Goat Tissues Research Articles

Identification and characterization of microRNAs in the pituitary of pubescent goats

BackgroundPuberty is the period during a female mammal’s life when it enters estrus and ovulates for the first time; this indicates that a mammal is capable of reproduction. The onset of puberty is a complex and tightly coordinated biological event; it has been reported that microRNAs (miRNAs) are involved in regulating the initiation of puberty.MethodsWe performed miRNA sequencing on pituitary tissue from prepubescent and pubescent goats to investigate differences in miRNA expression during the onset of puberty in female goats. The target genes of these miRNAs were evaluated by GO enrichment and KEGG pathway analysis to identify critical pathways regulated by these miRNAs during puberty in goats. Finally, we selected four known miRNA and one novel miRNAs to evaluate expression patterns in two samples via qRT-PCR to validate the RNA-seq data.ResultsIn this study, 476 miRNAs were detected in goat pituitary tissue; 13 of these were specifically expressed in the pituitary of prepubescent goats, and 17 were unique to the pituitary of pubescent goats. Additionally, 73 novel miRNAs were predicted in these two libraries. 20 differentially expressed miRNAs were identified in this study. KEGG pathway enrichment analysis revealed that the differentially expressed miRNA target genes were enriched in pathways related to ovary development during puberty, including the GABAergic synapse, oxytocin signaling pathway, the cAMP signaling pathway, progesterone-mediated oocyte maturation. In this study, differential miRNA expression in the pituitary tissue of prepubescent and pubescent goats were identified and characterized.ConclusionThese results provide important information regarding the potential regulation of the onset of goat puberty by miRNAs, and contribute to the elucidation of miRNA regulated processes during maturation and reproduction.

MiR-27b regulates myogenic proliferation and differentiation by targeting Pax3 in goat

This study found that miR-27 is expressed in muscle and regulates muscle proliferation and differentiation. We explored the function and regulatory mechanism of miR-27b in goat muscle proliferation and differentiation. Compared with the Boer goat, higher expression of miR-27b was observed in all of the collected muscle tissues of Anhuai goat, excluding the kidney, whereas the opposite expression pattern was observed for Pax3, which showed lower expression in Anhuai goat. Expression of miR-27b decreased gradually during the proliferation of skeletal muscle satellite cells in Anhuai goat and increased during differentiation; however, the expression pattern of Pax3 was opposite. The regulatory activity of miR-27b demonstrated that miR-27b inhibited the proliferation of skeletal muscle satellite cells, but promoted their differentiation. Moreover, function research demonstrated that Pax3 negatively regulated myogenic differentiation of goat skeletal muscle satellite cells, but accelerated their proliferation. The results of a dual-luciferase reporter analysis showed that miR-27b directly targeted the 3’-untranslated regions of Pax3 mRNA, and western blot and immunofluorescence staining analyses showed that miR-27b inhibited expression of the Pax3 protein. In goats, miR-27b can regulate myogenic proliferation and differentiation by targeting Pax3.

Novel alternative splicing variants of <i>ACOX1</i> and their differential expression patterns in goats

Abstract. As the first and rate-limiting enzyme of the peroxisomal β-oxidation pathway, acyl-coenzyme A oxidase 1 (ACOX1), which is regulated by peroxisome proliferator-activated alfa (PPARα), is vital for fatty acid oxidation and deposition, especially in the lipid metabolism of very long-chain fatty acids. Alternative splicing events of ACOX1 have been detected in rodents, Nile tilapia, zebra fish and humans but not in goats. Herein, we identified a novel splice variant of the ACOX1 gene, which was designated as ACOX1-SV1, in addition to the complete transcript, ACOX1, in goats. The length of the ACOX1-SV1 coding sequence was 1983 bp, which presented a novel exon 2 variation owing to alternative 5′-splice site selection in exon 2 and partial intron 1, compared to that in ACOX1. The protein sequence analysis indicated that ACOX1-SV1 was conserved across different species. Reverse-transcription quantitative real-time polymerase chain reaction (RT-qPCR) analysis showed that these two isoforms were expressed spatially and differently in different tissue types. ACOX1 and ACOX1-SV1 were expressed at high levels in liver, spleen, brain and adipose tissue in kid goats, and they were abundantly expressed in the fat, liver and spleen of adults. Interestingly, whether in kids or in adults, in fat, the mRNA level of ACOX1 was considerably higher than that of ACOX1-SV1. In contrast, in the liver, the expression of ACOX1-SV1 was considerably higher than that of ACOX1. This differential expression patterns showed the existence of a tissue-dependent splice regulation. These novel findings for ACOX1 should provide new insights for further studies on the function of ACOX1 and its variants that should aid in the breeding of goats with improved meat quality.

Molecular Detection and Isolation of Mycoplasma capricolum subsp. capripneumoniae in Pashmina and Local Goats in Five Districts of Kashmir, India

Contagious caprine pleuropneumonia (CCPP) is caused by Mycoplasma capricolum subsp. caprip-neumoniae (Mccp), which belongs to Mycoplasma mycoides cluster, a group of closely related Mycoplasmas that are pathogenic to ruminants. The clinical lesions of CCPP are restricted to alveolar tissues of infected goats, which distinguish it from other respiratory diseases of small ruminants caused by members of the Mycoplasma mycoides cluster. The outbreaks of CCPP in Pashmina goats have been reported in cold arid regions of Kashmir province, such as Leh and Kargil districts and northern parts of the Kashmir valley. In the current study, a total of 55 lung samples and 70 nasal swabs were collected from clinically sick goats showing respiratory symptoms prior to death and processed for Mccp isolation and identification. The Mccp was detected in 33 (60.0%) and 24 (34.2%) samples, respectively by PCR. All the PCR positive samples for Mccp were further processed for isolation but only five isolates were obtained due to their fastidious nature which were confirmed by typical colony morphology and DNA sequencing of 548 bp 16SrRNA gene fragment. The five isolates were isolated from lung samples of Pashmina goats of Leh district. The results of the current study reveal that Pashmina goats of Ladakh region are highly vulnerable to CCPP and is the major cause of mortality and morbidity among them particularly during the winter season. Hence, there is an immediate need to isolate and characterise the organism for the development of autogenous vaccine to combat this economically important disease of goats. This communication records the first ever report on molecular detection and isolation of Mccp from Pashmina goats in India and is the first step towards development of vaccine against CCPP.

Fatty Acid Composition of Muscles Tissues of Longdong Goat (Black and White Cashmere)

This study examined fatty acid (FA) composition of muscle tissue of Longdong black goat and Longdong white Cashmere goat breed under reared systems. Approximately 86.31% of fatty acids were observed. Oleic acid was extensively the most abundant ranged from 22.98% to 26.02% of Longdong black and white Cashmere goat respectively. The mean amount of SFA and MUFA were about 21.46% to 34.82% respectively. PUFA were 30.02% of total FA composition. C20:4 were founded 12.14% in all muscles tissues. Compare to C20:5, C22:6 16.43%, 4.88% respectively. The amount of SFA: MUFA ratio was low in Longdong goat muscles tissues. PUFA and MUFA had higher amount female black muscles tissues. Females and males of both had great amount of C16:0, C18:0, ranged from 0.86% to 29.16%. C18:2 were found in 15.01%. The study found fatty acids composition in both Longdong goat breed (black and white cashmere) has some significant effect (P<0:05) in fatty acids proportions. The greater ratios of unsaturated fatty acids MUFA and PUFA high amount than saturated fatty acids SFA in goat muscle tissue and this indicate that Longdong goats breed are healthier and have the same type of fatty acids composition.

High-grain diet feeding altered the composition and functions of the rumen bacterial community and caused the damage to the laminar tissues of goats

In the current intensive production system, ruminants are often fed high-grain (HG) diets. However, this feeding pattern often causes rumen metabolic disorders and may further trigger laminitis, the exact mechanism is not clear. This study investigated the effect of HG diet feeding on fermentative and microbial changes in the rumen and on the expression of pro-inflammatory cytokines and matrix metalloproteinases (MMPs) in the lamellar tissue. In all, 12 male goats were fed a hay diet (0% grain; n=6) or an HG diet (56.5% grain; n=6). On day 50 of treatment, samples of blood, rumen content, and lamellar tissue of hooves of goats were collected. The data showed that compared with the hay group, HG-fed goats had lower (P<0.05) rumen pH but higher (P<0.05) total volatile fatty acids and lactate in the rumen and higher (P<0.05) lipopolysaccharide (LPS) levels in the rumen and blood. HG diet feeding altered the composition of rumen bacterial community, and correspondingly, the results suggested that their functions in the HG group were also altered. HG diet feeding increased (P<0.05) the expression of interleukin-1β, interleukin-6, tumour necrosis factor-α and MMP-2 mRNA in the lamellar tissues compared with the hay group. Correlation analysis indicated that the expression of pro-inflammatory cytokines were positively correlated with MMP-2 expression in lamellar tissues. Overall, these results revealed that HG feeding altered the patterns of rumen fermentation and the composition and functions of rumen bacterial community, and lead to higher levels of LPS in the peripheral blood, and further activated the inflammatory response in lamellar tissues, which may progress to the level of laminar damage.

MiR-487b-3p Suppresses the Proliferation and Differentiation of Myoblasts by Targeting IRS1 in Skeletal Muscle Myogenesis.

MicroRNAs are endogenous, small non-coding RNAs that can play critical gene-regulatory roles during skeletal muscle development and are highly conserved. miR-487b-3p is expressed in muscle, and the detailed mechanism by which it regulates myoblast proliferation and differentiation has not been explored. Here, we found that miR-487b-3p expression was significantly higher in goat muscle tissues than in other tissues and was higher in fetal goat muscle tissues than in mature goat tissues, suggesting that miR-487b-3p has an important effect on skeletal muscle myogenesis. Functional studies showed that miR-487b-3p overexpression significantly suppressed C2C12 myoblast proliferation and differentiation, which was accompanied by the down-regulation of functional genes related to proliferation (MyoD, Pax7 and PCNA) and differentiation (Myf5, MyoG and Mef2c), whereas the inhibition of miR-487b-3p accelerated C2C12 myoblast proliferation and differentiation and was accompanied by the up-regulation of functional genes. Using Target-Scan and David, we found that miR-487b-3p targeted the 3'-UTR of IRS1, an essential regulator in the PI3K/Akt and MAPK/Erk pathways. We then confirmed the targeting of IRS1 by miR-487b-3p using dual-luciferase assays, RT-qPCR and western blotting. Furthermore, IRS1 silencing markedly inhibited proliferation and differentiation in cultured C2C12 myoblasts, confirming the important role of IRS1 in myogenesis. These results reveal an IRS1-mediated regulatory link between miR-487b-3p and the PI3K/Akt and MAPK/Erk pathways during skeletal muscle myogenesis.

Probing natural substitute for formalin: Comparing honey, sugar, and jaggery syrup as fixatives.

Background:Since its discovery in 1859, formalin has been considered as the “gold standard” in tissue fixation. As formalin is highly toxic and carcinogenic, the quest for its substitute has started recently. Literature search reveals very sparse studies on natural substitute for formalin. Here, it is an attempt to explore eco-friendly, economical, and readily available natural substance for formalin substitute.Aim:The aim of our study was to evaluate the efficacy of natural fixatives such as honey, sugar, jaggery, and water in comparison to the standard fixative used like formalin.Materials and Methods:Fresh goat tissues (tongue) were fixed separately with buffered 10% formalin (positive control), honey, sugar syrup, jaggery syrup, and distilled water (negative control). 24 h fixation was done at room temperature followed by conventional processing and routine H and E staining. The stained sections were assessed for cytoplasmic and nuclear detail by three pathologists under light microscope and were graded accordingly.Results:The results showed statistically significant differences between jaggery with other natural fixatives for both nuclear details and cytoplasmic staining.Conclusion:The preservation of tissue by honey, sugar, and jaggery syrup was comparable to that of formalin. Among the three natural fixatives, jaggery syrup excelled. Hence, it can be considered as an equally effective formalin substitute.

Laparoscopic ovarian biopsy pick-up method for goats

Biopsy pick-up (BPU) has been considered a safe method to harvest ovarian fragments from live animals. However, no studies have been reported on the use of BPU to collect in vivo ovarian tissue in goats. The goals of this study were: (i) to test different biopsy needle sizes to collect ovarian tissue in situ using the BPU method (Experiment 1), and (ii) to study ovarian tissue features such as preantral follicle density, morphology, class distribution, and stromal cell density in ovarian fragments obtained in vivo through a laparoscopic BPU method (Experiment 2). In Experiment 1, goat ovaries (n = 20) were collected in a slaughterhouse and subjected to in situ BPU. Three needles (16, 18, and 20G) were tested. In Experiment 2, the most efficient biopsy needle from Experiment 1 was used to perform laparoscopic BPU in goats (n = 8). In Experiment 1, the recovery rate was greater (P < 0.05; range 50–62%) with 16G and 18G needles than the 20G (17%) needle. The mean weight of ovarian fragments collected by the 16G needle was greater (P < 0.05) than the 18G and the 20G needle. In Experiment 2, 62 biopsy attempts were performed and 52 ovarian fragments were collected (90% success rate). Overall, 2054 preantral follicles were recorded in 5882 histological sections analyzed. Mean preantral follicular density was 28.4 ± 1.3 follicles per cm2. The follicular density differed (P < 0.05) among animals and ovarian fragments within the same animal. The mean stromal cell density in the ovarian fragments was 37.1 ± 0.5 cells per 2500 μm2, and differed (P < 0.05) among animals. Moreover, preantral follicle density and stromal cell density were associated (P < 0.001). The percentage of morphologically normal follicles was 70.1 ± 1.2, and differed (P < 0.05) among animals. The majority (79%) of the morphologically normal follicles was classified as primordial follicles, and differed (P < 0.05) among animals and between ovaries. In summary, a laparoscopic BPU method has been developed to harvest ovarian tissue in vivo with a satisfactory success rate in goats. Furthermore, this study described for the first time that goat ovarian biopsy fragments have a high heterogeneity in follicular density, morphology, class distribution, and stromal cell density.

Distinct expression profile and histological distribution of NLRP3 inflammasome components in the tissues of Hainan black goat suggest a site-specific role in the inflammatory response.

The NOD-like receptor protein 3 (NLRP3) inflammasome comprised of NLRP3, ASC and caspase-1 plays an important role in the inflammatory and innate immune response. However, little is known about the expression pattern and histological distribution of these genes in goat. Here, we first cloned the fulllength cDNAs of the NLRP3, ASC and caspase-1 genes of Hainan black goat and produced their polyclonal antibodies. Tissue-specific expression and histological distribution of these genes were analysed. Phylogenetic analysis revealed that these three goat genes had high homology with Bos taurus genes and low homology with avian or fish genes. After immunisations with these recombinant Histagged proteins, the titres of antiserum were higher than 1:1024 and purified IgG was obtained. These three genes were expressed in all examined tissues, the mRNA expression level of NLRP3 and caspase-1 was most abundant in the spleen and mesenteric lymph nodes (MLNs), while ASC was primary expressed in the liver, spleen and kidney. The histological distribution of NLRP3, ASC and caspase-1 was detected in myocardial cells, hepatocytes, focal lymphocytes, bronchiolar epithelial cells, renal tubular epithelial cells, cortical neurons and endothelial cells of the germinal centres in the MLNs. These results will be helpful in further investigations into the function of the NLRP3 inflammasome and in elucidating its role in caprine inflammatory diseases.

Comparative analysis of tissue expression and methylation reveal the crucial hypoxia genes in hypoxia resistant animals

Tibetan goat and Tibetan sheep are species peculiar to Qinghai–Tibetan Plateau which is the highest plateau in the world, and have high hypoxia resistance to the extremely bad environment. Unlike monogastrics, how the genes change responding to hypoxia in ruminants remains unclear. In this study, three healthy animals of Tibetan sheep, Tibetan goat, Chuanzhong goat, and Small-tailed Han sheep were selected. The expression of factor inhibiting hypoxia-inducible factor 1 (FIH 1) hypoxia-inducible factor 1α (HIF-1α), HIF-3α, and erythropoietin (EPO) in various tissues, including heart, liver, lungs, kidney, muscle, and brain, was investigated. The EPO was observed highly expressed in all the tissues of Tibetan goats and Tibetan sheep compared with low-altitude animals, respectively, implied that higher expression of EPO may give the explanation for the hypoxia resistance of plateau animals. Besides, we also cloned the promoters of FIH-1, HIF-1α, HIF-3α, and EPO in goats and sheep, analyzed their core regions...

MiR-182 aids in receptive endometrium development in dairy goats by down-regulating PTN expression.

Increasing evidence has shown that miRNAs play important roles in endometrium development during the menstrual cycle in humans and many other animals. Our previous data indicated that miR-182 levels increase 15.55-fold and pleiotrophin (PTN) levels decrease 20.97-fold in the receptive endometrium (RE, D15) compared with the pre-receptive endometrium (PE, D5) in dairy goats. The present study shows that miR-182 is widely expressed in different tissues of dairy goats and that its expression levels are regulated by E2 and P4 in endometrial epithelium cells (EECs). We confirmed that PTN is a target of miR-182 and that miR-182 regulates the protein levels of AKT, Bcl-2, FAS, MAPK, Caspase-3 and SP1 in EECs. Furthermore, miR-182 up-regulates or maintains the expression levels of osteopontin (OPN), cyclooxygenase-2 (COX-2) and prolactin receptor (PRLR) in EECs, suggesting that miR-182 is an important regulatory factor in the construction of endometrial receptivity in dairy goats. In conclusion, miR-182 participates in the development of endometrial receptivity by down-regulating PTN and affecting the expression of select apoptosis-related genes and increasing or maintaining the expression levels of OPN, COX-2 and PRLR in the EECs of dairy goats.

Comparative evaluation of FAT, IS900 PCR and microscopy vis-a-vis histopathology for the detection of Mycobacterium avium subsp paratuberculosis infection in tissues of goats naturally died in herds endemic for Johne’s disease

Study investigated infection of Mycobacterium avium subspecies paratuberculosis (MAP) in paired tissues viz. ileum and mesenteric lymph nodes (MLN) from 74 goats using histopathology, acid fast staining of tissue sections, fluorescent antibody test (FAT), IS900 tissue PCR and culture. Histopathology of H&amp;E stained tissue sections revealed variable grades of lesions of Johne's disease (JD) in 37 (50.0%) ileae and 21 (28.4%) MLN. In AFB staining, 16 (21.6%) intestine and 10 (13.5%) MLN revealed presence of acid fast bacilli indistinguishable to MAP. Thirteen (17.5%) intestine and 9 (12.1%) MLN were positive in FAT. Eleven (14.8%) intestine and 7 (9.4%) MLN were positive by IS900 tissue PCR, whereas 7 (9.5%) intestine and 8 (10.8%) MLN were positive in culture. The study showed that sensitivity of histopathology (H&amp;E staining) was highest as compared to other routine tests for the diagnosis of MAP infection in tissues of goats died (irrespective of cause of deaths) during study period from goat herds endemic for Johne's disease. Lower sensitivity of FAT than histology was due to the strong cell mediated immunity exhibited by MAP. Though difference was not significant between sensitivities of FAT and PCR, FAT and AFB staining, and agreement was very good; however, results of the study suggested, that FAT may be a useful test to improve the diagnostic efficacy of caprine Johne's disease and more effective in the detection of grade I lesions as compared to culture and PCR. Study also showed that it was essential to collect both tissues (intestines and MLNs) for the diagnosis of MAP infection at necropsy.

Identification and expression of X-linked inhibitor of apoptosis protein during follicular development in goat ovary

X-linked inhibitor of apoptosis protein (XIAP), an endogenous of inhibitor of caspases, plays crucial roles in regulating ovarian granulosa cell apoptosis during follicular atresia. The aim of the present study was to determine the presence and localization of XIAP in the goat ovary and its expression level during follicular development. The full length cDNA of XIAP from goat ovary cells was cloned using reverse transcription PCR. A total of 497 amino acid residues were encoded by open reading frame and had high identity with homologous sequences from other mammals. XIAP was widely expressed in adult goat tissues as determined by real-time PCR and it demonstrated higher expression in propagative organs. High level of XIAP was detected in large healthy follicles and corpus luteum in comparison with that in small antral follicles, which was in accordance with the immunohistochemistry results and atretic follicles had very low expression. XIAP was localized in both granulosa and theca cells in antral follicles but not in primordial follicles. Furthermore, luteinizing hormone stimulated the proliferation of mRNA encoding XIAP in granulosa cells in vitro. The present study demonstrated that XIAP was expressed in a follicular-stage–dependent manner in goat ovaries.

Isolation and characterization of Toxoplasma gondii from small ruminants (sheep and goats) in Chennai City, South India.

The present study aimed for the isolation and genotyping of Toxoplasma gondii from small ruminants (sheep and goats). 14 out of 193 tissue samples (either brain and heart) tested positive by MDAT for anti-T. gondii antibodies, were selected and bioassayed, which resulted 4 samples positive for T. gondii after 40days of post inoculation. Four samples consisting of 3 numbers of sheep and 1 number of goat tissues out of 14 samples detected by B1 PCR, were genotyped at SAG3 locus by nested polymerase chain reaction-restriction fragment length polymorphism technique (nPCR-RFLP). The results of the present study revealed that the four isolates designated as TgShIn19, TgShIn76, TgShIn77 and TgGtIn27 were circulating in small ruminants, were belonged to genotypes of type II (TgShIn19) and type III (TgShIn76, TgShIn77 and TgGtIn27) which are in concordance with the previously reported genotypes from other animal species and further this presumptive results indicating that the genotype II and III could be the predominant in different animal species including birds and humans in India.

Epigenetic Status of H19-Igf2 Imprinted Genes and Loss of 5-Hydroxymethylcytosine in the Brain of Cloned Goats.

In mammals, the imprinted genes play vital roles in development and are generally controlled by DNA methylation at imprinting control regions (ICRs). Recently, it was discovered that 5-hydroxymethylcytosine (5-hmC) is a stable epigenetic modification; however, its functions in cloned animal genomes have not yet been fully elucidated. In this study, we interrogated and quantified the 5-hmC levels in the brain of cloned goats and discovered upregulation of Uhrf1 (p < 0.001), Dnmt1 (p < 0.05), Dnmt3a (p < 0.05), Igf2 (p < 0.01), and H19 (p < 0.05) and downregulation of Dnmt3b (p < 0.001), Tet1 (p < 0.001), Tet2 (p < 0.05), Tet3 (p < 0.001), Mecp2 (p < 0.05), and Igf2r (p < 0.05) in deceased cloned goat tissues compared with the normal controls. We demonstrated that DNA methylation was increased at H19 ICR (51.33% ± 2.03% vs. 93.07% ± 3.06%; p < 0.01) and that DNA was hypomethylated at Igf2 ICR (4.57% ± 1.48% vs. 7.63% ± 1.83%; p > 0.05) in the brain of deceased cloned goats. Finally, we showed that within the cloned goat brain genome, the amount of genome-wide 5-hmC was significantly decreased (0.083% ± 0.026% vs. 0.024% ± 0.007%; p < 0.05), whereas the 5-hmC levels within H19 and Igf2 CCGG sites were not significantly altered (0.17% ± 0.09% vs. 0.03% ± 0.01%; p > 0.05) in the brain of deceased cloned goats. Our data bring further experimental evidence regarding the abnormalities in 5-hmC and advance our current understanding of the role of 5-hmC in cloned animals.

Effects of piroxicam on tissue distribution of sulfadimidine in West African Dwarf male and female goats.

Cases of Stevens-Johnson syndrome have been increasingly reported in Nigeria by individuals who consumed meat products of animals especially goats injected sulfonamides. Hence, tissue distribution and residues of intramuscular sulfadimidine were studied in West African Dwarf (WAD) goats. Twenty goats divided into two groups of 10 each (five males; five females) weighing 10.4 ± 1.63 kg were administered intramuscular sulfadimidine (100 mg/kg body weight), and the second group was coadministered 5 mg/kg of piroxicam via right and left thigh muscle, respectively. Samples of the liver, kidney, spleen, heart, lung, intestine, brain, and skeletal muscle were collected into sterile cellophane bags. Two untreated goats were killed and used for preparation of tissue standards. The tissue samples were stored frozen for analysis. High concentration of sulfadimidine residues was found in all the tissues of goats administered sulfadimidine as well as tissues of goats coadministered sulfadimidine/piroxicam for up to 30 days postdrug administration. Generally, residues of sulfadimidine were observed to be significantly higher than the acceptable limit (0.1 ppm). Hence, consumption of meats from WAD goats administered sulfadimidine may pose very high risk of Stevens-Johnson syndrome in sensitive humans. As such consumption of such meats should be avoided.

Postmortem recovery of proliferative fibroblasts up to three days in livestock ear skin stored at 35°C

Effect of elevated temperature (35°C) on postmortem survival of cells in goat, sheep and bovine tissues was studied. Skin explants (n = 30; 2-3 mm2) were cultured in petri dishes after 2, 24, 48, 72, 96 and 120 hours of postmortem interval in DMEM media supplemented with 10% FBS, 50 units/mL of penicillin, 50 µg/mL of streptomycin and 2.5 µg/mL of fungizone. Outgrowth of cells around the explants was observed up to 72 h in sheep, 48 h in bovine and 24 h in goats. In general, the number of explants exhibiting outgrowth as well as the level of confluence decreased with increasing postmortem time interval. Secondary cultures established from primary cells for 72 h postmortem interval show cytogenetically stable chromosomes with 54XX[17] normal female sheep karyotype, comparable cell morphology, and growth curve to that of fresh tissue derived cells. Till date these cells have been passaged 21 times and show normal growth. These results suggest that normal, proliferative cells can be recovered from skin tissues stored at an elevated temperature of 35°C in livestock from 24-72 h of postmortem interval. Reprogramming of these cells to clone the dead animals or their use for cell therapies remain to be seen in future.

Modulation of Host miRNAs Transcriptome in Lung and Spleen of Peste des Petits Ruminants Virus Infected Sheep and Goats

Peste des petits ruminants (PPR) is one of the highly contagious viral disease, characterized by fever, sore mouth, conjunctivitis, gastroenteritis, and pneumonia, primarily affecting sheep and goats. Reports suggested variable host response in goats and sheep and this host response vis-a-vis the expression of microRNAs (miRNAs) has not been investigated. Here, miRNAs were sequenced and proteomics data were generated to identify the role of differentially expressed miRNA (DEmiRNA) in PPR virus (PPRV) infected lung and spleen tissues of sheep and goats. In lungs, 67 and 37 DEmiRNAs have been identified in goats and sheep, respectively. Similarly, in spleen, 50 and 56 DEmiRNAs were identified in goats and sheep, respectively. A total of 20 and 11 miRNAs were found to be common differentially expressed in both the species in PPRV infected spleen and lung, respectively. Six DEmiRNAs—miR-21-3p, miR-1246, miR-27a-5p, miR-760-3p, miR-320a, and miR-363 were selected based on their role in viral infections, apoptosis, and fold change. The target prediction analysis of these six selected DEmiRNAs from the proteome data generated, revealed involvement of more number of genes in lung and spleen of goats than in sheep. On gene ontology analysis of host target genes these DEmiRNAs were found to regulate several immune response signaling pathways. It was observed that the pathways viz. T cell receptor signaling, Rap1 signaling, Toll-like receptor signaling, and B cell receptor signaling governed by DEmiRNAs were more perturbed in goats than in sheep. The data suggests that PPRV-induced miR-21-3p, miR-320a, and miR-363 might act cooperatively to enhance viral pathogenesis in the lung and spleen of sheep by downregulating several immune response genes. The study gives an important insight into the molecular pathogenesis of PPR by identifying that the PPRV—Izatnagar/94 isolate elicits a strong host response in goats than in sheep.

1693 Effects of feeding varying levels of deoiled distillers dried grains with solubles on fatty acid composition of subcutaneous adipose tissue in meat goats

1693 Effects of feeding varying levels of deoiled distillers dried grains with solubles on fatty acid composition of subcutaneous adipose tissue in meat goats