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• Deep learning pose estimation for multi-cattle lameness detection
  

  Sci Rep. 2023 Mar 18;13(1):4499. doi: 10.1038/s41598-023-31297-1.

  ABSTRACT

  The objective of this study was to develop a fully automated multiple-cow real-time lameness detection system using a deep learning approach for cattle detection and pose estimation that could be deployed across dairy farms. Utilising computer vision and deep learning, the system can analyse simultaneously both the posture and gait of each cow within a camera field of view to a very high degree of accuracy (94-100%). Twenty-five video sequences containing 250 cows in varying degrees of lameness were recorded and independently scored by three accredited Agriculture and Horticulture Development Board (AHDB) mobility scorers using the AHDB dairy mobility scoring system to provide ground truth lameness data. These observers showed significant inter-observer reliability. Video sequences were broken down into their constituent frames and with a further 500 images downloaded from google, annotated with 15 anatomical points for each animal. A modified Mask-RCNN estimated the pose of each cow to output 5 key-points to determine back arching and 2 key-points to determine head position. Using the SORT (simple, online, and real-time tracking) algorithm, cows were tracked as they move through frames of the video sequence (i.e., in moving animals). All the features were combined using the CatBoost gradient boosting algorithm with accuracy being determined using threefold cross-validation including recursive feature elimination. Precision was assessed using Cohen’s kappa coefficient and assessments of precision and recall. This methodology was applied to cows with varying degrees of lameness (according to accredited scoring, n = 3) and demonstrated that some characteristics directly associated with lameness could be monitored simultaneously. By combining the algorithm results over time, more robust evaluation of individual cow lameness was obtained. The model showed high performance for predicting and matching the ground truth lameness data with the outputs of the algorithm. Overall, threefold lameness detection accuracy of 100% and a lameness severity classification accuracy of 94% respectively was achieved with a high degree of precision (Cohen’s kappa = 0.8782, precision = 0.8650 and recall = 0.9209).

  PMID:36934125 | PMC:PMC10024686 | DOI:10.1038/s41598-023-31297-1

• Genome-wide identification and annotation of SNPs and their mapping in candidate genes related to milk production and fertility traits in Badri cattle
  

  Trop Anim Health Prod. 2023 Mar 16;55(2):117. doi: 10.1007/s11250-023-03535-1.

  ABSTRACT

  This study was conducted in Badri cattle using a double digest restriction-site associated DNA sequencing approach. The study aimed to identify and annotate high confidence single nucleotide polymorphisms (SNPs) and their mapping in candidate genes related to production and fertility in dairy cattle. A total of 7,168,552 genome-wide SNPs were initially identified in Badri cattle by alignment with the Bos indicus reference genome. After filtration of SNPs, 65,483 high confidence SNPs were retained and further used for downstream analysis. Annotation of high confidence SNPs revealed 99.197% SNPs had modifier impact, 0.326% SNPs were low impact, 0.036% were high impact, and 0.441% were moderate impact SNPs. Most SNPs in Badri cattle were found in intergenic, transcript and intronic regions. The candidate genes for milk production PRKCE, ABCG2, GHR, EPS8, CAST and NRXN1 were found to harbour maximum high confidence variants. Among candidate genes for fertility in cattle, ATP2B1, SOX5, WDR27, ARHGAP12, CACNA1D, ANKRD6, GRIA3, ZNF521 and CAST822 have maximum high confidence variants mapped in them. The SNPs found mapped in the candidate genes will be important genetic tools in the search for phenotype-modifying nucleotide changes and will aid in formulating relevant genetic improvement programmes for dairy cattle.

  PMID:36928332 | DOI:10.1007/s11250-023-03535-1

• Effects of propolis extract on growth performance and health condition of dairy calves
  

  Trop Anim Health Prod. 2023 Mar 16;55(2):115. doi: 10.1007/s11250-023-03542-2.

  ABSTRACT

  The aim of this study was to evaluate the effect of propolis ethanol extract, on performance, fecal structure, and general health status of calves during the preweaning phase. For this purpose, 24 Simmental calves that were given colostrum for the first 3 days after birth were enrolled in the study. The study started when the calves were 4 days old and finished when they were 60 days old. These calves were divided into two groups as propolis and control groups according to their weight and gender. Both groups were given the same ration and housed in individual calf huts under the same environmental conditions. Differently from the control group, the calves in the propolis group were given 4 mL of propolis extract (300 mg/mL) with an oral feeding syringe for 56 days after morning feeding. The amounts of feed intake and fecal scores of the calves were recorded daily. Health scores, rectal body temperature, fecal pH, body weight, and body measurements were recorded weekly. For the determination of the counts of Lactobacillus spp. and Bifidobacterium spp., fecal samples were collected at the beginning and at the end of the study. The results indicated that oral administration of propolis had positive effects in terms of some performance parameters, the number of days with diarrhea, and fecal scores. In this study, it was concluded that propolis extract had the potential to positively affect the growth performance and health status of preweaning calves.

  PMID:36928524 | DOI:10.1007/s11250-023-03542-2

• Bovine abortion storm highlights risk posed by Neospora-naive dogs
  

  Vet Rec. 2023 Mar;192(6):245-247. doi: 10.1002/vetr.2860.

  NO ABSTRACT

  PMID:36928961 | DOI:10.1002/vetr.2860

• Optimising colostrum management has long-term health and productivity benefits in beef herds
  

  Vet Rec. 2023 Mar;192(6):249-251. doi: 10.1002/vetr.2863.

  NO ABSTRACT

  PMID:36928962 | DOI:10.1002/vetr.2863

• Efficiency of conventional and nanoparticle oxytetracycline in treatment of clinical endometritis in postpartum dairy cows
  

  Trop Anim Health Prod. 2023 Mar 17;55(2):118. doi: 10.1007/s11250-023-03536-0.

  ABSTRACT

  The incidence of clinical endometritis in dairy cows postpartum is one of the important reasons for financial losses in the dairy industry. The costs of treatment, milk losses, infertility, repeated breeding, and high annual culling rate of dairy cows present immediate losses in case of treatment failure. The commonly used therapeutic methods for clinical endometritis have not been successful nor have given definitive solutions to overcome the complications of the disease in dairy cows. Therefore, it was necessary to propose an innovative treatment program to overcome the reasons for the failure and lack of effectiveness of the treatment of clinical endometritis. This was tackled in the current study; oxytetracycline with different concentrations, oxytetracycline 5% (OTCC5%), oxytetracycline 20% (OTCC20%), and oxytetracycline 20% nanoparticles (OTC-NPs) were used for the treatment of clinical endometritis. Diagnosis of clinical endometritis was based on the assessment of high serum concentration of pro-inflammatory cytokines, acute phase protein, increased endometrium thickness, and intrauterine discharges with different degrees of echogenicity monitored by ultrasonography. Application of OTC-NPs revealed a decrease in serum concentration of pro-inflammatory cytokines (IL-1, IL-6, and TNF-α) and acute phase proteins compared to OTCC20% and OTCC5% groups. The improvement achieved by OTC-NPs may be attributed to the reduction of OTC particles into nano size which facilitates its tissue bioavailability, dispersion, penetration power to deeper tissues, and its more broad-spectrum activities. These activities were clearly apparent after the evacuation of uterine contents using a single dose of PGF2α. The OTC-NPs revealed a reduction in serum concentration of cytokines compared to OTCC20% and OTCC5% groups arranged as follows: 10.11, 25.45, 35.56 for IL-1; 99, 300, 319 for IL-6; 1.01, 4.40, 8.06 for CRP; and 46, 183, 266 for TNF-α. Furthermore, an increase in serum concentration of albumin (3.34) was obtained by OTC-NPs compared to OTCC5% (1.70). This improvement can be taken as evidence of liver resumption functions and inflammatory reactions. On the other side, globulin concentration recorded an increase like albumin and total proteins in OTC-NPs compared to others. A reduction in the endometrium thickness in OTC-NPs with the disappearance of intrauterine discharges was monitored by ultrasonography. This confirmed the subsiding of clinical endometritis in OTC-NPs group. Moreover, a significant improvement in conception and pregnancy rate in OTC-NPs compared to other groups were observed.

  PMID:36930327 | PMC:PMC10023628 | DOI:10.1007/s11250-023-03536-0

• Repeat breeding: prevalence and potential causes in dairy cows at different milk pocket areas of Bangladesh
  

  Trop Anim Health Prod. 2023 Mar 17;55(2):120. doi: 10.1007/s11250-023-03537-z.

  ABSTRACT

  The objective of this study was to figure out the prevalence and probable causes of repeat breeding (RB) in dairy cows. Hence, a cross-sectional study was conducted on randomly selected 265 dairy farms in Sirajganj, Bogura, Rangpur, Satkhira, and Munshiganj districts of Bangladesh from December 2018 to February 2019. Data were collected through a direct interview method using a survey questionnaire. The reproductive organs of repeat breeder cows were examined for pathological, infectious, and functional reasons, and genital tract abnormalities. Additionally, the influence of nutrition, season, and age on the frequency of RB was recorded. The prevalence of RB was 28% among the 3824 cows investigated. Among the total repeat breeder cases, 72.54% of RB cases were found in Holstein-Friesian crossbred, 23.90% in Jersey crossbred, 1.50% in Sahiwal crossbred, and 2.06% in indigenous cows. The prevalence of RB was significantly highest (P < 0.01) in Satkhira (44.35%) and lowest in the Munshiganj district (15.87%). Data indicated that a major proportion of cows significantly (P < 0.05) faced RB problems due to functional causes (34.18%), followed by pathological causes (28.01%), genital tract abnormalities (21.32%), and infectious causes (16.49%). Furthermore, the cows were remarkably (P < 0.001) affected in RB during the summer season and nutritional deficient diseases like milk fever (70%). Age (3-7 years) had a significant (P < 0.001) effect on the RB occurrence (90%) in crossbred cows. However, particular focus should be given to systematic breeding, balanced nutrition, artificial inseminator efficiency, and hygienic inseminating tools to reduce RB incidence in high-yielding crossbred cows.

  PMID:36930420 | DOI:10.1007/s11250-023-03537-z

• Estimating genetic parameters of reproductive, carcass, and meat quality traits in Polled Nellore cattle
  

  Trop Anim Health Prod. 2023 Mar 17;55(2):119. doi: 10.1007/s11250-023-03541-3.

  ABSTRACT

  Considering the economic and commercial efficiency of the beef production chain, the yield and quality of the meat produced must also be included in breeding programs. For the Nellore breed, including the polled herd, these aspects have not been much studied. The aim of this study was to estimate genetic parameters for scrotal circumference adjusted to 365 (SC365) and 450 (SC450) days of age, age at first calving (AFC), accumulated productivity (AP), stayability (STAY), longissimus muscle area (LMA), thickness of subcutaneous fat over the 12th-13th ribs (BF), thickness of subcutaneous fat over the rump (RF), and shear force measured by Warner-Bratzler shear force (WBSF) of polled Nellore cattle. Bayesian analyses were performed by adopting a linear animal model, whereas STAY analyses used the linear threshold model. Heritability estimates were 0.31 (SC365), 0.37 (SC450), 0.16 (AFC), 0.25 (AP), 0.16 (STAY), 0.30 (LMA), 0.13 (BF), 0.24 (RF), and 0.15 (WBSF), indicating moderate response to selection. Genetic and residual correlations between SC365 and SC450 were high (0.91 and 0.74, respectively), as well as the genetic correlations of AP with SC365, SC450, AFC, and STAY (0.61, 0.62, – 0.69, and 0.83, respectively). Genetic and residual correlations of WBSF with reproductive and carcass characteristics exhibited high standard deviations, however favorable. Based on the results, it is expected that in the medium term, animals with greater sexual precocity will also have greater accumulated productivity and longer permanence of females in the herd, along with superior carcass traits. However, due to the low heritabilities and small genetic associations with reproductive traits, fat thickness characteristics (BF and RF) will still require direct selection.

  PMID:36930426 | DOI:10.1007/s11250-023-03541-3

• Expression of NFIL3 and CEBPA regulated by IFNT induced-PGE2 in bovine endometrial stromal cells during the pre-implantation period
  

  Front Endocrinol (Lausanne). 2023 Feb 21;14:1075030. doi: 10.3389/fendo.2023.1075030. eCollection 2023.

  ABSTRACT

  Prostaglandin E2 (PGE2) is considered as a luteoprotective factor, influencing the corpus luteum during the early pregnant period in the bovine species. Cyclic AMP (cAMP) is activated in response to PGE2 and plays a role in many physiological processes. The maternal recognition signal, interferon τ (IFNT), induces PGE2 secretion from the endometrial epithelial cells, the function of which in stroma cells has not been completely understood. In this study, PGE2 was found to activate cAMP in the bovine endometrial stromal cells (STRs). STRs were then treated with forskolin to activate the cAMP signaling, from which RNA extracted was subjected to global expression analysis. Transcripts related to transcription regulatory region nucleic acid binding of molecular function, nucleus of cellular component, and mitotic spindle organization of biological processes were up-regulated in cAMP-activated bovine STRs. An increase in the transcription factors, NFIL3, CEBPA, and HIF1A via the cAMP/PKA/CREB signaling pathway in the bovine STRs was also found by qPCR. Knockdown of NFIL3, CEBPA, or HIF1A blocked forskolin-induced PTGS1/2 and IGFBP1/3 expression. Moreover, NFIL3 and CEBPA were localized in endometrial stroma on pregnant day 17 (day 0 = estrous cycle), but not on cyclic day 17. These observations indicated that uterine PGE2 induced by conceptus IFNT is involved in the early pregnancy-related gene expression in endometrial stromal cells, which could facilitate pregnancy establishment in the bovine.

  PMID:36923228 | PMC:PMC10010167 | DOI:10.3389/fendo.2023.1075030

• Evaluation of the safety, immunogenicity and efficacy of a new live-attenuated lumpy skin disease vaccine in India
  

  Virulence. 2023 Dec;14(1):2190647. doi: 10.1080/21505594.2023.2190647.

  ABSTRACT

  Lumpy skin disease (LSD) was reported for the first time in India in 2019 and since then, it has become endemic. Since a homologous (LSD-virus based) vaccine was not available in the country, goatpox virus (GPV)-based heterologous vaccine was authorized for mass immunization to induce protection against LSD in cattle. This study describes the evaluation of safety, immunogenicity and efficacy of a new live-attenuated LSD vaccine developed by using an Indian field strain, isolated in 2019 from cattle. The virus was attenuated by continuous passage (P = 50) in Vero cells. The vaccine (50^th LSDV passage in Vero cells, named as Lumpi-ProVac^Ind) did not induce any local or systemic reaction upon its experimental inoculation in calves (n = 10). At day 30 post-vaccination (pv), the vaccinated animals were shown to develop antibody- and cell-mediated immune responses and exhibited complete protection upon virulent LSDV challenge. A minimum Neethling response (0.018% animals; 5 out of 26,940 animals) of the vaccine was observed in the field trials conducted in 26,940 animals. There was no significant reduction in the milk yield in lactating animals (n = 10108), besides there was no abortion or any other reproductive disorder in the pregnant animals (n = 2889). Sero-conversion was observed in 85.18% animals in the field by day 30 pv.

  PMID:36919498 | DOI:10.1080/21505594.2023.2190647

• Single-nuclei RNA sequencing (snRNA-seq) uncovers trophoblast cell types and lineages in the mature bovine placenta
  

  Proc Natl Acad Sci U S A. 2023 Mar 21;120(12):e2221526120. doi: 10.1073/pnas.2221526120. Epub 2023 Mar 13.

  ABSTRACT

  Ruminants have a semi-invasive placenta, which possess highly vascularized placentomes formed by maternal endometrial caruncles and fetal placental cotyledons and required for fetal development to term. The synepitheliochorial placenta of cattle contains at least two trophoblast cell populations, including uninucleate (UNC) and binucleate (BNC) cells that are most abundant in the cotyledonary chorion of the placentomes. The interplacentomal placenta is more epitheliochorial in nature with the chorion developing specialized areolae over the openings of uterine glands. Of note, the cell types in the placenta and cellular and molecular mechanisms governing trophoblast differentiation and function are little understood in ruminants. To fill this knowledge gap, the cotyledonary and intercotyledonary areas of the mature day 195 bovine placenta were analyzed by single nuclei analysis. Single-nuclei RNA-seq analysis found substantial differences in cell type composition and transcriptional profiles between the two distinct regions of the placenta. Based on clustering and cell marker gene expression, five different trophoblast cell types were identified in the chorion, including proliferating and differentiating UNC and two different types of BNC in the cotyledon. Cell trajectory analyses provided a framework for understanding the differentiation of trophoblast UNC into BNC. The upstream transcription factor binding analysis of differentially expressed genes identified a candidate set of regulator factors and genes regulating trophoblast differentiation. This foundational information is useful to discover essential biological pathways underpinning the development and function of the bovine placenta.

  PMID:36913592 | DOI:10.1073/pnas.2221526120

• The Effect of Supplementation Using a Mixture of Fish Oil and Linseed on the Level of Immunomodulatory Components in Bovine Colostrum
  

  Molecules. 2023 Feb 24;28(5):2154. doi: 10.3390/molecules28052154.

  ABSTRACT

  The aim of this study was to determine the effect of supplementing rations, with a mixture of fish oil and linseed, on the level of immunomodulatory components in colostrum. Twenty multiparous cows, that were three weeks before scheduled calving, had a body condition of 3-3.5, and had not been diagnosed with multiple pregnancies, were qualified for the experiment. The cows were divided into two groups: experimental (FOL) (n = 10) and control (CTL) (n = 10). The CTL group were individually given the standard food ration for dry cows for about 21 days before calving, while the FOL group received food rations that were enriched with 150 g of fish oil and 250 g of linseed (golden variety). Colostrum samples for testing were taken twice a day on the first and second days of lactation, and then once a day from the third to the fifth day of lactation. The experiment showed that the applied supplementation had an impact, in the form of increasing the fat, protein, IgG, IgA, IgM, vitamin A, C22:6 n-3 (DHA), and C18:2 cis9 trans11 (CLA) contents in colostrum; however, the C18: 2 n-6 (LA) and C20:4 n-6 (AA) contents decreased. Due to the lower quality of colostrum found in high-yield cows, and therefore in the Holstein-Friesian breed, it is possible to improve the quality by, among other things, introducing nutritional modifications during the second stage of the dry period.

  PMID:36903401 | PMC:PMC10004384 | DOI:10.3390/molecules28052154

• Genome-Wide Landscape of mRNAs, lncRNAs, and circRNAs during Testicular Development of Yak
  

  Int J Mol Sci. 2023 Feb 23;24(5):4420. doi: 10.3390/ijms24054420.

  ABSTRACT

  Testicular development is a tightly regulated process in mammals. Understanding the molecular mechanisms of yak testicular development will benefit the yak breeding industry. However, the roles of different RNAs, such as mRNA, lncRNA, and circRNA in the testicular development of yak, are still largely unclear. In this study, transcriptome analyses were performed on the expression profiles of mRNAs, lncRNAs, and circRNAs in testis tissues of Ashidan yak at different developmental stages, including 6-months-old (M6), 18-months-old (M18), and 30-months-old (M30). A total of 30, 23, and 277 common differentially expressed (DE) mRNAs, lncRNAs, and circRNAs were identified in M6, M18, and M30, respectively. Furthermore, functional enrichment analysis showed that the common DE mRNAs during the entire developmental process were mainly involved in gonadal mesoderm development, cell differentiation, and spermatogenesis processes. Additionally, co-expression network analysis identified the potential lncRNAs related to spermatogenesis, e.g., TCONS_00087394 and TCONS_00012202. Our study provides new information about changes in RNA expression during yak testicular development, which greatly improves our understanding of the molecular mechanisms regulating testicular development in yaks.

  PMID:36901865 | PMC:PMC10002557 | DOI:10.3390/ijms24054420

• Evaluation of serological diagnostic tests for bovine brucellosis in dairy cattle herds in an endemic area: a multicenter study
  

  Trop Anim Health Prod. 2023 Mar 1;55(2):104. doi: 10.1007/s11250-023-03519-1.

  ABSTRACT

  Brucellosis is known as one of the most common zoonotic diseases worldwide affecting both livestock and humans. It causes abortions, reduces milk production, and infertility in infected animals. The disease is routinely diagnosed through three serological techniques, such as rose bengal plate test (RBPT), standard agglutination test (SAT), and indirect enzyme-linked immunosorbent assay (I-ELISA). The aim of this study was to identify and compare the brucellosis seroprevalence among dairy cattle farms through these different serological tests. From 2112 sampled dairy cattle in different parts of Iran, RBPT, SAT, and I-ELISA led to 296 (14.02%), 215 (10.18%), and 297 (14.06%) positive results, respectively. Brucella abortus biovar 3 (62 cases) was identified as the most common cause of brucellosis in tested animals. Our results showed that the specificity and sensitivity of I-ELISA were higher than those obtained by RBPT and SAT. In this study, the overall agreement of RBPT and SAT with I-ELISA reached 95.21% and 94.12% in dairy cattle farms, respectively. Furthermore, Cohen’s kappa statistical analysis revealed that the best degree of agreement was seen between RBPT and I-ELISA (0.80), followed by RBPT and SAT (0.78) and finally SAT and I-ELISA (0.72), thereby indicating a strong agreement between RBPT and I-ELISA methods and good agreement between SAT and I-ELISA methods. The McNemar analysis also showed that a significant difference exists between positive and negative results determined by SAT and I-ELISA methods (p < 0.0001). However, the positive and negative results determined by I-ELISA and RBPT did not show a significant difference (p = 0.9207). Therefore, I-ELISA was a more specific and sensitive serological test when compared to RBPT and SAT and could remarkably decrease non-specific reaction by improving the serological screening specificity for an accurate brucellosis diagnosis in endemic areas.

  PMID:36856875 | DOI:10.1007/s11250-023-03519-1

• Predicting the development of T1D and identifying its Key Performance Indicators in children; a case-control study in Saudi Arabia
  

  PLoS One. 2023 Mar 1;18(3):e0282426. doi: 10.1371/journal.pone.0282426. eCollection 2023.

  ABSTRACT

  The increasing incidence of type 1 diabetes (T1D) in children is a growing global concern. It is known that genetic and environmental factors contribute to childhood T1D. An optimal model to predict the development of T1D in children using Key Performance Indicators (KPIs) would aid medical practitioners in developing intervention plans. This paper for the first time has built a model to predict the risk of developing T1D and identify its significant KPIs in children aged (0-14) in Saudi Arabia. Machine learning methods, namely Logistic Regression, Random Forest, Support Vector Machine, Naive Bayes, and Artificial Neural Network have been utilised and compared for their relative performance. Analyses were performed in a population-based case-control study from three Saudi Arabian regions. The dataset (n = 1,142) contained demographic and socioeconomic status, genetic and disease history, nutrition history, obstetric history, and maternal characteristics. The comparison between case and control groups showed that most children (cases = 68% and controls = 88%) are from urban areas, 69% (cases) and 66% (control) were delivered after a full-term pregnancy and 31% of cases group were delivered by caesarean, which was higher than the controls (χ2 = 4.12, P-value = 0.042). Models were built using all available environmental and family history factors. The efficacy of models was evaluated using Area Under the Curve, Sensitivity, F Score and Precision. Full logistic regression outperformed other models with Accuracy = 0.77, Sensitivity, F Score and Precision of 0.70, and AUC = 0.83. The most significant KPIs were early exposure to cow’s milk (OR = 2.92, P = 0.000), birth weight >4 Kg (OR = 3.11, P = 0.007), residency(rural) (OR = 3.74, P = 0.000), family history (first and second degree), and maternal age >25 years. The results presented here can assist healthcare providers in collecting and monitoring influential KPIs and developing intervention strategies to reduce the childhood T1D incidence rate in Saudi Arabia.

  PMID:36857368 | PMC:PMC9977054 | DOI:10.1371/journal.pone.0282426

• Hair cortisol concentrations in bovine neonates born to healthy and ill cows
  

  Schweiz Arch Tierheilkd. 2023 Mar;165(3):181-0. doi: 10.17236/sat00388.

  ABSTRACT

  The goal of this study was to investigate the hair cortisol concentration (HCC) in healthy and ill cows and their newborn calves. A total of 40 cows and their 42 newborn calves were divided into two groups: group 1 consisted of 19 clinically healthy cows and their 20 newborn calves, and group 2 comprised 21 cows that had had a chronic illness in the third trimester of gestation and their 22 newborn calves. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) system was used to measure the HCC in hair samples that were collected from the cows and calves on the day the calves were born. In both groups, the mean HCCs of the calves was significantly higher than that of the cows (group 1, 31,0 vs. 0,6 pg/mg; group 2, 19,4 vs. 0,8 pg/mg; P.

  PMID:36852871 | DOI:10.17236/sat00388

• Repeated injection of multivitamins and multiminerals during the transition period enhances immune response by suppressing inflammation and oxidative stress in cows and their calves
  

  Front Immunol. 2023 Feb 9;14:1059956. doi: 10.3389/fimmu.2023.1059956. eCollection 2023.

  ABSTRACT

  Periparturient dairy cows undergo major physiological and metabolic changes as well as immunosuppression, associated with decrease in plasma concentrations of various minerals and vitamins. The present study was conducted to investigate effects of repeated injections of vitamins and minerals on oxidative stress, innate and adaptive immune response in periparturient dairy cows and their offspring. Experiment was carried out on 24 peripartum Karan-Fries cows, randomly divided into four groups (n=6): control, Multi-mineral (MM), Multi-vitamin (MV) and Multi-minerals and Multi-vitamin (MMMV). Five ml of MM (Zinc 40 mg/ml, Manganese 10 mg/ml, Copper 15 mg/ml, Selenium 5 mg/ml) and five ml of MV (Vitamin E 5 mg/ml, Vitamin A 1000 IU/ml, B-Complex 5 mg/ml, and Vitamin D₃ 500 IU/ml) were injected intramuscularly (IM) to the MM and MV groups. MMMV group cows were injected with both. In all treatment groups, injections and blood sampling were carried out on 30^th, 15^th, 7^th days before and after expected date of parturition and at calving. In calves, blood was collected at calving and on 1, 2, 3, 4, 7, 8, 15, 30 and 45 days post-calving. Colostrum/milk were collected at calving and at days 2, 4, and 8 post-calving. A lower percentage of total neutrophils and immature neutrophils, higher percentage of lymphocytes together with increased phagocytic activity of neutrophils and proliferative capacity of lymphocytes found in blood of MMMV cows/calves. Lower relative mRNA expression of TLRs and CXCRs and higher mRNA expression of GR-α, CD62L, CD11b, CD25 and CD44 found in blood neutrophils of MMMV groups. Total antioxidant capacity was higher, activity of antioxidant enzymes (SOD and CAT), TBARS levels were lower in the blood plasma of treated cows/calves. In both cows/calves, plasma pro-inflammatory cytokines (IL-1α, IL-1β, IL-6, IL-8, IL-17A, IFN-γ and TNF-α) increased, whereas anti-inflammatory cytokines (IL-4 and IL-10) decreased in MMMV groups. Total immunoglobulins increased in colostrum/milk of MMMV injected cows and plasma of their calves. Results indicate that repeated injections of multivitamins and multiminerals to peripartum dairy cows could be a major strategy to improve immune response and decrease in inflammation and oxidative stress in transition dairy cows and their calves.

  PMID:36845154 | PMC:PMC9950815 | DOI:10.3389/fimmu.2023.1059956

• Glutamine, proline, and isoleucine support maturation and fertilisation of bovine oocytes
  

  Theriogenology. 2023 Apr 15;201:59-67. doi: 10.1016/j.theriogenology.2023.02.019. Epub 2023 Feb 18.

  ABSTRACT

  Successful in-vitro production of bovine embryos relies on meiotic maturation of oocytes in vitro (IVM) before they can be fertilised. High levels of IVM are currently achieved using a complex medium that contains all 20 common amino acids, namely TCM199, but can also be achieved using a simple inorganic salt solution containing non-essential amino acids, proline, and glutamine. Further simplification of the amino acid content of medium used for IVM could lead to a more defined medium that provides reproducible IVM. The aim of this study was, therefore, to determine the minimal amino acid requirements for bovine oocyte nuclear maturation, as measured by progression to metaphase II (MII) of meiosis. Supplementation of a simple medium composed of inorganic salts (M1 medium) with multiple amino-acid combinations showed that M1 containing glutamine, proline, and isoleucine resulted in nuclear maturation comparable to that of TCM199 (57.4 ± 3.4% vs 67% ± 1.7%, respectively) but was reduced when cystine (Cys2) to that seen with M1 alone (38.0 ± 2.2%). Viability of oocytes matured in this simplified medium was equal to those matured in TCM199 since the same proportion of zygotes with 2 pronuclei were observed following fertilisation in medium containing no amino acids (33.9 ± 6.5% vs 33.3 ± 3.6%, respectively). Addition of glutamine, proline and isoleucine to fertilisation medium also increased the proportion of zygotes but did not increase blastocyst development rates. Thus, a defined medium containing only glutamine, proline and isoleucine is sufficient for oocyte maturation and successful fertilisation.

  PMID:36842262 | DOI:10.1016/j.theriogenology.2023.02.019

• Effect of subclinical endometritis and flunixin meglumine administration on pregnancy in embryo recipient beef cows
  

  Theriogenology. 2023 Apr 15;201:76-82. doi: 10.1016/j.theriogenology.2023.02.020. Epub 2023 Feb 20.

  ABSTRACT

  Fertility of recipient beef cows with subclinical endometritis (SCE) that did or did not receive flunixin meglumine (FM) treatment were compared following transfer of d 7 embryo. The study population comprised of 600 Angus cross cows that expressed estrus following Select-Synch + CIDR (Controlled Internal Drug Release) estrus synchronization protocol. At the time of embryo transfer, approximately 3 wk after sampling for subclinical endometritis, cows were randomly allocated either to receive FM treatment (500 mg of Banamine®; n = 300) or not (Control; n = 300). The effect of subclinical endometritis (at ≥ 1% PMN on endometrial cytology by cytobrush method) and FM treatment on pregnancy/embryo transfer (P/ET, %) were evaluated by mixed model. Of the 600 cows, 323 (53.8%) became pregnant; 55.0% (165/300) cows that received FM treatment vs. 52.7% (158/300) control cows (P > 0.1), and 55.9% (266/476) normal vs. 46.0% (57/124) subclinical endometritis cows (P < 0.05). There was a trend for treatment by subclinical endometritis for P/ET (P = 0.09). Pregnancy was recorded in 55.3% (134/242) of normal and 53.4% (31/58) of subclinical endometritis cows that received FM treatment, and in 56.4% (132/234) of normal and 39.4% (26/66) of subclinical endometritis cows that did not receive FM treatment (P = 0.09). In conclusion, subclinical endometritis in recipient beef cows resulted in lower P/ET. Though not significant in cows with subclinical endometritis, FM treatment resulted in 14.0% points more pregnancy compared with control.

  PMID:36842264 | DOI:10.1016/j.theriogenology.2023.02.020

• mRNA Signatures in Peripheral White Blood Cells Predict Reproductive Potential in Beef Heifers at Weaning
  

  Genes (Basel). 2023 Feb 15;14(2):498. doi: 10.3390/genes14020498.

  ABSTRACT

  Reproductive failure is a major contributor to inefficiency within the cow-calf industry. Particularly problematic is the inability to diagnose heifer reproductive issues prior to pregnancy diagnosis following their first breeding season. Therefore, we hypothesized that gene expression from the peripheral white blood cells at weaning could predict the future reproductive potential of beef heifers. To investigate this, the gene expression was measured using RNA-Seq in Angus-Simmental crossbred heifers sampled at weaning and retrospectively classified as fertile (FH, n = 8) or subfertile (SFH, n = 7) after pregnancy diagnosis. We identified 92 differentially expressed genes between the groups. Network co-expression analysis identified 14 and 52 hub targets. ENSBTAG00000052659, OLR1, TFF2, and NAIP were exclusive hubs to the FH group, while 42 hubs were exclusive to the SFH group. The differential connectivity between the networks of each group revealed a gain in connectivity due to the rewiring of major regulators in the SFH group. The exclusive hub targets from FH were over-represented for the CXCR chemokine receptor pathway and inflammasome complex, while for the SFH, they were over-represented for immune response and cytokine production pathways. These multiple interactions revealed novel targets and pathways predicting reproductive potential at an early stage of heifer development.

  PMID:36833425 | PMC:PMC9957530 | DOI:10.3390/genes14020498

• Testis-Specific Protein Y-Encoded (TSPY) Is Required for Male Early Embryo Development in <em>Bos taurus</em>
  

  Int J Mol Sci. 2023 Feb 8;24(4):3349. doi: 10.3390/ijms24043349.

  ABSTRACT

  TSPY is a highly conserved multi-copy gene with copy number variation (CNV) among species, populations, individuals and within families. TSPY has been shown to be involved in male development and fertility. However, information on TSPY in embryonic preimplantation stages is lacking. This study aims to determine whether TSPY CNV plays a role in male early development. Using sex-sorted semen from three different bulls, male embryo groups referred to as 1Y, 2Y and 3Y, were produced by in vitro fertilization (IVF). Developmental competency was assessed by cleavage and blastocyst rates. Embryos at different developmental stages were analyzed for TSPY CN, mRNA and protein levels. Furthermore, TSPY RNA knockdown was performed and embryos were assessed as per above. Development competency was only significantly different at the blastocyst stage, with 3Y being the highest. TSPY CNV and transcripts were detected in the range of 20-75 CN for 1Y, 20-65 CN for 2Y and 20-150 CN for 3Y, with corresponding averages of 30.2 ± 2.5, 33.0 ± 2.4 and 82.3 ± 3.6 copies, respectively. TSPY transcripts exhibited an inverse logarithmic pattern, with 3Y showing significantly higher TSPY. TSPY proteins, detected only in blastocysts, were not significantly different among groups. TSPY knockdown resulted in a significant TSPY depletion (p < 0.05), with no development observed after the eight-cell stage in male embryos, suggesting that TSPY is required for male embryo development.

  PMID:36834761 | PMC:PMC9959854 | DOI:10.3390/ijms24043349

• Dairy calves are exposed to isoflavones during the developmentally most sensitive period of their life
  

  Theriogenology. 2023 Apr 15;201:53-58. doi: 10.1016/j.theriogenology.2023.02.007. Epub 2023 Feb 10.

  ABSTRACT

  Isoflavones represent a class of phytoestrogens present in plants. In dairy cows, dietary isoflavones have been shown to negatively affect reproductive performance. To the best of our knowledge, no studies have yet been conducted to determine if calves are pre- or neonatally confronted with isoflavones and their metabolites. In the present study, we hypothesize that isoflavones are passed on from the dam to the offspring in utero. Twenty-three pregnant Holstein Friesian dams and their calves, originating from three commercial dairy farms in Belgium, were included. Heparin blood samples were collected during the first, second, and third trimester of gestation from all pregnant dams. Heparin blood and hair samples were obtained from the offspring within 24 h after parturition. Colostrum samples were collected from a subset of eight dams to determine the concentration of isoflavones and their metabolites. During the first and second trimester of gestation, the dams were fed either a youngstock (nulliparous dams) or a lactation (multiparous dams) diet. During the third trimester, both groups received a similar dry cow diet. Genistein and daidzein levels were unaffected by diet type, while their metabolite [equol, dihydrodaidzein (DHD), and o-desmethylangolensin (ODMA)] concentrations were significantly higher in the lactation group. Furthermore, metabolite concentrations decreased significantly during gestation. Isoflavones and their metabolites were detected in all colostrum samples. No correlation could be found between levels in colostrum and blood of pregnant dams or calves. Peripheral levels of isoflavones and their metabolites were significantly lower in newborn calves in comparison to their dams. Genistein and daidzein concentrations were found to be significantly higher in the calves’ hair versus blood samples, suggesting prenatal exposure to isoflavones for an extended period of time. In contrast, no isoflavone metabolites were detected in the calves’ hair samples. This is the first study to demonstrate that dairy calves are exposed to isoflavones during the developmentally most sensitive period of their lives. Results obtained pave the way for more extensive research to examine which effects isoflavones might have on developing organ systems like the reproductive system.

  PMID:36841122 | DOI:10.1016/j.theriogenology.2023.02.007

• Short-term physiological responses to moderate heat stress in grazing dairy cows in temperate climate
  

  Animal. 2023 Mar;17(3):100718. doi: 10.1016/j.animal.2023.100718. Epub 2023 Jan 25.

  ABSTRACT

  Even in temperate climate regions, an increase in ambient temperature and exposure to solar radiation can cause heat stress in lactating dairy cows. We hypothesised that grazing dairy cows exhibit short-term physiological changes due to increasing heat load under moderate climate conditions. Over two consecutive summers, 38 lactating Holstein dairy cows were studied in a full-time grazing system. Data were collected in 10 experimental periods of up to three consecutive days with a moderate comprehensive climate index (CCI). The individual animals’ vaginal temperature (VT), heart rate, and locomotor activity data were automatically monitored with sensors. Blood samples and proportional whole milk samples were collected at afternoon milking. The concentrations of beta-hydroxybutyrate, glucose, non-esterified fatty acids, urea nitrogen, plasma thyroxine and triiodothyronine were analysed in blood plasma, and fat, protein, lactose, urea nitrogen, cortisol, Na⁺, K⁺, and Cl^– concentrations were analysed in milk. The daily distribution of VT recordings greater than 39 °C showed a circadian rhythm with a proportion of recordings of 2% and lower during the night and a percentage of 10% or higher in the afternoon. The cows’ maximal daily vaginal temperature (VT_MAX) between 0830 and 1430 h was positively related to the mean daily CCI in the same time period (CCI_MEAN; mean and SD 23.6 ± 5.4 °C). Cows with greater VT_MAX had an increased mean heart rate, plasma glucose and milk cortisol concentrations and decreased concentrations of plasma thyroxine and triiodothyronine. The concentration of Na⁺ in milk was lower, and the concentration of K⁺ in milk tended to be higher in cows with increased VT_MAX. For beta-hydroxybutyrate, non-esterified fatty acids and urea nitrogen concentrations in plasma and fat and lactose concentrations in milk no relationships were found in terms of increasing VT. For milk urea nitrogen and protein concentrations, the proportion of total variance explained by inter-individual or -period variance was high. In conclusion, changes observed in milk and blood likely reflected short-term physiological responses to moderate heat stress. In particular, milk cortisol and Na⁺ may be useful traits for timely monitoring of heat stress in individual cows because their inter-individual variances were relatively small and samples can be collected non-invasively.

  PMID:36827848 | DOI:10.1016/j.animal.2023.100718

• Nitrogen and energy losses and methane emissions from beef cattle fed diets with gradual replacement of maize silage and concentrate with grass silage and corn-cob mix
  

  Animal. 2023 Mar;17(3):100722. doi: 10.1016/j.animal.2023.100722. Epub 2023 Jan 30.

  ABSTRACT

  Diets based on large proportions of grassland-based feed are uncommon in forage-based intensive beef production, thus contradicting governmental or commercial strategies to promote the use of grassland-based feed in ruminant production systems. Compared with typical maize silage/concentrate diets, grassland-based diets are associated with impaired nitrogen (N) and energy utilisation because of the comparably lower energy and higher CP content of these feeds. However, quantitative studies concerning the effects of increased dietary proportions of grassland-derived feeds on N and energy losses and utilisation and on methane emissions are missing and the compensation potential of using a limited proportion of an energy-rich forage is unknown. Therefore, we tested five diets with varying types and proportions of forage and concentrate. Three diets consisted of grass silage, maize silage, and concentrate in ratios of, g/kg DM, 100:600:300 (G100; control), 300:500:200 (G300), and 500:300:200 (G500), respectively. Two diets were composed of grass silage, corn-cob mix (CCM), and concentrate in ratios of, g/kg DM, 500:300:200 (G500_CCM), and 750:150:100 (G750_CCM), respectively. A high-protein concentrate (270 g CP/kg DM) was fed to G100, whereas a low-protein concentrate (140 g CP/kg DM) was used in the remaining diets. Diets were fed throughout the entire fattening period to groups of six Limousin-crossbred bulls each. When weighing 246 ± 18 kg, each animal underwent a 7-day total daily faeces and urine collection, which was followed by measuring methane emissions in respiration chambers for 48 h. Total DM intake was similar across all diets, whereas the N intake varied (P < 0.05). Urinary N loss (g/day) was the highest for G750_CCM (28.2) and G100 (26.6) and lowest for G500_CCM (15.2) and G300 (16.9) (P < 0.001). Energy utilisation was comparable among all groups. Metabolisable energy intake decreased numerically only with increasing proportions of grass silage in the diet. Substituting maize silage with CCM counteracted the loss in metabolisable energy intake. Absolute methane emissions were not different across the groups, but methane emission intensity (mg/g body protein retention) varied (P < 0.05), being numerically lower for G100 (349) and G500_CCM (401) compared with the other groups (488 on average). In conclusion, the results show that the grass silage proportion in beef cattle diets can be substantially increased when strategically combined with energy-dense forages, such as CCM. This also limits the need for concentrate and additional protein sources; in addition, the associated urinary N emissions, which are potentially noxious to the environment, are avoided.

  PMID:36827849 | DOI:10.1016/j.animal.2023.100722

• The Effects of Incremental Doses of Aflatoxin B₁ on In Vitro Ruminal Nutrient Digestibility and Fermentation Profile of a Lactating Dairy Cow Diet in a Dual-Flow Continuous Culture System
  

  Toxins (Basel). 2023 Jan 18;15(2):90. doi: 10.3390/toxins15020090.

  ABSTRACT

  Aflatoxin B₁ (AFB₁) is a mycotoxin known to impair human and animal health. It is also believed to have a deleterious effect on ruminal nutrient digestibility under in vitro batch culture systems. The objective of this study was to evaluate the effects of increasing the dose of AFB₁ on ruminal dry matter and nutrient digestibility, fermentation profile, and N flows using a dual-flow continuous culture system fed a diet formulated for lactating dairy cows. Eight fermenter vessels were used in a replicated 4 × 4 Latin square design with 10 d periods (7 d adaptation and 3 d sample collection). Treatments were randomly applied to fermenters on diet DM basis: (1) 0 μg of AFB₁/kg of DM (Control); (2) 50 μg of AFB₁/kg of DM (AF50); (3) 100 μg of AFB₁/kg of DM (AF100); and (4) 150 μg of AFB₁/kg of DM (AF150). Treatments did not affect nutrient digestibility, fermentation, and N flows. Aflatoxin B₁ concentration in ruminal fluid increased with dose but decreased to undetectable levels after 4 h post-dosing. In conclusion, adding incremental doses of AFB₁ did not affect ruminal fermentation, digestibility of nutrients, and N flows in a dual-flow continuous culture system fed diets formulated for lactating dairy cows.

  PMID:36828405 | PMC:PMC9964898 | DOI:10.3390/toxins15020090

• Sperm capacitation and transcripts levels are altered by in vitro THC exposure
  

  BMC Mol Cell Biol. 2023 Feb 23;24(1):6. doi: 10.1186/s12860-023-00468-3.

  ABSTRACT

  BACKGROUND: Delta-9-tetrahydrocannabinol (THC) is the primary phytocannabinoid responsible for the psychoactive properties of cannabis and is known to interact with the endocannabinoid system, which is functionally present in the male reproductive system. Since cannabis consumption is the highest among reproductive aged males, the current study aimed to further investigate the effects of THC exposure to phenotypical, physiological, and molecular parameters in sperm. Bull sperm of known fertility were used as a translational model for human sperm and subjected to in vitro treatment with physiologically relevant experimental doses of THC. Sperm parameters, capacitation, apoptosis, and transcript levels were evaluated following treatment.

  RESULTS: Motility, morphology, and viability of bovine sperm was unaltered from THC exposure. However, 0.32µM of THC caused an increased proportion of capacitating sperm (p < 0.05) compared to control and vehicle group sperm. Transcriptome analysis revealed that 39 genes were found to be differentially expressed by 0.032µM THC exposure, 196 genes were differentially expressed by 0.32µM THC exposure, and 33 genes were differentially expressed by 3.2µM THC. Secondary analysis reveals pathways involving development, nucleosomes, ribosomes and translation, and cellular metabolism to be significantly enriched.

  CONCLUSION: Phytocannabinoid exposure to sperm may adversely affect sperm function by stimulating premature capacitation. These findings also show for the first time that spermatozoal transcripts may be altered by THC exposure. These results add to previous research demonstrating the molecular effects of cannabinoids on sperm and warrant further research into the effects of cannabis on male fertility.

  PMID:36823609 | PMC:PMC9951432 | DOI:10.1186/s12860-023-00468-3

• Effect of carvacrol antioxidant capacity on oocyte maturation and embryo production in cattle
  

  Zygote. 2023 Apr;31(2):173-179. doi: 10.1017/S0967199422000673. Epub 2023 Feb 20.

  ABSTRACT

  Carvacrol (C₁₀H₁₄O), an efficient phenolic antioxidant substance for several cell types, may become a useful antioxidant for female germ cells and embryo culture. This study investigates the effects of carvacrol supplementation on bovine oocytes in in vitro maturation (IVM) and embryo production. In total, 1222 cumulus-oocyte complexes were cultured in TCM-199⁺ alone (control treatment) or supplemented with carvacrol at the concentrations of 3 µM (Carv-3), 12.5 µM (Carv-12.5), or 25 µM (Carv-25). After IVM, the oocytes were subjected to in vitro fertilization and embryo production, and the spent medium post-IVM was used for evaluating the levels of reactive oxygen species and the antioxidant capacity (2,2-diphenyl-1-picryl-hydrazyl-hydrate and 2,2′-azinobis-3-ethyl-benzothiozoline-6-sulphonic acid quantification). A greater (P < 0.05) antioxidant potential was observed in the spent medium of all carvacrol-treated groups compared with the control medium. Moreover, the addition of carvacrol to the maturation medium did not affect (P > 0.05) blastocyst production on days 7 and 10 of culture; however, the total number of cells per blastocyst was reduced (P < 0.05) in two carvacrol-treated groups (Carv-3 and Carv-25). In conclusion, carvacrol demonstrated a high antioxidant capacity in the spent medium after oocyte maturation; however, although embryo production was not affected, in general, carvacrol addition to IVM medium reduced the total number of cells per blastocyst. Therefore, due to the high antioxidant capacity of carvacrol, new experiments are warranted to investigate the beneficial effects of lower concentrations of carvacrol on embryo production in cattle and other species.

  PMID:36804925 | DOI:10.1017/S0967199422000673

• Rumen-protected glucose stimulates the secretion of reproductive hormones and the mTOR/AKT signaling pathway in the ovaries of early postpartum
  

  Sci Rep. 2023 Feb 20;13(1):2940. doi: 10.1038/s41598-023-30170-5.

  ABSTRACT

  This study was conducted to determine the response of the reproductive hormones and the mTOR/AKT/PI3K pathway in the ovaries of postpartum dairy cows with dietary rumen-protected glucose (RPG). Twelve Holstein cows were randomly assigned to two groups (n = 6/group): the control group (CT) and the RPG group. Blood samples were collected on d 1, 7, and 14 after calving for the gonadal hormone assay. The expression of the gonadal hormones receptors and PI3K/mTOR/AKT pathways were detected using RT-PCR and Western blot. The RPG addition increased the plasma LH, E2, and P4 concentrations on d 14 after calving and upregulated the mRNA and protein expressions of the ERα, ERβ, 17β-HSD, FSHR, LHR, and CYP17A1 but downregulated StAR expression. Immunohistochemical analysis identified higher expressions of the FSHR and LHR in the ovaries of RPG-fed cows compared to CT cows. Furthermore, the protein expressions of p-AKT/AKT and p-mTOR/mTOR were significantly increased in the ovaries of RPG-fed cows compared to the CT group, but the addition of RPG did not alter the protein expression of p-PI3K/PI3K. In conclusion, the current results indicated that dietary RPG supplementation regulated gonadotropin secretion and stimulated expression of hormone receptors and the mTOR/AKT pathway in the ovaries of early postpartum dairy cows. RPG may be beneficial for the recovery of ovarian activity in post-calving dairy cows.

  PMID:36808140 | PMC:PMC9941576 | DOI:10.1038/s41598-023-30170-5

• Direct cleavage during the first mitosis is a sign of abnormal fertilization in cattle
  

  Theriogenology. 2023 Apr 1;200:96-105. doi: 10.1016/j.theriogenology.2023.01.028. Epub 2023 Feb 4.

  ABSTRACT

  Direct cleavage, a type of abnormal cleavage in which one zygote divides into three or more blastomeres, has been reported in mammals. The incidence of direct cleavage increases in zygotes with three or more pronuclei (multi-PN) and those showing abnormal pronuclei migration. However, there are few reports on the relationship between pronuclei and direct cleavage, and the effects of these relationships on subsequent embryogenesis have not been clarified. It is difficult to observe pronuclei under visible light, especially in bovine zygotes, because of abundant dark lipid droplets in the cytoplasm. We visualized pronuclei by removing lipid droplets from bovine zygotes and analyzed the relationship between the number of pronuclei and direct cleavage using time-lapse cinematography. The direct cleavage rate of multi-PN zygotes was 78.6%, which was significantly higher than that of zygotes with one pronucleus (1 PN, 0.0%) and two pronuclei (2 PN, 8.2%). Observation of pronuclei migration in 2 PN zygotes showed that 3.1% of 2 PN zygotes had non-apposed pronuclei. The direct cleavage rate of zygotes with non-apposed pronuclei was 66.7%, which was significantly higher than that of zygotes with apposed pronuclei (6.4%). Among multi-PN zygotes, the proportions of zygotes with apposed pronuclei and non-apposed pronuclei were 37.5% and 64.3%, respectively. The direct cleavage rate of multi-PN zygotes with non-apposed pronuclei was 100.0%, which was significantly higher than that of zygotes with apposed pronuclei (40.0%). Three-dimensional live-cell imaging of bovine zygotes injected with the mRNA-encoding histone H2B-mCherry showed that the direct cleavage rates of 2 PN and multi-PN zygotes bypassing syngamy were 63.2% and 75.5%, respectively. These rates were significantly higher than that of 2 PN and multi-PN zygotes that underwent syngamy (5.6% and 20.0%, respectively). Regardless of the number of pronuclei, a high frequency of direct cleavage was observed in zygotes in which the pronuclei did not migrate inward the cytoplasm and bypassed syngamy. These results suggest that abnormal fertilization such as multi-PN and migration error of pronuclei in cattle is the primary reason for direct cleavage during the first mitosis. Assessment of direct cleavage during the first mitosis allows exclusion of embryos with abnormal fertilization and may contribute to in vitro produced embryo transfer success.

  PMID:36805250 | DOI:10.1016/j.theriogenology.2023.01.028

• Effect of prepartum dietary energy sources on productive and reproductive performance in Nili Ravi buffaloes
  

  Trop Anim Health Prod. 2023 Feb 18;55(2):88. doi: 10.1007/s11250-023-03510-w.

  ABSTRACT

  Feeding of dietary energy sources has been extensively studied in dairy cows but not well described in dairy buffaloes. The objective of this study was to evaluate the effect of prepartum dietary energy sources on productive and reproductive performance in Nili Ravi buffaloes (n = 21). The buffaloes were offered isocaloric (1.55 Mcal/kg DM NE_L (net energy for lactation)) glucogenic (GD), lipogenic (LD), and mixed diet (MD) during 63 days prepartum and maintained (1.27 Mcal/kg DM NE_L) at lactation diet (LCD) during 14 weeks postpartum. Effects of dietary energy sources and week on animals were analyzed with the mixed model. The DMI, BCS, and body weights remained similar during the pre- and postpartum periods. The prepartum diets did not affect birth weight, blood metabolites, milk yield, and composition. The GD tended to early uterine involution, more follicle numbers, and early follicle formation. The prepartum feeding of dietary energy source had a similar effect on first estrus expression, days open, conception rate, pregnancy rate, and calving interval. So, it could be concluded that prepartum feeding of an isocaloric dietary energy source had a similar effect on the performance of buffaloes.

  PMID:36808487 | DOI:10.1007/s11250-023-03510-w

• The development of sleep-like posture expression with age in female Holstein calves
  

  Anim Sci J. 2023 Jan;94(1):e13816. doi: 10.1111/asj.13816.

  ABSTRACT

  Sleep plays an important role in cattle production and welfare. Therefore, this study aimed to investigate the development of sleep-like posture (SLP) expression in dairy calves from birth to the first calving as an indicator of their sleep behavior. Fifteen female Holstein calves were subjected. Daily SLP was measured eight times (0.5, 1, 2, 4, 8, 12, and 18 months and 23 months or 1 month before the first calving) using an accelerometer. Calves were kept in individual pens until weaning at 2.5 months old and then kept in the group. The daily SLP time decreased rapidly in early life; however, the rate of decrease gradually slowed and eventually reached a stable level of approximately 60 min/day after 12 months old. The daily SLP bout frequency showed the same change as the SLP time. In contrast, the averaged SLP bout duration slowly decreased with age. Longer daily SLP time during early life in female Holstein calves might be related to brain development. The individual expression of daily SLP time differs before and after weaning. Some external and/or internal factors associated with weaning may be involved SLP expression.

  PMID:36802332 | DOI:10.1111/asj.13816

• Identification and profiling of microRNAs during yak's testicular development
  

  BMC Vet Res. 2023 Feb 20;19(1):53. doi: 10.1186/s12917-023-03602-7.

  ABSTRACT

  BACKGROUND: Normal testicular development is highly crucial for male reproduction and is a precondition for spermatogenesis that is the production of spermatozoa in the testes. MiRNAs have been implicated in several testicular biological processes, including cell proliferation, spermatogenesis, hormone secretion, metabolism and reproductive regulation. In the present study, we used deep sequencing data to study the functions of miRNAs during testicular development and spermatogenesis, by analyzing the expression patterns of small RNAs in 6-, 18- and 30-month-old yak testis tissues.

  RESULTS: A total of 737 known and 359 novel miRNAs were obtained from 6-, 18- and 30-month-old yak testes. In all, we obtained 12, 142 and 139 differentially expressed (DE) miRNAs in 30- vs. 18-, 18- vs. 6-, and 30- vs. 6-month-old testes, respectively. Gene Ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of all DE miRNA target genes revealed BMP2, TGFB2, GDF6, SMAD6, TGFBR2 and other target genes as participants in different biological processes, including TGF-β, GnRH, Wnt, PI3K-Akt, MAPK signaling pathways and several other reproductive pathways. In addition, quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) was used to detect the expression of seven randomly selected miRNAs in 6-, 18- and 30-month-old testes, and the results were consistent with the sequencing data.

  CONCLUSIONS: The differential expression of miRNAs in yak testes at different development stages was characterized and investigated using deep sequencing technology. We believe that the results will contribute to further understanding the functions of miRNAs in regulating the development of yak testes and improving the reproductive performance of male yaks.

  PMID:36803968 | PMC:PMC9940382 | DOI:10.1186/s12917-023-03602-7

• Flow Cytometric Identification and Enumeration of Monocyte Subsets in Bovine and Water Buffalo Peripheral Blood
  

  Curr Protoc. 2023 Feb;3(2):e676. doi: 10.1002/cpz1.676.

  ABSTRACT

  Monocytes are innate immune system key players with pivotal roles during infection and inflammation. They migrate into tissues and differentiate into myeloid effect cells (macrophages, dendritic cells) which orchestrate inflammatory processes and are interfaces between the innate and adaptive immune responses. Their clinical relevance to health and disease of cattle (Bos taurus) and water buffalo (Bubalus bubalis), two of the most important livestock species, has been highlighted in physiologic (pregnancy) and pathologic (mastitis, metritis, and viral infections) conditions. The existence of three different monocyte subsets in cattle was established by flow cytometry (FC), as follows: classical (cM; CD14⁺⁺ CD16^-/low ), intermediate (intM; CD14^++/+ CD16⁺ ), and non-classical (ncM; CD14^-/low CD16⁺⁺ ) monocytes. FC applications for studying the immune system of cattle and water buffalo still have significant limitations. In this article, we describe some practical approaches to overcome these limitations and, in particular, allow the identification and enumeration of cM, intM, and ncM subpopulations in cattle and buffalo peripheral blood. Indeed, we propose the new procedure lyse/wash/no-centrifugation (L/W/NC) that can be combined with the FC absolute counting procedures and can overcome specific issues of the lyse/no-wash protocols (L/NW). Finally, for the first time, we demonstrated the existence of cM, intM, and ncM monocyte subsets also in the water buffalo, showing some interesting differences with cattle, such as the bubaline cM are mainly CD14^+/++ /CD16⁺ . These subtle differences may influence inflammatory disease regulation in, for example, mastitis and metritis. The upregulation of CD16 expression on cM may reveal different monocyte priming, leading to different functional features of macrophages/dendritic cells in tissues after infection. © 2023 Wiley Periodicals LLC. Basic Protocol: Absolute count of cM, intM, and ncM without compensation Alternate Protocol: Absolute count of cM, intM, and ncM for single laser platform Support Protocol 1: In-house monoclonal antibody labeling using a Pacific Blue^™ kit Support Protocol 2: In-house monoclonal antibody labeling using an Alexa Fluor^® 647 kit Support Protocol 3: Titration of fluorochrome-conjugated antibodies.

  PMID:36799694 | DOI:10.1002/cpz1.676

• Performance and milk fatty acid profile of beef cows with a different energy status with short nutrient restriction and refeeding
  

  J Anim Sci. 2023 Jan 3;101:skad053. doi: 10.1093/jas/skad053.

  ABSTRACT

  Our study objective was to determine the effect of a short feed restriction (4 d) and subsequent refeeding (4 d) on the performance and metabolism of beef cows with a different nutritional status by particularly focusing on their milk fatty acid (FA) profile, to consider its potential use as biomarker of metabolic status. Thirty-two Parda de Montaña multiparous lactating beef cows were individually fed a diet based on the average cow’s net energy (NE) and metabolizable protein requirements. At 58 d in milk (DIM, day 0), cows underwent a 4 d feed restriction (55% requirements, restriction period). Before and after the restriction, diets met 100% of their requirements (basal and refeeding periods). Cow performance, milk yield and composition, and plasma metabolites, were determined on day -2, 1, 3, 5, 6, and 8. Cows were classified into two status clusters according to their pre-challenge performance and energy balance (EB) (Balanced vs. Imbalanced). All traits were statistically analyzed considering the fixed effect of status cluster and feeding period or day, with cow as a random effect. Imbalanced cows were heavier and had a more negative EB (P < 0.001), but similar milk yield, milk composition, and circulating metabolites (except for greater urea) than Balanced cows (P > 0.10). Milk contents of C18:1 cis-9, monounsaturated FA (MUFA), and mobilization FA were greater (P < 0.05), whereas saturated FA (SFA) and de novo FA were lesser in Imbalanced than Balanced cows (P < 0.05). Restriction decreased body weight (BW), milk yield, and milk protein compared to the basal period, but increased milk urea and plasma nonesterified fatty acids (NEFA) (P < 0.001). Milk contents of SFA, de novo, and mixed FA decreased immediately during the restriction, while MUFA, polyunsaturated FA and mobilization FA increased (P < 0.001). Basal milk FA contents were recovered on day 2 of refeeding, and all their changes strongly correlated with differences in EB and NEFA (P < 0.05). The general lack of interactions between status clusters and feeding periods implied that the response mechanisms to diet changes did not differ between cows with a different pre-challenge nutritional status.

  PMID:36795068 | PMC:PMC9996621 | DOI:10.1093/jas/skad053

• Hypoxia-inducible factor-1α is involved in the response to heat stress in lactating dairy cows
  

  J Therm Biol. 2023 Feb;112:103460. doi: 10.1016/j.jtherbio.2023.103460. Epub 2023 Jan 6.

  ABSTRACT

  Hypoxia-inducible factor-1α (HIF-1α) is important in maintaining cellular oxygen homeostasis and cellular heat tolerance. To explore the role of HIF-1α in the response to heat stress (HS) in dairy cows, 16 Chinese Holstein cows (milk yield: 32 ± 4 kg/d, days in milk: 272 ± 7 d, parity: 2-3) were used to collect coccygeal vein blood and milk samples when cows were under mild (temperature-humidity index = 77) and moderate HS (temperature-humidity index = 84), respectively. Compared to cows under mild HS, the respiratory rate (P < 0.01), rectal temperature (P < 0.01), and blood concentrations of heat shock protein (HSP)70 (P < 0.01) and HSP27 (P < 0.01) were higher, but oxygen saturation (P = 0.02) and hemoglobin (P < 0.01) were lower in cows under moderate HS. Blood HIF-1α concentration was greater (P < 0.01) during moderate HS, indicating that HIF-1α is involved in lactating cows' response to HS. To confirm these findings, we collected coccygeal vein blood and milk samples from 59 dairy cows under moderate HS. The HIF-1α levels were correlated with the levels of heat shock transcription factor (HSF) (r = 0.7857, P < 0.01), HSP70 (r = 0.4543, P < 0.01) and HSP27 (r = 0.8782, P < 0.01). A comparison of 15 cows with higher HIF-1α (>482 ng/L) and 15 cows with lower HIF-1α levels (<439 ng/L) showed that reactive oxidative species were higher (P = 0.02), but superoxide dismutase (P < 0.01), total antioxidation capacity (P = 0.02) and glutathione peroxidase (P < 0.01) were lower in higher HIF-1α cows. These results suggested that HIF-1α may be indicative of the risk of oxidative stress in heat-stressed cows and may participate in the response of cows to HS by synergistically activating the expression of the HSP family with HSF.

  PMID:36796905 | DOI:10.1016/j.jtherbio.2023.103460

• Toxoplasma gondii in lactating animals: potential risk to milk consuming population in Khyber Pakhtunkhwa
  

  Braz J Biol. 2023 Feb 13;83:e267369. doi: 10.1590/1519-6984.267369. eCollection 2023.

  ABSTRACT

  Toxoplasma gondii is an intracellular zoonotic protozoan parasite usually infects human and animal worldwide. This study aimed to analyze the sero-prevalence of T. gondii in blood of lactating animals and human living in close proximity and also to detect Toxoplasma DNA in unpasteurized milk of the studied animals. A total of 233 blood and milk samples were collected from lactating animals, and 735 blood samples were taken from humans in District Upper Dir, Khyber Pakhtunkhwa, Pakistan. The blood samples were analyzed through ELISA while the milk samples were analyzed by PCR for the presence of T. gondii DNA. A standard questionnaire was introduced to collect the data from the participants. In animals, the reported sero-prevalence was 32.18% for IgM, 17.16% for IgG, and 6.4% for both IgM and IgG. The reported positivity for T. gondii DNA in milk was 14.44%, 34.8%, 20%, and 26% in sheep, goats, cows, and buffaloes, respectively. In the human blood samples, 9.8% were found positive for IgM and 11.2% for IgG while none of the samples was found positive for both IgM and IgG. Overall sero-prevalence reported in females was significantly higher than the male (p<0.05) poor hygiene condition (p < 0.0001) were the significant risk factors associated with T. gondii infections in animals. In conclusion, T. gondii infection is prevalent in lactating animals and humans using their raw milk in the study area. It is suggested that raw milk should be considered as a vehicle for the transmission of T. gondii to humans. Proper pasteurization of milk is very useful in limiting the transmission of infection. Awareness and control programs should be implemented to prevent the infection.

  PMID:36790276 | DOI:10.1590/1519-6984.267369

• Bimodal milk flow and overmilking in dairy cattle: risk factors and consequences
  

  Animal. 2023 Mar;17(3):100716. doi: 10.1016/j.animal.2023.100716. Epub 2023 Jan 20.

  ABSTRACT

  To maximise the return on capital invested in the milking parlour, the largest number of cows should be milked gently and completely in the shortest possible time. Bimodal milk flow and overmilking negatively influence the efficiency of the milk removal process and teat health. This observational study had the objective of investigating the prevalence of bimodal milk flow and overmilking, determining which individual and farm-related variables are associated with these occurrences, and determining the association of overmilking and bimodal milk flow with milk yield and with short- and long-term teat changes. Twenty-one farms were visited once during the study period, wherein the milking routine was timed, the teat condition was assessed, and dynamic evaluation of the milking vacuum was performed. A total of 606 vacuum graphic records were obtained, with an average of 29 ± 3 records per farm, in order to indirectly evaluate the milk flow and thus determine the occurrence of bimodal milking and overmilking time. The average percentage of bimodality per farm was 41.7%. The median overmilking time was 59 seconds, and on average, 78.3% of the cows in a herd were overmilked longer than 30 seconds. An association was found at cow level between the occurrence of bimodal milk flow and days in milk, the total stimulation time, parity, and the preparation lag time. The increase in the mean total stimulation time and the number of passes during preparation were associated with a decrease in the proportion of bimodality in the herd. Parity, reattachment of the milking unit and milking in manual mode were associated with an increase in overmilking time of an individual cow. The presence of a clogged air bleed hole in the claw and the reduction of the cluster removal milk flow threshold were associated with an increase in the herd’s median overmilking time. The average milk flow decreased with the increase in overmilking time and with the occurrence of bimodal milk flow. An association was also found between the occurrence of bimodal milk flow and decreased milk yield. A mean of 78.4% of cows per farm had short-term teat changes in at least one teat, and 33.6% of evaluated cows per farm displayed at least one teat with hyperkeratosis. These results emphasise the association of bimodality and overmilking on milking efficiency and reinforce the importance of the milkers’ actions and the functioning of the milking parlour for its prevention.

  PMID:36791490 | DOI:10.1016/j.animal.2023.100716

• Abortion outbreak in pregnant goats and cows with coinfection of 'Candidatus Mycoplasma haemobos' and HoBi-like pestivirus
  

  Vet Microbiol. 2023 Apr;279:109690. doi: 10.1016/j.vetmic.2023.109690. Epub 2023 Feb 10.

  ABSTRACT

  New emerging diseases, including ‘Candidatus Mycoplasma haemobos’ (‘Ca. M. haemobos’) and HoBi-like pestivirus in central China has been a huge challenge for ruminant production. From July to September 2022, an abortion outbreak affected 14 goat farms and 11 cattle farms in this area. To investigate whether the two pathogens are associated with the disease, samples were collected: Group 1 included 55 goat blood samples with foetal tissue samples and ticks on the skin; Group 2 included nine healthy goat blood samples; Group 3 included 36 cow blood samples with foetal tissue samples and ticks on the skin; and Group 4 included seven healthy cow blood samples. Then, these samples were analysed by serology, PCR, sequence analysis, and identification. A total of 339 Rhipicephalus (Boophilus) microplus and 61 Haemaphysalis longicornis ticks were identified in Group 1 and Group 3. By molecular detection, 32 ‘Ca. M. haemobos’-positive amplicons, and 27 HoBi-like pestivirus-positive amplicons were amplified from goats in Group 1; meanwhile, 20 ‘Ca. M. haemobos’-positive amplicons, and 18 HoBi-like pestivirus-positive amplicons were amplified from foetuses. Statistical significance (P = 0.002) and association (OR=7.556) between the ‘Ca. M. haemobos’ PCR results of foetus and goat samples were observed, and statistical significance (P = 0.017) and association (OR=4.271) between the HoBi-like pestivirus PCR results of foetus and goat samples were observed. These significances and associations were also observed in Group 3. In addition, coinfections were detected in Group 1 and Group 3. ‘Ca. M. haemobos’ was detected in both tick species. Further serological results revealed that the frequency of HoBi-like pestivirus was 20.0 % (11/55) in Group 1 % and 30.6 % (11/36) in Group 3 in central China. No ticks, pathogens, or neutralizing antibodies were observed in Group 2 or Group 4. This is the first molecular evidence of ‘Ca. M. haemobos’ and HoBi-like pestivirus natural coinfections in goats and cows with abortion in China. HoBi-like pestivirus and ‘Ca. M. haemobos’ can be transferred from goats/cows to their foetuses.

  PMID:36791616 | DOI:10.1016/j.vetmic.2023.109690

• Histone remodeling reflects conserved mechanisms of bovine and human preimplantation development
  

  EMBO Rep. 2023 Mar 6;24(3):e55726. doi: 10.15252/embr.202255726. Epub 2023 Feb 13.

  ABSTRACT

  How histone modifications regulate changes in gene expression during preimplantation development in any species remains poorly understood. Using CUT&Tag to overcome limiting amounts of biological material, we profiled two activating (H3K4me3 and H3K27ac) and two repressive (H3K9me3 and H3K27me3) marks in bovine oocytes, 2-, 4-, and 8-cell embryos, morula, blastocysts, inner cell mass, and trophectoderm. In oocytes, broad bivalent domains mark developmental genes, and prior to embryonic genome activation (EGA), H3K9me3 and H3K27me3 co-occupy gene bodies, suggesting a global mechanism for transcription repression. During EGA, chromatin accessibility is established before canonical H3K4me3 and H3K27ac signatures. Embryonic transcription is required for this remodeling, indicating that maternally provided products alone are insufficient for reprogramming. Last, H3K27me3 plays a major role in restriction of cellular potency, as blastocyst lineages are defined by differential polycomb repression and transcription factor activity. Notably, inferred regulators of EGA and blastocyst formation strongly resemble those described in humans, as opposed to mice. These similarities suggest that cattle are a better model than rodents to investigate the molecular basis of human preimplantation development.

  PMID:36779365 | PMC:PMC9986824 | DOI:10.15252/embr.202255726

• Medium chain fatty acid supplementation improves animal metabolic and immune status during the transition period: A study on dairy cattle
  

  Front Immunol. 2023 Jan 27;14:1018867. doi: 10.3389/fimmu.2023.1018867. eCollection 2023.

  ABSTRACT

  The transition period is the stage of the high incidence of metabolic and infectious diseases in dairy cows. Improving transition dairy cows’ health is crucial for the industry. This study aimed to determine the effects of dietary supplementation medium-chain fatty acids (MCFAs) on immune function, metabolic status, performance of transition dairy cows. Twenty multiparous Holstein cows randomly assigned to two treatments at 35 d before calving. 1) CON (fed the basal 2) MCFA treatment (basal diet was supplemented at an additional 20 g MCFAs mixture every day) until 70 d after calving. The results showed that the serum amyloid A myeloperoxidase concentrations in the blood of cows in MCFA treatment significantly decreased during the early lactation (from 1 d to 28 d after calving) 0.03, 0.04, respectively) compared with the CON, while the tumor necrosis factor concentration was significantly decreased at 56 d after calving (P = 0.02). In addition, the concentration of insulin in the pre-calving (from 21 d before calving to calving) blood of cows in MCFA treatment was significantly decreased (P = 0.04), and concentration of triglyceride also showed a downward trend at 28 d after calving 0.07). Meanwhile, MCFAs supplementation significantly decreased the concentrations of lithocholic acid, hyodeoxycholic acid, and hyocholic acid in the blood at 1 d calving (P = 0.02, < 0.01, < 0.01, respectively), and the level of hyocholic acid taurocholic acid concentrations (P < 0.01, = 0.01, respectively) decreased dramatically at 14 d after calving. However, compared with the CON, the pre-calving dry matter intake and the early lactation milk yield in MCFA treatment were significantly decreased (P = 0.05, 0.02, respectively). In conclusion, MCFAs supplementation transition diet could improve the immune function and metabolic status of dairy cows, and the health of transition cows might be beneficial from the endocrine status.

  PMID:36776875 | PMC:PMC9911908 | DOI:10.3389/fimmu.2023.1018867

• Effects of the inclusion of coffee pulp silage in the diet on the performance and profitability of crossbred milk cows in the middle tropics
  

  Trop Anim Health Prod. 2023 Feb 13;55(2):78. doi: 10.1007/s11250-023-03451-4.

  ABSTRACT

  The objective of this study was to evaluate the inclusion of coffee pulp silage (CPS) on the performance and profitability of crossbred cows in the middle tropics. The research took place at the Villa Marina Experimental Farm of the University of Pamplona, Pamplonita, Colombia. Four lactating multiparous bovines with a body weight of 380 ± 10 kg, 6 ± 0.03 years of age and a body condition of 3.8 on a scale of 1 to 5. The animals were distributed in a balanced 4 × 4 Latin square for residual effect. The experiment consisted of four subperiods of 21 days each, being the first 14 days of adaptation to the experimental diet and 7 days for taking samples. The base diet consisted of Pennisetum sp. forage, concentrated in a proportion of 0.2% of body weight, water, and mineralized salt at will. The inclusion of the CPS was included in levels of 4, 8, and 12% of the dry matter (DM) intake. DM intake parameters, nutrient digestibility, milk production and composition, blood parameters, and economic analysis were evaluated. Treatments were compared using orthogonal contrasts; contrasts were constructed in order to evaluate the effects of inclusion of CPS, and the linear and quadratic effects of level inclusion in the DM of the diet. For the variables which did not present inclusion of CPS effect but a linear or quadratic effect was significant, a Dunnett’s test was performed to identify whether a supplemented treatment differed from the control. Significance was considered at P < 0.05 and tendencies when 0.05 < P ≤ 0.10. The inclusion of CPS in the diet increases (P < 0.05) the voluntary intake in kg day^-1 of DM, organic matter (OM), CP, ethereal extract (EE), non-fiber carbohydrates (NFC), digested OM (DM), neutral detergent fiber corrected for ash and protein (NDFap), DNDF and dietary NTD concentration, the total digestibility of DM and OM, and dietary DOM content. The production and chemical composition of milk was not affected (P > 0.10) by the inclusion of EPC in the diet. The inclusion of CPS increased (P < 0.05) the concentration of albumin and globulins in the blood. The inclusion of CPS in the diet improved the productive efficiency of dairy cattle. The inclusion of 4% CPS in the DM of the diet is a nutritional strategy, which improves the nutritional characteristics and the concentration of albumin and globulins in the blood of crossed dairy cows in the middle tropics and benefit: cost ratio without affecting milk production.

  PMID:36780029 | PMC:PMC9925550 | DOI:10.1007/s11250-023-03451-4

• Safety and Immunogenicity of a Chimeric Subunit Vaccine against Shiga Toxin-Producing <em>Escherichia coli</em> in Pregnant Cows
  

  Int J Mol Sci. 2023 Feb 1;24(3):2771. doi: 10.3390/ijms24032771.

  ABSTRACT

  Shiga toxin-producing Escherichia coli (STEC) is a zoonotic pathogen that causes gastroenteritis and Hemolytic Uremic Syndrome. Cattle are the main animal reservoir, excreting the bacteria in their feces and contaminating the environment. In addition, meat can be contaminated by releasing the intestinal content during slaughtering. Here, we evaluated the safety and immunogenicity of a vaccine candidate against STEC that was formulated with two chimeric proteins (Chi1 and Chi2), which contain epitopes of the OmpT, Cah and Hes proteins. Thirty pregnant cows in their third trimester of gestation were included and distributed into six groups (n = 5 per group): four groups were administered intramuscularly with three doses of the formulation containing 40 µg or 100 µg of each protein plus the Quil-A or Montanide™ Gel adjuvants, while two control groups were administered with placebos. No local or systemic adverse effects were observed during the study, and hematological parameters and values of blood biochemical indicators were similar among all groups. Furthermore, all vaccine formulations triggered systemic anti-Chi1/Chi2 IgG antibody levels that were significantly higher than the control groups. However, specific IgA levels were generally low and without significant differences among groups. Notably, anti-Chi1/Chi2 IgG antibody levels in the serum of newborn calves fed with colostrum from their immunized dams were significantly higher compared to newborn calves fed with colostrum from control cows, suggesting a passive immunization through colostrum. These results demonstrate that this vaccine is safe and immunogenic when applied to pregnant cows during the third trimester of gestation.

  PMID:36769094 | PMC:PMC9917558 | DOI:10.3390/ijms24032771

• Effect of canola oil supplementation level on total tract digestion, ruminal fermentation, and methane emissions of cows grazing Urochloa sp. supplemented with a fixed amount of concentrate
  

  Trop Anim Health Prod. 2023 Feb 11;55(2):77. doi: 10.1007/s11250-023-03485-8.

  ABSTRACT

  Four rumen-cannulated cows (Bos taurus × Bos indicus, 657 ± 92 kg body weight, BW) in a rotational grazing (Urochloa sp.) system were assigned to different canola oil (CO) inclusion levels, 0.0, 0.40, 0.80, and 1.2 g/kg according to shrunk body weight (SBW, BW adjusted for gastrointestinal filling) in a 4 × 4 Latin Square design to evaluate CO on the CH₄ emissions and dietary energy intake. CH₄ emissions were estimated using an infrared analyzer methodology (Sniffer method). Grass intake and fecal production were estimated using Cr₂O₃ as an external marker. CO supplementation increased (linear effect, P ≤ 0.05) total dry matter and gross energy intake with a linear increase (P = 0.09) in neutral detergent fiber (NDF) intake. While digestible energy (Mcal/kg) linearly increased with increasing CO supplementation level (linear effect, P < 0.05), total tract digestion of organic matter, NDF, and CP was comparable (P > 0.05) between levels. Maximal CO supplementation (1.2 g/kg SBW) significantly decreased total ruminal protozoa population, acetate:propionate ratio, and enteric methane production (g/kg DMI) by 9, 5.3, and 17.5%, respectively. This study showed that, for cows grazing tropical forages, CO can be supplemented up to 1.2 g/kg SBW (5.8% of the total diet) without negatively affecting intake and nutrient digestion while reducing ruminal fermentation efficiency and enteric methane emission (≤ 17.5%).

  PMID:36773073 | PMC:PMC9922234 | DOI:10.1007/s11250-023-03485-8

• Evidence that elevation of maternal somatic cell count could lead to production of offspring with inferior reproductive and productive performance in dairy cows during the first lactation period
  

  Theriogenology. 2023 Apr 1;200:79-85. doi: 10.1016/j.theriogenology.2023.02.002. Epub 2023 Feb 4.

  ABSTRACT

  Although the effect of mastitis on reproduction and production of lactating dairy cows has been vastly studied, little information is available about the association of maternal udder health status with offspring reproduction and production. Therefore, the present research was conducted to study the association between maternal average monthly somatic cell count (SCC) with reproduction, anti-Müllerian hormone (AMH) concentration, udder health status and milk production in the offspring. Based on maternal average monthly SCC (MSCC), offspring were classified into five categories including MSCC1 (SCC <200,000; n = 3005), MSCC2 (200,000 ≤ SCC <400,000; n = 252), MSCC3 (400,000 ≤ SCC <600,000; n = 103), MSCC4 (600,000 ≤ SCC <800,000; n = 40) and MSCC5 (SCC ≥800,000; n = 61). Data associated with reproduction, production and udder health status of offspring were retrieved from the herd database. In addition, blood samples were collected from a subset of offspring (n = 136) for measurement of serum AMH, as a reliable marker of ovarian reserves. Offspring in MSCC5 category had more services per conception and longer calving to conception interval than offspring in MSCC1 and MSCC2 categories (P < 0.05). The average number of SCC and risk of clinical mastitis in the offspring were not associated with MSCC (P > 0.05). But offspring in MSCC5 category produced less milk, fat and protein than offspring in MSCC1 category (P < 0.05). In addition, AMH concentration was lower in MSCC5 than MSCC1 category (P < 0.05). In conclusion, the present study showed that elevated maternal average monthly SCC could culminate in birth of offspring with inferior reproductive performance, smaller size of ovarian reserves and lower level of milk production during the first lactation period.

  PMID:36773383 | DOI:10.1016/j.theriogenology.2023.02.002

• Serological testing of an equal-volume milk sample – a new method to estimate the seroprevalence of small ruminant lentivirus infection?
  

  BMC Vet Res. 2023 Feb 10;19(1):43. doi: 10.1186/s12917-023-03599-z.

  ABSTRACT

  BACKGROUND: In cattle attempts to evaluate within-herd prevalence of various infectious and parasitic diseases by bulk-tank milk (BTM) testing with ELISA have been made with moderate success. The fact that BTM is composed of variable and unknown volumes of milk from individual lactating animals weakens the relationship between numerical result of the ELISA and the within-herd prevalence. We carried out a laboratory experimental study to evaluate if a pooled milk sample created by mixing an equal volume of individual milk samples from seropositive and seronegative goats, henceforth referred to as an equal-volume milk sample (EVMS), would allow for accurate estimation of within-herd seroprevalence of caprine arthritis-encephalitis (CAE) using 3 different commercial ELISAs. By mixing randomly selected milk samples from seronegative and seropositive goats, 193 EVMS were created – 93 made of seronegative samples and 100 with the proportion of seropositive individual milk samples (EVMS_%POS) ranging from 1 to 100%. EVMS_%POS could be considered as a proxy for the within-herd seroprevalence. Then, OD of EVMS (OD_EVMS) of the 193 EVMS was measured using 3 commercial ELISAs for CAE – 2 indirect and 1 competitive.

  RESULTS: The cut-off values of OD_EVMS indicating SRLV infection were determined. The regression functions were developed to link OD_EVMS with EVMS_%POS. A significant monotonic relationship between OD_EVMS measured with 2 commercial indirect ELISAs and EVMS_%POS was identified. Two regression models developed on this basis described approximately 90% of variability and allowed to estimate EVMS_%POS, when it was below 50%. High OD_EVMS indicated EVMS_%POS of > 50%.

  CONCLUSION: Our study introduces the concept of serological testing of EVMS as a method of detecting SRLV-infected herds and estimating the proportion of strongly seropositive goats. Further field studies are warranted to assess practical benefits of EVMS serological testing.

  PMID:36759821 | PMC:PMC9912530 | DOI:10.1186/s12917-023-03599-z

• Associations between insulin-like factor 3, scrotal circumference and semen characteristics in young Norwegian Red bulls
  

  Animal. 2023 Mar;17(3):100713. doi: 10.1016/j.animal.2023.100713. Epub 2023 Jan 20.

  ABSTRACT

  With the integration of genomic selection in the cattle artificial insemination (AI) industry, bulls are selected for their semen production capacity and fertility at a younger age than previously. Norwegian Red bull calves selected as candidates to become future AI bulls based on their genomic breeding value are kept in a performance testing station from around the age of 3-12 months, allowing for sample collection and analysis of different parameters during their pre- and peripubertal period. Insulin-like factor 3 (INSL3) is a small peptide hormone specifically secreted by the mature Leydig cells of the testes. In the foetus, it induces the first phase of testicular descent and is considered to reflect Leydig cell development during puberty; it could therefore be an interesting early indicator of future semen production capacity. The main objective of our study was to evaluate the relationship between INSL3, scrotal circumference (SC), and semen characteristics. This is the first time INSL3 was measured in the Norwegian Red population. We collected blood samples for analysis of INSL3 from 142 Norwegian Red bulls at the performance testing station and measured their SC on the same day. Altogether, measurements were made at four time points: upon arrival at the performance testing station (quarantine (Q): 2-5 months) and later at approximately 6, 9 and 12 months of age. Information on season and place of birth were made available from the database of the breeding company Geno, together with data on semen characteristics from the test station and the AI station. The median SCs for age groups Q, 6, 9, and 12 were 15, 21.5, 29, and 34 cm, respectively. INSL3 was shown to be positively correlated with SC (R = 0.4) but not with any of the semen characteristics. Similarly, we found no correlation between SC and sperm characteristics from data on ejaculates analysed at the performance testing station and AI station. The mean sperm volume for the 31 selected bulls with at least 10 ejaculates produced in the AI station increased from 2.3 ml at the performance testing station to 6.4 ml at the AI station. The corresponding increase in mean sperm concentration was from 497 million/ml to 1 049 million/ml. We conclude that INSL3 exhibits high inter-individual variability in the Norwegian Red bull population, which cannot be explained by the parameters measured in this study. At present, INSL3 cannot be used as a biomarker of sperm production in this breed.

  PMID:36764017 | DOI:10.1016/j.animal.2023.100713

• Comparison of the 7-day CO-Synch and 8-day estradiol-based protocols for estrus synchronization and timed artificial insemination in suckled Bos taurus cows
  

  Theriogenology. 2023 Apr 1;200:70-76. doi: 10.1016/j.theriogenology.2023.02.003. Epub 2023 Feb 4.

  ABSTRACT

  Two experiments were conducted to compare, follicle diameter (FD) on Day -1, corpus luteum (CL) area on Day 7, progesterone (P4) concentration on Day 7 and 18, pregnancy per timed artificial insemination (TAI) on Day 30, and pregnancy loss (PL) between Days 30 and 60 after TAI (TAI, Day 0) using two different synchronization protocols. In Experiment 1, Angus cows (n = 1148) were randomly assigned to either 7-d progesterone CO-Synch (7-d CO-Synch) or 8-d progesterone + estradiol (8-d P + ES) synchronization protocols for TAI. On Day -10, cows in the 7-d CO-Synch treatment group (n = 574) received a progesterone-releasing intravaginal device (PIVD; 0.5 g P4) and GnRH (0.105 mg), on Day -3 the PIVD was removed and cows received cloprostenol (0.150 mg), then, on Day 0 (64 h after PIVD removal), cows received GnRH (0.105 mg) and were TAI. On Day -10, cows in the 8-d P + ES treatment group (n = 574) received a PIVD (0.5 g P4) and estradiol benzoate (2.0 mg), on Day -2 the PIVD was removed, and cows received cloprostenol (0.150 mg) and estradiol cypionate (0.5 mg), then, on Day 0 (48 h after PIVD removal), cows were TAI. Pregnancy per TAI was determined on Days 30 and 60. In a subset of cows (7-d CO-Synch, n = 41; 8-d P + ES, n = 40), serum P4 concentration was evaluated on Day 18. In Experiment 2, anestrus (n = 34) and cyclic (n = 34) suckled beef cows were selected and submitted at random on Day -10, to either 7-d CO-Synch or 8-d P + ES treatment groups. Follicle diameter on Day -1, CL area, and serum P4 concentration on Day 7 were determined. In Experiment 1, pregnancy per TAI on Day 30 did not differ (7-d CO-Synch = 48.9%; 8-d P + ES = 45.6%) between treatments but it was greater for cows with BCS ≥5 (P < 0.01). Pregnancy loss between Days 30 and 60 did not differ between treatment groups but tended to be greater in cows with BCS <5.0 (P < 0.1). In a subset of cows, serum P4 concentration on Day 18 did not differ between treatment groups but tended to be lower (P < 0.1) in cows that had PL between Days 30 and 60 compared to cows that had no PL. In Experiment 2, FD tended to be greater (P < 0.1) and CL area was greater (P = 0.05) in anestrus cows from 7-d CO-Synch treatment. In cyclic cows, the treatment did not affect the FD or CL area. In conclusion, there was no difference in pregnancy per TAI on Day 30 and PL between Days 30 and 60 between cows using 7-d CO-Synch + PIVD or 8-d estradiol-based + PIVD protocols for estrus synchronization and TAI.

  PMID:36764187 | DOI:10.1016/j.theriogenology.2023.02.003

• Proteomic analysis of milk fat globule membranes from small-sized milk fat globules and their function in promoting lipid droplet fusion in bovine mammary epithelial cells
  

  Food Funct. 2023 Mar 6;14(5):2304-2312. doi: 10.1039/d2fo03476j.

  ABSTRACT

  In mammary epithelial cells, milk fat is synthesized as lipid droplets and secreted in the form of globules. Milk fat globules (MFGs) are covered by a lipid-protein membrane known as the milk fat globule membrane (MFGM). We randomly divided 12 Holstein cows into control and conjugated linoleic acid (CLA) groups. The control group was fed a basal diet, while the CLA group was fed the basal diet + CLA (15 g per kg DM) for 10 days. Cow performance, milk composition, and MFG size were measured daily. On day 10, we extracted MFGM proteins (n = 3) and identified them via quantitative proteomic analysis. We investigated the effects of the MFGM proteins from control and CLA-treated milk on the lipid droplet formation in MAC-T cells. Compared with the control group, the CLA group had reduced milk fat content (3.39 g/100 mL vs. 2.45 g/100 mL) and MFG size parameters (D_[4,3] of 3.85 μm vs. 3.37 μm; D_[3,2] of 3.24 μm vs. 2.83 μm). The specific surface area (SSA) increased in the CLA group. A total of 361 differentially expressed proteins were identified in the CLA group by iTRAQ quantitative proteomic analysis. Among these proteins, 100 were upregulated and 251 were downregulated (p < 0.05). In MAC-T cells, CLA-MFGM proteins increased the diameter of the lipid droplets to 1.32 μm. CLA-MFGM proteins decreased the proportion of the small lipid droplets (15.33% vs. 47.78%) and increased the proportion of the large lipid droplets (25.04% vs. 11.65%). CLA-MFGM proteins promoted lipid droplet fusion. Therefore, MFGM proteins play an important role in the regulation of the lipid droplet size.

  PMID:36752527 | DOI:10.1039/d2fo03476j

• Machine-learning methods applied to integrated transcriptomic data from bovine blastocysts and elongating conceptuses to identify genes predictive of embryonic competence
  

  FASEB J. 2023 Mar;37(3):e22809. doi: 10.1096/fj.202201977R.

  ABSTRACT

  Early pregnancy loss markedly impacts reproductive efficiency in cattle. The objectives were to model a biologically relevant gene signature predicting embryonic competence for survival after integrating transcriptomic data from blastocysts and elongating conceptuses with different developmental capacities and to validate the potential biomarkers with independent embryonic data sets through the application of machine-learning algorithms. First, two data sets from in vivo-produced blastocysts competent or not to sustain a pregnancy were integrated with a data set from long and short day-15 conceptuses. A statistical contrast determined differentially expressed genes (DEG) increasing in expression from a competent blastocyst to a long conceptus and vice versa; these were enriched for KEGG pathways related to glycolysis/gluconeogenesis and RNA processing, respectively. Next, the most discriminative DEG between blastocysts that resulted or did not in pregnancy were selected by linear discriminant analysis. These eight putative biomarker genes were validated by modeling their expression in competent or noncompetent blastocysts through Bayesian logistic regression or neural networks and predicting embryo developmental fate in four external data sets consisting of in vitro-produced blastocysts (i) competent or not, or (ii) exposed or not to detrimental conditions during culture, and elongated conceptuses (iii) of different length, or (iv) developed in the uteri of high- or subfertile heifers. Predictions for each data set were more than 85% accurate, suggesting that these genes play a key role in embryo development and pregnancy establishment. In conclusion, this study integrated transcriptomic data from seven independent experiments to identify a small set of genes capable of predicting embryonic competence for survival.

  PMID:36753406 | DOI:10.1096/fj.202201977R

• Assessing pregnancy outcomes in cow-calf operations after administration of modified-live or killed virus vaccinations at the initiation of synchronization for fixed-time AI
  

  Theriogenology. 2023 Apr 1;200:43-48. doi: 10.1016/j.theriogenology.2023.01.027. Epub 2023 Feb 1.

  ABSTRACT

  The objective of this study was to compare the reproductive outcomes (artificial insemination [AI] pregnancy rates, season pregnancy rates, AI pregnancy losses) and calf traits (birth and weaning weights) after vaccination of suckled beef cows against bovine herpesvirus 1 and bovine viral diarrhea virus using commercially-available modified-live virus (MLV) or killed virus (KV) vaccine at the initiation of a fixed-time AI program. Previously-vaccinated cows (n = 2138) on 14 farms throughout Virginia were enrolled in the study during the Fall 2017 and Spring 2018 breeding seasons. Animals received a single vaccination injection at 10 d pre-breeding, corresponding with time of CIDR insertion at initiation of the 7-d CO-Synch + CIDR synchronization protocol. Cows were inseminated at a fixed time (60-66 h after removal of the CIDR insert) and subsequently turned out with bulls approximately 1 wk after insemination for a natural service. Cows treated with the MLV vaccine had greater AI pregnancy rates than cows treated with the KV vaccine during the fall (P = 0.008; 54% vs. 46%, respectively), but not during the spring breeding season (P = 0.62; 48 vs. 49%). Season pregnancy rates were greater (P = 0.01) in the fall (95-96%) than in the spring breeding season (89-90%), but were not affected by vaccine treatment (P = 0.49) or treatment by season (P = 0.30) interactions. Percentage of AI pregnancy losses was not affected by season (P = 0.85), vaccine treatment (P = 0.83), or treatment by season interactions (P = 0.68). The number of cycles it took for cows to become pregnant by natural service differed by season (P = 0.006) but not treatment (P = 0.87) or treatment by season interaction (P = 0.997). Cows treated with the MLV vaccine gave birth earlier in the calving season (8.36 ± 0.6 d) than those treated with the KV vaccine (10.31 ± 0.6 d; P = 0.02). There was a main effect of season on birth weights (P = 0.008), weaning weights (P < 0.001), and ADG at weaning (P < 0.001), but no effects of treatment (P ≥ 0.26) or treatment by season interaction (P ≥ 0.10) on any of these parameters. Overall, this study demonstrated that the administration of an MLV vaccine at 10 d before fixed-time AI did not have any adverse effects on pregnancy or calf outcomes compared with KV vaccine administration.

  PMID:36753795 | DOI:10.1016/j.theriogenology.2023.01.027

PubMedに掲載されている雑誌の牛の繁殖学に関する最新文献をrssで自動収集して表示しております。

PubMedの検索式は以下です。適宜改変してご利用ください。2016/8/12
“Reproductive Physiological Phenomena”[Mesh] AND “Cattle”[mesh] AND “English”[language]