RESUMO
Bedding materials are important for suckling buffalo calves. Treated dung has been used as a bedding material for dairy cows but the lack of an appropriate safety assessment limits its application. In this study, we evaluated the feasibility of treated dung (TD) as a bedding material for suckling calves by comparing TD with rice husk (RH) and rice straw (RS) bedding materials. The TD was prepared through high-temperature composting by Bacillus subtilis. Thirty-three newborn suckling buffalo calves (Bubalus bubalis, 40.06 ± 5.79 kg) were randomly divided into 3 bedding material groups (TD, RH, and RS) and bedded with 1 of the 3 bedding materials for 60 d. We compared cost, moisture content, bacterial counts, and microbial composition of the 3 bedding materials, and investigated growth performance, health status, behavior, rumen fermentation, and blood parameters of bedded calves. The results showed that TD contained the fewest gram-negative bacteria and coliforms on d 1 and 30 and the lowest relative abundance of Staphylococcus throughout the experiment. The RH and TD bedding materials had the lowest cost. Calves in the TD and RS groups showed a higher dry matter intake, and final body weight and average daily gain in the TD and RS groups tended to be higher than in the RH group. Calves in the TD and RS groups had a lower disease incidence (diarrhea and fever), fewer antibiotic treatments, and lower fecal score than calves in the RH group. Higher contents of IgG, IgA, and IgM were observed in calves of the TD and RS groups than in calves of the RH group on d 10, indicating higher immune ability in TD and RS groups. Furthermore, TD bedding increased the butyric acid content in the calf's rumen, whereas RS bedding increased the acetate content, which might be attributed to the longer time and higher frequency of eating bedding material in the RS group. Considering all of the above indicators, we concluded that TD is the optimal bedding material for calves based on economics, bacterial count, microbial diversity, growth performance, and health status. Our findings provide a valuable reference for bedding material choice and calf farming.
Assuntos
Compostagem , Feminino , Animais , Bovinos , Búfalos , Temperatura , Peso Corporal , Roupas de Cama, Mesa e Banho , Nível de Saúde , Dieta/veterinária , Ração Animal/análise , Desmame , RúmenRESUMO
Objective: To determine the effects of ω-3 polyunsaturated fatty acids from different sources on glucolipid metabolism in type 2 diabetic patients with dyslipidemia. Methods: We recruited participants from the diabetes specialist clinic at the Guanlin hospital in Yixing city, Jiangsu Province from February 2017 to March 2017. A total of 180 subjects were randomly assigned to 3 g/day fish oil (FO), perilla oil (PO), or fish oil mixed with linseed oil (FLO) for 6 months. The basic conditions and fasting venous blood sample were obtained from each study subject at baseline, after 6 months of intervention. Serum glucose and lipid metabolism were investigated. Results: A total of 156 subjects aged (62.6±8.6) years completed the final follow-up after 6 months (FO,54 subjects; PO,52 subjects; FLO,50 subjects). Among them,59 patients (37.8%) were male. Serum glucose, glycated hemoglobin, C peptide, insulin and homeostasis model assessment-insulin resistance were not significantly different among the three groups after 6 months. Serum triglyceride decreased, whereas high-density lipoprotein cholesterol increased in FO [1.33 (1.05,1.93) mmol/L, (1.36±0.29) mmol/L, respectively] compared with PO [1.71 (1.23, 2.17) mmol/L, (1.23±0.22) mmol/L, respectively] and FLO [1.51 (1.12, 2.22) mmol/L, (1.29±0.30) mmol/L, respectively] (P<0.05). Serum low-density lipoprotein cholesterol and apolipoprotein B decreased in PO [(2.60±0.57) mmol/L,(0.96±0.23) g/L, respectively] compared with FO [(2.89±0.76) mmol/L, (1.07±0.30) g/L, respectively] (P<0.05). Serum lipoprotein(a) decreased in FLO [130.7 (63.3,270.6) mg/L] compared with FO [137.4 (58.7,333.2) mg/L] (P<0.05). Serum free fatty acid decreased in FLO [(0.43±0.15) mmol/L] compared with PO [(0.53±0.22) mmol/L] (P<0.05). Conclusion: The effects of ω-3 PUFA from different sources on glucose metabolism in type 2 diabetic patients with dyslipidemia are similar. Each of them has a good application prospect in improving lipid metabolism.
Assuntos
Diabetes Mellitus Tipo 2 , Dislipidemias , Ácidos Graxos Ômega-3 , Idoso , Glicemia , HDL-Colesterol , Diabetes Mellitus Tipo 2/tratamento farmacológico , Dislipidemias/tratamento farmacológico , Ácidos Graxos Ômega-3/uso terapêutico , Feminino , Óleos de Peixe , Humanos , Masculino , Pessoa de Meia-Idade , TriglicerídeosRESUMO
Water buffalo is the second largest resource of milk supply around the world, and it is well known for its distinctive milk quality in terms of fat, protein, lactose, vitamin, and mineral contents. Understanding the genetic architecture of milk production traits is important for future improvement by the buffalo breeding industry. The advance of genome-wide association studies (GWAS) provides an opportunity to identify potential genetic variants affecting important economical traits. In the present study, GWAS was performed for 489 buffaloes with 1,424 lactation records using the 90K Affymetrix Buffalo SNP Array (Affymetrix/Thermo Fisher Scientific, Santa Clara, CA). Collectively, 4 candidate single nucleotide polymorphisms (SNP) in 2 genomic regions were found to associate with buffalo milk production traits. One region affecting milk fat and protein percentage was located on the equivalent of Bos taurus autosome (BTA)3, spanning 43.3 to 43.8 Mb, which harbored the most likely candidate genes MFSD14A, SLC35A3, and PALMD. The other region on the equivalent of BTA14 at 66.5 to 67.0 Mb contained candidate genes RGS22 and VPS13B and influenced buffalo total milk yield, fat yield, and protein yield. Interestingly, both of the regions were reported to have quantitative trait loci affecting milk performance in dairy cattle. Furthermore, we suggest that buffaloes with the C allele at AX-85148558 and AX-85073877 loci and the G allele at AX-85106096 locus can be selected to improve milk fat yield in this buffalo-breeding program. Meanwhile, the G allele at AX-85063131 locus can be used as the favorable allele for improving milk protein percentage. Genomic prediction showed that the reliability of genomic estimated breeding values (GEBV) of 6 milk production traits ranged from 0.06 to 0.22, and the correlation between estimated breeding values and GEBV ranged from 0.23 to 0.35. These findings provide useful information to understand the genetic basis of buffalo milk properties and may play a role in accelerating buffalo breeding programs using genomic approaches.
Assuntos
Búfalos/fisiologia , Cromossomos/genética , Variação Genética/genética , Estudo de Associação Genômica Ampla/veterinária , Leite/metabolismo , Locos de Características Quantitativas/genética , Animais , Cruzamento , Búfalos/genética , Feminino , Genômica , Proteínas do Leite/análise , Fenótipo , Polimorfismo de Nucleotídeo Único/genética , Reprodutibilidade dos TestesRESUMO
This study was conducted to determine the effect of immunization with inhibin DNA vaccine delivered by attenuated Salmonella choleraesuis on ovarian responses and fertility in cross-bred buffaloes. A total of 134 cross-bred buffaloes were divided into four groups: groups T1 (n = 34), T2 (n = 35) and T3 (n = 31) were nasal immunized twice a day with 10 ml of 1 × 1010 CFU/ml of the C501 (pVAX-asd-IS) vaccine for 5, 3 and 1 day, respectively. Group C (n = 34) was nasal immunized with 10 ml PBS for 5 days. All animals were immunized twice with an interval of 14 days and administered with 200 µg of a GnRH analogue on day 28, 0.5 mg PGF2α on day 35 and 200 µg of the same GnRH analogue on day 37. TAI was performed at 18 and 24 hr after the second GnRH treatment. Fourteen days after primary immunization, C501 (pVAX-asd-IS) elicited significant immune responses, and anti-inhibin IgG antibody titres in group T1 were significantly higher (p < .01) than groups T3 and C. After the second GnRH treatment, the growth speed of the dominant follicles in group T1 was significantly faster (p < .05) than groups T3 and C. The number and diameter of large follicles (≥10 mm) as well as ovulatory follicles in group T1 were the greatest in all groups, resulting in a greater conception rate in buffaloes with positive anti-inhibin antibodies. These results demonstrate that immunization with the C501 (pVAX-asd-IS) vaccine, coupled with the Ovsynch protocol, could be used as an alternative approach to improve reproductive performance in cross-bred buffaloes.
Assuntos
Búfalos/fisiologia , Inibinas/imunologia , Folículo Ovariano/efeitos dos fármacos , Ovulação/efeitos dos fármacos , Administração Intranasal , Animais , Búfalos/genética , Sincronização do Estro , Feminino , Ovulação/imunologia , Ovulação/fisiologia , Gravidez , Salmonella/genética , Vacinas de DNARESUMO
Spermatogenesis, is a complex process of precisely regulated intracellular events, where it is affected by many factors. Long-distance transport of animals is one of the stressors that may influence spermatogenesis and sperm quality. The present study chose luteinizing hormone receptor (LHR), androgen receptor (AR), and heat-shock protein 70 (HSP70) as our target genes to investigate their mRNA and protein expression in the testes of long-distance transported (about 1000 km) mice. Histological analysis showed that there was a reduction in the thickness of the seminiferous epithelium in the transported mice, and a significant decrease in body weight and sperm count in the epididymis was also observed. mRNA expression was determined by QPCR in the testis of transported and control mice. The levels for AR decreased significantly in transported mice. LHR and HSP70 expression in the testes of the transported mice was slightly higher than that of control mice but did not reach a significant level. A similar tendency of protein expression was also observed by Western blot analysis. The levels of LHR and HSP70 increased slightly after transportation. However, none of the changes were statistically significant compared with the control mice. In conclusion, long-distance transport has an adverse effect on reproductive organs and spermatozoa in adult mice.
Assuntos
Regulação da Expressão Gênica , Proteínas de Choque Térmico HSP70/genética , Receptores Androgênicos/genética , Receptores do LH/genética , Testículo/metabolismo , Animais , Peso Corporal , Masculino , Camundongos , Tamanho do Órgão , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Contagem de Espermatozoides , Espermatogênese/genética , Estresse Fisiológico/genética , Testículo/anatomia & histologiaRESUMO
Mastitis is the most important disease in the global dairy industry, and causes large economic losses. Staphylococcus aureus is one of most common pathogens that cause bovine mastitis. CXCR1 has been implicated as a prospective genetic marker for mastitis resistance in dairy cows; CXCR1 expression significantly increases when cows have mastitis. To investigate the mechanisms involved in its increased expression, bisulfite sequencing polymerase chain reaction (PCR) was used to detect the methylation status of CXCR1 CpG island, and quantitative fluorescence PCR was used to detect CXCR1 expression in bovine mammary tissue induced with S. aureus in three Chinese Holstein cows. No CpG island was found for bovine CXCR1 in the upstream 2-kb region, whereas one CpG island that contained 13 CpG sites was found in exon 1 of CXCR1. All of the CpG sites were under hypermethylation from 90 to 100% in the mammary tissues. When the mammary gland mRNA expression of CXCR1 was 12.10-fold higher in infected cow quarters than in uninfected quarters, the methylation levels of the CpG site at position 519 were significantly lower in the infected quarters than in the uninfected quarters. Pearson correlation analysis showed that the methylation level at position 519 was significantly negatively correlated with the CXCR1 mRNA expression level (P < 0.05). These results indicate that the methylation of the CpG site at position 519 may regulate CXCR1 expression in cows with mastitis induced by S. aureus, but further studies are needed to elucidate the mechanisms involved.
Assuntos
Metilação de DNA , Glândulas Mamárias Animais/metabolismo , Mastite Bovina/genética , Receptores de Interleucina-8A/genética , Infecções Estafilocócicas/veterinária , Staphylococcus aureus/isolamento & purificação , Animais , Bovinos , Ilhas de CpG , Feminino , Mastite Bovina/metabolismo , Mastite Bovina/microbiologia , Reação em Cadeia da Polimerase , Estudos Prospectivos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores de Interleucina-8A/metabolismo , Infecções Estafilocócicas/metabolismoRESUMO
We demonstrate experimentally Manchester (MC) coding based W-band (75 - 110 GHz) radio-over-fiber (ROF) system to reduce the low-frequency-components (LFCs) signal distortion generated by two independent low-cost lasers using spectral shaping. Hence, a low-cost and higher performance W-band ROF system is achieved. In this system, direct-beating of two independent low-cost CW lasers without frequency tracking circuit (FTC) is used to generate the millimeter-wave. Approaches, such as delayed self-heterodyne interferometer and heterodyne beating are performed to characterize the optical-beating-interference sub-terahertz signal (OBIS). Furthermore, W-band ROF systems using MC coding and NRZ-OOK are compared and discussed.
Assuntos
Tecnologia de Fibra Óptica/instrumentação , Lasers , Telecomunicações/instrumentação , Tecnologia sem Fio/instrumentação , Desenho de EquipamentoRESUMO
This study aimed to evaluate the efficacy and safety of an oral DNA vaccine against somatostatin (SS) (pGS/2SS-asd, encoding two copies of somatostatin genes) mediated by attenuated Salmonella choleraesuis C500 without antibiotic resistance gene on piglets growth. A total of 50 piglets were uniformly divided into five groups. The animals in the first three groups were orally given vaccine in dose of either 5 9 1010, 5 9 109 or 5 9 108 colony-forming units (CFU).The remaining two groups were orally administered with either bacteria C500(containing pVAX-asd plasmid without somatostatin gene) or phosphate buffered saline (PBS) as controls. The results indicated that the vaccine induced SS-specific antibodies in a dose-dependent pattern. Compared with the PBS control, animals in the high-dose group showed lower SS levels and higher growth hormone (GH) levels in sera. Average daily gain of animals in the high dose group was increased by 32.88% and 26.46% during 4 and 8 weeks,respectively. Anti-SS antibodies were positively correlated with either GH levels or average daily gain at week 8 after primary immunization (P < 0.05). Faecal,soil and water samples originating from immunized piglets and surrounding environment were collected. The target gene (the fusion gene GS/2SS) of C500(pGS/2SS-asd) was not detected by PCR amplification in these samples,indicating that the surrounding environment was not contaminated by residual recombinant bacteria. In conclusion, the vaccine without antibiotic resistance gene is attributable to improve growth performance of piglets through an influence on GH secretion. Moreover, the immunization did not contaminate the surrounding environment of animals.
Assuntos
Hormônio do Crescimento/metabolismo , Salmonella arizonae/genética , Somatostatina/antagonistas & inibidores , Somatostatina/imunologia , Suínos/crescimento & desenvolvimento , Vacinas de DNA/administração & dosagem , Administração Oral , Animais , Anticorpos Antinucleares/imunologia , Resistência Microbiana a Medicamentos/genética , Vetores Genéticos/genética , Hormônio do Crescimento/antagonistas & inibidores , Hormônio do Crescimento/sangue , Reação em Cadeia da Polimerase , Somatostatina/genética , Vacinação , Vacinas de DNA/efeitos adversos , Vacinas de DNA/genéticaRESUMO
Follicle-stimulating hormone receptor (FSHR), which mediates the functioning of FSH, plays a central role in reproduction. We investigated bovine FSHR gene polymorphisms and analyzed their relationships with pregnancy rates after embryo transfer and with hormone concentrations on the day of embryo transfer. One reported SNP of FSHR, G-278A, located in the 5'-upstream region, was analyzed and three genotypes (GG, GA and AA) were detected in 132 Luxi cattle recipients. Statistical analysis revealed that recipients with the GG genotype had significantly higher estrogen levels on the day of embryo transfer than did GA and AA genotypes. There were no significant differences in pregnancy rates among genotypes, after embryo transfer. We conclude that variation at these loci of the FSHR gene has no significant effect on pregnancy rates in Luxi cattle.
Assuntos
Transferência Embrionária , Estrogênios/sangue , Receptores do FSH/genética , Reprodução/genética , Animais , Bovinos , Feminino , Genótipo , Polimorfismo de Nucleotídeo Único , Gravidez , Taxa de Gravidez , Receptores do FSH/sangueRESUMO
Inhibin is a major regulator of secretion of follicle-stimulating hormone, which is involved in follicular development and regulation of steroidogenesis in females. The objectives of this study were to detect polymorphisms of the bovine inhibin beta-A subunit (INHßA) gene and to evaluate its associations with superovulatory responses in 171 Chinese Holstein cows treated for superovulation. Polymerase chain reaction-restricted fragment length polymorphism revealed a C>T transition determining the StyI polymorphism at position 7639 in intron I of the bovine INHßA gene, and three genotypes (CC, CT, and TT) were detected. The frequencies of the three genotypes showed a tendency for CT > TT > CC, and this polymorphism was in Hardy-Weinberg equilibrium. Statistical analysis revealed no significant differences of least square means for superovulation traits among the three genotypes (P > 0.05). These results demonstrate, for the first time, that the detected loci of the INHßA gene have no significant effects on superovulation performance in Chinese Holstein cows.
Assuntos
Bovinos/genética , Subunidades beta de Inibinas/genética , Polimorfismo Genético , Superovulação/genética , Animais , Estudos de Casos e Controles , Bovinos/fisiologia , FemininoRESUMO
Bone morphogenetic protein 15 (BMP-15) expression has been detected in the testis, but its roles in this organ has not been well elucidated. We evaluated polymorphisms of the BMP-15 gene by PCR-SSCP and PCR-RFLP in 212 Chinese Holstein bulls, and investigated possible associations with sperm quality traits, including semen volume per ejaculate, sperm density, fresh sperm motility, thawed sperm motility, acrosome integrity rate, and abnormal sperm rate. A single nucleotide polymorphism (C5697T) in intron 1 of the BMP-15 gene was identified in these bulls. Age was found to have significant effects on both fresh sperm motility and abnormal sperm rate. A significant effect of genotype on fresh sperm motility was also observed. Least square analysis showed that CT genotype bulls had significantly lower fresh sperm motility than CC or TT genotype bulls. In conclusion, BMP-15 should be considered as a potential genetic marker for sperm quality, based on its association with fresh sperm motility.
Assuntos
Proteína Morfogenética Óssea 15/genética , Motilidade dos Espermatozoides/genética , Espermatozoides , Animais , Bovinos , Marcadores Genéticos , Genótipo , Masculino , Polimorfismo de Nucleotídeo ÚnicoRESUMO
DNA vaccination has been studied intensively as a potential vaccine technology. We evaluated the effect of an attenuated Salmonella choleraesuis-mediated inhibin DNA vaccine in rats. First, 15 rats were treated with different doses of an inhibin vaccine to evaluate vaccine safety. Next, 30 rats were divided into 3 groups and injected intramuscularly with the inhibin vaccine two (T1) or three times (T2) or with control bacteria (Con) at 4-week intervals. The inhibin antibody levels increased [positive/negative well (P/N) value: T1 vs Con = 2.39 ± 0.01 vs 1.08 ± 0.1; T2 vs Con = 2.36 ± 0.1 vs 1.08 ± 0.1, P < 0.05] at week 2 and were maintained at a high level in T1 and T2 until week 8, although a small decrease in T2 was observed at week 10. Rats in the T1 group showed more corpora lutea compared with the Con group (10.50 ± 0.87 vs 7.4 ± 0.51, P < 0.05). Estradiol (0.439 ± 0.052 vs 0.719 ± 0.063 ng/mL, P < 0.05) and progesterone (1.315 ± 0.2 vs 0.737 ± 0.11 ng/mL, P < 0.05) levels differed significantly at metestrus after week 10 between rats in the T1 and Con groups. However, there were no significant differences in body, ovary, uterus weights, or pathological signs in the ovaries after immunization, indicating that this vaccine is safe. In conclusion, the attenuated S. choleraesuis-mediated inhibin vaccine may be an alternative to naked inhibin plasmids for stimulating ovarian follicular development to increase the ovulation rate in rats.
Assuntos
Inibinas/genética , Inibinas/imunologia , Salmonella/genética , Vacinas Atenuadas/imunologia , Vacinas de DNA/imunologia , Animais , Anticorpos/sangue , Anticorpos/imunologia , Estradiol/sangue , Feminino , Imunização , Folículo Ovariano/imunologia , Folículo Ovariano/patologia , Ovulação , Progesterona/sangue , Ratos , Vacinas Atenuadas/administração & dosagem , Vacinas Atenuadas/efeitos adversos , Vacinas de DNA/administração & dosagem , Vacinas de DNA/efeitos adversosRESUMO
In this work, we propose and demonstrate a stable and wavelength-tunable erbium-doped fiber (EDF) ring laser. Here, a silicon-on-insulator (SOI)-based silicon-micro-ring-resonator (SMRR) is used as the wavelength selective element inside the fiber ring cavity. A uniform period grating coupler (GC) is used to couple between the SMRR and single mode fiber (SMF) and serves also as a polarization dependent element in the cavity. The output lasing wavelength of the proposed fiber laser can be tuned at a tuning step of 2 nm (defined by the free spectral range (FSR) of the SMRR) in a bandwidth of 35.2 nm (1532.00 to 1567.20 nm), which is defined by the gain of the EDF. The optical-signal-to-noise-ratio (OSNR) of each lasing wavelength is larger than 42.0 dB. In addition, the output stabilities of power and wavelength are also discussed.
Assuntos
Lasers de Estado Sólido , Lasers , Silício/química , Transdutores , Desenho de Equipamento , Análise de Falha de Equipamento , MiniaturizaçãoRESUMO
Growth differentiation factor 9 (GDF9) belongs to the transforming growth factor ß superfamily and plays a critical role in ovarian follicular development and ovulation rate. We examined the bovine GDF9 gene polymorphism and analyzed its association with superovulation performance. Based on the sequence of the bovine GDF9 gene, six pairs of primers were designed to detect single nucleotide polymorphisms of two exons and intron 1 of GDF9 using polymerase chain reaction-single-strand conformation polymorphism. Only the products amplified by primer 3-1 displayed polymorphisms. Sequencing revealed two mutations of A485T and A625T in intron 1 of the GDF9 gene in 171 Chinese Holstein cows treated for superovulation. Association analysis showed that these two single nucleotide polymorphisms of A485T and A625T had significant effects on the number of transferable embryos (P < 0.05), and the A625T polymorphism was significantly associated with the total number of ova (P < 0.05). In addition, a significant additive effect on the number of transferable embryos was detected in polymorphisms of A485T (P < 0.05). This study is the first to identify two polymorphisms in bovine GDF9 and describe their correlation with superovulation traits in Chinese Holstein cows.
Assuntos
Bovinos/genética , Fator 9 de Diferenciação de Crescimento/genética , Superovulação/genética , Animais , Feminino , Frequência do Gene , Estudos de Associação Genética , Genótipo , Íntrons , Polimorfismo de Nucleotídeo ÚnicoRESUMO
Members of the transforming growth factor-ß (TGFß) superfamily are critical regulators of germ cell development that act as extracellular ligands of the signal transduction pathways regulating proliferation, differentiation, apoptosis, and other aspects of cell behavior. Growth differentiation factor 9 (GDF9) is a member of the TGFß superfamily that plays a critical role in ovarian follicular development and ovulation rate in females; however, its role in the testis has not been well elucidated. Therefore, in this study we investigated the effects of GDF9 mutations on the quality of fresh and frozen semen of Holstein bulls. Two reported single nucleotide polymorphisms of GDF9, A485TA and A625C, were analyzed in 129 Holstein bulls. Analysis of variance revealed that the A485T polymorphism had significant effects on the acrosome integrity rate (P < 0.05), whereas the A625T polymorphism was significantly associated with sperm concentration (P < 0.05). In addition, a significant additive effect on sperm concentration was detected for the A485T polymorphism (P < 0.05), whereas the polymorphisms A485TA and A625C had significant dominant effects on acrosome integrity rate and sperm motility in frozen semen, respectively (P < 0.05). This study is the first to show a significant association of GDF9 with sperm quality traits, and the results implied that GDF9 is involved in the initiation or maintenance of spermatogenesis; however, further verification is needed.
Assuntos
Acrossomo/fisiologia , Fator 9 de Diferenciação de Crescimento/genética , Polimorfismo de Nucleotídeo Único , Motilidade dos Espermatozoides/genética , Animais , Animais Endogâmicos , Bovinos , Masculino , Contagem de EspermatozoidesRESUMO
The proportion of type A spermatogonia in the isolated testis cells is a prerequisite for conducting experiments and the manipulation of these germ cells. Thus, this study was designed to examine the wide range of strategies for the isolation, identification and enrichment of type A spermatogonia in pre-pubertal buffalo calves (3-6 months). Histological findings revealed the presence of maximum number of type A spermatogonia at 5 months, which was further confirmed by DBA immunohistochemistry. In a newly modified strategy for the isolation of testis tissues, mincing followed by trituration and two rounds of digestion with collagenase, hyaluronidase and DNase yielded more than 95% testis cell population. Differential plating with laminin, poly-l-lysine and gelatin significantly (p < 0.05) affected the purity of type A spermatogonia. Among these extracellular matrix (ECMs) molecules, laminin and gelatin performed well and reached at a purity of 39.38 ± 1.21% and 32.15 ± 1.60%, respectively. In addition, combination of laminin and gelatin followed by Percoll centrifugation performed the best and yielded >90% type A spermatogonial purity. Moreover, viability of the cells was not affected (p > 0.05) irrespective of different enrichment methods. In conclusion, type A spermatogonia isolation and enrichment system was developed using different ECM molecules in buffaloes, which will aid in solving wide range of problems especially fertility-related problems and transgenic animal production in buffaloes.
Assuntos
Búfalos/genética , Búfalos/fisiologia , Espermatogônias/citologia , Espermatogônias/fisiologia , Testículo/citologia , Envelhecimento , Animais , MasculinoRESUMO
Porcine reproductive and respiratory syndrome virus (PRRSV), Haemophilus parasuis and Pseudorabies become a widespread problem causing great economic losses associated with reproductive disturbance, respiratory diseases, neonatal mortality, fibrinous polyserositis, meningitis and arthritis in the pig industry. The important candidate genes are assumed to play crucial roles in host defense against the diseases. The aims of this study were to evaluate the variants in HLA-B associated transcript 2 (BAT2), CXCL12, myxovirus resistance protein 1 (Mx1) and EHMT2 genes and their effects on the risk of infection PRRSV and H. parasuis in a case-control (diseased-healthy pigs) population of Duroc × Landrace × LargeWhite. The results showed that the mutations in BAT2, Mx1 and EHMT2 genes were significantly associated with the antibody and the reisk of infection PRRSV and H. parasuis. Those individuals with AA genotype of BAT2 had significantly higher Pseudorabies virus antibody than that with GG and GA (P < 0.05), and the individuals with TT genotype of EHMT2 generated higher Hog Cholera and Pseudorabies virus antibody than that wtih GG and GA (P < 0.01). These results indicated that the polymorphisms in Mx1, BAT2 and EHMT2 genes changed the diseases susceptibility and could be the potential markers assisting the pig breeding selection and disease resistance.
Assuntos
Quimiocina CXCL12/genética , Proteínas de Ligação ao GTP/genética , Histona-Lisina N-Metiltransferase/genética , Polimorfismo de Nucleotídeo Único , Doenças dos Suínos/genética , Animais , Anticorpos Antibacterianos/imunologia , Anticorpos Antivirais/imunologia , Estudos de Casos e Controles , Resistência à Doença/genética , Resistência à Doença/imunologia , Predisposição Genética para Doença , Infecções por Haemophilus/veterinária , Incidência , Proteínas de Resistência a Myxovirus , Síndrome Respiratória e Reprodutiva Suína/genética , Síndrome Respiratória e Reprodutiva Suína/imunologia , Pseudorraiva/genética , Pseudorraiva/imunologia , Suínos , Doenças dos Suínos/epidemiologia , Doenças dos Suínos/imunologiaRESUMO
It is necessary that genetic markers or biomarkers can be used to predict resistance towards a wide range of infectious diseases. In the present study, we estimated the potential markers and measured their relationship with heritabilities of a wide range of immune traits. Polymorphisms in exon 13 of Mx1, intron 25 of BAT2 and intron 3 of CXCL12 were identified by sequencing, and the genotypes were analyzed by PCR-RFLP in a resource population composed of 352 pure breed Landrace piglets at days 0, 17 and 32 after birth. Associations of single-nucleotide polymorphisms (SNPs) in these genes with a variety of immunological traits and antibody levels for pig reproduction and porcine respiratory syndrome virus (PRRSV), pseudorabies virus (PRV) and classical swine fever virus (CSFV) were performed. The performance of GG genotype of BAT2 on hemoglobin concentration (HBG) and hematocrit (HCT) of piglets at day 0 was significantly higher than that of the AA and AG individuals. For Mx1, compared with CT genotype, the pigs with TT or CC generated more PRRS antibody at day 0. The piglets with CT genotype had highly significant difference of PRV antibody from those with CC and TT genotypes at day 0. And the piglets with CC genotype had higher level red blood cell count (RBC), hemoglobin concentration (HBG) and hematocrit (HCT) than those with CT and TT genotypes at day 17. For the C7462G SNP in the intron 3 of CXCL12, the PRV antibody level of piglets with the CG genotype were higher than that of piglets with CC and GG genotypes at day 17, and the mean corpuscular volume (MCV) of GG piglets were larger than that of CC and CG individuals at day 0. At the locus 7331 bp in the intron 3 of CXCL12, there were significantly differences of mean corpuscular hemoglobin concentrations (MCHC) at day 0 and white blood cell count (WBC) at day 32, which showed the trend GG or AG>AA, AA>AG or GG, respectively. The pigs with AA or GG genotype had more platelet distribution width (PDW), mean platelet volume (MPV) and platelet-large cell ratio (PLR) at day 17 than those with AG. The results of this study indicated that polymorphisms in Mx1, BAT2 and CXCL12 genes were significantly associated with the immunological traits in Landrace piglets and had potential application value for marker-assisted selection of pig breeding with disease resistance.
Assuntos
Quimiocina CXCL12/genética , Resistência à Doença/genética , Proteínas de Ligação ao GTP/genética , Marcadores Genéticos/genética , Polimorfismo Genético/genética , Sus scrofa/genética , Análise de Variância , Animais , Anticorpos Antivirais/sangue , Sequência de Bases , Plaquetas/citologia , Cruzamento/métodos , Primers do DNA/genética , Resistência à Doença/imunologia , Estudos de Associação Genética/veterinária , Genótipo , Hematócrito , Hemoglobinas/análise , Dados de Sequência Molecular , Proteínas de Resistência a Myxovirus , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Polimorfismo de Nucleotídeo Único/genética , Análise de Sequência de DNA/veterinária , Sus scrofa/imunologiaRESUMO
Alpha-(1,2)-fucosyltransferase (FUT1) gene has been identified as a candidate gene for regulating the expression of Escherichia coli F18 receptor gene (ECF18R) which promotes adherence of Enterotoxigenic (ETEC) and Verotoxigenic (VTEC) Escherichia coli (E. coli) via F18 fimbriae. In order to illustrate the polymorphisms of FUT1 and their effects on resistance to natural infection by Porcine Respiratory and Reproductive Symdrome Virus (PRRSV) and Haemophilus parasuis, the distributions of different genotypes and the relative risks of disease incidence in pigs were investigated. A total of 1,041 pigs representing three European breeds (Duroc, Landrace and LargeWhite), five Chinese local breeds (Wild pig, Small MeiShan, QinPing, JinHua, and JianLi) and three commercial populations (LargeWhite × JianLi, Duroc × Landrace × LargeWhite and Duroc × wild pig) were selected to analyze the genotype of the FUT1 gene by PCR-RFLP. Only the GG genotype associated with susceptibility to ECF18 bacteria was detected in Chinese local pig breeds and a population of LargeWhite × JianLi, while the AA genotype which confers resistance to ECF18 was detected in two European breeds (Duroc and LargeWhite), two populations of Duroc × wild pig and Duroc × Landrace × LargeWhite. Regarding relative risk of incidence, Duroc × Landrace × LargeWhite with genotypes GG or AG showed greater relative risk (OR = 2.040, P = 0.025; OR = 1.750, P = 0.081, respectively) than those with genotype AA during natural infection by both PRRSV and Haemophilus parasuis. It can be concluded that the mutation of FUT1 gene might play a role in pig infection by multi-pathogens, and that AA may be a favourable genotype for increasing the resistance to disease.