Animals from the T-6.6 and T-9.9 groups showed 30% and 27.6% reduced bacterial activity in the rumen compared to the control group, respectively. Steers from the T-6.6 and T-9.9 groups fed with GAA showed a 20% and 37% increase in short-chain fatty acids (SCFAs) compared to the control group, respectively. A higher concentration of acetic, propionic, and butyric acids in the ruminal fluid of cattle T-9.9 group was observed at day 60. The two highest doses of GAA showed lower fat levels in the meat, just as the cattle that received 9.9 g/d showed higher levels of total polyunsaturated fatty acids. Complementary data results draw attention to the dose of 9.9 g/d GAA in cattle diets, as anti-inflammatory action can be seen and combined with a higher concentration of SCFAs, consequently increases weight gain. We concluded that consuming this GAA increases the concentration of some unsaturated fatty acids (omegas) in the meat, which adds quality to the product for the consumer. [ABSTRACT FROM AUTHOR]

Simple Summary: Low fertility is the main cause of low productivity in beef cattle, and the non-conception of cows after fertilization is an important factor contributing to this phenomenon. miRNAs may exert unique metabolic and immune effects on the early gestation process in cattle. In this study, serum exosomes on the 0 th, 14 th, and 21 st days of the early gestation of cattle were obtained by simultaneous estrus technology, and the expression profile of exosomal miRNA was detected by RNA-seq technology. Among them, bta-miR-3604, bta-miR-2889, bta-miR-3432a, and bta-miR-409b are newly discovered miRNAs. This study can provide a theoretical reference for the screening of early pregnancy establishment and maternal pregnancy recognition (MRP) molecular markers in cattle. Low fertility is the main cause of the low productivity in beef cattle and is mainly associated with a lack of conception after fertilization. The establishment of early pregnancy in cattle is a complex physiological process, and embryo implantation is crucial for the successful establishment of pregnancy. Exosomal miRNAs play an important role in regulating mammalian embryo implantation and development. This study used synchronous estrus technology to extract exosomes from bovine serum at 0, 14, and 21 days of early pregnancy and analyzed the expression profile of exosomal miRNAs through RNA-seq technology. We identified 472 miRNA precursor sequences and 367 mature miRNA sequences in the three sample groups, with the majority of the miRNAs having high abundance. Differentially expressed miRNAs (DEmiRNAs) were screened, and 20 DEmiRNAs were obtained. The differential expression analysis results show that compared to day 0, there were 15 DEmiRNAs in the serum on day 14 and 5 on day 21 of pregnancy. Compared to the 14th day of pregnancy, there were eight DEmiRNAs in the serum on the 21st day of pregnancy. Bioinformatics analysis shows that the target genes of DEmiRNAs regulated the signaling pathways closely related to early pregnancy, including the VEGF, NF-κB, and MAPK signaling pathways. In addition, the newly discovered miRNAs were bta-miR-3604, bta-miR-2889, bta-miR-3432a, and bta-miR-409b. These results provide a theoretical reference for screening the molecular markers for early pregnancy establishment and maternal recognition of pregnancy (MRP) in cattle and new ideas for shortening the calving interval in cows. [ABSTRACT FROM AUTHOR]

Despite the cytotoxicity and embryotoxicity previously reported artesunate is a recommended drug to treat malaria for adults, children, and women in the first trimester of pregnancy. To address the putative effects of artesunate on female fertility and preimplantation embryo development, when the pregnancy is not detectable yet, artesunate was added to the oocyte in vitro maturation and in vitro embryo development of bovine. Briefly, in experiment 1 the cumulus-oocyte complexes (COCs) were in vitro matured for 18 h with 0.5, 1, or 2 µg/mL of artesunate or not (negative control) and then checked for nuclear maturation and subsequent embryo development. In experiment 2, the COCs were in vitro matured and fertilized without artesunate, which was added (0.5, 1, or 2 µg/mL) from the 1st to the 7th day of embryo culture along with a negative and a positive control group with doxorubicin. As a result, the use of artesunate on oocyte in vitro maturation did not differ from the negative control (p > 0.05) regarding nuclear maturation, cleavage, and blastocyst formation. Also, artesunate on in vitro embryo culture did not differ from negative control (p > 0.05) regarding cleavage and blastocyst formation, except for positive control, with doxorubicin (p < 0.05). In conclusion, under the conditions investigated, there was no evidence of artesunate toxicity on oocyte competence and the preimplantation period of in vitro embryo development in the bovine model, however, artesunate use still should be taken carefully as the outcome of implantation after oocytes and blastocysts exposure to artesunate remains unknown. HIGHLIGHTS: Artesunate added to in vitro maturation did not impair oocyte competence in bovine. Artesunate added to in vitro culture did not affect cleavage and blastocyst formation. No evidence of artesunate toxicity in oocytes and embryos of bovine was found. [ABSTRACT FROM AUTHOR]

Ragworts like tansy ragwort (J. vulgaris Gaertn., syn. Senecio jacobaea L.) contain hepatotoxic and cancerogenic pyrrolizidine alkaloids (PA) and their corresponding pyrrolizidine alkaloid N-oxides (PANO). Due to increasing spread of ragworts (Jacobaea spp.) PA/PANO may pose a health risk to animals and humans consuming contaminated feed and food. Therefore, the aim of the present study was to investigate the transfer of individual PA/PANO originating from a well-defined PA/PANO extract into the milk of dairy cows. For this objective, 16 German Holstein cows were assigned to four treatment groups (n = 4) in a 28-day dose-response study. Administration into the reticulorumen was performed daily by gavage after the morning milking. Three groups received different amounts of the J. vulgaris extract resulting in a PA/PANO exposure of 0.47, 0.95, or 1.91 mg PA/PANO/kg body weight/day, respectively. Furthermore, a control group received molasses to account for the sugar content of the used PA/PANO extract. While the composition of the PA/PANO extract was more diverse, the PA/PANO pattern in milk was dominated by the PA in their free base form. It was shown that mainly PA considered stable in the rumen environment were transferred into the milk. The main compounds in milk were jacoline (74.3 ± 2.4% of the PA/PANO sum), jaconine (11.2 ± 1.3%), and jacobine (7.2 ± 0.6%) with concentrations up to 29.7, 4.65 µg/l, or in the highest exposed group, 3.44 µg/l. There was no dose-dependent effect on the total PA/PANO transfer rate into the milk. The average transfer rate was 0.064 ± 0.005% of the administered content. [ABSTRACT FROM AUTHOR]

Context: The overproduction of reactive oxygen species (ROS) during in vitro culture of ovarian tissues impairs follicular development and survival. Aims: To evaluate the effects of punicalagin on the development and survival of primordial follicles, stromal cell and collagen fibres, as well as on the levels of mRNA for nuclear factor erythroid 2-related factor 2 (NRF2), superoxide dismutase 1 (SOD1), catalase (CAT) , glutathione peroxidase 1 (GPX1) and perirredoxin 6 (PRDX6), and activity of antioxidant enzymes in cultured bovine ovarian tissues. Methods: Bovine ovarian cortical tissues were cultured for 6 days in α-MEM+ alone or with 1.0, 10.0, or 100.0 μM punicalagin at 38.5°C with 5% CO2. Follicle morphology and growth, stromal cell density, and collagen fibres were evaluated by classical histology, while the expression of mRNA was evaluated by real-time PCR. The activity of enzymes was analysed by the Bradford method. Key results: Punicalagin improved follicle survival and development, reduced mRNA expression for SOD1 and CAT , but did not influence stromal cells or collagen fibres. Punicalagin (10.0 μM) increased the levels of thiol and activity of SOD1, CAT , and GPX1 enzymes. Conclusions: Punicalagin (10.0 μM) promotes follicle survival and development and activates SOD1, CAT , and GPX1 enzymes in bovine ovarian tissues. Implications: Punicalagin improves follicle development and survival in cultured ovarian tissues. The in vitro overproduction of reactive oxygen species (ROS) impairs follicular growth and survival. This study evaluated the effects of punicalagin on growth and survival of primordial follicles, stromal cells, collagen fibres, levels of mRNA and activity of antioxidant enzymes in cultured bovine ovarian tissues. The results showed that punicalagin improved follicle survival and development, and reduced mRNA levels for SOD1 and CAT. Punicalagin 10.0 μM increased the levels of thiol and activity of SOD1, CAT and GPX1 enzymes. Image by V. S. Bezerra. [ABSTRACT FROM AUTHOR]

There is a lack of information about transforming growth factor beta-1 (TGF-β1) and cytokines contained in pure platelet-rich plasma (P-PRP) and release from pure-platelet-rich gel supernatants (P-PRGS) might be affected by the temperature and time factors; P-PRP from 6 heifers was activated with calcium gluconate. Thereafter, P-PRG and their supernatants (P-PRGS) were maintained at −80, −20, 4, 21, and 37 °C and collected at 3, 6, 12, 24, 48, 96, 144, 192, 240, and 280 h for subsequent determination of TGF-β1, tumor necrosis factor alfa (TNF-α), interleukin (IL)-2, and IL-6; TGF-β1 concentrations were significantly (p < 0.05) higher in PRGS maintained at 21 and 37 °C when compared to PRGS maintained at 4, −20, and −80 °C; PRGS TNF-α concentrations were not influenced by temperature and time factors. However, PRGS maintained at 4 °C showed significantly (p < 0.05) higher concentrations when compared to PRGS maintained at −20, and −80 °C at 144, and 192 h. IL-6 concentrations were significantly (p < 0.05) higher in PRGS stored at −20, and −80 over the first 48 h and at 10 days when compared to PRGS stored at 4, 21, and 37 °C. These results could suggest that P-PRP/P-PRGS could be maintained and well preserved for at least 12 days at room temperature for clinical use in bovine therapeutic massive protocols. [ABSTRACT FROM AUTHOR]

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Livestock farming is vital to a country's economy, and technological innovations in animal genetics and reproductive biotechnologies are key for environmental and socioeconomic development. This study aimed to evaluate the antioxidant capacity of Coffea arabica grounds (CAG) extract on the semen parameters of Fleckvieh cattle. CAG was processed, and its antioxidant capacity was assessed using DPPH, FRAP, and total phenols assays. Semen was collected from a pedigree bull and analysed macroscopically and microscopically. Different concentrations of CAG (0, 1, 1.5, and 2 mg/ml) were tested at 4, 12, 24, and 36 h post-refrigeration. The study used a completely randomised design with ten replications, and variables such as motility, kinetic parameters, membrane functionality, and acrosomal integrity were analysed. The normal distribution of the variables -- motility, kinetic parameters, membrane functionality, and acrosomal integrity -- was analysed using the Shapiro-Wilk statistical test (P > 0.05). An analysis of variance was then performed with a significance level of P < 0.05 to compare the means, using InfoStat software. Results showed that 1 mg of CAG significantly improved total motility and progressive motility at 4 h, along with kinetic parameters and membrane integrity. Overall, CAG exhibits antioxidant properties that enhance sperm characteristics, particularly at 1 mg/ml concentration after 4 h of refrigeration. However, further studies are needed to understand better the mechanisms of action and the overall efficacy of CAG as an antioxidant agent in this specific context. [ABSTRACT FROM AUTHOR]

Optimising the feed composition, especially the forage choice, and reducing costs are essential for improving dairy production efficiency. Different forage sources and proportions were used to formulate rations containing equal energy and protein, and their effects on rumen microbiota and milk production performance of dairy cows were evaluated in two experiments. In experiment 1, thirty lactating cows (235 ±13 d postpartum; milk production 29.1 ± 1.0 kg/day) were divided into Pangola and Bermuda groups. In experiment 2, twenty lactating cows (79.9 ± 8.1 d postpartum; milk production 34.7 ± 0.6 kg/day) were divided into Pangola and oat groups. In experiment 1, the Simpson index for rumen microbiota of the Pangola group was significantly higher than in the Bermuda group (P < 0.05). Analysis of the weighted unique fraction (UniFrac) distances indicated significant differences in the beta diversity of the community composition of rumen microbiota between Pangola, Bermuda and oat groups in both experiments (P < 0.001). The relative abundance of Prevotella brevis was significantly higher in the Pangola group than in the oat group in experiment 2 (P < 0.05). The somatic cell counts (SCCs), C18:0, and C18:1 in milk were significantly higher in the Bermuda group than in the Pangola group (P < 0.05) in experiment 1. On the other hand, milk crude protein (CP) and solids-not-fat (SNF) were significantly higher in the Oat group than in the Pangola group (P < 0.05) in experiment 2; however, milk urea nitrogen (MUN) was significantly higher in the Pangola group (P < 0.05). In conclusion, a switch of forage (Pangola vs Bermuda) at a lower proportion of the diet under the high forage level condition (experiment 1) caused only minor changes in rumen microbiota diversity (Simpson index, beta diversity) and milk production performance (milk SCCs, C18:0 and C18:1). On the other hand, a switch of forage (Pangola vs oat) at a higher proportion of the diet under the low forage level condition (experiment 2) resulted in greater changes in rumen microbiota diversity (beta diversity, relative abundances of bacterial taxa, P. brevis relative abundance) and milk production performance (milk CP, SNF, and MUN). [ABSTRACT FROM AUTHOR]

Objective: This study aimed to determine the prevalence of hydatid cysts and assess their direct economic impact. Methods: The study utilized a cross-sectional design to examine cattle slaughtered in the Torra Municipal Abattoir, in which 384 bovine species were systematically selected. Results: Out of the 384 slaughtered animals, 66 were found to have bovine hydatid cysts. A total of 112 cysts were collected, 44 (39.29%) of which were viable and 68 (60.71%) calcified. All examined organs showed hydatid cysts, but livers and lungs were most commonly affected, while tongues had the least amount. In terms of viable cysts, lung and liver contained the highest proportions, followed by heart and tongue. Consequently, the lung was the organ that was often impacted and had the greatest number of cysts. It was calculated that US$ 2,909 was lost annually as a direct economic loss. The animal's origin and sex did not significantly affect the infection rate statistically (p > 0.05). Similarly, there was no significant difference with age (p > 0.05). Conclusions: The study revealed a notable prevalence and economic impact of bovine hydatidosis in the study area. Therefore, it is crucial to implement effective control and preventive measures to mitigate the impact of disease's impact on public health. [ABSTRACT FROM AUTHOR]