水牛受精卵胞质内注射转基因的初步研究(英文)     

陈自洪  崔奎青  孟凡丽  刘玉兵  王丹  陆凤花  石德顺 《农业科学与技术》2012,(6):1167-1170,1174

[目的]旨在探讨通过向水牛受精卵胞质内注射外源DNA实现转基因的可行性。[方法]水牛卵母细胞体外成熟20～22h后随机分为2组,①在体外受精7～10或18～20h后向卵胞质内注入约7.5pL50μg/ml含线性EGFP片段的DNA溶液;②则分别注入单个精子与约7.5pL50μg/ml含线性EGFP片段的DNA混合物,观察外源基因在胚胎发育过程中的表达情况。[结果]受精卵胞质内注射的早期胚胎基因表达率、囊胚基因表达率与ICSI-Tr差异不显著(P>0.05),且IVF7～10h时注射的分裂率、早期胚胎基因表达率均显著高于18～20h(P<0.05)。[结论]水牛IVF受精卵胞质内注射外源基因能获得转基因胚胎,且IVF后7～10h注射的效果优于IVF后18～20h注射。  相似文献   

Milk progesterone enzyme-linked immunosorbent assay as a tool to investigate ovarian cyclicity of water buffaloes in relation to body condition score and milk production     

Turgish A Banu  Mohammed Shamsuddin  Jayonta Bhattacharjee  Mohammad F Islam  Saiful I Khan  Jalal U Ahmed 《Acta veterinaria Scandinavica》2012,54(1):30

Background

Application of assisted reproductive technologies in buffaloes is limited to some extent by farmers’ inability to detect oestrus because of its poor expression. The present study aimed at investigating reliability of a milk progesterone enzyme-linked immunosorbent assay (ELISA) to assess the ovarian cyclicity during post partum, oestrus and post-breeding periods in water buffaloes.

Methods

Progesterone concentrations were measured by an ELISA in milk of 23 postpartum buffaloes in relation to oestrus, pregnancy, body condition score (BCS) and milk production. Two milk samples were taken at 10 days intervals, every month starting from day 30 and continued to day 150 post partum. BCS and milk production were recorded during sample collection. Milk samples from bred buffaloes were collected at Day 0 (day of breeding), Days 10–12 and Days 22–24. Defatted milk was preserved at −80°C until analysis. Pregnancy was confirmed by palpation per rectum on Days 70–90.

Results

Seventeen buffaloes had 47 ovulatory cycles, one to four in each, 13 were detected in oestrus once (28 % oestrus detection rate). Progesterone concentration ≥1 ng/ml in one of the two 10-day-interval milk samples reflected ovulation and corpus luteum formation. The intervals between calving to first luteal activity and to first detected oestrus varied from 41 to 123 days (n = 17) and 83 to 135 (n = 13) days, respectively. Eight buffaloes were bred in the course of the study and seven were found pregnant. These buffaloes had a progesterone profile of low (<1 ng/ml), high (≥ 1 ng/ml) and high (≥ 1 ng/ml) on Day 0, Days 10–12 and Days 22–24, respectively. Buffaloes cycling later in the postpartum period had fewer missed oestruses (P < 0.05). Buffaloes with a superior BCS had a shorter calving to oestrus interval and produced more milk (P < 0.05).

Conclusions

Milk progesterone ELISA is a reliable tool for monitoring ovarian cyclicity and good BCS may be an indicator of resuming cyclicity in water buffalo.  相似文献   

FSH up-regulates angiogenic factors in luteal cells of buffaloes     

L.A. Fátima  M.C. Evangelista  R.S. Silva  A.P.M. Cardoso  P.S. Baruselli  P.C. Papa 《Domestic animal endocrinology》2013

Follicle-stimulating hormone has been widely used to induce superovulation in buffaloes and cows and usually triggers functional and morphologic alterations in the corpus luteum (CL). Several studies have shown that FSH is involved in regulating vascular development and that adequate angiogenesis is essential for normal luteal development. Angiogenesis is regulated by many growth factors, of which vascular endothelial growth factor (VEGF) and fibroblast growth factor 2 (FGF2) have an established central role. Therefore, we have used a combination of in vitro and in vivo studies to assess the effects of FSH on the expression of VEGF and FGF2 and their receptors in buffalo luteal cells. The in vivo model consisted of 12 buffalo cows, divided into control (n = 6) and superovulated (n = 6) groups, and CL samples were collected on day 6 after ovulation. In this model, we analyzed the gene and protein expression of FGF2 and its receptors and the protein expression of VEGFA systems with the use of real-time PCR, Western blot analysis, and immunohistochemistry. In the in vitro model, granulosa cells were collected from small follicles (diameter, 4–6 mm) of buffaloes and cultured for 4 d in serum-free medium with or without FSH (10 ng/mL). To induce in vitro luteinization, LH (250 ng/mL) and fetal bovine serum (10%) were added to the medium, and granulosa cells were maintained in culture for 4 d more. The progesterone concentration in the medium was measured at days 4, 5, and 8 after the beginning of cell culture. Cells were collected at day 8 and subjected to real-time PCR, Western blot analysis, and immunofluorescence for assessment of the expression of FGF2, VEGF, and their receptors. To address the percentage of steroidogenic and growth factor-expressing cells in the culture, flow cytometry was performed. We observed that in superovulated buffalo CL, the FGF2 system mRNA expression was decreased even as protein expression was increased and that the VEGF protein was increased (P < 0.05). In vitro experiments with granulosa cells showed an increase in the mRNA expression of VEGF and FGF2 and its receptors 1 and 2 and protein expression of VEGF, kinase insert domain receptor, FGF receptor 2, and FGF receptor 3 in cells treated with FSH (P < 0.05), in contrast to the in vivo experiments. Moreover, the progesterone production by FSH-treated cells was elevated compared with untreated cells (P < 0.05). Our findings indicate that VEGF, FGF2, and their receptors were differentially regulated by FSH in vitro and in vivo in buffalo luteal cells, which points toward a role of CL environment in modulating cellular answers to gonadotropins.  相似文献   

Cryptosporidiosis in buffalo calves (<Emphasis Type="Italic">Bubalus bubalis</Emphasis>): Prevalence and potential risk factors   总被引：1，自引：0，他引：1  

El-Khodery SA  Osman SA 《Tropical animal health and production》2008,40(6):419-426

The objective of the present study was to describe the prevalence and risk factors associated with cryptosporidiosis in buffalo calves in Middle Egypt. During one year, 458 fecal samples were collected from buffalo calves less than 3 month age in 55 small scale herds and examined for the presence of Cryptosporidium oocysts. Data describing age, gender, season, and herd management practices were gathered to assess potential risk factors. Fecal examination showed that 14.19% of the examined calves were positive for Cryptosporidium spp. Calves at 1-15 days were at the highest risk (P < 0.001), and a significant relationship between season and infection (P < 0.05) was recorded. A significant association between infection and hygiene (P < 0.001), type of floor (P < 0.01) and source of water (P < 0.01) was also recorded. Statistical analysis concerning the clinical signs and fecal characteristics revealed a significant association with fecal consistency (P < 0.001), presence of blood (P < 0.01) and mucous (P < 0.01). Moreover, a significant association was found between infection and the desire for suckling (P < 0.05) and tenesmus (P < 0.05). The results of the present study demonstrated the strong relation between infections by Cryptosporidium spp. and diarrhea in buffalo calves.  相似文献   

Selenium and vitamin E increases polymorphonuclear cell phagocytosis and antioxidant levels during acute mastitis in riverine buffaloes   总被引：1，自引：0，他引：1  

Reena Mukherjee 《Veterinary research communications》2008,32(4):305-313

Antioxidant, antiinflammatory and phagocytic activities were studied in milk polymorphonuclear cells (PMNs) isolated from healthy buffaloes (group I) and during clinical mastitis with the treatment of Enrofloxacin alone (group II) and combined treatment with Enrofloxacin and Vitamin E plus selenium (group III). On days 0,3, 8 and 15 the milk Somatic cell count (SCC) were significantly higher in mastitic milk than in milk obtained from healthy buffaloes. In group II SCC decreased significantly on day 3 and day 8, however in group III reduction in SCC was observed on day 3, day 8 and day 15 (P < 0.05). The antiinflammatory activity was evaluated by determining nitrite plus nitrate (NOx) production in the milk PMNs before treatment and on day 8. NOx activity was significantly higher in mastitic milk than from healthy controls, both before and after treatment (P < 0.05). In group II and group III the activity decreased significantly on day 8 (P < 0.05). The Glutathione peroxidase (GSH-Px) activity was estimated in the milk polymorphonuclear cell (PMNs) supernatant. GSH-Px activity was significantly lower in mastitic buffaloes than in healthy controls, both before and after treatment (P < 0.05). In group II levels did not change in response to treatment, whereas in group III levels had increased significantly on day 8 (P < 0.05). The phagocytic activity (PA) (percentage of neutrophil that had phagocytosed 1–6 bacteria) and phagocytic index (PI) (average number of bacteria/ leukocytes counted in 100 cells) of the milk PMNs was significantly lower in mastitic buffaloes (P < 0.05). In group II the PA and PI did not change in response to treatment, whereas in group III both the parameters had increased significantly on day 8 (P < 0.05). The results of the present experiment indicated enhancement of antioxidative and cellular defense and reduction of somatic cell count in the mastitic animals treated with Enrofloxacin and Vitamin E plus Selenium as compared to the Enrofloxacin treatment alone. Hence Vitamin E plus selenium therapy may be added along with the antibiotics for effective amelioration of intramammary infection in buffaloes.  相似文献   

Effects of Saccharomyces cerevisiae on in vivo organic matter digestibility and milk yield in buffalo cows     

Giuseppe Campanile  Fabio Zicarelli  Domenico Vecchio  Corrado Pacelli  Gianluca Neglia  Anna Balestrieri  Rossella Di Palo  Federico Infascelli 《Livestock Science》2008,114(2-3):358-361

The influence of a dietary supplementation with Saccharomyces cerevisiae (SC) during the first and the second phase of lactation on dry matter (DM) intake, organic matter digestibility, milk yield and quality and haematological profile was evaluated in buffalo cows. Lactating buffaloes (n = 190), 118.7 days in milk (DIM), were randomly divided into Group C (control, n = 95) and Group T (fed diet supplemented with 98 billion CFU of S. cerevisiae, n = 95). Eight buffaloes for each group (Groups T1 and C1), 85.4 DIM, were used to study the in vivo digestibility and the haematological profile. No differences were found for DM intake (16.5 kg·day^− 1) and haematological profile. The SC supplementation increased milk yield (7.9 ± 0.2 vs. 7.4 ± 0.2; P < 0.01) but did not affect milk fat and protein. SC supplementation increased OM digestibility, mainly, in the first phase of lactation (< 135 days), thus allowing a higher energy availability for milk yield and reduced fat mobilization.  相似文献   

Progesterone and pregnancy status of buffaloes treated with a GNRH agonist     

G. Campanile  D. Vecchio  G. Neglia  R. Di Palo  A. Prandi  M.J. D''Occhio 《Livestock Science》2008,115(2-3):242-248

The primary aim of the present study was to establish whether the treatment with a GnRH agonist on Day 5 after AI may result in the formation of an accessory corpus luteum, greater progesterone secretion, and the increased likelihood of pregnancy success in buffaloes. The study was conducted during a period of increasing daylight length when progesterone secretion is suppressed and embryonic mortality is relatively high in buffaloes. In Experiment 1, treatment with a GnRH agonist (buserelin, 12 μg) on Day 5 after AI induced acute increases in circulating concentrations of LH, FSH and oestradiol-17β. Pregnant buffaloes (n = 14) at Day 40 following AI showed an increase (P < 0.01) in milk whey progesterone concentration between Day 5 (310 ± 55 pg/ml) and Day 15 (424 ± 50 pg/ml). The non-pregnant buffaloes (n = 7) showed a decrease (P < 0.01) in progesterone level from Day 5 (410 ± 87 pg/ml) to Day 15 (188 ± 30 pg/ml) following AI. In Experiment 2, the treatment with buserelin (12 μg) on Day 5 after AI induced ovulation in 62% of the buffaloes (31/50) and these buffaloes showed a progressive increase in milk whey progesterone concentration on Day 10, 15 and 20 of pregnancy. Buffaloes that did not ovulate, recorded a relatively constant milk whey progesterone level from Day 10 to Day 20 following AI. Milk whey progesterone concentrations increased after the administration of the GnRH agonist in 97% of the pregnant buffaloes and 68% of the non-pregnant buffaloes. The diameter of the largest follicle in buffaloes that ovulated (ovulated n = 31) (8.9 ± 0.04 mm; range 4.2 – 13.0 mm) did not differ significantly from the diameter of the largest follicle in buffaloes that did not ovulate (not ovulate n = 19) (8.7 ± 0.04 mm; range: 4.0 – 12.0 mm). The latter observation suggested that notional ovulatory size follicles in buffaloes are heterogeneous with respect to stage of follicle maturation and capacity to respond to plasma LH. The present study showed that treatment with a GnRH agonist on Day 5 following AI provides a strategy to increase progesterone secretion and the likelihood of pregnancy in buffaloes mated during periods of increasing daylight length.  相似文献   

An outbreak of neonatal enteritis in buffalo calves associated with astrovirus     

Paolo Capozza  Vito Martella  Gianvito Lanave  Cristiana Catella  Georgia Diakoudi  Farzad Beikpour  Michele Camero  Barbara Di Martino  Giovanna Fusco  Anna Balestrieri  Giuseppe Campanile  Krisztian Banyai  Canio Buonavoglia 《Journal of veterinary science (Suw?n-si, Korea)》2021,22(6)

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水牛乳β-酪蛋白基因多态性与消化性能及抗氧化性能的关联研究     

赵烜影  郭鸰  任大喜  李楠  李玲  刘振民 《中国奶牛》2022,(1):12-16

本研究以水牛乳β-酪蛋白(β-CN)基因多态性分析结果为依据,从水牛乳中分离并纯化出具有活性的β-酪蛋白亚型.采用体外消化模型分析其在胃肠道中的消化性能;根据水解液的抗氧化活性来分析生物活性肽的释放.高效液相色谱法(HPLC)结果显示,水牛乳中β-CN有A和B两种等位基因.消化性能结果显示A等位基因更利于β-酪蛋白在肠...  相似文献   

Detection of Coxiella burnetii in buffaloes aborted fetuses by IS111 DNA amplification: A preliminary report   总被引：1，自引：0，他引：1  

A.G. Perugini  F. Capuano  A. Esposito  C. Marianelli  A. Martucciello  G. Iovane  G. Galiero 《Research in veterinary science》2009,87(2):189-191

The aim of this study was to evaluate by PCR analyses the presence of Coxiella burnetii infection in fetuses of water buffalo (Bubalus bubalis) in the Campania region (Southern Italy). Samples were collected only from aborted fetuses and C. burnetii presence was evaluated by one-tube nested PCR amplification of the IS111 repetitive element. Of the 164 fetuses examined 14 (17.5%) were positive after DNA amplification, showing that C. burnetii occurs in this population of water buffaloes.However, more extensive prevalence studies need to be carried out to define the role of buffaloes as reservoirs for this pathogen and also the role of C. burnetii as an abortive agent in this animal.  相似文献