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Abstract Arthropod vectors play a crucial role in the transmission of hemotropic mycoplasmas, small bacteria that infect red blood cells in a wide range of animals and humans globally, leading to intravascular infections. Traditional Giemsa-stained thin blood smears, used for diagnosing hemotropic mycoplasmas through microscopic examination, have low sensitivity and are effective only when bacteremia levels are high. This study aimed to employ molecular methods to detect and genetically characterize hemotropic mycoplasmas in goats as well as investigate the potential role of fleas as vectors. Blood and flea samples were collected concurrently from goats on 16 farms across seven provinces in Thailand from January 2017 to October 2023. The 16 S rRNA, 23 S rRNA, and rnpB genes of hemoplasmas were amplified and sequenced. All fleas were identified morphologically and molecularly through DNA barcoding of the cytochrome oxidase I gene. A total of 78 out of 500 goats (15.6%), three pooled flea samples (3/6, 50%), and one individual flea (1/49, 2.04%) tested positive for hemoplasmas and all fleas were identified as Ctenocephalides orientis. BLASTN searches utilizing the three genetic markers revealed that the hemoplasmas detected in this study showed 97.81–100% similarity to Mycoplasma ovis and Candidatus Mycoplasma haemovis, which have been previously reported in sheep, goats, and humans, suggesting their zoonotic potential. The sequences were grouped into 28 unique nucleotide sequence types (ntSTs) based on minor variations in the 16 S rRNA gene. Hemotropic mycoplasma infection was significantly associated with farm locations and seasonality of sample collection (p

Abstract Background Giardia duodenalis (G. duodenalis) is a globally distributed zoonotic protozoan that parasitizes the small intestines of humans and various mammals, such as goats and sheep. The objective of this study was to establish a convenient, accurate, and specific method based on restriction fragment length polymorphism (RFLP) for genotyping assemblages A, B and E of G. duodenalis in goats. The β-giardin gene was amplified using primer pairs bgF1, bgR1, bgF2 and bgR2 by nested PCR. The PCR products were digested with the restriction enzymes Hinf I and Bgl I. The established PCR-RFLP method was used to detect and analyze the genetic subtypes of G. duodenalis in 130 fecal samples from goats and compared simultaneously with microscopic examination and nucleic acid sequencing for G. duodenalis. Results Genetic sequencing confirmed that the PCR-RFLP method accurately distinguished G. duodenalis assemblages A, B and E, as well as different combinations of mixed infections of these three assemblages. Among the 130 samples tested by PCR-RFLP, a total of 26 samples (20.00%) tested positive for G. duodenalis, a higher sensitivity than microscopic examination at 13.85% (18/130). Sequence alignment analysis revealed that among the 26 PCR-positive samples, two were identified as assemblage AI, while the remaining 24 were identified as assemblage E or E12. Conclusions This study established an accurate, efficient and rapid PCR-RFLP genotyping method using the bg sequence of G. duodenalis, enabling accurate identification and effective differentiation of goat-derived G. duodenalis assemblages without requiring sequencing.

Abstract Background Circular RNAs (circRNAs) function as essential regulatory elements with pivotal roles in various biological processes. However, their expression profiles and functional regulation during the differentiation of goat myoblasts have not been thoroughly explored. This study conducts an analysis of circRNA expression profiles during the proliferation phase (cultured in growth medium, GM) and differentiation phase (cultured in differentiation medium, DM1/DM5) of skeletal muscle satellite cells (MuSCs) in goats. Results A total of 2,094 circRNAs were identified, among which 84 were differentially expressed as determined by pairwise comparisons across three distinct groups. Validation of the expression levels of six randomly selected circRNAs was performed using reverse transcription PCR (RT-PCR) and quantitative RT-PCR (qRT-PCR), with confirmation of their back-splicing junction sites. Enrichment analysis of the host genes associated with differentially expressed circRNAs (DEcircRNAs) indicated significant involvement in biological processes such as muscle contraction, muscle hypertrophy, and muscle tissue development. Additionally, these host genes were implicated in key signaling pathways, including Hippo, TGF-beta, and MAPK pathways. Subsequently, employing Cytoscape, we developed a circRNA-miRNA interaction network to elucidate the complex regulatory mechanisms underlying goat muscle development, encompassing 21 circRNAs and 47 miRNAs. Functional assays demonstrated that circTGFβ2 enhances myogenic differentiation in goats, potentially through a miRNA sponge mechanism. Conclusion In conclusion, we identified the genome-wide expression profiles of circRNAs in goat MuSCs during both proliferation and differentiation phases, and established that circTGFβ2 plays a role in the regulation of myogenesis. This study offers a significant resource for the advanced exploration of the biological functions and mechanisms of circRNAs in the myogenesis of goats.

Background and Aim: Goats are valuable livestock because they can generate meat and milk for human consumption. Goat husbandry is becoming more intensive due to the growing demand for goat products, which may impact animal welfare and natural behavior. This study aimed to investigate the impact of natural rubber (para rubber)-based scratchers as an environmental enrichment on scratching behavior, cortisol levels, and semen quality in stable bucks (male goats/goats). Materials and Methods: Nine male goats were used in this study. Scratching behavior and cortisol levels were used as welfare indicators, whereas semen quality was evaluated as an indicator of reproductive potential. These indicators were analyzed before and after scratcher installation. Results: After installing the scratchers, the goats showed a significant increase in scratching behavior and a notable decrease in cortisol levels (p < 0.001). Notably, the goats exhibited a marked preference for scratching against the scratcher (p < 0.001) compared to the stable. They significantly preferred using their heads for scratching (p < 0.001) instead of other body parts. In addition, goats preferred to scratch on the softest rubber scratchers at specific installation locations (p < 0.001). Although there was a slight improvement in semen quality, there was no statistically significant difference (p > 0.05). Conclusion: A natural rubber-based scratcher can increase scratching behavior and reduce cortisol levels, indicating its potential to improve the welfare of farm goats. Selecting an appropriate hardness and preferred location is essential to ensure that the scratcher effectively encourages animals to use it.

Milk microbiome contributes substantially to the formation of specific organoleptic and physicochemical characteristics of dairy products. The assessment of the composition and abundance of milk microbiota is a challenging task strongly influenced by many environmental factors. Specific dairy products may be designated by the Protected Designation of Origin (PDO) and Protected Geographical Indication (PGI) labeling, which however, occasionally fail to differentiate them according to specific quality characteristics, which are defined by different microbiota-driven reactions. Combining the above limitations, the scope of the present study, was to summarize the existing information toward three main issues. First, to assess the influence level of the diet type and grazing to rumen–GI tract, mammary gland, and udder microbiome formation in ruminants. Second, to discuss the factors affecting milk microbiota, as well as the effect of the endo-mammary route on milk microbial taxa. Lastly, to evaluate “milk microbiome” as a tool for product differentiation, according to origin, which will contribute to a more robust PDO and PGI labeling. Although the limitations are still a matter of fact (especially considering the sample collection, process, evaluation, and avoidance of its contamination), significant progress has been made, regarding the identification of the factors affecting dairy products’ microbiota and its core composition. In conclusion, although so far not totally efficient in dairy products molecular identification, with the progress in soil, water, plant, and animal host’s microbiota assembly’s characterization, microbiomics could provide a powerful tool for authentication and traceability of dairy products.

Abstract After cryopreservation, reactive oxygen species (ROS) can damage sperm. Antioxidants are the primary defense against oxidative damage. Berberine is a bioactive alkaloid found in Berberis vulgaris, Curcuma longa, and Ergon grape, and is a potent antioxidant. Due to the negative effects of free radicals in oxidative stress processes, antioxidant chemicals are required to protect sperm. However, berberine has low bioavailability, making it less effective. Loading techniques on nanoparticles and nanotechnology can help overcome this limitation. Selenium nanoparticles were synthesized with barberry extract, and berberine was loaded on them. Berberine nanoparticles were then synthesized using anti-solvent precipitation with a syringe pump technique. The synthesis of nanoparticles was confirmed by EDX, UV-visible, FE-SEM, Zeta-Potential, and FTIR tests. In this experiment, we aim to investigate the impact of nano-berberine and berberine loaded on Se-NPs on goat sperm parameters after freeze-thawing. We assessed the generation of reactive oxygen species (ROS), in vitro fertility, and the subsequent embryo development of zygote with treated sperm after determining the optimal concentration of various chemicals on sperm parameters. The study found that all treatments had significant differences from the control group in terms of motility, viability, DNA and membrane integrity, ROS level, lipid peroxidation, in vitro fertility ability, and the capacity to develop inseminated oocytes (p

Abstract Background Unveiling genetic diversity features and understanding the genetic mechanisms of diverse goat phenotypes are pivotal in facilitating the preservation and utilization of these genetic resources. However, the total genetic diversity within a species can’t be captured by the reference genome of a single individual. The pan-genome is a collection of all the DNA sequences that occur in a species, and it is expected to capture the total genomic diversity of the specific species. Results We constructed a goat pan-genome using map-to-pan assemble based on 813 individuals, including 723 domestic goats and 90 samples from their wild relatives, which presented a broad regional and global representation. In total, 146 Mb sequences and 974 genes were identified as absent from the reference genome (ARS1.2; GCF_001704415.2). We identified 3,190 novel single nucleotide polymorphisms (SNPs) using the pan-genome analysis. These novel SNPs could properly reveal the population structure of domestic goats and their wild relatives. Presence/absence variation (PAV) analysis revealed gene loss and intense negative selection during domestication and improvement. Conclusions Our research highlights the importance of the goat pan-genome in capturing the missing genetic variations. It reveals the changes in genomic architecture during goat domestication and improvement, such as gene loss. This improves our understanding of the evolutionary and breeding history of goats.

Interbody fusion devices are critical in spinal surgery to restore spinal stability, reduce pain and improve function. Polyetheretherketone (PEEK) has become a commonly used alternative material for fusion cages owing to its excellent mechanical properties and biocompatibility, but its biological inertness limits bone regeneration and may lead to poor fusion. In this study, a novel strategy for preparing bioactive biomineralized PEEK cages was developed using a unique combination of osteoinductive CaP bioceramic fillings in the cage window, acid sulfonation and simulated body fluid incubation. In vitro experiments showed that biomineralized PEEK cages and CaP bioceramics regulate immunity and promote angiogenesis and bone integration via activation of hypoxia-inducible factor 1-alpha and cyclic guanosine monophosphate/protein kinase G signaling pathways. In vivo goat spinal fusion experiments demonstrated that PEEK cages filled with CaP bioceramics resulted in good bone growth and spinal fusion. Therefore, the high mechanical strength and good biocompatibility of biomineralized PEEK cages, together with the excellent bioactivity and degradation properties of CaP bioceramics, provide an ideal microenvironment for bone fusion. The development of this composite material not only addresses some of the limitations of existing fusion devices but also will facilitate the development of spinal fusion technology.

Phospholipase A (PLA) in goat semen aggregates with egg yolk in semen diluent, leading to sperm death. The aim of this study is to address the issue of sperm death caused by the interaction between PLA and egg yolk, and to explore the protective effect and metabolic regulation mechanism of the combination of dexamethasone (DXMS) and azithromycin (AZM) on goat sperm under low temperature conditions. At a low temperature of 4 °C, different concentrations of DXMS were added to semen diluents containing 30 μg/mL AZM to detect the quality of goat sperm. The optimal concentration of DXMS was determined to be 20 μg/mL. On the 5th day of storage, antioxidant capacity, total cholesterol (TC) levels, energy metabolism, and metabolomics analysis were performed on the sperm of the 20 μg/mL DXMS group. The results showed that there was no aggregation caused by the interaction between PLA and egg yolk in the group containing 30 μg/mL AZM at 4 °C. 20 μg/mL DXMS significantly improved sperm motility, plasma membrane integrity, acrosome integrity, glutathione peroxidase (GPX) (P < 0.05), catalase (CAT) (P < 0.01), and superoxide dismutase (SOD) activity (P < 0.01). The content of reactive oxygen species (ROS) and Fe2+ significantly decreased (P < 0.01), while the content of ATP (P < 0.01) and TC (P < 0.05) significantly increased. Through metabolomics analysis, a total of 56 differential metabolites (P < 0.05) were screened, including 5a, 6-Anhydrotetracycline, Betamethasone, and 11-Dehydrocorticosterone, mainly enriched in 8 metabolic pathways (P < 0.05), including steroid hormone biosynthesis, glycerophospholipid metabolism, and choline metabolism in cancer. Among them, 5 metabolic pathways are related to lipid metabolism. The results indicate that AZM effectively inhibits the aggregation of PLA and yolk, and the combination of AZM and DXMS enhances the preservation quality of goat sperm during low-temperature preservation by regulating lipid metabolism. [ABSTRACT FROM AUTHOR]

Background: Ammonium chloride is the most common urine acidifier used for urolithiasis treatment and prevention in goats. Aim: The objective of this study was to determine a guideline for ammonium chloride administration to acidify urine in goats. Methods: A three-period, three-treatment crossover study was conducted in three groups, with 10 female goats in each group. Three dosages of ammonium chloride were used (250, 350, or 450 mg/kg). In each experimental period, ammonium chloride was orally administered for three consecutive days or until the urine pH dropped below 6.5. Urine samples were collected to measure the pH before each administration and one day after the last administration. The Cox proportional hazards model was conducted to identify factors associated with the success rate in reducing the urine pH below 6.5 at a time. Results: The dosage of ammonium chloride and period of administration were significantly associated with the success rate (p < 0.001). The success rates during a unit of time for goats receiving 350 mg/kg and 450 mg/kg were greater than that of goats receiving 250 mg/kg. The second and third periods of ammonium chloride administration provided higher success rates during a unit of time compared to the first period. The success rate was also associated with the pretreatment urine pH (p = 0.044). There was a higher success rate for goats with a lower pretreatment urine pH. Conclusion: The recommended ammonium chloride dosage was 350 mg/kg. Periodic use can be considered to achieve better outcomes. The pretreatment urine pH should be measured to anticipate the clinical response. [Open Vet J 2024; 14(9.000): 2310-2314]

Abstract Background Milk is essential for mammalian nutrition because it provides vital nutrients for growth and development. Milk composition, which is influenced by genetic and environmental factors, supports lactation, a complex process crucial for milk production and quality. Recent research has focused on noncoding RNAs, particularly microRNAs (miRNAs), which are present in body fluids and regulate gene expression post-transcriptionally. This study comprehensively characterizes miRNAs in milk of four livestock species, namely Bubalus bubalis, Capra hircus, Equus asinus, and Ovis aries and identifies potential target genes. Results High-throughput sequencing of milk RNA resulted in distinct read counts across species: B. bubalis (8,790,441 reads), C. hircus (12,976,275 reads), E. asinus (9,385,067 reads), and O. aries (7,295,297 reads). E. asinus had the highest RNA mapping rate (94.6%) and O. aries the lowest (84.8%). A substantially greater proportion of miRNAs over other small RNAs was observed for the donkey milk sample (7.74%) compared to buffalo (0.87%), goat (1.57%), and sheep (1.12%). Shared miRNAs, which included miR-200a, miR-200b, miR-200c, and miR-23a among others, showed varying expression levels across species, confirmed by qPCR analysis. Functional annotation of predicted miRNA target genes highlighted diverse roles, with an enrichment in functions linked to metabolism and immunity. Pathway analysis identified immune response pathways as significant, with several miRNAs targeting specific genes across species, suggesting their regulatory function in milk. Conclusions Both conserved and species-specific miRNAs were detected in milk of the investigated species. The identified target genes of these miRNAs have important roles in neonatal development, adaptation, growth, and immune response. Furthermore, they influence milk and meat production traits in livestock.

Abstract Background Adipose tissue affects not only the meat quality of domestic animals, but also human health. Adipocyte differentiation is regulated by a series of regulatory genes and cyclins. Four and half-LIM protein (FHL2) is positively correlated with the hypertrophy of adipocytes and can cause symptoms such as obesity and diabetes. Result In the transcriptome sequencing analysis of intramuscular adipocytes after three days of differentiation, the differentially expressed gene FHL2 was found. To further explore the biological significance of the differentially expressed gene FHL2, which was downregulated in the mature adipocytes. We revealed the function of FHL2 in adipogenesis through the acquisition and loss of function of FHL2. The results showed that the overexpression of FHL2 significantly increased the expression of adipogenic genes (PPARγ, C/EBPβ) and the differentiation of intramuscular and subcutaneous adipocytes. However, silencing FHL2 significantly inhibited adipocyte differentiation. The overexpression of FHL2 increased the number of adipocytes stained with crystal violet and increased the mRNA expression of proliferation marker genes such as CCNE, PCNA, CCND and CDK2. In addition, it significantly increased the rate of EdU positive cells. In terms of apoptosis, overexpression of FHL2 significantly inhibited the expression of P53 and BAX in both intramuscular and subcutaneous adipocytes, which are involved in cell apoptosis. However, overexpression of FHL2 promoted the expression of BCL, but was rescued by the silencing of FHL2. Conclusions In summary, FHL2 may be a positive regulator of intramuscular and subcutaneous adipocyte differentiation and proliferation, and acts as a negative regulator of intramuscular and subcutaneous adipocyte apoptosis. These findings provide a theoretical basis for the subsequent elucidation of FHL2 in adipocytes.

Abstract Background Exploring the physiological and molecular mechanisms underlying goat sexual maturation can enhance breeding practices and optimize reproductive efficiency and is therefore substantially important for practical breeding purposes. As an essential neuroendocrine organ in animals, the hypothalamus is involved in sexual development and other reproductive processes in female animals. Although microRNAs (miRNAs) have been identified as significant regulators of goat reproduction, there is a lack of research on the molecular regulatory mechanisms of hypothalamic miRNAs that are involved in the sexual development of goats. Therefore, we examined the dynamic changes in serum hormone profiles and hypothalamic miRNA expression profiles at four developmental stages (1 day (neonatal, D1, n = 5), 2 months (prepubertal, M2, n = 5), 4 months (sexual maturity, M4, n = 5), and 6 months (breeding period, M6, n = 5)) during sexual development in Jining grey goats. Results Transcriptome analysis revealed 95 differentially expressed miRNAs (DEMs) in the hypothalamus of goats across the four developmental stages. The target genes of these miRNAs were significantly enriched in the GnRH signalling pathway, the PI3K-Akt signalling pathway, and the Ras signalling pathway (P

Abstract Objectives The biuret method is frequently used to determine serum total protein. On the other hand refractometer, a quicker and less expensive option, is available to determine serum total protein. However, there is no study conducted in Ethiopia to compare serum total protein measurement in veterinary settings. Therefore, this study was conducted to compare the serum total protein concentration measurement in cattle and goats obtained by the biuret method and refractometer. Results Serum samples from 60 cattle and 60 goats were assayed by both methods and data were analyzed with a paired t-test, Pearson’s correlation, and Bland-Altman plots. There was a strong positive correlation between the total protein values determined with the refractometer and the biuret method in cattle (r = 0.93) and goats (r = 0.97). There were no significant differences (p > 0.05) in the protein values measured with the refractometer and those evaluated with the biuret method in both species. Bland-Altman plots showed that biases indicating the analytic and user error were 8.33% in both species which is below the acceptable total error (

Abstract MicroRNAs (miRNAs) are important regulators of gene expression and are involved in bacterial pathogenesis and host–pathogen interactions. In this study, we investigated the function of miRNAs in the regulation of host responses to Pasteurella multocida infection. Using next-generation sequencing, we analyzed miRNA expression pattern and identified differentially expressed miRNAs in Pasteurella multocida-infected goat lungs. In addition, we investigated the function of differentially expressed miRNAs andtheir targeted signaling pathways in bacterial infection processes. The results showed that Pasteurella multocida infection led to 69 significantly differentially expressed miRNAs, including 28 known annotated miRNAs with miR-497-3p showing the most significant difference. Gene target prediction and functional enrichment analyses showed that the target genes were mainly involved in cell proliferation, regulation of the cellular metabolic process, positive regulation of cellular process, cellular senescence, PI3K-Akt signaling pathway, FoxO signaling pathway and infection-related pathways. In conclusion, these data provide a new perspective on the roles of miRNAs in Pasteurella multocida infection.

Abstract Scrapie is a transmissible spongiform encephalopathy affecting sheep and goats. The prion protein-encoding gene (PRNP) plays a crucial role in determining susceptibility and resistance to scrapie. At the European level, surveillance of scrapie is essential to prevent the spread of the disease to livestock. According to the Regulation EU 2020/772 polymorphisms K222, D/S146 could function as resistance alleles in the genetic management of disease prevention. In Italy, a breeding plan for scrapie eradication has not been implemented for goats. However, surveillance plans based on the PRNP genotype have been developed as a preventive measure for scrapie. This research aimed to describe the polymorphisms at 7 positions within the PRNP gene in 956 goats of the Alpine, Saanen and mixed populations farmed in the Lombardy Region in Italy. PRNP polymorphisms were detected using single nucleotide polymorphism markers included in the Neogen GGP Goat 70 k chip. The K222 allele occurred in all populations, with frequencies ranging from 2.1 to 12.7%. No animals carried the S/D146 resistance allele. However, it has been demonstrated that polymorphisms in the other positions analysed could influence resistance or susceptibility to scrapie outbreaks in different ways. Ten potentially distinct haplotypes were found, and the most prevalent of the three populations was H2, which differed from the wild type (H1) in terms of mutation (S vs P) at codon 240. This study provided additional information on the genetic variability of the PRNP gene in these populations in the Lombardy region of Italy, contributing to the development of genetic control measures for disease prevention.

Abstract Background Diabetes mellitus is a disease that may result from interaction between environmental factors and a strong genetic component. The current study is aimed at exploring three single nucleotide polymorphisms to identify the associated ones with type 2 diabetes in the Egyptian society. The studied single nucleotide polymorphisms (rs10096097 in GOAT, rs6740584 in CREB1, and rs62521874 in MAFA) were examined via genotyping cases (n = 98) and irrelevant healthy subjects (n = 82). Results Associations were checked using dominant, recessive, genotypic, allelic, and Cochran–Armitage trend models. By comparing diabetic patients with controls, rs6740584 was associated with type 2 diabetes by employing all used models except the recessive model. Rs10096097 was connected with type 2 diabetes using the genotypic association, Cochran–Armitage trend test, and recessive model and not any other model. Rs62521874 was not linked with type 2 diabetes in all models. Moreover, haplotype association for rs10096097 and rs62521874 was conducted as these two single nucleotide polymorphisms were located on the same chromosome. The haplotype pattern rs10096097:G—rs62521874:A was identified as a biomarker for type 2 diabetes susceptibility in the Egyptian community. Conclusions The GOAT and CREB1 polymorphisms showed susceptibility to type 2 diabetes. Moreover, MAFA had no role in the disease except through the haplotype with GOAT polymorphism.

The study investigated the detection of caprine theileriosis using Giemsa-stained direct blood smears and conventional PCR techniques, with a sample size of 150 goats of both gender and various ages were collected between July 7, 2023, and October 20, 2023, in Erbil governorate, Iraq. The infection rates of theileriosis were determined as 29.3% and 65.3%, respectively. Clinical symptoms in infected goats included dullness, depression, fever, increased respiratory rate, anorexia, pale conjunctival mucous membrane, lethargy, rough hairy skin with tick infestations, cough, nasal discharge, and prescapular lymph node enlargement. Gender-based analysis showed no significant difference in infection rates between females 17.76% and males 11.81%. However, a higher infection rate 21.8% was observed in goats aged 1-5 years compared to other age groups. Imported goats had the highest infection rate 50%, followed by imported black goats 27.3% and Merzi goats 21.4%. PCR analysis using 'catch-all' primers detected a 1098 bp band, indicating Theileria spp. infection. Specific primers for T. ovis revealed a 520 bp band, confirming T. ovis infection. All goats identified as positive through microscopic examination were also positive in PCR. Phylogenetic analysis based on 18S rRNA sequences indicated a 98-100% similarity to T. ovis. Sequences were isolated from Iraq, Turkey, and China. In conclusion, the study provides insights into the prevalence, clinical manifestations, and molecular characteristics of caprine theileriosis. The findings contribute valuable information for understanding and managing theileriosis in goat populations.

Abstract Background Breeding polled goats is a welfare-friendly approach for horn removal in comparison to invasive methods. To gain a comprehensive understanding of the genetic basis underlying polledness in goats, we conducted whole-genome sequencing of 106 Xinong Saanen dairy goats, including 33 horned individuals, 70 polled individuals, and 3 polled intersexuality syndrome (PIS) individuals. Methods The present study employed a genome-wide association study (GWAS) and linkage disequilibrium (LD) analysis to precisely map the genetic locus underlying the polled phenotype in goats. Results The analysis conducted in our study revealed a total of 320 genome-wide significant single nucleotide polymorphisms (SNPs) associated with the horned/polled phenotype in goats. These SNPs exhibited two distinct peaks on chromosome 1, spanning from 128,817,052 to 133,005,441 bp and from 150,336,143 to 150,808,639 bp. The present study identified three genome-wide significant SNPs, namely Chr1:129789816, Chr1:129791507, and Chr1:129791577, as potential markers of PIS-affected goats. The results of our LD analysis suggested a potential association between MRPS22 and infertile intersex individuals, as well as a potential association between ERG and the polled trait in goats. Conclusion We have successfully identified three marker SNPs closely linked to PIS, as well as several candidate genes associated with the polled trait in goats. These results may contribute to the development of SNP chips for early prediction of PIS in goats, thereby facilitating breeding programs aimed at producing fertile herds with polled traits.

Abstract The Yunshang black goat is a renowned mutton specialist breed mainly originating from China that has excellent breeding ability with varying litter sizes. Litter size is an important factor in the economics of goat farming. However, ruminal microbiome structure might be directly or indirectly regulated by pregnancy-associated factors, including litter sizes. Therefore, the current experiment aimed to evaluate the association of different litter sizes (low versus high) with ruminal microbiome structure by 16S rRNA gene sequencing and metabolomic profiling of Yunshang black does. A total of twenty does of the Yunshang Black breed, approximately aged between 3 and 4 years, were grouped (n = 10 goats/group) into low (D-l) and high (D-h) litter groups according to their litter size (the lower group has ≤ 2 kids/litter and the high group has ≧ 3 kids/litter, respectively). All goats were sacrificed, and collected ruminal fluid samples were subjected to 16S rRNA sequencing and LC–MS/MC Analysis for ruminal microbiome and metabolomic profiling respectively. According to PCoA analysis, the ruminal microbiota was not significantly changed by the litter sizes among the groups. The Firmicutes and Bacteroidetes were the most dominant phyla, with an abundance of 55.34% and 39.62%, respectively. However, Ruminococcaceae_UCG-009, Sediminispirochaeta, and Paraprevotella were significantly increased in the D-h group, whereas Ruminococcaceae_UCG-010 and Howardella were found to be significantly decreased in the D-l group. The metabolic profiling analysis revealed that litter size impacts metabolites as 29 and 50 metabolites in positive and negative ionic modes respectively had significant differences in their regulation. From them, 16 and 24 metabolites of the D-h group were significantly down-regulated in the positive ionic mode, while 26 metabolites were up-regulated in the negative ionic mode for the same group. The most vibrant identified metabolites, including methyl linoleate, acetylursolic acid, O-desmethyl venlafaxine glucuronide, melanostatin, and arginyl-hydroxyproline, are involved in multiple biochemical processes relevant to rumen roles. The identified differential metabolites were significantly enriched in 12 different pathways including protein digestion and absorption, glycerophospholipid metabolism, regulation of lipolysis in adipocytes, and the mTOR signaling pathway. Spearman’s correlation coefficient analysis indicated that metabolites and microbial communities were tightly correlated and had significant differences between the D-l and D-h groups. Based on the results, the present study provides novel insights into the regulation mechanisms of the rumen microbiota and metabolomic profiles leading to different fertility in goats, which can give breeders some enlightenments to further improve the fertility of Yunshang Black goats.

Oogenesis involves two phases: initial volumetric growth driven by nutrient accumulation and subsequent nuclear maturation. While melatonin (MLT) has been employed as a supplement to enhance the quality of fully grown oocytes during nuclear maturation phase, its impact on oocyte growth remains poorly studied. Here, we provide in vivo evidence demonstrating that follicle‐stimulating hormone increases MLT content in ovary. Administration of MLT improves oocyte growth and quality in mice and goats by enhancing nutrient reserves and mitochondrial function. Conversely, MLT‐deficient mice have smaller oocytes and dysfunctional mitochondria. Exploring the clinical implications of MLT in promoting oocyte growth, we observe that a brief 2‐day MLT treatment enhances oocyte quality and reproductive performance in older mice. These findings highlight the role of MLT in regulating oocyte growth and provide a specific treatment window for optimizing oocyte quality and reproductive performance in female animals. [ABSTRACT FROM AUTHOR]

The growth rate of young ruminants has been associated with production performance in later life, with recent studies highlighting the importance of rumen microbes in supporting the health and growth of ruminants. However, the specific role of rumen epithelium bacteria and microbiota-host interactions in influencing the early life growth rate of ruminants remains poorly understood. In this study, we investigated the rumen fermentation pattern, microbiota characteristics, and global gene expression profiles of the rumen epithelium in 6-month-old goats with varying growth rates. Our results showed that goats with high average daily gain (HADG) exhibited higher rumen propionate concentrations. Goats with low average daily gain (LADG) had the higher relative abundances of rumen epithelium bacteria genera U29-B03 and Quinella, while exhibiting a lower relative abundance of Lachnospiraceae UCG-009. In the rumen fluid, the relative abundances of bacteria genus Alloprevotella were lower and Desulfovibrio were higher in LADG goats compared to HADG goats. Additionally, the relative abundance of fungal genus Symmetrospora was lower in LADG goats compared to HADG goats. Transcriptome analysis showed that 415 genes were differentially expressed between LADG and HADG goats, which were enriched in functions related to cell junction and cell adhesion, etc. Correlation analysis revealed that rumen epithelium bacteria genera UCG-005 and Candidatus Saccharimonas were negatively associated, while Lachnospiraceae NK3A20 group and Oscillospiraceae NK4A214 group were positively associated with average daily gain (ADG) and genes related to barrier function. The rumen fluid bacteria genus Alloprevotella was positively correlated, while Desulfovibrio was negatively correlated with rumen propionate and ammoniacal nitrogen (NH3-N) concentrations, as well as genes related to barrier function and short chain fatty acids (SCFAs) transport. In summary, our study reveals that the higher ruminal fermentation efficiency, improved rumen epithelial barrier functions, and enhanced SCFAs transport in HADG goats could be attributed to the rumen microbiota, particularly the rumen epithelium bacteria, such as Lachnospiraceae and Oscillospiraceae NK4A214 group. [ABSTRACT FROM AUTHOR]

Simple Summary: Genomic selection (GS) is defined as a marker-assisted selection method that uses high-density molecular markers covering the entire genome combined with phenotypic data to estimate genomic estimated breeding values (GEBV). This paper reviews the principles, methods, and influencing factors of GS for breeding by using phenotypic information and genetic relationships simultaneously. It also covers the research progress of GS in meat, fiber, dairy, and reproductive traits in sheep and goats. Compared to other livestock, especially cattle, sheep and goat breeding is relatively small-scale and has lagged behind. GS combined with genomic information can help promote the breeding process and development in commercial farming of sheep and goats. We highlight the potential for phenotypic information and genetic relationships to work together to drive the breeding process, providing valuable insights for implementing GS in sheep and goat breeding. The advancement of sequencing technology and molecular breeding methods has provided technical support and assurance for accurate breeding. Genomic Selection (GS) utilizes genomic information to improve livestock breeding, and it is more accurate and more efficient than traditional selection methods. GS has been widely applied in domestic animal breeding, especially in cattle. However, there are still limited studies on the application and research of GS in sheep and goats. This paper outlines the principles, analysis methods, and influential factors of GS and elaborates on the research progress, challenges, and prospects of applying GS in sheep and goat breeding. Through the review of these aspects, this paper is expected to provide valuable references for the implementation of GS in the field of sheep and goat breeding. [ABSTRACT FROM AUTHOR]

The ovary is a crucial reproductive organ in mammals, and its development directly influences an individual's sexual maturity and reproductive capacity. To comprehensively describe ovarian sexual maturation in goats, we integrated phenotypic, hormonal, metabolomic, and transcriptomic data from four specific time points: after birth (D1), at 2 months old (M2), at 4 months old (M4), and at 6 month old (M6). The study showed that during the early stage (D1–M2), ovarian growth was the most rapid, with weight and morphology increasing by 284% and 65%, respectively, and hormone levels rose significantly, with estradiol increasing by 57%. Metabolomic analysis identified 1231 metabolites, primarily lipids, lipid molecules, and organic acids, which can support hormone balance and follicle development by providing energy and participating in signaling transduction. Transcriptomic analysis identified 543 stage-specific differentially expressed genes, mainly enriched in steroid biosynthesis, amino acid metabolism, and the PI3K/AKT pathway, which are key factors influencing ovarian cell proliferation, apoptosis, hormone secretion, and metabolism. The integrated analysis revealed the key processes in the ovarian steroid hormone biosynthesis pathway and gene/metabolite networks associated with ovarian phenotypes and hormone levels, ultimately highlighting scavenger receptor class B type 1 (SCARB1), Cytochrome P450 Family 1 Subfamily A Member 1 (CYP11A1), 3beta-hydroxysteroid dehydrogenase (3BHSD), progesterone, estradiol, and L-phenylalanine as key regulators of ovarian morphological and functional changes at different developmental stages. This study is the first to reveal the metabolic changes and molecular regulatory mechanisms during ovarian sexual maturation in goats, providing valuable insights for understanding reproductive system development and optimizing reproductive performance and breeding efficiency. [ABSTRACT FROM AUTHOR]

Skeletal muscle development is spotlighted in mammals since it closely relates to animal health and economic benefits to the breeding industry. Researchers have successfully unveiled many regulatory factors and mechanisms involving myogenesis. However, the effect of N6-methyladenosine (m6A) modification, especially demethylase and its regulated genes, on muscle development remains to be further explored. Here, we found that the typical demethylase FTO (fat mass- and obesity-associated protein) was highly enriched in goats' longissimus dorsi (LD) muscles. In addition, the level of m6A modification on transcripts was negatively regulated by FTO during the proliferation of goat skeletal muscle satellite cells (MuSCs). Moreover, a deficiency of FTO in MuSCs significantly retarded their proliferation and promoted the expression of dystrophin-associated protein 1 (DAG1). m6A modifications of DAG1 mRNA were efficiently altered by FTO. Intriguingly, the results of DAG1 levels and its m6A enrichment from FB23-2 (FTO demethylase inhibitor)-treated cells were consistent with those of the FTO knockdown, indicating that the regulation of FTO on DAG1 depended on m6A modification. Further experiments showed that interfering FTO improved m6A modification at site DAG1-122, recognized by Insulin-like growth factor 2 mRNA-binding protein 1 (IGF2BP1) and consequently stabilized DAG1 transcripts. Our study suggests that FTO promotes the proliferation of MuSCs by regulating the expression of DAG1 through m6A modification. This will extend our knowledge of the m6A-related mechanism of skeletal muscle development in animals. [ABSTRACT FROM AUTHOR]

Goats are natural hosts of Mycobacterium (M.) bovis, and affected herds can be the cause of significant economic losses. Similarites in disease course and lesions of M. bovis infections in goats and M. tuberculosis in humans make goats good models for human tuberculosis. The aim of this investigation was to characterize M. bovis challenge models in goats. For this, goats were endobronchially inoculated with three doses of M. bovis or culture medium. Clinical signs, shedding, and immune responses were monitored until 146 days post inoculation (dpi). At necropsy, lesions were examined by computed tomography, histology, and bacteriological culture. Infected goats did not develop clinical signs. M. bovis was cultured from feces, but never from nasal swabs. IGRAs were positive from 28 dpi onwards, antibodies at 140 dpi, and SICCT at 146 dpi. The increase in CD25+, IFN-γ+, and IFN-γ-releasing T-cell subpopulations was time-related, but not dose-dependent. All infected goats developed paucibacillary granulomas in the lungs and regional lymph nodes. M. bovis was regularly cultured. Dose-dependent effects included the size of pulmonary lesions, caverns, intestinal lesions, and early generalization in the high-dose group. In summary, reproducible challenge models with dose-dependent differences in lesions were established, which may serve for testing vaccines for veterinary or medical use. [ABSTRACT FROM AUTHOR]

Background: Milk is essential for mammalian nutrition because it provides vital nutrients for growth and development. Milk composition, which is influenced by genetic and environmental factors, supports lactation, a complex process crucial for milk production and quality. Recent research has focused on noncoding RNAs, particularly microRNAs (miRNAs), which are present in body fluids and regulate gene expression post-transcriptionally. This study comprehensively characterizes miRNAs in milk of four livestock species, namely Bubalus bubalis, Capra hircus, Equus asinus, and Ovis aries and identifies potential target genes. Results: High-throughput sequencing of milk RNA resulted in distinct read counts across species: B. bubalis (8,790,441 reads), C. hircus (12,976,275 reads), E. asinus (9,385,067 reads), and O. aries (7,295,297 reads). E. asinus had the highest RNA mapping rate (94.6%) and O. aries the lowest (84.8%). A substantially greater proportion of miRNAs over other small RNAs was observed for the donkey milk sample (7.74%) compared to buffalo (0.87%), goat (1.57%), and sheep (1.12%). Shared miRNAs, which included miR-200a, miR-200b, miR-200c, and miR-23a among others, showed varying expression levels across species, confirmed by qPCR analysis. Functional annotation of predicted miRNA target genes highlighted diverse roles, with an enrichment in functions linked to metabolism and immunity. Pathway analysis identified immune response pathways as significant, with several miRNAs targeting specific genes across species, suggesting their regulatory function in milk. Conclusions: Both conserved and species-specific miRNAs were detected in milk of the investigated species. The identified target genes of these miRNAs have important roles in neonatal development, adaptation, growth, and immune response. Furthermore, they influence milk and meat production traits in livestock. [ABSTRACT FROM AUTHOR]

Since the first mouse induced pluripotent stem cells (iPSCs) was derived, the in vitro culture of domestic iPSCs functionally and molecularly comparable with mouse iPSCs has been a challenge. Here, we established dairy goat iPSCs (giPSCs) from goat ear fibroblast cells with mouse iPSCs morphology, the expression of pluripotent markers and differentiation ability in vitro delivered by piggyBac transposon with nine Dox-inducible exogenous reprogramming factors. These reprogramming factors were bOMSK (bovine OCT4 , CMYC , SOX2 , and KLF4), pNhL (porcine NANOG and human LIN28), hRL (human RARG and LRH1), and SV40 Large T. Notably, AF-giPSCs (induced in activin A and bFGF condition) were capable of differentiation in embryoid bodies in vitro and could contribute to interspecies chimerism in mouse E6.5 embryos in vitro , demonstrating that AF-giPSCs have the developmental capability to generate some embryonic cell lineages. Moreover, Wnt/β-catenin signaling has an important role in driving goat induced trophoblast-like stem cells (giTLSCs) from Dox-independent giPSCs. This study will support further establishment of the stable giPSC lines without any integration of exogenous genes. • The goat induced pluripotent stem cells (giPSCs) expressed pluripotent markers and have differentiation ability in vitro. • The AF-giPSCs contributed to mouse E6.5 embryos in vitro. • The goat induced trophoblast like stem cells (giTLSCs) were firstly derived from Dox-independent giPSCs. • We found Sox2 co-expressed with Cdx2, and as well as with Eomes in the nucleus indicated the potency of giTLSC. [ABSTRACT FROM AUTHOR]

Background: Adipose tissue affects not only the meat quality of domestic animals, but also human health. Adipocyte differentiation is regulated by a series of regulatory genes and cyclins. Four and half-LIM protein (FHL2) is positively correlated with the hypertrophy of adipocytes and can cause symptoms such as obesity and diabetes. Result: In the transcriptome sequencing analysis of intramuscular adipocytes after three days of differentiation, the differentially expressed gene FHL2 was found. To further explore the biological significance of the differentially expressed gene FHL2, which was downregulated in the mature adipocytes. We revealed the function of FHL2 in adipogenesis through the acquisition and loss of function of FHL2. The results showed that the overexpression of FHL2 significantly increased the expression of adipogenic genes (PPARγ, C/EBPβ) and the differentiation of intramuscular and subcutaneous adipocytes. However, silencing FHL2 significantly inhibited adipocyte differentiation. The overexpression of FHL2 increased the number of adipocytes stained with crystal violet and increased the mRNA expression of proliferation marker genes such as CCNE, PCNA, CCND and CDK2. In addition, it significantly increased the rate of EdU positive cells. In terms of apoptosis, overexpression of FHL2 significantly inhibited the expression of P53 and BAX in both intramuscular and subcutaneous adipocytes, which are involved in cell apoptosis. However, overexpression of FHL2 promoted the expression of BCL, but was rescued by the silencing of FHL2. Conclusions: In summary, FHL2 may be a positive regulator of intramuscular and subcutaneous adipocyte differentiation and proliferation, and acts as a negative regulator of intramuscular and subcutaneous adipocyte apoptosis. These findings provide a theoretical basis for the subsequent elucidation of FHL2 in adipocytes. [ABSTRACT FROM AUTHOR]

Background: Exploring the physiological and molecular mechanisms underlying goat sexual maturation can enhance breeding practices and optimize reproductive efficiency and is therefore substantially important for practical breeding purposes. As an essential neuroendocrine organ in animals, the hypothalamus is involved in sexual development and other reproductive processes in female animals. Although microRNAs (miRNAs) have been identified as significant regulators of goat reproduction, there is a lack of research on the molecular regulatory mechanisms of hypothalamic miRNAs that are involved in the sexual development of goats. Therefore, we examined the dynamic changes in serum hormone profiles and hypothalamic miRNA expression profiles at four developmental stages (1 day (neonatal, D1, n = 5), 2 months (prepubertal, M2, n = 5), 4 months (sexual maturity, M4, n = 5), and 6 months (breeding period, M6, n = 5)) during sexual development in Jining grey goats. Results: Transcriptome analysis revealed 95 differentially expressed miRNAs (DEMs) in the hypothalamus of goats across the four developmental stages. The target genes of these miRNAs were significantly enriched in the GnRH signalling pathway, the PI3K-Akt signalling pathway, and the Ras signalling pathway (P < 0.05). Additionally, 16 DEMs are common among the M2 vs. D1, M4 vs. D1, and M6 vs. D1 comparisons, indicating that the transition from D1 to M2 represents a potentially critical period for sexual development in Jining grey goats. The bioinformatics analysis results indicate that miR-193a/miR-193b-3p-Annexin A7 (ANXA7), miR-324-5p-Adhesion G protein-coupled receptor A1 (ADGRA1), miR-324-3p-Erbb2 receptor tyrosine kinase 2 (ERBB2), and miR-324-3p-Rap guanine nucleotide exchange factor 3 (RAPGEF3) are potentially involved in biological processes such as hormone secretion, energy metabolism, and signal transduction. In addition, we further confirmed that miR-324-3p targets the regulatory gene RAPGEF3. Conclusion: These results further enrich the expression profile of hypothalamic miRNAs in goats and provide important insights for studying the regulatory effects of hypothalamic miRNAs on the sexual development of goats after birth. [ABSTRACT FROM AUTHOR]

Background: Goats (Capra hircus) have always been a source of fiber for human use and hold an important place in international high-end textiles. Fiber diameter is the most concerning economic indicator for producers. Understanding the formation mechanism of fiber diameter and its related key proteins can help optimize and control the production of cashmere. Methods: Cashmere goats (n = 36) of the Alpas (n = 18) and Alxa (n = 18) breeds, with a similar age (2 years old) and live weight (25–26 kg), were selected from the Yiwei White Cashmere Goat Breeding Farm, Erdos, Inner Mongolia. Using phenotypic indicators, we evaluated the diameter of the cashmere fibers in Alxa and Alpas goats. We also used electron microscopy to examine the cashmere fiber's structure and label-free liquid chromatography–tandem mass spectrometry to determine the protein content of the two cashmere fibers. The proteins affecting fiber diameter were identified and analyzed by Western blot, Co-Immunoprecipitation, and bioinformatics analysis. Results: The average diameter of the Alxa breed was smaller (p < 0.05) than that of the Alpas breed (Alxa's cashmere vs. Alpas' cashmere). Proteomics technology enabled the highly confident detection of 171 proteins. A total of 68 differentially expressed proteins were identified in the two types of cashmere; 131 proteins were specifically expressed in Alpas goats, and 40 proteins were specifically expressed in Alxa goats. A key protein group that could cause variations in fiber diameter was found using the protein–protein interaction network. To ascertain the reason for the variation in fiber diameter, a structural study of the major protein groups was carried out. Conclusions: KRT10, KRT14, KRT17, and KRT82 are the main proteins impacting the diameter difference, and they have a substantial effect on the average fiber diameter. [ABSTRACT FROM AUTHOR]

Background: Orbital volume refers to the sum of bulbus oculi volume and orbital cavity volume. Scientists use orbital volume as a guide to diagnose, treat and prognose various eye diseases such as thyroid eye disease and enophthalmos. Objective: This study aims to examine the orbital volumes of sheep, goats and gazelles to demonstrate homotypic variations and interspecific differences between them. Methods: Crania of six male tuj sheep, five male hair goats and five male gazelle were used in this study. The crania were scanned using a 64‐detector multi‐detector computed tomography with a slice thickness of 0.625 mm at 80 kV, 200 MA and 639 mGy. The cross‐sectional images were stored in DICOM format and then transferred to MIMICS 20.1 software. Afterwards, models were made by establishing the orbital borders on the cross‐sections. Volume values were calculated based on the models. Results and Conclusion: No homotypic variation was found between the right and left volumes within the species (p > 0.05). A statistically significant difference was observed in terms of interspecific right and left orbital volumes between sheep and gazelles (p < 0.05). It was concluded that the orbital parameters of the sheep, goats and gazelles in the same family help reveal their differences in the visual anatomy of these species and provide reference data for clinical diagnosis, treatment and the prognosis follow‐up of ocular diseases in animals. [ABSTRACT FROM AUTHOR]

Background: Widely regarded as one of the chief causes of diseases in cattle population, bovine herpesvirus‐1 (BoHV‐1) has the potential to infect sheep and goat, making them potential reservoirs or hosts for this virus. Thus, preventive measures against BoHV‐1 in cattle should not overlook the ability of this virus to infect other animals. Aims: Therefore, the focal point of this study was to ascertain the seroprevalence of BoHV‐1 in 300 healthy goats, the relationship between host and the environmental determinants of infection, and the contributing role of goats in the epidemiology of the BoHV‐1. Materials & Methods: In order to pinpoint the existing antibodies to BoHV‐1, the obtained sera were analyzed by Virus Neutralization test. Results: According to this test, the seroprevalence of BoHV‐1 appeared to be 64.33% in southwestern Iran. What logistic regression disclosed was that the odds ratio between age and infection with BoHV‐1 was 0.83 (p = 0.01), representing a decrease of 17% as goats grew one year older. In addition, females manifested a higher relative frequency of infection compared to males, with the odds of infection in female goats being registered at 1.88, compared to those in males (p = 0.2). Moreover, contrasted with goats lacking any history of abortion, those with a history of abortion featured 1.1 as the odds ratio (p = 0.87). The seroprevalence in Hendijan, Ahvaz, Shushtar and Dasht e Azadegan was detected to stand at 73.24, 71.30, 55.56 and 47.06 percent, respectively, with 6% of fluctuation in the infection rates being attributed to various geographical locations under the scrutiny of this study (p = 0.003). Discussion and Conclusion: Having attested the marked seroprevalence of BoHV‐1, the definitive role of goats in the epidemiology of this virus as a secondary host or reservoir was confirmed by the present study, necessitating the strict monitoring of BoHV‐1 in goats by animal health authorities in areas where BoHV‐1 abounds in cattle. [ABSTRACT FROM AUTHOR]

Prime editor, an editing tool based on the CRISPR/Cas9 system, allows for all 12 types of nucleotide exchanges and arbitrary indels in genomic sequences without the need for inducing DNA double-strand breaks. Despite its flexibility and precision, prime editing efficiency is still low and hindered by various factors such as target sites, editing types, and the length of the primer binding site. In this study, we developed a prime editing system by incorporating an RNA motif at the 3′ terminal of the pegRNA and integrating all twin prime editor factors into a single plasmid. These two strategies enhanced prime editing efficiency at target sites by up to 3.58-fold and 2.19-fold, respectively. Subsequently, enhanced prime editor was employed in goat cells and embryos to efficiently insert a 38 bp attB sequence into the Gt(ROSA)26Sor (Rosa26) and C-C motif chemokine receptor 5 (CCR5) loci. The enhanced prime editor can mediate 11.9% and 6.8% editing efficiency in parthenogenetic activation of embryos through embryo microinjection. In summary, our study introduces a modified prime editing system with improved editing and transfection efficiency, making it more suitable for inserting foreign sequences into primary cells and embryos. These results broaden the potential applications of prime editing technologies in the production of transgenic animals. [ABSTRACT FROM AUTHOR]

Simple Summary: Investigating the proliferation and differentiation of skeletal muscle satellite cells (MuSCs) in vitro is crucial for elucidating the underlying mechanisms of skeletal muscle development in goats. Real-time quantitative PCR (RT-qPCR) is a widely utilized technique for quantifying the expression levels of target genes. A significant challenge associated with this method is the identification of optimal reference genes for accurate normalization. In this study, we evaluated ten candidate reference genes for the standardization of gene expression. Our results indicated that RPL14 and RPS15A constituted the most stable reference gene combination during the proliferation and differentiation of goat MuSCs. The process of skeletal muscle development is intricate and involves the regulation of a diverse array of genes. Accurate gene expression profiles are crucial for studying muscle development, making it essential to choose the right reference genes for real-time quantitative PCR (RT-qPCR). In the present study, eight candidate reference genes were identified from our previous transcriptome sequencing analysis of caprine skeletal muscle satellite cells (MuSCs), and two traditional reference genes (ACTB and GAPDH) were assessed. The quantitative levels of the candidate reference genes were determined through the RT-qPCR technique, while the stability of their expression was evaluated utilizing the GeNorm, NormFinder, BestKeeper, and RefFinder programs. Furthermore, the chosen reference genes were utilized for the normalization of the gene expression levels of PCNA and Myf5. It was determined that conventional reference genes, including ACTB and GAPDH, were not appropriate for normalizing target gene expression. Conversely, RPL14 and RPS15A, identified through RNA sequencing analysis, exhibited minimal variability and were identified as the optimal reference genes for normalizing gene expression during the proliferation and differentiation of goat MuSCs. Our research offers a validated panel of optimal reference genes for the detection of differentially expressed genes in goat muscle satellite cells using RT-qPCR. [ABSTRACT FROM AUTHOR]

OBJECTIVE To determine the accuracy of a continuous glucose monitoring system (CGMS) device by comparing glucose concentrations measured over time as determined by the CGMS to those of the chemistry analyzer (reference method). ANIMALS 7 healthy goats and 7 dairy calves. METHODS A randomized, crossover design with 3 treatments: control, hypoglycemia, and hyperglycemia. The CGMS device was applied to the neck. Hypoglycemia and hyperglycemia were induced by insulin and xylazine, respectively. Glucose concentrations were measured by the chemistry analyzer CGMS, point-of-care glucometer, and intensive care unit machine at 0 (before treatment), 2, 4, 6, 8, 10, and 12 hours. Agreement between the CGMS and the chemistry analyzer was determined by Bland-Altman plots. The analytical and clinical accuracy of the CGMS was determined using the International Organization for Standardization (ISO) 15197:2013 criteria and the Parkes error grid analysis. RESULTS In goats, the CGMS overestimated glucose concentrations during the hypoglycemic, normoglycemia, and hyperglycemia treatments. In calves, the CGMS underestimated glucose concentrations during the hypoglycemic treatment but overestimated glucose concentrations in normoglycemia and hyperglycemic treatments. The CGMS met the ISO clinical accuracy criteria for goats and calves, with > 99% of the glucose measurements in zones A and B of the Parkes grid. However, the CGMS did not meet the ISO 15197:2013 criteria for analytical accuracy. CLINICAL RELEVANCE The CGMS evaluated in our study only met the ISO 15197:2013 clinical accuracy criteria, not the analytical accuracy. Therefore, the device might be considered for clinical use. [ABSTRACT FROM AUTHOR]

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ORF virus (ORFV) causes contagious ecthyma ("ORF"), a disease of sheep and goats characterized by lesions ranging from vesicles and pustules to atypical papilloma-like and angiomatous lesions in the skin and mucosae. The authors investigated the molecular factors leading to the ORF-associated atypical tumor-like changes. Fifteen lambs, 15 kids, and an adult ram clinically affected by natural ORFV infection were enrolled in the study and examined by several methods. ORFV was detected by viral culture or real-time polymerase chain reaction (RT-PCR) in the lesioned tissues and in the blood of the clinically affected sheep and goats. Surprisingly, ORFV was also detected in the blood of healthy goats from an affected herd. Microscopically, they found a pseudo-papillomatous proliferation of the epithelium, while the dermis and lamina propria were expanded by a proliferating neovascular component that highly expressed the viral vascular endothelial growth factor (vVEGF) and its host receptor vascular endothelial growth factor receptor 2 (VEGFR2). Immunohistochemistry, immunofluorescence, and in situ hybridization for mRNA showed that epidermal growth factor receptor (EGFR) was expressed in the fibrovascular component, in the infiltrating CD163+ macrophages, and in the basal stratum of the epidermis. Confocal immunofluorescence microscopy demonstrated that CD163+ macrophages were associated with VEGF and VEGFR2. Finally, they found by quantitative RT-PCR the overexpression of the interleukin-6 and VEGFR2 genes in the lesioned tissues. These findings suggest that ORFV activates an inflammatory reaction characterized by CD163+ macrophages expressing EGFR and VEGFR2, which might play an oncogenic role through synergistic action with vVEGF signaling. [ABSTRACT FROM AUTHOR]

Objectives: The study aimed to detect Neospora caninum by nested PCR (nPCR) in aborted fetuses of cattle, sheep, and goats in Bangladesh. Materials and Methods: The head portion of each aborted fetus (111) was dissected at each sampling site and transferred to the laboratory in an ice box. Data on risk factors associated with N. caninum infection were simultaneously collected. Deoxyribonucleic acid was extracted from brain tissue to perform nPCR targeting the internal transcribed spacer 1 (ITS1) ribosomal DNA (rDNA) gene of N. caninum and sequencing was performed from the representative positive samples. Results: By nPCR, N. caninum was found in 16.0% of aborted fetuses of cattle, followed by sheep (14.81%) and goats (11.78%). The highest prevalence was found in aborted fetuses of animals during the second trimester (27.78%) of pregnancy aged 2 to 4 years (18.75%). Obtained sequences showed they were completely matched with N. caninum ITS1 rDNA gene deposited in GenBank. Univariate analysis demonstrated that pregnancy stages (trimesters), abortion history of the animals, and access to dogs in animal farms were significantly (p = 0.05) correlated with N. caninum infection. Conclusion: This study represents the first investigation into the molecular detection, phylogenetic characterization, and analysis of risk factors associated with N. caninum in livestock in Bangladesh. According to the research findings, N. caninum infection may have a role in abortion cases and the ensuing financial losses in the nation's livestock industry. [ABSTRACT FROM AUTHOR]

This study aims to describe socio-economic aspects, farming practices and production limitations of goats by surveying 53 farms holding 5507 head in the regions of Tataouine-Tunisia. It was revealed that goats represented a major source of income for most of the producers (66.1%). Approximately 35.8% of the farmers had not received formal education, with the majority (56.6%) having been engaged in goat breeding for over 10 years. Additionally, 50.9% of stockholders relied on technical knowledge for animal breeding. The investigated herds were mostly (97.4%) comprised of local goat breeds. The most common management system was semi-intensive (49.1%) based on grazing in communal rangelands and supplementation with agricultural by-products and concentrates. The fertility, fecundity and prolificacy rates were 91.3 ± 8.3, 109.8 ± 19.5 and 127.0 ± 15.4%, respectively. The main reasons for keeping goats were their major contribution to the income by the sale of male kids and the provision of milk for home consumption. Sale of male kids at an age of 7.7 months was practiced mostly via intermediaries (94.3%) and rarely directly to butchers and markets (5.7%). The total amount of milk produced on each farm was on average 15.1 l/day, and more than half of farmers (54.7%) consumed goat milk as part of their daily diet. The majority (71.7%) of respondents locally processed milk into Leben, Rayeb and cheese because the goat milk market faced constraints in the Tataouine regions, mainly in terms insufficient milk production and low consumer acceptability. Goats contribute to the income source in rural zones of southern Tunisia, where their breeding is mainly for the purpose of meat sale and the provision of milk for home consumption. The development of goat milk market can represent an added value to this sector. [ABSTRACT FROM AUTHOR]

Simple Summary: Paratuberculosis (PTB) is a chronic disease that affects domestic and wild ruminants worldwide. This study was conducted in 12 diary caprine farms on the Canary Islands. The region counts with the fourth largest goat population in Spain and has "officially free" bovine tuberculosis status. Two sampling sessions were conducted, and 2774 serum samples were tested by an enzyme-linked immunosorbent assay. In the first session, a prevalence of 18.4% was obtained, varying from 2.5% up to 61.1%. In the second session, the effect of PTB vaccination was evaluated and both non-vaccinated (nV) and vaccinated (V) were included. Variable tendencies in antibody development were registered in farms with different initial seroprevalences. In farms in which up to 10% of the animals were positive, more adult goats had antibodies against PTB after vaccination. In farms with more than 10% of ELISA-positive animals, a heterogeneous response to vaccination was reported. We observed that in farms with higher initial prevalence, fewer goats that were V developed antibodies. Our work characterizes the caprine PTB situation on the Canary Islands and gives new insights on the effect of farm prevalence on the immune response to PTB vaccination, although further studies on a greater scale are needed. Paratuberculosis (PTB), caused by Mycobacterium avium subspecies paratuberculosis (MAP), is a chronic disease with economic impact on ruminant farming worldwide. The Canary Islands count with the fourth largest goat population in Spain and are "officially free" of bovine tuberculosis. Twelve farms were included with 2774 serum samples tested by an enzyme-linked immunosorbent assay (ELISA) for detection of anti-MAP antibodies in two sessions. In the first session, an overall apparent prevalence of 18.4% (2.5% up to 61.1%) was obtained. Farms with prevalences (0–10%], (10–20%] and >20% were identified, with differences in seroconversion in the same prevalence group between farms and age ranges. Non-vaccinated (nV) and vaccinated (V) animals were included in the second sampling session. Higher levels of antibodies were detected in V animals older than 12 months, with considerable variations between age ranges and farms. Our results describe the current PTB status of the Canary Islands' goat farming. Furthermore, new insights on the effect of the farm prevalence on seroconversion in V animals are provided, although further studies are needed to evaluate the multiple factors affecting the immune response to anti-MAP vaccination. [ABSTRACT FROM AUTHOR]

Background: Theileriosis is a tick-borne disease caused by protozoon species in the Theileria genus of the Theileriidae family. The biochemical changes induced by infection are considered to be an important understanding of the pathophysiology of caprine theileriosis. In this study, it was aimed to determine oxidative stress, thyroid hormones, trace elements, and biochemical parameters in theileriosis infection. Materials and Methods: A sample of 14 goat was used for this purpose, of which 7 were healthy and 7 were infected with Theileria ovis. Theileria infection was diagnosed from the polymerase chain reaction (PCR). Sera from blood samples was tested for total antioxidant capacity (TAC), total oxidant capacity (TOC), oxidative stress index (OSI), alanine aminotransferase (ALT), aspartate transaminase (AST), gamma glutamyl transferase (GGT), total protein, albumin, triglyceride, cholesterol, high density lipoprotein (HDL), low density lipoprotein (LDL), urea, blood urea nitrogen (BUN), creatinine, triiodothyronine (T3), thyroxine (T4), copper (Cu), zinc (Zn), cobalt (Co), manganese (Mn), selenium (Se), iron (Fe). Result: TOC, OSI, AST, ALT and GGT values were higher in the patient group than in the healthy group (P < 0.05). On the other hand, there were decreases in TAC, T3, T4, total protein, albumin, creatinine, Cu, Zn, Se, and Co values (P < 0.05). However, there was not found to be a statistical difference between the healthy and patient groups in terms of triglyceride, cholesterol, HDL, LDL, urea, BUN, Mn, and Fe values (P > 0.05). Conclusions: It can be stated that oxidative stress is a complication of caprine theileriosis and it may be accompanied with hypothyroidism and deficits in trace minerals. [ABSTRACT FROM AUTHOR]

Listeriosis is highly prevalent in the animal farming industry, with Listeria monocytogenes as the causative pathogen. To identify potential therapeutic targets for LM infection, we investigated the mechanisms of LM infection in goat uteri. We inoculated a group of goats with LM via jugular vein injection, isolated and raised them, and subsequently collected sterile samples of their uterine tissue after they exhibited clinical symptoms of LM infection. We used Giemsa staining, immunohistochemical staining, real-time qPCR, and Western blotting as experimental methods.First, we investigated the mechanism of Listeria monocytogenes (LM) infection in the goat uterus by examining the expression levels of listeriolysin O, E-cadherin, and tyrosine kinase c-Met in the uterus. Furthermore, we investigated the impact of LM infection on uterine autophagy and cell apoptosis. The results indicate that the injection of LM into the goats' jugular veins leads to LM infection in the goats' uteri. During LM survival inside the goat uterine cells, there is a significant increase in the expression levels of LLO, E-cadherin, and c-Met in the host uterine tissue. This suggests that LM may potentially infect goat uteri through the InlA/E-cadherin and InlB/c-Met pathways. Furthermore, LM infection increases the levels of apoptosis and autophagy in goat uteri. Apoptosis genes Bcl-2 and Bax, as well as autophagyrelated genes LC3B, PINK1, and Parkin, exhibit varying degrees of changes in localization and expression in goat uteri, mediating the occurrence of apoptotic and autophagic responses. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs) are important regulators of gene expression and are involved in bacterial pathogenesis and host–pathogen interactions. In this study, we investigated the function of miRNAs in the regulation of host responses to Pasteurella multocida infection. Using next-generation sequencing, we analyzed miRNA expression pattern and identified differentially expressed miRNAs in Pasteurella multocida-infected goat lungs. In addition, we investigated the function of differentially expressed miRNAs andtheir targeted signaling pathways in bacterial infection processes. The results showed that Pasteurella multocida infection led to 69 significantly differentially expressed miRNAs, including 28 known annotated miRNAs with miR-497-3p showing the most significant difference. Gene target prediction and functional enrichment analyses showed that the target genes were mainly involved in cell proliferation, regulation of the cellular metabolic process, positive regulation of cellular process, cellular senescence, PI3K-Akt signaling pathway, FoxO signaling pathway and infection-related pathways. In conclusion, these data provide a new perspective on the roles of miRNAs in Pasteurella multocida infection. [ABSTRACT FROM AUTHOR]

Objectives: The biuret method is frequently used to determine serum total protein. On the other hand refractometer, a quicker and less expensive option, is available to determine serum total protein. However, there is no study conducted in Ethiopia to compare serum total protein measurement in veterinary settings. Therefore, this study was conducted to compare the serum total protein concentration measurement in cattle and goats obtained by the biuret method and refractometer. Results: Serum samples from 60 cattle and 60 goats were assayed by both methods and data were analyzed with a paired t-test, Pearson's correlation, and Bland-Altman plots. There was a strong positive correlation between the total protein values determined with the refractometer and the biuret method in cattle (r = 0.93) and goats (r = 0.97). There were no significant differences (p > 0.05) in the protein values measured with the refractometer and those evaluated with the biuret method in both species. Bland-Altman plots showed that biases indicating the analytic and user error were 8.33% in both species which is below the acceptable total error (< 10%). Thus, refractometer can be used in place of biuret method since it is valid enough to measure serum total protein in cattle and goats. [ABSTRACT FROM AUTHOR]

Introduction: Gut microbiota are associated with the health and performance of ruminant species, and they are affected by altitude, host genetics, and sex. However, there has been little research on comparing the fecal microbiota of indigenous small ruminants such as sheep and goats in Guizhou province, China. In the present study, we revealed the effect of altitude, genetics, and sex on fecal microbiota profiles and enterotypes in indigenous small ruminants of Guizhou province, China. Methods: Fecal samples were collected from Hei and Qianbei Ma goats and Weining sheep in the Chinese province of Guizhou. 16S rRNA gene sequencing targeting the V3-V4 region was performed using the Illumina MiSeq platform. Sequences were processed using QIIME2, and the qualified sequences were processed using the plugin DADA2 to generate amplicon sequence variants (ASVs). The statistical analysis was performed using R studio. Results: The fecal microbial profile was found to vary by herd (influenced by genetics/altitude) and sex. All samples were categorized into two enterotypes. The first enterotype is dominated by UCG-005, and the second enterotype is dominated by the Christensenellaceae_R-7_group, which may be highly driven by the host's genetics (breed). The predicted functional profiles of the fecal microbiota were also assigned to two clusters that corresponded exactly to the enterotypes. Cluster 1 of the functional profiling was characterized by biosynthesis pathways, and cluster 2 was characterized by energy metabolism pathways. Discussion: Our findings may provide new insights into the fecal microbial community and enterotypes in small ruminants by herds, offering clues for understanding the mechanisms by which the fecal microbiota contribute to divergent host phenotypes in indigenous small ruminants in Guizhou. [ABSTRACT FROM AUTHOR]

Simple Summary: Hemp (Cannabis sativa L.), traditionally used for textile fiber, has gained interest as animal feed due to its nutritional properties. This study investigated the impact of supplementing goats' diets with hemp hay on the milk yield, composition, and fatty acid profile of 20 dairy goats. The milk yield was higher in the hemp group, though the milk composition remained unchanged. The fatty acid profile of the hemp group's milk showed lower levels of certain saturated fatty acids (C11:0, C12:0, C13:0, C14:0, C15:0, and C17:0) and higher levels of C16:0 and C18:0. Additionally, the hemp group's milk had lower levels of some polyunsaturated fatty acids (C18:2 n6 and C20:4) but higher C20:5 n3 levels. Hemp hay may represent an advantageous alternative as feed supplementation in grazing goats' diet. Also, in terms of eco-sustainability and the efficiency of agricultural production, the hemp hay is a by-product of the supply chain, which may represent an additional benefit of its use in ruminant nutrition. Hemp (Cannabis sativa L.) is a cosmopolitan annual herbaceous plant used in the past as a source of textile fiber. Currently, hemp is receiving great interest as animal feed due to its chemical and nutritional properties. The aim of this study was to explore the effects of supplementing goats' diets with hemp hay on the milk yield, chemical composition, and fatty acid profile. Twenty multiparous goats, immediately after kidding, were divided into two homogenous groups (C: control vs. H: hemp); the goats had free access to the pasture, and both groups received a supplement of 500 g/head/day of a barley and corn meal mixture (50/50). In addition, group H was given 250 g/head/day of hemp hay while group C received the same amount of alfalfa hay. The milk yield was measured daily, and milk samples were collected monthly 4 times to evaluate the milk composition and fatty acid profile. The milk yield was significantly (p < 0.05) higher in the experimental group, while no differences were found in the milk chemical composition. Concerning the fatty acid profile, the milk from group H was characterized by significantly lower concentrations of C11:0, C12:0, C13:0, C14:0, C15:0, and C17:0 and higher C16:0 and C18:0. Among the polyunsaturated FA, C18:2 n6 and C20:4 were significantly (p < 0.001) lower, and C20:5 n3 was significantly (p < 0.05) higher in the milk from group H than that from group C. The n6/n3, LA/ALA and AA/EPA ratios were significantly (p < 0.001) lower in the milk from group H, while the CLAs were unaffected by the treatment. [ABSTRACT FROM AUTHOR]