[Objective] The experiment aimed to set up a method for isolating and culturing endometrial stromal cells (BESC) and endometrial glandular epithelial cells(BEGEC) of buffalo as well as laid foundation for studying bio...[Objective] The experiment aimed to set up a method for isolating and culturing endometrial stromal cells (BESC) and endometrial glandular epithelial cells(BEGEC) of buffalo as well as laid foundation for studying biological mechanism of embryo implantation and uterine diseases. [Method] The enzymatic digestion method, scraping method, serial filtration and differential velocity adherent technique were used to isolate BESC and BEGEC, then immunocytochemical method and TRYPAN-Blue assay were used to determine the purity and survival rate of isolated cells. [Result] The BESC and BEGEC were successfully isolated and cultured while immunocytochemical method and cell count method demonstrated that the purity was over 90%. The result of TRYPAN-Blue assay shown that survival rate of BESC and BEGEC was 91% and 78% respectively. [Conclusion] The enzymatic digestion method, scraping method, serial filtration and differential velocity adherent technique could isolate BESC and BEGEC with high purity.展开更多
To study the association of oxytocin (OT)'s distribution in hypothalamatic,pituitary and ovary,and understand how the OT secrete releasing in hypothalamus,pituitary and ovaries,the paraffin section immunohistochem...To study the association of oxytocin (OT)'s distribution in hypothalamatic,pituitary and ovary,and understand how the OT secrete releasing in hypothalamus,pituitary and ovaries,the paraffin section immunohistochemistry SuperPicTureTM two step method was used to detect the distribution of OT in hypothalamatic-pituitary-ovary axis of five femal Guangxi local buffalo. The test results could provide morphology according to study the OT's synthesis and mechanism of action,and could play reference and directions part in breeding Guangxi local buffalo. The test results display:oxytocin immuno reactive (OT-IR) neuronsw eremainly distributed arcuate nucleus,supraoptic nucleus and paraventricular nucleus,and OT-IR neurons was also found in ventromedial nucleus,ventrolateralis nucleus,suprachiasmaticus nucleus,dorsomedial nucleus,mamillary body,anterior hypothalamic nucleus and so on. The OT immunoactive production was found in pituitary and few OT-IR nerve fibers extended to post pituitary from hypophyseal stalk and medium eminence. In ovaries,OT immunoactive productions were only distributed in germinal epithelium cells,granulosa cells and lutein cells. The OT was first discovered in singulorum link of hypothalamatic-pituitary-ovary axis of Guangxi local buffalo. The OT immunoactive neurons were first discovered in every main nucleus of Guangxi local buffalo hypothalamus,especially distributed in arcuate nucleus,supraoptic nucleus and paraventricular nucleus.展开更多
[Objective] This study aimed to investigate the feasibility of transgenesis by injecting exogenous DNA into zygote cytoplasm of Buffalo. [Method] Buffalo oocytes were randomly divided into two groups 20-22 h after in ...[Objective] This study aimed to investigate the feasibility of transgenesis by injecting exogenous DNA into zygote cytoplasm of Buffalo. [Method] Buffalo oocytes were randomly divided into two groups 20-22 h after in vitro maturation. One group of oocytes was introduced with about 7.5 pl of 50 μg/ml DNA solution containing linear EGFP fragment by cytoplasmic injection 7-10 h or 18-20 h after in vitro fertilization （IVF）; the other group of oocytes was introduced with mixture of a single buffalo sperm and about 7.5 pl of 50 μg/ml DNA solution containing linear EGFP fragment by cytoplasmic injection （generally called ICSI-Mediated Gene Transfer, ICSI-Tr）. Expression of exogenous DNA was observed and recorded during the process of embryonic development. [Result] Early embryonic gene expression efficiency and blastocyst gene expression efficiency in IVF injection group showed no significant difference compared with that in ICSI-Tr group （P0.05）. In addition, the cleavage rate and early embryonic gene expression efficiency in IVF injection group were significantly higher with injection at 7-10 h post IVF than that at 18-20 h post IVF （P0.05）. [Conclusion] These results indicate that transgenic buffalo embryos can be generated by injecting exogenous DNA into cytoplasm of IVF oocytes, and the optimal injection time is 7-10 h post IVF.展开更多
The aim of this study was to investigate the feasibility of stimulating ovarian fol icle development in order to improve fertility in water buffalo cows by immunization against inhibin. The experiment was carried out ...The aim of this study was to investigate the feasibility of stimulating ovarian fol icle development in order to improve fertility in water buffalo cows by immunization against inhibin. The experiment was carried out in early summer (May) and included 24 multi-parity crossbred Murrah-Swamp buffaloes that were divided into immunized (n=11) and control (n=13) groups. Each immunized cow was administered with a 2-mL immunogen of mineral oil adjuvant containing 2 mg of recombinant inhibinα-subunit fusion protein. The controls were treated with the adjuvant only. Al animals received Ovsynch protocol treatment, starting on the day of the antigen administration, and they were artiifcial y inseminated upon behavioral estrus. As a result, al of the immunized buffaloes generated antibodies against inhibin during the experimental period and had higher plasma concentrations of fol icle-stimulating hormone (FSH), activin, and estradiol (E2) related to estrous expression. A higher proportion of immunized animals expressed estrus behavior than did the controls (72%vs. 30%, P<0.05). On aver-age, inhibin-immunized buffaloes had signiifcantly more large fol icles (≥9 mm in diameter) than the controls (mean±SEM;1.2±0.1 vs. 0.84±0.1, respectively;P<0.05) and a slightly higher mean total number of fol icles (≥2 mm;11.4±0.7 vs. 9.0±1.1, respectively;P=0.09) and smal (2–4 mm) fol icles (8.81±0.6 vs. 6.84±1.0, respectively;P=0.12). A higher percentage of cows ovulated in the immunized group than in the control group (91%(10/11) vs. 54%(7/13), respectively;P<0.05). Moreover, inhibin-immunized cows had slightly larger corpus luteum (CL) than the controls 9 days after ovulation and signiifcantly higher (P<0.01) post-ovulation peak plasma progesterone (P4) concentrations. Immunization against inhibin also mar-ginal y increased the conception rate 42 days after insemination (45.8%vs. 15.4%;P>0.05). These results demonstrate that immunization against inhibin, coupled with the treatment with the Ovsynch protocol, can constitute a new technique to increase fertility in water buffalo cows.展开更多
Changes of concentrations were studied in water buffaloes and goats infected with Fasciola he-patica on blood NO(nitric oxide) and TNF-α(tumor necrosis factor-α). Twenty healthy male castrated water buffaloes of 2 -...Changes of concentrations were studied in water buffaloes and goats infected with Fasciola he-patica on blood NO(nitric oxide) and TNF-α(tumor necrosis factor-α). Twenty healthy male castrated water buffaloes of 2 - 3 years old and weighing 300 - 500 kg as well as six goats were confirmed free of fasciolosis by fecal examination and Dot-ELISA. Two studies were conducted using the water buffaloes. In the first experiment, 8 water buffaloes were randomly divided into control group (n = 3) and infection group (n = 5). Each buffalo in the infected group received orally 60 metacercariae of F. hepatica per day for 20 days (total 1 200 metacercariae) to produce a chronic infection. In the second experiment, 12 water buffaloes were randomly divided into infected (n=9) and control group (n = 3). Each buffalo in the infected group was given a single oral dose of 1 600 metacercariae to produce an acute infection. The 6 goats were randomly divided into two infected groups and a control group. The sheep in two infections received a single oral dose of 200 and 500 metacercariae respectively, the control group remained uninfected. Blood NO and TNF-a concentrations of the test animals were measured by a reductive enzyme assay and RIA, respectively. Blood NO concentration in both acutely and chronically infected water buffaloes progressively increased from week 3 post-infection and was significantly greater than that of the control group (P<0. 05) at the 5th week (acute infection) and 7th week (chronic infection), and remaining at higher concentration for the remaining period of the studies. Blood TNF-αconcentrations in both chronically and acutely infected water buffaloes also increased after infection. In the goat experiment, plasma NO concentrations in both infection groups increased from week 3 after infection, and remained higher than that of the control group until the end of the experiment. TNF-α concentrations in goats in infection group 1 and 2 gradually increased after infection and were significantly greater than those of the control group from the 9th to 11th week and from the 11th to 15th week respectively.展开更多
Objective:To improvein vitro embryo production in buffalo by supplementation of L-ascorbic acid during maturation and development (experiment 1) and combination with another antioxidant as cysteamine (experiment 2).Me...Objective:To improvein vitro embryo production in buffalo by supplementation of L-ascorbic acid during maturation and development (experiment 1) and combination with another antioxidant as cysteamine (experiment 2).Methods:Two experiments were performed, the first one aimed to evaluate the different concentrations (0, 25, 50, 100 μM) of L-ascorbic acid on embryo developmental rate of buffalo oocytes. The L-ascorbic acid was added to the maturation and culture media. In the second experiment, oocytes were cultured in media with two type of antioxidant (ascorbic acid + cysteamine) or ascorbic acid only.Results:There was a significant increase in cleavage rate at 25, 50 μM than 100 μM and control group. But, the blastocyst rate was higher at 50 μM ascorbic acid than other concentrations (0, 25, 100 μM). Supplementation of ascorbic acid and cysteamine to maturation and cultured media improved embryo development than ascorbic acid alone.Conclusions: Using of 50 μM L-ascorbic acid duringin vitro maturation and development improve the developmental competence of buffalo oocytes, this effect was increase with the presence of cysteamine.展开更多
A comparative study on the prevalence of Anaplasma parasite was conducted on ticks carrying buffaloes and cattle. Five hundred blood samples of both animals (250 of each) were collected during February, March and Apri...A comparative study on the prevalence of Anaplasma parasite was conducted on ticks carrying buffaloes and cattle. Five hundred blood samples of both animals (250 of each) were collected during February, March and April. Thin blood smears on glass slides were made, fixed in 100% methyl alcohol and examined. Microscopic examination revealed that 205 (41%) animals had Anaplasma parasites, out of which 89, 44 and 72 animals had Anaplasma marginale, Anaplasma centrale and mixed infection respectively. Infected buffaloes and cattle were 75 and 130 respectively. The infection in female was 53 and 92 in buffaloes and cattle respectively. Twenty-two and 92 blood samples of male were found positive in buffaloes and cattle respectively. Com- parative study revealed that the cattle were 26.82% more susceptible than buffaloes. The parasite prevailing percentage in female of both animals was slightly higher than that of the male. This investigation was aimed at studying the comparative prevalence of Anaplasma parasite in tick carrying buffaloes and cattle.展开更多
The insulin-like growth factor 1（IGF1） gene is a member of the group of somatotropin axis genes that play a significant role in cell proliferation and growth of muscles. Here, we searched for polymorphisms in buffal...The insulin-like growth factor 1（IGF1） gene is a member of the group of somatotropin axis genes that play a significant role in cell proliferation and growth of muscles. Here, we searched for polymorphisms in buffalo IGF1 and found two novel single nucleotide polymorphisms（SNPs）, G64 A and G280A, in the noncoding sequences of exon 1 and exon 4, respectively. Statistical analysis of different genotypes showed that the individuals with GG genotypes had significantly（P〈0.05） higher body weight（BW） and average daily gain（ADG） than those with other genotypes at ages of 3–6 months in G64A SNP and 6–9 months in G280A SNP. The combined genotypes of these two SNPs produced three haplotypes, GG/GG, AG/AG, and AA/AA, which were significantly associated（P〈0.0001） with BW and ADG at an age from 3 to 12 months. Buffaloes with the homozygous GG/GG haplotype showed higher growth performance than other buffaloes. The two SNPs were correlated with m RNA levels of IGF1 and IGF1 receptor（IGF1 R） in semitendinosus muscle as well as with the serum concentration level of IGF1. Also, buffaloes with GG/GG haplotype showed higher m RNA and serum concentration levels. The data revealed that these two SNPs could be valuable genetic markers for selection of Egyptian buffaloes for better performance in the population.展开更多
Objective:To assess the effects of melatonin and/or cysteamineonin vitro maturation, culturing and post-warming of buffalo embryos.Methods: Buffalo oocytes were classified into control, cysteamine (50 μM), melatonin ...Objective:To assess the effects of melatonin and/or cysteamineonin vitro maturation, culturing and post-warming of buffalo embryos.Methods: Buffalo oocytes were classified into control, cysteamine (50 μM), melatonin (10 ng/mL) and cysteamine (50 μM) + melatonin (10 ng/mL) treatment groups. In experiment 1, previous treatments were added duringin vitromaturation and culturing of buffalo oocytes.Results:Cleavage and blastocyst rates were significantly (P<0.05) increased in melatonin treated group (70.5±0.9 and 12.8±1.0, respectively). However this effect was potentiated when combined with cysteamine (74.0±1.7 and 14.8±1.7, respectively). In experiment 2, the treatements were added in maturtaion, culturing as well as post-warming culture media. Embryos at 7 d were vitrified.Viability assessement directly after warming showed significant increase (P<0.05) in cysteamine, melatonin and their combination groups (76.8±2.8, 80.0±2.1 and 83.3±1.7, respectively) than control (65.8±2.4);but the viability after 24 h post-warming was the best in cysteamine + melatonin combination group (61.4±2.1).Conculsions: Enriching maturation, culturing and post-warming media of buffalo oocytes and embryos with melatonin and/or cysteamine have significantly beneficial effects on oocyte developmental competence as well as embryos vitrification procedure outcomes which in turn resulting in enhancement of commercial buffalo embryo production.展开更多
[ Objectivel The paper aimed to investigate the expression pattern of bbu-miR-103-1 in buffalo （Bubalus bubalis） at lactation and non-lactation periods, and to predict its target gene and function. [ Method] Express...[ Objectivel The paper aimed to investigate the expression pattern of bbu-miR-103-1 in buffalo （Bubalus bubalis） at lactation and non-lactation periods, and to predict its target gene and function. [ Method] Expression pattern of bbu-miR-103-1 at lactation and non-lactation periods were detected by qRT-PCR. The precursor expression plasmid of bbu-miR-103-1 was constructed and named LpEZX-pre-miR-103-1. It was packaged and propagated to produce high-titer lenti- virus in 293T cell lines, which could be used to infect buffalo mammary epithelial cells （BMECs） and over express bbu-miR-103-1. The inhibitor of bbu-miR- 103-1 was chemically synthesized and transfected into BMECs to suppress bbu-miR-103-1 at the same time. The relative expression of pantothenate kinase 3 （ PANK3 ） and milk fat metabolism related genes were detected by qRT-PCR. [ Result] The relative expression of bbu-miR-103-1 at lactation period was 5.29 times higher than that at non-lactation period in buffalo （ P 〈 0.01 ）. The LpEZX-pre-miR-103-1 had been successfully constructed and packaged with the infection titer of 3.47×10^6 PFU/mL. Overexpress or suppress of bbu-miR-103-1 extremely down-regulated or up-regulated the expression level of PANK3 in BMECs （ P 〈 0.01 ）. Over expression of bbu-miR-103~l extremely enhanced the expression of Acetyl-CoA carboxylase alpha（ACACA）, Glycerol-3-phosphate acyhransferase 1 mitochon- drial （GPAM）, Diacylglycerol Oacyhransferase l （DGAT1） and Pyrnvate dehydrogenase lipoamide kinase isozyme 4 （PDK4） （P 〈0.01 ）, and also significantly up-regulated the expression of sterol regulatory element binding protein-1 c （SREBPI c）, Adipose differentiation-related protein （ADFP）, Cluster of differentiation 36 （ CD36）, Acetyl-CoA synthetase short-chain subfamily member 1 （ACSS1） （P 〈0.05）. Over expression of bbu-miR-103-1 down-regulated the expression of PANK3, and improved the mRNA level of SREBPlc by feedback regulation, finally promoting the de novo synthesis of fatty acid beginning with ACACA. [ Conclusion] bbu-miR-103-1 plays an important role in enhancing milk fatty acid synthesis, which provides a molecular base for revealing formation and regulatory mechanism of high-level milk fat in buffalo.展开更多
基金Supported by the Innovation Foundation For Postgraduate of Guangxi University(2008105930905D001) the Tackle Key Program in Science and Technology of Science and Technology Bureau of Guangxi Province(0815008-2-4)~~
文摘[Objective] The experiment aimed to set up a method for isolating and culturing endometrial stromal cells (BESC) and endometrial glandular epithelial cells(BEGEC) of buffalo as well as laid foundation for studying biological mechanism of embryo implantation and uterine diseases. [Method] The enzymatic digestion method, scraping method, serial filtration and differential velocity adherent technique were used to isolate BESC and BEGEC, then immunocytochemical method and TRYPAN-Blue assay were used to determine the purity and survival rate of isolated cells. [Result] The BESC and BEGEC were successfully isolated and cultured while immunocytochemical method and cell count method demonstrated that the purity was over 90%. The result of TRYPAN-Blue assay shown that survival rate of BESC and BEGEC was 91% and 78% respectively. [Conclusion] The enzymatic digestion method, scraping method, serial filtration and differential velocity adherent technique could isolate BESC and BEGEC with high purity.
基金Supported by Guangxi Scientific Fund Project (Guikezi0991042, Guikezi 0640015 and Guikezi 0832043)Guangxi Area Education Department Educational and Scientific Layout Project (C, 2006C3)+1 种基金Guangxi Education Department Scientific Research Fund (200709LX075)Guangxi Large Apparatus Collaborated Sharing Net~~
文摘To study the association of oxytocin (OT)'s distribution in hypothalamatic,pituitary and ovary,and understand how the OT secrete releasing in hypothalamus,pituitary and ovaries,the paraffin section immunohistochemistry SuperPicTureTM two step method was used to detect the distribution of OT in hypothalamatic-pituitary-ovary axis of five femal Guangxi local buffalo. The test results could provide morphology according to study the OT's synthesis and mechanism of action,and could play reference and directions part in breeding Guangxi local buffalo. The test results display:oxytocin immuno reactive (OT-IR) neuronsw eremainly distributed arcuate nucleus,supraoptic nucleus and paraventricular nucleus,and OT-IR neurons was also found in ventromedial nucleus,ventrolateralis nucleus,suprachiasmaticus nucleus,dorsomedial nucleus,mamillary body,anterior hypothalamic nucleus and so on. The OT immunoactive production was found in pituitary and few OT-IR nerve fibers extended to post pituitary from hypophyseal stalk and medium eminence. In ovaries,OT immunoactive productions were only distributed in germinal epithelium cells,granulosa cells and lutein cells. The OT was first discovered in singulorum link of hypothalamatic-pituitary-ovary axis of Guangxi local buffalo. The OT immunoactive neurons were first discovered in every main nucleus of Guangxi local buffalo hypothalamus,especially distributed in arcuate nucleus,supraoptic nucleus and paraventricular nucleus.
基金Supported by National High Technology Research and Development (863) Program of China (2011AA100607)National Transgenic Major Project of China (2010ZX08007003)~~
文摘[Objective] This study aimed to investigate the feasibility of transgenesis by injecting exogenous DNA into zygote cytoplasm of Buffalo. [Method] Buffalo oocytes were randomly divided into two groups 20-22 h after in vitro maturation. One group of oocytes was introduced with about 7.5 pl of 50 μg/ml DNA solution containing linear EGFP fragment by cytoplasmic injection 7-10 h or 18-20 h after in vitro fertilization （IVF）; the other group of oocytes was introduced with mixture of a single buffalo sperm and about 7.5 pl of 50 μg/ml DNA solution containing linear EGFP fragment by cytoplasmic injection （generally called ICSI-Mediated Gene Transfer, ICSI-Tr）. Expression of exogenous DNA was observed and recorded during the process of embryonic development. [Result] Early embryonic gene expression efficiency and blastocyst gene expression efficiency in IVF injection group showed no significant difference compared with that in ICSI-Tr group （P0.05）. In addition, the cleavage rate and early embryonic gene expression efficiency in IVF injection group were significantly higher with injection at 7-10 h post IVF than that at 18-20 h post IVF （P0.05）. [Conclusion] These results indicate that transgenic buffalo embryos can be generated by injecting exogenous DNA into cytoplasm of IVF oocytes, and the optimal injection time is 7-10 h post IVF.
基金supported by the National Key Technology R&D Program of China (2011BAD19B02-6)the Open Grant of Guangxi Provincial Key Laboratory of Water Buffalo Genetics, Breeding and Reproduction, China (SNKF-2012-04)
文摘The aim of this study was to investigate the feasibility of stimulating ovarian fol icle development in order to improve fertility in water buffalo cows by immunization against inhibin. The experiment was carried out in early summer (May) and included 24 multi-parity crossbred Murrah-Swamp buffaloes that were divided into immunized (n=11) and control (n=13) groups. Each immunized cow was administered with a 2-mL immunogen of mineral oil adjuvant containing 2 mg of recombinant inhibinα-subunit fusion protein. The controls were treated with the adjuvant only. Al animals received Ovsynch protocol treatment, starting on the day of the antigen administration, and they were artiifcial y inseminated upon behavioral estrus. As a result, al of the immunized buffaloes generated antibodies against inhibin during the experimental period and had higher plasma concentrations of fol icle-stimulating hormone (FSH), activin, and estradiol (E2) related to estrous expression. A higher proportion of immunized animals expressed estrus behavior than did the controls (72%vs. 30%, P<0.05). On aver-age, inhibin-immunized buffaloes had signiifcantly more large fol icles (≥9 mm in diameter) than the controls (mean±SEM;1.2±0.1 vs. 0.84±0.1, respectively;P<0.05) and a slightly higher mean total number of fol icles (≥2 mm;11.4±0.7 vs. 9.0±1.1, respectively;P=0.09) and smal (2–4 mm) fol icles (8.81±0.6 vs. 6.84±1.0, respectively;P=0.12). A higher percentage of cows ovulated in the immunized group than in the control group (91%(10/11) vs. 54%(7/13), respectively;P<0.05). Moreover, inhibin-immunized cows had slightly larger corpus luteum (CL) than the controls 9 days after ovulation and signiifcantly higher (P<0.01) post-ovulation peak plasma progesterone (P4) concentrations. Immunization against inhibin also mar-ginal y increased the conception rate 42 days after insemination (45.8%vs. 15.4%;P>0.05). These results demonstrate that immunization against inhibin, coupled with the treatment with the Ovsynch protocol, can constitute a new technique to increase fertility in water buffalo cows.
文摘Changes of concentrations were studied in water buffaloes and goats infected with Fasciola he-patica on blood NO(nitric oxide) and TNF-α(tumor necrosis factor-α). Twenty healthy male castrated water buffaloes of 2 - 3 years old and weighing 300 - 500 kg as well as six goats were confirmed free of fasciolosis by fecal examination and Dot-ELISA. Two studies were conducted using the water buffaloes. In the first experiment, 8 water buffaloes were randomly divided into control group (n = 3) and infection group (n = 5). Each buffalo in the infected group received orally 60 metacercariae of F. hepatica per day for 20 days (total 1 200 metacercariae) to produce a chronic infection. In the second experiment, 12 water buffaloes were randomly divided into infected (n=9) and control group (n = 3). Each buffalo in the infected group was given a single oral dose of 1 600 metacercariae to produce an acute infection. The 6 goats were randomly divided into two infected groups and a control group. The sheep in two infections received a single oral dose of 200 and 500 metacercariae respectively, the control group remained uninfected. Blood NO and TNF-a concentrations of the test animals were measured by a reductive enzyme assay and RIA, respectively. Blood NO concentration in both acutely and chronically infected water buffaloes progressively increased from week 3 post-infection and was significantly greater than that of the control group (P<0. 05) at the 5th week (acute infection) and 7th week (chronic infection), and remaining at higher concentration for the remaining period of the studies. Blood TNF-αconcentrations in both chronically and acutely infected water buffaloes also increased after infection. In the goat experiment, plasma NO concentrations in both infection groups increased from week 3 after infection, and remained higher than that of the control group until the end of the experiment. TNF-α concentrations in goats in infection group 1 and 2 gradually increased after infection and were significantly greater than those of the control group from the 9th to 11th week and from the 11th to 15th week respectively.
文摘Objective:To improvein vitro embryo production in buffalo by supplementation of L-ascorbic acid during maturation and development (experiment 1) and combination with another antioxidant as cysteamine (experiment 2).Methods:Two experiments were performed, the first one aimed to evaluate the different concentrations (0, 25, 50, 100 μM) of L-ascorbic acid on embryo developmental rate of buffalo oocytes. The L-ascorbic acid was added to the maturation and culture media. In the second experiment, oocytes were cultured in media with two type of antioxidant (ascorbic acid + cysteamine) or ascorbic acid only.Results:There was a significant increase in cleavage rate at 25, 50 μM than 100 μM and control group. But, the blastocyst rate was higher at 50 μM ascorbic acid than other concentrations (0, 25, 100 μM). Supplementation of ascorbic acid and cysteamine to maturation and cultured media improved embryo development than ascorbic acid alone.Conclusions: Using of 50 μM L-ascorbic acid duringin vitro maturation and development improve the developmental competence of buffalo oocytes, this effect was increase with the presence of cysteamine.
文摘A comparative study on the prevalence of Anaplasma parasite was conducted on ticks carrying buffaloes and cattle. Five hundred blood samples of both animals (250 of each) were collected during February, March and April. Thin blood smears on glass slides were made, fixed in 100% methyl alcohol and examined. Microscopic examination revealed that 205 (41%) animals had Anaplasma parasites, out of which 89, 44 and 72 animals had Anaplasma marginale, Anaplasma centrale and mixed infection respectively. Infected buffaloes and cattle were 75 and 130 respectively. The infection in female was 53 and 92 in buffaloes and cattle respectively. Twenty-two and 92 blood samples of male were found positive in buffaloes and cattle respectively. Com- parative study revealed that the cattle were 26.82% more susceptible than buffaloes. The parasite prevailing percentage in female of both animals was slightly higher than that of the male. This investigation was aimed at studying the comparative prevalence of Anaplasma parasite in tick carrying buffaloes and cattle.
基金Project supported by the Science Technology Development Fund(STDF,No.2585),Ministry of Scientific Research,Egypt
文摘The insulin-like growth factor 1（IGF1） gene is a member of the group of somatotropin axis genes that play a significant role in cell proliferation and growth of muscles. Here, we searched for polymorphisms in buffalo IGF1 and found two novel single nucleotide polymorphisms（SNPs）, G64 A and G280A, in the noncoding sequences of exon 1 and exon 4, respectively. Statistical analysis of different genotypes showed that the individuals with GG genotypes had significantly（P〈0.05） higher body weight（BW） and average daily gain（ADG） than those with other genotypes at ages of 3–6 months in G64A SNP and 6–9 months in G280A SNP. The combined genotypes of these two SNPs produced three haplotypes, GG/GG, AG/AG, and AA/AA, which were significantly associated（P〈0.0001） with BW and ADG at an age from 3 to 12 months. Buffaloes with the homozygous GG/GG haplotype showed higher growth performance than other buffaloes. The two SNPs were correlated with m RNA levels of IGF1 and IGF1 receptor（IGF1 R） in semitendinosus muscle as well as with the serum concentration level of IGF1. Also, buffaloes with GG/GG haplotype showed higher m RNA and serum concentration levels. The data revealed that these two SNPs could be valuable genetic markers for selection of Egyptian buffaloes for better performance in the population.
文摘Objective:To assess the effects of melatonin and/or cysteamineonin vitro maturation, culturing and post-warming of buffalo embryos.Methods: Buffalo oocytes were classified into control, cysteamine (50 μM), melatonin (10 ng/mL) and cysteamine (50 μM) + melatonin (10 ng/mL) treatment groups. In experiment 1, previous treatments were added duringin vitromaturation and culturing of buffalo oocytes.Results:Cleavage and blastocyst rates were significantly (P<0.05) increased in melatonin treated group (70.5±0.9 and 12.8±1.0, respectively). However this effect was potentiated when combined with cysteamine (74.0±1.7 and 14.8±1.7, respectively). In experiment 2, the treatements were added in maturtaion, culturing as well as post-warming culture media. Embryos at 7 d were vitrified.Viability assessement directly after warming showed significant increase (P<0.05) in cysteamine, melatonin and their combination groups (76.8±2.8, 80.0±2.1 and 83.3±1.7, respectively) than control (65.8±2.4);but the viability after 24 h post-warming was the best in cysteamine + melatonin combination group (61.4±2.1).Conculsions: Enriching maturation, culturing and post-warming media of buffalo oocytes and embryos with melatonin and/or cysteamine have significantly beneficial effects on oocyte developmental competence as well as embryos vitrification procedure outcomes which in turn resulting in enhancement of commercial buffalo embryo production.
基金Supported by National Natural Science Foundation of China(31260552)National High-tech Research and Development Plan(863 plan)(2011AA100607)+1 种基金Selection of Excellent Ecological Forage Grass Varieties and Research and Demonstration of Carbon and Nitrogen Source of Fruit-grass Coupling System(GKH 14125008-2-13)Breeding and Popularization of National Approval New Forage Variety Pennisetum purpureum(GYMK 201453057)
文摘[ Objectivel The paper aimed to investigate the expression pattern of bbu-miR-103-1 in buffalo （Bubalus bubalis） at lactation and non-lactation periods, and to predict its target gene and function. [ Method] Expression pattern of bbu-miR-103-1 at lactation and non-lactation periods were detected by qRT-PCR. The precursor expression plasmid of bbu-miR-103-1 was constructed and named LpEZX-pre-miR-103-1. It was packaged and propagated to produce high-titer lenti- virus in 293T cell lines, which could be used to infect buffalo mammary epithelial cells （BMECs） and over express bbu-miR-103-1. The inhibitor of bbu-miR- 103-1 was chemically synthesized and transfected into BMECs to suppress bbu-miR-103-1 at the same time. The relative expression of pantothenate kinase 3 （ PANK3 ） and milk fat metabolism related genes were detected by qRT-PCR. [ Result] The relative expression of bbu-miR-103-1 at lactation period was 5.29 times higher than that at non-lactation period in buffalo （ P 〈 0.01 ）. The LpEZX-pre-miR-103-1 had been successfully constructed and packaged with the infection titer of 3.47×10^6 PFU/mL. Overexpress or suppress of bbu-miR-103-1 extremely down-regulated or up-regulated the expression level of PANK3 in BMECs （ P 〈 0.01 ）. Over expression of bbu-miR-103~l extremely enhanced the expression of Acetyl-CoA carboxylase alpha（ACACA）, Glycerol-3-phosphate acyhransferase 1 mitochon- drial （GPAM）, Diacylglycerol Oacyhransferase l （DGAT1） and Pyrnvate dehydrogenase lipoamide kinase isozyme 4 （PDK4） （P 〈0.01 ）, and also significantly up-regulated the expression of sterol regulatory element binding protein-1 c （SREBPI c）, Adipose differentiation-related protein （ADFP）, Cluster of differentiation 36 （ CD36）, Acetyl-CoA synthetase short-chain subfamily member 1 （ACSS1） （P 〈0.05）. Over expression of bbu-miR-103-1 down-regulated the expression of PANK3, and improved the mRNA level of SREBPlc by feedback regulation, finally promoting the de novo synthesis of fatty acid beginning with ACACA. [ Conclusion] bbu-miR-103-1 plays an important role in enhancing milk fatty acid synthesis, which provides a molecular base for revealing formation and regulatory mechanism of high-level milk fat in buffalo.
