11,835 results on '"Goat"'
Search Results
2. Analysis of the zoonotic tick-borne encephalitis virus (TBEV) in raw milk and dairy products in mountain pastures of the Lombardy region, Italy.
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Scarazzato, Annalisa, Righi, Francesco, Sommariva, Marco Pietro, Bertoletti, Irene, Sala, Giovanni, Paterlini, Franco, Daminelli, Paolo, Finazzi, Guido, Losio, Marina-Nadia, and Pavoni, Enrico
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TICK-borne encephalitis viruses ,TICK-borne encephalitis ,GOAT milk ,GOATS ,CENTRAL nervous system ,RAW milk - Abstract
Over the last few decades, tick-borne encephalitis (TBE) has become a growing public health problem in Europe. The tick-borne encephalitis virus (TBEV) is a zoonotic virus that affects the central nervous system (CNS). TBEV has been detected in 27 European countries, and the rise in TBE cases is mainly due to environmental and ecological factors, and factors that increase the risk of human exposure to infected ticks. The infection via the alimentary route is the second most common means of TBEV transmission to humans. Raw milk from infected goats, sheep, or cows has been identified as a source of human food-borne infections. This study aims to gather new information on the prevalence of tick-borne encephalitis virus (TBEV) in raw goat's and cow's milk and related raw products in the Lombard Alps (Italy). This is important due to the close proximity of Lombardy to the Triveneto region, where TBE is endemic, and southern Switzerland, where numerous TBEVpositive mammals have been found. Throughout 2023, a passive monitoring plan was implemented on samples delivered for TBEV analyses from the Alpine pastures. In total, 248 specimens including raw milk, raw milk cheese, and butter were tested. This is the first monitoring of food at risk of TBEV transmission in a non-endemic region with evidence of TBEV circulation. Despite testing a wide range of dairy products, no sample tested positive for RNA-TBEV by real-time RT-PCR. Preliminary results suggest that raw milk and raw dairy products do not pose a significant risk of TBEV transmission to humans in the territory of Lombardy. [ABSTRACT FROM AUTHOR]
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- 2024
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3. The Goat Cytotoxic T Lymphocyte-Associated Antigen-4 Gene: mRNA Expression and Association Analysis of Insertion/Deletion Variants with the Risk of Brucellosis.
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Wang, Congliang, Liu, Xiaoyu, Ren, Zhaofei, Du, Xiaomin, Li, Na, Song, Xiaoyue, Wu, Weiwei, Qu, Lei, Zhu, Haijing, and Hua, Jinlian
- Abstract
The cytotoxic T lymphocyte-associated antigen-4 (CTLA4) gene, a member of the immunoglobulin superfamily, is crucial for maintaining immune homeostasis and preventing autoimmune diseases. Studies have shown that polymorphisms in the CTLA4 gene are linked to an increased risk of brucellosis in humans, but its association with brucellosis in goats remains unexplored. In this study, the tissue expression profile of CTLA4 in goats was investigated, and the correlation between InDel polymorphisms in the CTLA4 gene and susceptibility to brucellosis in goats was examined. The findings reveal the widespread expression of CTLA4 in goat tissues, particularly in the spleen and testes. The tested goat populations presented genotypes insertion/insertion (II), insertion/deletion (ID), and deletion/deletion (DD) at both the P1 and P2 loci, and an association analysis revealed significant differences in the distribution of genotypes and allele frequencies at the P1 and P2 loci of the CTLA4 gene between the Brucella goat case and the control groups (p < 0.05). Specifically, compared with the II genotype, the P1 and P2 loci were significantly associated with an elevated risk of brucellosis development in goats under both the codominant (ID/II) and dominant (ID + DD/II) models (P1, p = 0.042, p = 0.016; P2, p = 0.011, p = 0.014). Additionally, haplotype analysis indicated that haplotypes IP1DP2, DP1IP2, and DP1DP2 were significantly associated with an increased risk of brucellosis in goats compared to the reference haplotype IP1IP2 (p = 0.029, p = 0.012, p = 0.034). Importantly, the Lipopolysaccharide (LPS) stimulation of peripheral blood monocytes and/or macrophages from goats with the II, ID, and DD genotypes resulted in increased CTLA4 expression levels in the II genotype, leading to a robust LPS-induced inflammatory response. Through bioinformatic analysis, the observed effect of the InDel locus on Brucella pathogenesis risk in goats could be attributed to the differential binding of the transcription factors nuclear factor kappaB (NF-κB) and CCAAT/enhancer-binding protein α (C/EBPα). These findings offer potential insights for breeding strategies against brucellosis. [ABSTRACT FROM AUTHOR]
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- 2024
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4. Effects of nano-berberine and berberine loaded on green synthesized selenium nanoparticles on cryopreservation and in vitro fertilization of goat sperm.
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Piri, Mehrangiz, Mahdavi, Amir Hossein, Hajian, Mehdi, Nasr-Esfahani, Mohammad Hossein, Soltani, Leila, and Vash, Nima Tanhaei
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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 < 0.05). The most significant outcomes were observed with berberine loaded on Se-NPs and the combination of selenium nanoparticles with berberine nanoparticles. [ABSTRACT FROM AUTHOR]
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- 2024
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5. Colonization profiles of gut microbiota in goat kids from neonatal to weaning period.
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Yuqin Wu, Dunying Hou, Siyuan Zhan, Linjie Wang, Jiazhong Guo, Li Li, Hongping Zhang, Lili Niu, and Tao Zhong
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COLONIZATION (Ecology) ,AMINO acid metabolism ,GUT microbiome ,ANIMAL weaning ,BACTERIAL colonies - Abstract
Understanding the colonization and change patterns of gut microbiota is pivotal for comprehending host health. As a newly cultured breed, the studies on the gut microbiota of Tianfu goats remain limited. This study aimed to address this gap by analyzing the microbial composition and colonization patterns of fecal samples collected from goat kids from birth to weaning. Fecal samples were collected on days 0, 7, 14, 21, 28, 35, 42, 49, 53, 55, 57, and 64, and the changes and colonization patterns of microorganisms were analyzed through high-throughput 16S rRNA sequencing. The results showed that the abundance of fecal microbiota in goat kids gradually increased over time, followed by a decrease after weaning and stabilization, with reduced individual differences. The colonization of fecal microorganisms mainly presented three different stages: days 0-14, days 21-49, and days 53-64. During the suckling period, the relative abundance of Proteobacteria (72.34%) was the highest, followed by Firmicutes (21.66%). From 21 days old, the microbiota in goat kids gradually to be diverse, with Lachnospiraceae and Ruminococcaceae being dominant. During post-weaning, Ruminococcaceae (30.98-33.34%) was becoming prominence which helpful for cellulose decomposition. LEfSe analyzed three important time points (d0 vs. d7, d7 vs. d14, d49 vs. d53, LDA score > 4 and p < 0.05), 53 microbial communities with stage differences were identified. Functional prediction using PICRUSt revealed that differential microbial communities are mainly related to carbohydrate and amino acid metabolism pathways. Overall, this study addresses the intricate relationship between ages, diets, and microbiota compositions in Tianfu goat kids, and also offering insights into microorganisms-host interactions. [ABSTRACT FROM AUTHOR]
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- 2024
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6. The goat pan-genome reveals patterns of gene loss during domestication.
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Liu, Jiaxin, Shi, Yilong, Mo, Dongxin, Luo, Lingyun, Xu, Songsong, and Lv, Fenghua
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GOAT breeds , *GERMPLASM conservation , *GENETIC variation , *PAN-genome , *GOATS - 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. [ABSTRACT FROM AUTHOR]
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- 2024
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7. Environmental contamination of arsenic: pathway analysis through water-soil-feed-livestock in Nadia District (India) and potential human health risk.
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Yadav, Sushil Kumar, Dutta, Tapas Kumar, Chatterjee, Anupam, Dutta, Sneha, Mohammad, Asif, and Das, Arun Kumar
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FEED analysis ,GOATS ,GOAT milk ,WELLS ,CONCENTRATE feeds ,PLANT cell walls ,ERECTOR spinae muscles - Abstract
This study investigated arsenic (As) concentrations in diverse environmental components and their potential impact on the health risks faced by residents of the arsenic (As)-contaminated Nadia district in West Bengal, India. A random selection of 182 cattle and 255 goats from 40 livestock farmers in the district revealed that both animals and humans were naturally exposed to elevated arsenic levels through contaminated drinking water, foods, grasses, concentrate feeds, various fodder tree leaves, and other food/feed resources. The mean As concentration in roughages (483.18 µg/kg DM) was significantly higher (p < 0.001) than in tree leaves (391.53 µg/kg DM), and concentrate feed/ingredients (186.66 µg/kg DM). Pond water exhibited higher arsenic levels (106.11 µg/L) compared to shallow tube well water (47.96 µg/L) and deep tube well water/tap water (10.64 µg/L and 10.04 µg/L, respectively). The mean arsenic concentration in soils DM of fodder fields, crop fields, and grassland was 10.25, 10.58, and 10.20 mg/kg, respectively. It was observed that protein-rich feeds had lower levels of arsenic accumulation (p < 0.048), while fiber-rich feeds containing more cellulose, hemicellulose, and lignin had higher arsenic levels (p < 0.017). Goats consumed 73.46% more arsenic per kg body weight compared to dairy cows. Although chronic and sub-chronic arsenic exposure in the district did not typically manifest symptoms or visible signs in ruminant animals, concentrations in the hair and feces of both cattle and goats exceeded normal values. Cattle feces had significantly higher arsenic (410.43 µg/kg DM) levels (p < 0.001) than goat feces (227.00 µg/kg DM), and arsenic concentration in cattle hair (1917.74 µg/kg DM) was also significantly greater (p < 0.001) than goat hair (1435.74 µg/kg DM). Arsenic levels in milk samples from both species were below 10 µg/kg. Liver (356.02 µg/kg DM) and kidney (317.22 µg/kg DM) contained significantly higher (p < 0.001) levels of arsenic compared to muscle (204.23 µg/kg DM), and bone (161.98 µg/kg DM) in local meat-type adult male goats. The skin accumulated the highest amount of arsenic (576.24 µg/kg DM) among the non-edible parts of the goat carcass. The cumulative cancer risk value for adults was 4.96 × 10
−3 , exceeding the threshold value (1 × 10−6 ). This suggests a significant risk of cancer development for the population in arsenic-affected areas. Non-cancer risks (hazard indexes) were estimated at 11.01 for adults. Our observations revealed that the highest bioaccumulation of arsenic occurred in the hair of cows, and goats in the examined localities. The biotransformation factor (BTF) for hair was much higher compared to other excreted samples from both species. The calculated BTF followed the order: hair > feces > milk for cows and goats. Livestock farmers in Nadia district are advised to carefully select feed resources, prioritizing those high in crude protein and low in neutral detergent fiber, and they should provide drinking water from deep aquifers to ensure the safety of milk and meat for human consumption. [ABSTRACT FROM AUTHOR]- Published
- 2024
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8. Multiomic meta‐analysis suggests a correlation between steroid hormone‐related genes and litter size in goats.
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Liu, Cheng‐Li, Mou, Hui‐Long, Na, Ri‐Su, Wang, Xiao, Hu, Peng‐Fei, Ceccobelli, Simone, Huang, Yong‐Fu, and E, Guang‐Xin
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STEROID synthesis , *GENE expression , *GOAT breeds , *GENE regulatory networks , *PEARSON correlation (Statistics) , *OVARIAN follicle - Abstract
Litter size is a key indicator of production performance in livestock. However, its genetic basis in goats remains poorly understood. In this work, a genome‐wide selection sweep analysis (GWSA) on 100 published goat genomes with different litter rates was performed for the first time to identify candidate genes related to kidding rate. This analysis was combined with the public RNA‐sequencing data of ovary tissues (follicular phase) from high‐ and low‐yielding goats. A total of 2278 genes were identified by GWSA. Most of these genes were enriched in signaling pathways related to ovarian follicle development and hormone secretion. Moreover, 208 differentially expressed genes between groups were obtained from the ovaries of goats with different litter sizes. These genes were substantially enriched in the cholesterol and steroid synthesis signaling pathways. Meanwhile, the weighted gene co‐expression network was used to perform modular analysis of differentially expressed genes. The results showed that seven modules were reconstructed, of which one module showed a very strong correlation with litter size (r = −0.51 and p‐value <0.001). There were 51 genes in this module, and 39 hub genes were screened by Pearson's correlation coefficient between core genes > 0.4, correlation coefficient between module members > 0.80 and intra‐module connectivity ≥5. Finally, based on the results of GWSA and hub gene Venn analysis, seven key genes (ACSS2, HECW2, KDR, LHCGR, NAMPT, PTGFR and TFPI) were found to be associated with steroid synthesis and follicle growth development. This work contributes to understanding of the genetic basis of goat litter size and provides theoretical support for goat molecular breeding. [ABSTRACT FROM AUTHOR]
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- 2024
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9. ADAR1 Promotes Myogenic Proliferation and Differentiation of Goat Skeletal Muscle Satellite Cells.
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Zhao, Zihao, Xiao, Miao, Xu, Xiaoli, Song, Meijun, Dai, Dinghui, Zhan, Siyuan, Cao, Jiaxue, Guo, Jiazhong, Zhong, Tao, Wang, Linjie, Li, Li, and Zhang, Hongping
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ADENOSINE deaminase , *SATELLITE cells , *MUSCLE growth , *ANIMAL culture , *DOMESTIC animals , *PSOAS muscles , *SKELETAL muscle - Abstract
As one of the most important economic traits for domestic animal husbandry, skeletal muscle is regulated by an intricate molecular network. Adenosine deaminase acting on RNA (ADAR1) involves various physiological processes and diseases, such as innate immunity and the development of lung adenocarcinoma, breast cancer, gastric cancer, etc. However, its role in skeletal muscle growth requires further clarification. Here, we explored the functions of ADAR1 in the myogenic process of goat skeletal muscle satellite cells (MuSCs). The ADAR1 transcripts were noticeably enriched in goat visceral tissues compared to skeletal muscle. Additionally, its levels in slow oxidative muscles like the psoas major and minor muscles were higher than in the fast oxidative glycolytic and fast glycolytic muscles. Among the two common isoforms from ADAR1, p110 is more abundant than p150. Moreover, overexpressing ADAR1 enhanced the proliferation and myogenic differentiation of MuSCs. The mRNA-seq performed on MuSCs' knockdown of ADAR1 obtained 146 differentially expressed genes (DEGs), 87 upregulated and 59 downregulated. These DEGs were concentrated in muscle development and process pathways, such as the MAPK and cAMP signaling pathways. Furthermore, many DEGs as the key nodes defined by protein–protein interaction networks (PPI), including STAT3, MYH3/8, TGFβ2, and ACTN4, were closely related to the myogenic process. Finally, RNA immunoprecipitation combined with qPCR (RIP-qPCR) showed that ADAR1 binds to PAX7 and MyoD mRNA. This study indicates that ADAR1 promotes the myogenic development of goat MuSCs, which provides a useful scientific reference for further exploring the ADAR1-related regulatory networks underlying mammal skeletal muscle growth. [ABSTRACT FROM AUTHOR]
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- 2024
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10. Impact of semen cryopreservation season on in vitro embryo production of prepubertal goat oocytes.
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Ferrer‐Roda, Mònica, Catalán, Jaime, Izquierdo, Dolors, Oliveira, Maria Emilia Franco, and Paramio, Maria‐Teresa
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LIFE sciences , *ACROSOME reaction , *SPRING , *INTRACELLULAR calcium , *AUTUMN , *SPERMATOZOA , *FROZEN semen , *SEMEN - Abstract
Goat production is affected by reproductive seasonality. In vitro embryo production (IVEP) could overcome this effect. This study aimed to evaluate the impact of the season of semen collection/freezing on IVEP of prepubertal goat oocytes and on sperm quality and functionality concerning capacitation. Semen from six fertile bucks was collected, pooled and cryopreserved in spring and autumn and used for IVEP of oocytes recovered during the breeding season. Oocytes were IVM in TCM‐199 with hormones, EGF and cysteamine; fertilized and cultured in BO‐IVF and BO‐IVC media (IVF Bioscience, UK). Semen samples were assessed at 0 and 3 h after culture in capacitating (BO‐IVF, CAP) and non‐capacitating conditions for sperm plasma membrane and acrosome integrity, mitochondrial membrane potential (MMP), intracellular calcium and plasma membrane lipid disorder. Blastocyst production was higher with spring sperm compared to autumn (12.0% vs. 2.1%, respectively; p <.05). After CAP, acrosome reaction and intracellular calcium were higher (p <.05) in spring than autumn sperm. No differences were found in other sperm parameters. In conclusion, seasonal variations in the IVEP of prepubertal goats could be linked to differences in sperm ability to undergo in vitro capacitation. [ABSTRACT FROM AUTHOR]
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- 2024
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11. Parasite diversity among domestic goats of tropical monsoon climatic zone in India.
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Dittrich, Sonja Pia, Adithya, Sasi, Ajith, Y., Athira, N., Athira, K. S., Safeer, M. Saifudeen, Preena, P., Aishwarya, A., Athira, K., Nisha, A. R., Devi, Gopinath, Mäder, Mia, Beena, V., and Jacob, Siju Susan
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Among different climatic zones in India, the tropical monsoon region comprises a diverse ecosystem characterized by the endemic nature of several parasites including certain emerging and re-emerging vector-borne pathogens of humans, whereas a systematic investigation of the occurrence of different parasites among domestic goats in this area is not yet explored. The goal of the present study is to explore the parasite diversity focusing on molecular identification of vector-borne hemoparasites and its health impacts on domestic goats reared in the tropical monsoon climate zone of Kerala, India. Among 227 goats presented to the Teaching Veterinary Clinical Complex (TVCC) in the monsoon months of 2023, thirty animals were recruited for the study. The animals were screened for the presence of different hemoparasites (Anaplasma spp., Theileria spp., and Babesia spp.), ectoparasites (ticks, lice, and fleas), and gastrointestinal (GI) parasites (hookworms, threadworms, tapeworms, whipworms, and coccidia). The isolated hemoparasites were further characterized by sequencing and phylogenetic analysis. The correlation studies to elucidate the association between the occurrence of different parasites and clinical manifestations (hyperthermia, pallor of mucous membrane, circulatory failure, respiratory signs, neurological instability, and GI signs), blood picture (anemia, leukopenia, thrombocytopenia), demographics (sex and age), and treatment history (hemoparasitic therapy, ectoparasiticidal application, and prophylactic deworming) were conducted. The co-infection status of these parasites was also evaluated. A substantial portion of the goats in the study group was found to be affected by vector-borne hemoparasitic diseases and their arthropod vectors or GI parasites or both. This can be attributed to the constantly warm and humid climate of the region, which is favorable for the survival and growth of different life cycle stages of these parasites and vectors. A strategic parasitic disease surveillance-cum-control program is the need of the hour for ensuring climate resilience and profitable goat farming in the region. [ABSTRACT FROM AUTHOR]
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- 2024
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12. Unconventional Ingredients from the Industrial Oilseed By-Products in Dairy Goat Feeding: Effects on the Nutritional Quality of Milk and on Human Health.
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Tristan Asensi, Marta, Pagliai, Giuditta, Napoletano, Antonia, Lotti, Sofia, Dinu, Monica, Mannelli, Federica, Invernizzi, Guido, Sofi, Francesco, Colombini, Barbara, and Buccioni, Arianna
- Abstract
Oilseed by-products (Cynara cardunculus and Camelina sativa) (CCCS) are rich in bioactive compounds. This study aimed to evaluate the health effects of consuming yogurt made from goat milk fed with CCCS industrial residues in adults. In this randomized, crossover clinical trial, 20 healthy adults (14F; 37.7 ± 14.2 years) consumed either yogurt made from goat milk fed with CCCS or regular goat yogurt (C) daily for 1 month in each phase. Anthropometric parameters and blood samples were collected at the beginning and end of each phase. CCCS yogurt consumption resulted in a significant fat mass reduction (−1.8% and −1.1 kg) and fat-free mass increase (+1.5% and +0.9 kg). Regarding blood parameters, a non-significant decrease in triglycerides, total cholesterol, and LDL cholesterol was observed, particularly after the CCCS intervention. Moreover, creatinine levels exhibited an opposite trend (p = 0.023) after CCCS, decreasing in subjects aged ≤30 years (−0.03 mg/dL) and increasing in older subjects (+0.05 mg/dL). Regarding inflammatory parameters, a non-significant trend in increased IL-1ra levels was observed especially after CCCS yogurt consumption compared to the C yogurt (+56.9 vs. +19.1 pg/mL, respectively). The use of unconventional feed derived from oilseed by-products for dairy goat feeding may have potential possible beneficial effects on human health. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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13. Molecular characterization of Peste des petits ruminants virus (PPRV) in sheep and goats and risk factors associated with it in selected districts of Khyber Pakhtunkhwa-Pakistan.
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Awaz, Saira, Maqsood, Iram, Rahman, Hanif Ur, Ali, Muhammad Ijaz, Khan, Baitullah, Muhammad, Gul, Shah, Imtiaz Ali, Azam, Asima, Hidayat, Ayesha, and Nizam, Almas Faryal
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Background: Peste des Petits Ruminants (PPR) is an economically significant transboundary viral disease of sheep and goats caused by the PPRV virus, affecting annual losses of 1.45–2.10 billion US dollars globally. We designed the current study to evaluate the positive cases, molecular characterization, phylogenetic analysis, and risk factors correlated with the disease in various districts of Khyber Pakhtunkhwa, Pakistan, with the aim of contributing to these strategies. Methods and results: A total of 384 samples from three selected districts, i.e., Peshawar, Charsadda and Chitral (n = 128 each), were collected, and the virus was investigated by using the sandwich ELISA, while the N gene of the virus was used as a target for molecular detection via RT-PCR. The confirmed samples were then sequenced, and phylogenetic analysis was performed. According to our findings, the highest positive cases was found in district Peshawar (50.87%), followed by Charsadda and Chitral (24.56%), respectively, while risk factor analysis showed that certain categories, such as species, sex, and age less than two years, have higher risk (P < 0.05) in contrast to their respective categories. Furthermore, sequencing and phylogenetic analysis of representative samples showed that the PPRV strains in the current study clustered in lineage IV, which is circulating in the small ruminant population of Asia, the Middle East, and African countries. Comparative residue analysis highlighted the mutation by representing 242 variable sites out of 371 locations. Conclusions: PPRV has foremost importance in Pakistan because the virus was detected in a considerable number of samples, and most of which were sourced from subsidiary areas where veterinary services are not prioritized. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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14. Recombinant 3AB3 nonstructural protein-based indirect ELISA for detection of foot-and-mouth disease virus infection-elicited antibodies in goat.
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Rout, M., Pandey, L. K., Prusty, B. R., Samanta, R., Mohapatra, J. K., and Singh, R. P.
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Foot-and-mouth disease (FMD) is one of the most important animal diseases of economic significance globally. It is a highly infectious and contagious disease of cloven-hoofed animals including sheep and goat. For sero-diagnosis of FMD, recombinant antigen-based assays are considered as alternatives to conventional approaches such as the liquid phase blocking ELISA (LPBE). The early interventions towards control measures cannot be implemented unless the disease gets promptly diagnosed. It is relatively difficult to clinically diagnose FMD in goat due to the usual milder form or unapparent nature of symptoms. Under such situations where clinical samples are not available, demonstration of infection-specific FMD virus (FMDV) antibodies in serum sample may help identifying the animals exposed to the virus in retrospect. Antibody to 3AB nonstructural protein (NSP) has been considered to be the most reliable indicator for FMD diagnosis. The current study extended the earlier designed recombinant 3AB3 protein-based indirect ELISA originally validated on bovine serum samples to testing serum samples of goat. The performance of the indirect ELISA was validated using internationally accepted PrioCHECK
® FMDV NS kit. The overall diagnostic sensitivity (DSn) of the indirect ELISA was estimated to be 95.52% (619/648), while the diagnostic specificity (DSp) on naïve and vaccinated animals varied at 98.06% (557/568) and 94.15% (435/462), respectively. In India, where FMD is prevalent and the goat population is so high, this 'in-house' optimized assay can be considered to be an adjunct in sero-epidemiological investigation of FMD in goat. [ABSTRACT FROM AUTHOR]- Published
- 2024
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15. Melatonin facilitates oocyte growth in goats and mice through increased nutrient reserves and enhanced mitochondrial function.
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He, Changjiu, Wu, Hao, Liu, Ruiyan, Liao, Jianning, Wang, Xiaodong, Shi, Hongru, Hou, Fuqin, Reiter, Russel J., Liu, Guoshi, and Li, Xiang
- Abstract
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]
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- 2024
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16. Multi-omics analysis reveals the effects of host-rumen microbiota interactions on growth performance in a goat model.
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Juncai Chen, Xiaoli Zhang, Xuan Chang, Bingni Wei, Yan Fang, Shanshan Song, Daxiang Gong, Deli Huang, Yawang Sun, Xianwen Dong, Yongju Zhao, and Zhongquan Zhao
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SHORT-chain fatty acids ,CELL junctions ,GENE expression profiling ,GOATS ,MULTIOMICS - Abstract
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]
- Published
- 2024
- Full Text
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17. The African introgression of Murciano Granadina goats has a Moroccan origin and displays remarkable levels of inter‐individual variability.
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Petretto, E., Luigi‐Sierra, M. G., Vacca, G. M., Martínez, A., Delgado, J. V., Fernández Álvarez, J., Castelló, A., Pazzola, M., Jordana, J., Dettori, M. L., and Amills, M.
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LIVESTOCK breeding , *LIVESTOCK breeds , *GOATS , *SUDANESE , *SAMPLE size (Statistics) - Abstract
There is evidence that Murciano Granadina (MG), the most important caprine dairy breed in Spain, has been introgressed by African goats, but the precise geographic origin of such introgression has not been identified yet. Moreover, an accurate estimate of the magnitude of this African introgression is lacking, since current estimates are based on small numbers of sampled individuals. The aim of our work was to tackle these two issues by genotyping 500 MG goats with the Goat SNP50 BeadChip and comparing their genotypes with those of reference populations from Spain (Bermeya), France (Saanen), Morocco (Barcha, Draa, Ghazalia, Noire de Atlas, Nord, Moroccan), Egypt (Barki, Oasis, Saidi), Algeria (Arabia, Makatia, M'Zabite, Kabyle), Tunisia (Tunisian native breeds) and Sudan (Desert, Nilotic, Taggar). The population of 500 MG goats was subdivided into 10 datasets of 50 individuals to ensure that sample sizes of the target (MG) and reference populations are balanced. Performance of an unsupervised ADMIXTURE analysis demonstrated that MG goats have a North African ancestry, with an average proportion of 4.4 ± 2.3%. Next, we did a supervised ADMIXTURE analysis that revealed that the Moroccan genetic component reaches a proportion of 4.01 ± 3.9% in MG goats, while the Algerian (0.001 ± 0.001%), Egyptian (0.2 ± 0.1%), Sudanese (0.1 ± 0.1%) and Tunisian (0.3 ± 0.4%) components are present in extremely small proportions. The historical circumstances of this introgression event are currently unknown, but several plausible scenarios are outlined. Moreover, our results show considerable inter‐individual heterogeneity regarding the magnitude of the Moroccan introgression of MG goats (0%– 12% depending on the MG data set under analysis). This result implies that reliable estimates about the introgression of autochthonous livestock by exotic breeds can only be obtained by extensively sampling target populations. [ABSTRACT FROM AUTHOR]
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- 2024
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18. Caprine and Ovine Genomic Selection—Progress and Application.
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Zhang, Linyun, Duan, Yixin, Zhao, Shengnan, Xu, Naiyi, and Zhao, Yongju
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LIVESTOCK breeding , *ANIMAL breeds , *GOAT breeds , *LIVESTOCK breeds , *SHEEP farming - Abstract
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]
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- 2024
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19. Metabolomic and Transcriptomic Analyses Reveal the Potential Mechanisms of Dynamic Ovarian Development in Goats during Sexual Maturation.
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Wang, Yanyan, Chao, Tianle, Li, Qing, He, Peipei, Zhang, Lu, and Wang, Jianmin
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AMINO acid metabolism , *PI3K/AKT pathway , *STEROID hormones , *GENITALIA , *CYTOCHROME P-450 - Abstract
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]
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- 2024
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20. Fat Mass- and Obesity-Associated Protein (FTO) Promotes the Proliferation of Goat Skeletal Muscle Satellite Cells by Stabilizing DAG1 mRNA in an IGF2BP1-Related m 6 A Manner.
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Yao, Jiangzhen, Xu, Liang, Zhao, Zihao, Dai, Dinghui, Zhan, Siyuan, Cao, Jiaxue, Guo, Jiazhong, Zhong, Tao, Wang, Linjie, Li, Li, and Zhang, Hongping
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SOMATOMEDIN A , *MUSCLE growth , *SKELETAL muscle , *SATELLITE cells , *RNA methylation - Abstract
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]
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- 2024
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21. Challenge Dose Titration in a Mycobacterium bovis Infection Model in Goats.
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Liebler-Tenorio, Elisabeth M., Wedlich, Nadine, Figl, Julia, Köhler, Heike, Ulrich, Reiner, Schröder, Charlotte, Rissmann, Melanie, Grode, Leander, Kaufmann, Stefan H. E., and Menge, Christian
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MYCOBACTERIUM bovis , *MYCOBACTERIAL diseases , *VETERINARY vaccines , *VACCINE trials , *COMPUTED tomography , *GOAT diseases - Abstract
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]
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- 2024
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22. In vitro generation of trophoblast like stem cells from goat pluripotent stem cells.
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Na, Qin, Zhang, Siyu, Shao, Peng, Jia, Yu, Wang, Yanqiu, Wei, Mengyi, Chen, Yanglin, Chen, Chen, Zhao, Lixia, Wang, Zixin, Song, Yongli, Wu, Baojiang, Bao, Siqin, and Li, Xihe
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PLURIPOTENT stem cells , *TROPHOBLAST , *STEM cells , *GOATS , *INDUCED pluripotent stem cells - Abstract
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]
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- 2024
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23. Analysis of miRNAs in milk of four livestock species.
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Cendron, Filippo, Rosani, Umberto, Franzoi, Marco, Boselli, Carlo, Maggi, Flavio, De Marchi, Massimo, and Penasa, Mauro
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GENE expression , *GOATS , *SHEEP , *WATER buffalo , *NON-coding RNA , *DONKEYS , *LACTATION - 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 FROM AUTHOR]
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- 2024
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24. Effects of the FHL2 gene on the development of subcutaneous and intramuscular adipocytes in goats.
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Li, An, Wang, Youli, Wang, Yong, Xiong, Yan, Li, Yanyan, Liu, Wei, Zhu, Jiangjiang, and Lin, Yaqiu
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GENE expression , *REGULATOR genes , *ADIPOSE tissues , *GENTIAN violet , *FAT cells , *ADIPOGENESIS - 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 FROM AUTHOR]
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- 2024
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25. Transcriptome analysis reveals miRNA expression profiles in hypothalamus tissues during the sexual development of Jining grey goats.
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Li, Qing, Chao, Tianle, Wang, Yanyan, Xuan, Rong, Guo, Yanfei, He, Peipei, Zhang, Lu, and Wang, Jianmin
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GENE expression , *GUANINE nucleotide exchange factors , *PHYSIOLOGY , *HYPOTHALAMIC hormones , *PROTEIN-tyrosine kinases , *G protein coupled receptors - 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 < 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]
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- 2024
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26. Identification of Key Proteins Related to Cashmere Fiber Diameter by Integrated Proteomics and Bioinformatic Analyses in the Alpas and Alxa Goat Breeds.
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Zhang, Chongyan, Qin, Qing, Wang, Yichuan, Wang, Zhixin, and Liu, Zhihong
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GOAT breeds , *GOATS , *PROTEOMICS , *GOAT farming , *ELECTRON microscopy , *TANDEM mass spectrometry - Abstract
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]
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- 2024
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27. Determining orbital volumes of sheep, goats and gazelles via computed tomography.
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Koçyiğit, Ali, Demircioğlu, İsmail, Demiraslan, Yasin, Karaavci, Funda Aksünger, and Dayan, Mustafa Orhun
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GAZELLES , *COMPUTED tomography , *ANIMAL diseases , *SHEEP , *GOATS , *THYROID eye disease - Abstract
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]
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- 2024
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28. Seroepidemiology of bovine herpesvirus‐1 in goats in south‐western Iran.
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Pourmahdi Borujeni, Mahdi, Abbasi, Amir Hossein, Haji Hajikolaei, Mohammad Rahim, and Seifi Abad Shapouri, Masoud Reza
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NEUTRALIZATION tests , *CATTLE diseases , *ODDS ratio , *ANIMAL health , *LOGISTIC regression analysis - Abstract
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]
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- 2024
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29. Precise Insertion of AttB Sequences in Goat Genome Using Enhanced Prime Editor.
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Li, Aicong, Zhu, Zhenliang, Yang, Jing, Liu, Yayi, Zhang, Yong, and Liu, Jun
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DOUBLE-strand DNA breaks , *PRIME factors (Mathematics) , *GENOME editing , *TRANSGENIC animals , *BINDING sites - Abstract
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]
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- 2024
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30. Evaluation of Reference Gene Stability in Goat Skeletal Muscle Satellite Cells during Proliferation and Differentiation Phases.
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Zhan, Siyuan, Zhang, Lufei, Zhong, Tao, Wang, Linjie, Guo, Jiazhong, Cao, Jiaxue, Li, Li, and Zhang, Hongping
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GENE expression , *CHOICE (Psychology) , *SATELLITE cells , *GENE expression profiling , *MUSCLE growth - Abstract
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]
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- 2024
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31. Evaluation of a continuous glucose monitoring system in healthy dairy calves and adult goats.
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Brobst, Megan N., Abi-Nader, Bailey A., Blasczynski, Sarah J., and Chigerwe, Munashe
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CONTINUOUS glucose monitoring , *INTENSIVE care units , *BLAND-Altman plot , *GOATS , *HYPOGLYCEMIA , *HYPERGLYCEMIA - Abstract
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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- 2024
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32. Evaluation of Some Biochemical Parameters In Saanen Goats Naturally Infected With Mycoplasma agalactiae.
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ARSLAN, Sezai
- Abstract
Copyright of Journal of Agriculture & Nature / Kahramanmaraş Sütçü İmam Üniversitesi Tarım & Doğa Dergisi is the property of Kahramanmaras Sutcu Imam Universitesi and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
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- 2024
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33. ORF virus causes tumor-promoting inflammation in sheep and goats.
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Pintus, Davide, Cancedda, Maria G., Puggioni, Giantonella, Scivoli, Rosario, Rocchigiani, Angela M., Maestrale, Caterina, Coradduzza, Elisabetta, Bechere, Roberto, Silva-Flannery, Luciana, Bullock, Hannah A., Macciocu, Simona, Montesu, Maria A., Marras, Vincenzo, Dore, Simone, Ritter, Jana M., and Ligios, Ciriaco
- Subjects
VASCULAR endothelial growth factor receptors ,EPIDERMAL growth factor receptors ,VASCULAR endothelial growth factors ,POLYMERASE chain reaction ,SHEEP diseases - Abstract
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]
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- 2024
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34. Socio-economic aspects and farming practices of goats in Southern Tunisia.
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Chniter, Mohamed, Dhaoui, Amel, Houidheg, Achref, Atigui, Moufida, and Hammadi, Mohamed
- Abstract
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]
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- 2024
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35. Caprine Paratuberculosis Seroprevalence and Immune Response to Anti- Mycobacterium avium Subspecies paratuberculosis Vaccination on the Canary Islands, Spain.
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Stefanova, Elena Plamenova, Paz-Sánchez, Yania, Quesada-Canales, Óscar, Quintana-Montesdeoca, María del Pino, Espinosa de los Monteros, Antonio, Ramírez, Ana Sofía, Fernández, Antonio, and Andrada, Marisa
- Subjects
MYCOBACTERIUM avium paratuberculosis ,AGRICULTURE ,TUBERCULOSIS in cattle ,ENZYME-linked immunosorbent assay ,ECONOMIC impact of disease ,PARATUBERCULOSIS - Abstract
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]
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- 2024
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36. Evaluation of Oxidative Stress, Thyroid Hormones, Trace Elements and Some Biochemical Markers in Goats Naturally Infected with Theileria ovis.
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Caglar, Yunus Emre, Deger, Yeter, and Oguz, Bekir
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OXIDANT status ,HIGH density lipoproteins ,BLOOD urea nitrogen ,TRACE elements ,COPPER ,ALANINE aminotransferase - Abstract
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]
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- 2024
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37. Listeria monocytogenes: possible mechanism of infection of goat uterus and its effects on uterine autophagy and cell apoptosis.
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Hailong Hong, Yunhai Hu, Siyuan Shi, Ben Liu, Wenya Zheng, Ruonan Bo, Zhongjie Xu, Yifan Wu, and Yu Cao
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LISTERIA monocytogenes ,CADHERINS ,UTERUS ,GOATS ,AUTOPHAGY ,APOPTOSIS ,PESTE des petits ruminants - Abstract
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]
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- 2024
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38. miRNA expression signatures induced by pasteurella multocida infection in goats lung.
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Xu, Feng, Zheng, Hao, Dong, Xia, Zhou, Ao, and Emu, Quzhe
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GENE expression , *PASTEURELLA multocida , *LUNGS , *LUNG infections , *IMMUNOSENESCENCE , *CELLULAR aging , *REGULATOR genes , *GOAT diseases - 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 FROM AUTHOR]
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- 2024
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39. Comparison of biuret and refractometery method for serum total protein measurements in cattle and goat.
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Mesa, Sileshi Mesele and Megerssa, Yoseph Cherinet
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PEARSON correlation (Statistics) , *BLOOD proteins , *BLAND-Altman plot , *REFRACTOMETERS , *CATTLE - 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 (< 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]
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- 2024
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40. Survey of the fecal microbiota of indigenous small ruminants living in different areas of Guizhou.
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Wei Guo, Tingmei Liu, Weiwei Wang, Yinshu Yu, Alves Neves, André Luis, Mi Zhou, and Xiang Chen
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GUT microbiome ,ENTEROTYPES ,GENE targeting ,ENERGY metabolism ,RUMINANTS - Abstract
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]
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- 2024
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41. The Effects of Hemp Hay (Canapa sativa L.) in the Diets of Grazing Goats on Milk Production and Fatty Acid Profile.
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Iommelli, Piera, Zicarelli, Fabio, Amato, Ruggero, Musco, Nadia, Sarubbi, Fiorella, Bailoni, Lucia, Lombardi, Pietro, Di Bennardo, Federica, Infascelli, Federico, and Tudisco, Raffaella
- Subjects
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SATURATED fatty acids , *UNSATURATED fatty acids , *TEXTILE fibers , *GOATS , *ALFALFA as feed - Abstract
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]
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- 2024
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42. Detection of caprine paratuberculosis (Johne's disease) in pre- and post-vaccinated herds: morphological diagnosis, lesion grading, and bacterial identification.
- Author
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Plamenova Stefanova, Elena, Sierra, Eva, Fernández, Antonio, Quesada-Canales, Oscar, Paz-Sánchez, Yania, Colom-Rivero, Ana, Espinosa de los Monteros, Antonio, Herráez, Pedro, Domínguez, Lucas, Bezos, Javier, Pérez-Sancho, Marta, Moreno, Inmaculada, Risalde, María A., and Andrada, Marisa
- Subjects
MYCOBACTERIUM avium paratuberculosis ,PARATUBERCULOSIS ,CROHN'S disease ,DIAGNOSIS ,GOAT diseases ,DIAGNOSTIC use of polymerase chain reaction - Abstract
Samples from the mesenteric lymph nodes (MS LNs) and ileocecal valves (ICV) of 105 goats, comprising 61 non-vaccinated and 44 vaccinated against Mycobacterium avium subspecies paratuberculosis (MAP), were collected at slaughter from a farm with a confirmed history of paratuberculosis (PTB). These goats had subclinical infections. PTB-compatible lesions in the MS LNs, ICV lamina propria (LP), and Peyer's patches (PPs) were graded separately. Furthermore, the load of acid-fast bacilli was quantified using Ziehl-Neelsen staining (ZN), MAP antigens by immunohistochemistry (IHC), and MAP DNA by PCR targeting the IS900 sequence. Gross PTB-compatible lesions were found in 39% of the goats, with 31.72% vaccinated (V) and 68.29% non-vaccinated (nV). Histopathological lesions inducedMAP were observed in 58% of the animals, with 36.07% vaccinated and 63.93% non-vaccinated. The inclusion of histopathology as a diagnostic tool led to a 28% increase in diagnosed cases in MS LNs and 86.05% in ICV. Grade IV granulomas with central mineralization and necrosis were the most common lesions in MS LNs. In the ICV, mild granulomatous enteritis with multifocal foci of epithelioid macrophages was predominant, occurring more frequently in the PPs than in the LP. Furthermore, statistical differences in the presence of histopathological lesions between vaccinated and non-vaccinated goats were noted in MS LNs, ICV LPs, and ICV PPs. Non- vaccinated animals showed higher positivity rates in ZN, IHC, and PCR tests, underscoring the benefits of anti-MAP vaccination in reducing PTB lesions and bacterial load in target organs. Our findings emphasize the necessity of integrating gross and histopathological assessments with various laboratory techniques for accurate morphological and etiological diagnosis of PTB in both vaccinated and non-vaccinated goats with subclinical disease. However, further studies are required to refine sampling protocols for subclinical PTB in goats to enhance the consistency of diagnostic tools. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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43. Exploring the reticulo-ruminal motility pattern in goats through medical barium meal imaging technology.
- Author
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Yang Song, Xinyi Lan, Lei Liu, Fachun Wan, Weijun Shen, and Zuo Wang
- Subjects
GASTROINTESTINAL motility ,BARIUM ,BODY weight ,GOATS ,CENTROID - Abstract
The motility pattern of the reticulo-rumen is a key factor affecting feed intake, rumen digesta residence time, and rumen fermentation. However, it is difficult to study reticulo-ruminal motility using general methods owing to the complexity of the reticulo-ruminal structure. Thus, we aimed to develop a technique to demonstrate the reticulo-ruminal motility pattern in static goats. Six Xiangdong black goats (half bucks and half does, body weight 29.5 ± 1.0 kg) were used as model specimens. Reticulo-ruminal motility videos were obtained using medical barium meal imaging technology. Videos were then analyzed using image annotation and the centroid method. The results showed that reticulo-ruminal motility was divided into primary (stages I, II, III, and IV) and secondary contraction, and the movements of ruminal digesta depended on reticulo-ruminal motility. Our results indicated common motility between the ruminal dorsal sac and ruminal dorsal blind sac. We observed that stages I (3.92 vs. 3.21 s) (P < 0.01), II (4.81 vs. 4.23 s) (P < 0.01), and III (5.65 vs. 5.15 s) (P < 0.05); interval (53.79 vs. 50.95 s); secondary contraction time (10.5 vs. 10 s); and were longer, whereas stage IV appeared to be shorter in the bucks than in the does (7.83 vs. 14.67 s) (P < 0.01). The feasibility of using barium meal imaging technology for assessing reticulo-ruminal and digesta motility was verified in our study. We determined the duration of each stage of reticulo-ruminalmotility and collected data on the duration and interval of each stage of ruminal motility in goats. This research provides new insights for the study of gastrointestinal motility and lays a solid foundation for the study of artificial rumen. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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44. Exploring the genetic variability of the PRNP gene at codons 127, 142, 146, 154, 211, 222, and 240 in goats farmed in the Lombardy Region, Italy.
- Author
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Ferrari, Carlotta, Punturiero, Chiara, Milanesi, Raffaella, Delledonne, Andrea, Bagnato, Alessandro, and Strillacci, Maria G.
- 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 FROM AUTHOR]
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- 2024
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45. Identification and Full-Length Sequence Analysis of a Novel Recombinant Goat Astrovirus Genotype in Guangxi, China.
- Author
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Zhou, Yulu, Xu, Pengju, Huang, Yanhua, Wang, Jie, Cui, Chang, Wang, Yanglin, Luo, Yuhang, Wang, Xiaoling, Xie, Jiang, Li, Fengmei, Wei, Zuzhang, Chen, Ying, Ouyang, Kang, Qin, Yifeng, Pan, Yan, and Huang, Weijian
- Subjects
- *
WHOLE genome sequencing , *NUCLEOTIDE sequence , *SEQUENCE analysis , *AMINO acids , *ASTROVIRUSES - Abstract
Astroviruses are single-stranded, positive-sense RNA viruses capable of infecting humans as well as a wide range of mammalian and avian species, with a length of approximately 6.6–7.7 kb. In this study, 139 goat fecal samples collected from the Guangxi province were used for the RT-PCR detection, and two of these were positive for goat astrovirus, with a positivity rate of 1.44% (2/139). The complete genome sequence of an astrovirus strain and the partial genome sequence of a strain astrovirus, named GX WZ 2023 and GX HC 2023, were amplified and sequenced, and their sequence lengths were 6284 nt and 6213 nt, respectively. Among them, the capsid protein of goat astrovirus GX HC 2023 showed the highest amino acid identity of 95.9% with ovine astrovirus GX, which belonged to the MAstV-2 genotype. However, the closest relative of the GX WZ 2023 strain was found to be the caprine astrovirus Sichuan, with a nucleotide sequence identity of 76.8%. The ORF1ab nonstructural protein of this strain showed the highest amino acid identities of 89.2 and 95.8% with the ovine astrovirus S5.1 and caprine astrovirus G5.1 strains, respectively. However, its ORF2 capsid protein has 68.4% amino acid identity with the bovine astrovirus (BAstV) 16 2021 CHN strain and only 21.9–64% amino acid identity with all available strains of goat astrovirus. The GX WZ 2023 strain was recombined with the Chinese (BAstV 16 2021 CHN) and Japanese bovine strains (BAstV JPN 2015) in the ORF2 region. Therefore, the goat astrovirus GX WZ 2023 is proposed as a new member of the family goat astroviridae based on the species classification criteria of the International Committee on Taxonomy of Viruses. These findings enhance our understanding of the prevalence and genetic evolution of goat astrovirus and provide a scientific basis for future studies of these viruses in other animals. [ABSTRACT FROM AUTHOR]
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- 2024
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46. Thyroid dysgenesis associated with dwarfism, osteoporosis and spontaneous fractures in a goat.
- Author
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Pastor, Felipe M., Amaral, Camila I., de Melo Ocarino, Natália, Dias, Lucas A., da Silveira Xavier, Andressa B., Castelo Branco, Eloá C., and Serakides, Rogéria
- Subjects
SPINAL cord compression ,SPONTANEOUS fractures ,CEREBRAL cortical thinning ,SPINAL canal ,METABOLIC bone disorders ,THYROID gland - Abstract
An 11-month-old female Saanen goat, weighing 12.7 kg, was taken to the Veterinary Hospital of the Federal University of Minas Gerais because of sternal recumbency. On clinical examination, the animal was much smaller than expected and had hair similar to that of puppies and areas of hyperpigmentation on the head and dorsocervical and dorsothoracic cranial regions. Radiographic examination revealed fractures in both femurs and severe generalized osteoporosis. Given the unfavourable prognosis, the animal was euthanized. Necropsy revealed generalized pallor, muscular atrophy of the pelvic limbs and little reserve of subcutaneous adipose tissue. Both femurs had complete and closed diaphyseal fractures. The second lumbar vertebra was severely reduced in length as a result of a fracture, with dorsal displacement of the vertebral body towards the vertebral canal and compression of the spinal cord. Long bones and vertebrae had severe cortical thinning, enlargement of the medullary canal and reduced resistance. The thyroid gland was not in its normal anatomical location. A pale red nodule (1.0 × 0.4 cm) in the serosa of the middle third of the trachea, close to the thoracic entrance, was confirmed as ectopic thyroid tissue. Microscopically, the bones had evidence of growth arrest and severe osteoporosis. The ectopic thyroid nodule was hyperplastic with severe hypertrophy of follicular cells. The spinal cord was compressed by vertebral fractures and had focally extensive and severe myelomalacia. Based on the pathological features, the case was diagnosed as thyroid dysgenesis characterized by eutopic thyroid agenesis and ectopic thyroid tissue, associated with interruption of bone growth with dwarfism, osteoporosis and spontaneous secondary fractures with compression of the lumbar spinal cord. [ABSTRACT FROM AUTHOR]
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- 2024
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47. Malignant skin neoplasms in goats in Sicily, Italy: clinical, virological and pathological investigations.
- Author
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Mignacca, Sebastian A., Agnello, Stefano, Castiglione, Silvana, Guercio, Annalisa, Purpari, Giuseppa, and Capucchio, Maria Teresa
- Subjects
SKIN tumors ,SQUAMOUS cell carcinoma ,POLYMERASE chain reaction ,NASAL polyps ,VIRUS diseases - Abstract
Neoplasms in small ruminants are considered uncommon and their reported incidence is variable. The aims of this investigation were to characterize malignant skin neoplasms in adult goats reared in Sicily, Italy, and to evaluate potential correlations between gross and histopathology features of the tumours and signalment, tumour location and/or viral infections. A total of 75 malignant skin masses were examined. In selected animals with perineal masses (n = 28) virological and serological investigations on tissues and blood were also conducted. According to the histological features, the lesions were classified as 67 squamous cell carcinomas (SCCs) (of which 65 were located in the perineum), six melanomas and two fibrosarcomas. In three cases, neoplasms at the base of the horn were associated with nasal polyps. Among the selected perineal SCCs, papillomaviruses (PVs), caprine herpesvirus 1 and parapoxvirus were not detected on polymerase chain reaction or on serological examination. However, further investigation on a larger sample size is required to evaluate the potential role of PVs in the pathogenesis of skin tumours in goats. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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48. Mutational and Evolutionary Analysis of Interleukin-2 Gene in Pakistani Goat Breeds.
- Author
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Hussain, Tanveer, Wajid, Abdul, Khan, Jabbar, Nadeem, Asif, Hussain, Misbah, ul-Ain, Qurat, and Babar, Masroor
- Abstract
Interleukin 2 (IL-2) is produced by activated T cells and play important role in immune response against antigen. It acts in both autocrine and paracrine manner. It can stimulate B cells and various other phagocytic cells like monocytes, lymphokine-activated killer cells and natural killer cells. Acting in autocrine fashion, IL-2 protein plays a crucial role in proliferation of T cells. IL-2 triggers the release of pro and anti- inflammatory cytokines by activating several pathways. In present study, exon 1 of IL-2 gene of four local Pakistani breeds (Dera Din Panah, Beetal, Nachi and Kamori) was amplified by using reported ovine IL-2 primers. Amplified products of 4 breeds of goat were bidirectionally sequenced to decipher polymorphisms. Only a single substitution (T→A) was found in non-coding region of IL-2 gene. Comparison of IL-2 gene sequence of all four breeds with other goat breeds showed high similarity in sequence. Phylogenetic analysis of our local breeds with other mammals showed that IL-2 was highly variable. This high substitution rate could be due to changed selective pressure. These rapid changes may also lead to the changes in the functions of immune system. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
49. A Simple and Versatile Method for Ex Vivo Monitoring of Goat Vaginal Mucosa Transduction by Viral Vector Vaccines.
- Author
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Minesso, Sergio, Odigie, Amienwanlen Eugene, Franceschi, Valentina, Cotti, Camilla, Cavirani, Sandro, Tempesta, Maria, and Donofrio, Gaetano
- Subjects
GENETIC vectors ,WESTERN immunoblotting ,GENE expression ,VIRAL vaccines ,GOAT diseases - Abstract
Goat may represent a valid large animal model for human pathogens and new vaccines testing. Appropriate vaccine administration is a critical component of a successful immunization program. The wrong route of administration may reduce the efficacy of the vaccine, whereas the proper administration strategy can enhance it. Viral vectors have been employed successfully for goat and sheep immunization; however, no data concerning the vaginal route are available. A viral vector's ability to transduce the site of inoculation is of primary interest. In this study, a fast and reliable ex vivo assay for testing the transduction capability of an Ad5-based vector when intravaginally administered was developed. An Ad5 vector delivering an expression cassette with a bicistronic reporter gene, Ad5-CMV-turboGFP-IRES-Luc2, was constructed. We demonstrated Ad5-CMV-turboGFP-IRES-Luc2's ability to transduce caprine vaginal mucosa by ex vivo bioluminescent imaging (BLI) employing a simple CCD camera apparatus for chemiluminescence western immunoblotting. These data, though simple, provide valuable insights into developing a vaginal immunization strategy using a viral vector-based vaccine to protect against pathogens causing genital diseases. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
50. Identification of genomic regions under positive selection that control the type traits in different goat breeds.
- Author
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Mohammadi, H.
- Abstract
Introduction: Molecular markers that reveal polymorphisms at the DNA level now play a key role in animal genetics. However, the selection of molecular markers is crucial depending on the purpose, viz. this depends on different molecular biology techniques and their effects. Over the last decade, interest in identifying genes or genomic regions targeted by selection has grown. Identifying selection signatures can provide valuable insights into the genes or genomic regions that are or have been under selection pressure, which in turn leads to a better understanding of genotype-phenotype relationships. Type characteristics are important for breed identification and classification and are also positively correlated with body weight. This study aimed to identify effective genes and genomic regions under positive selection signatures in different goat breeds using selection signature methods. For this purpose, FST and hapFLK analyses were performed using the genome-wide single nucleotide polymorphisms (SNPs). Materials and methods: In this research, the information from 728 goats of four different breeds was used to identify genomic regions associated with type traits. To determine the genotype of the samples, Illumina caprine Bead Chip 50K was used. The genomic information of goat breeds was extracted from the Figshare database. Quality control was performed using the Plink software. The markers or individuals were excluded from further study based on the following criteria: unknown chromosomal or physical location, call rate <0.95, missing genotype frequency >0.05, minor allele frequency (MAF) <0.05, and a P-value for Hardy-Weinberg equilibrium test less than 10
-3 . After quality control, 36861 SNPs from goat SNP chip 50K from 691 goats remained for further analysis. To identify the signatures of selection, two statistical methods of FST and hapFLK were used under the software packages FST and hapFLK, respectively. Candidate genes were identified using the Plink v1.9 software and the Illumina gene list in R by SNPs located in the highest FST and hapFLK values. In addition, the latest published version of the animal genome database was used to define QTLs associated with economically important traits at identified loci. The GeneCards (http://www.genecards.org) and UniProtKB (http://www.uniprot.org) databases were also used to interpret the function of the obtained genes. Results and discussion: The FST and hapFLK statistics were used to identify genomic regions subjected to positive selection associated with type traits in four goat breeds. Using the FST approach, we identified eight genomic regions on chromosomes 3, 4, 7, 13, 15, 18, 20, and 29. The identified candidate genes associated with type traits in these genomic regions included TGFBR3, CALCR, ACAD8, BCAR1, and ADAMTS6. Some of the genes located in the identified selection regions were directly and indirectly related to cell differentiation and proliferation, skeletal muscle growth and development, body length, calcium channel regulation, muscle fiber homeostasis, protein synthesis, and muscle cell size. Some of these genes in the selected regions were consistent with previous studies. The results of the reported QTLs in the selected regions and the bovine orthologous regions were QTLs located in the identified regions that were related to average daily gain, body weight, trunk width, and metabolic body weight. Furthermore, the results of the hapFLK statistics in this research led to the identification of five genomic regions on chromosomes 1, 5, 6, 13, and 30, and they were in the 99.9th percentile of all hapFLK values. The identified candidate genes associated with the type trait in these genomic regions included FNDC3B, STAB2, and CCNY. They were found to have different functions in fibroblast proliferation and bone cell differentiation. Conclusions: Various/different genes that emerged in studied regions can be considered candidates for selection based on their function. By the way, various genes found in these regions can be considered candidates for selection based on their function. Most of the selected genes were found to be consistent with some previous studies and to be involved in production traits. A survey of extracted QTLs also found that these QTLs are involved in some economically important traits in goats, such as average daily gain and body weight in yearlings. However, further association and functional studies are required to demonstrate the importance of the genes obtained from association analyses. Leveraging these findings can accelerate genetic progress in breeding programs and help understand the genetic mechanism that controls these traits. [ABSTRACT FROM AUTHOR]- Published
- 2024
- Full Text
- View/download PDF
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