Objective: To evaluate the effects of reduced glutathione on the quality of cryopreserved Boer buck spermatozoa. Methods: The current study was carried out on five Boer bucks from which semen samples were collected by...Objective: To evaluate the effects of reduced glutathione on the quality of cryopreserved Boer buck spermatozoa. Methods: The current study was carried out on five Boer bucks from which semen samples were collected by artificial vagina. After microscopical evaluation at 37 ℃, semen samples that fulfill the ideal requirements for extension were diluted in a tris–based extender including different concentrations of reduced glutathione (2, 5, 7 and 10 mM) and those without glutathione served as a control. Sperm motility, viability, acrosome integrity, DNA integrity, total antioxidant capacity and lipid peroxidation were assessed post-thawing. Results: The current results revealed that post-thawing motility, viability and acrosomal integrity were significantly improved [(66.67±5.50)%, 168.30±18.59 and (12.75±2.45)%, respectively] when 5 mM glutathione was added to semen extender;especially as compared with the control ((40.00 ±2.88)%, 95.00±8.90 and (25.75±3.46)%, respectively)Similarly, at this concentration (5 mM) sperm DNA damage, tail length and tail moment of cryopreserved semen were significantly (P<0.05) reduced [(2.32±0.27)%, (1.64±0.49) μm and 3.55±0.63, respectively] compared with the control extender ((6.66±0.84)%, (4.09±0.47) μm and 26.47±0.51, respectively)Moreover, addition of 5 mM glutathione to buck semen extender significantly (P<0.05) increased total antioxidant capacity [0.51±0.07) mμ/mL] and decreased lipid peroxidation of cryopreserved spermatozoa [(8.68±2.72) nmol/mL] compared with the control ((0.18±0.02) mμ/mL and (24.92±5.80) nmol/mL, respectively)Conclusions: The addition of 5 mM glutathione to semen diluent improve freezability of Boer buck spermatozoa through DNA protection from deterioration and oxidative stress reduction. Moreover, 10 mM of glutathione exerts cytotoxic effects on Boer buck semen.展开更多
Mycoplasma species(spp.)are bacteria that are difficult to detect.Currently,the polymerase chain reaction(PCR)is considered the most effective diagnostic tool to detect these microorganisms in both human and veterinar...Mycoplasma species(spp.)are bacteria that are difficult to detect.Currently,the polymerase chain reaction(PCR)is considered the most effective diagnostic tool to detect these microorganisms in both human and veterinary medicine.There are 13 known species of human Mycoplasma and 15 species of canine Mycoplasma.Owing to the difficulties in identifying the individual species of Mycoplasma,there is a lack of information regarding which species are saprophytic and which are pathogenic.The prevalence of the individual species is also unknown.In addition,in both humans and dogs,the results of some studies on the impact of Mycoplasma are conflicting.The presence of Mycoplasma spp.on the epithelium of reproductive tract is often associated with infertility,although they are also detected in healthy individuals.The occurrence of Mycoplasma spp.is more common in dogs(even 89%)than in humans(1.3%–4%).This is probably because the pH of a dog's genital is more conducive to the growth of Mycoplasma spp.than that of humans.Phylogenetically,human and canine Mycoplasma are related,and majority of them belong to the same taxonomic group.Furthermore,40%of canine Mycoplasma spp.are placed in common clusters with those of human.This suggests that species from the same cluster can play a similar role in the canine and human reproductive tracts.This review summarizes the current state of knowledge about the impact of Mycoplasma on canine and human male fertility as well as the prospects of further development in this field.展开更多
Objective:To evaluate the cryoprotective effect of butylated hydroxytoluene on buck frozen semen.Methods:Semen was collected from Boer(n=6)and Zaraibi(n=6)bucks by electroejaculator for 5 weeks.Semen aliquots were dil...Objective:To evaluate the cryoprotective effect of butylated hydroxytoluene on buck frozen semen.Methods:Semen was collected from Boer(n=6)and Zaraibi(n=6)bucks by electroejaculator for 5 weeks.Semen aliquots were diluted at 38℃in Tris-buffer with egg yolk 15.0%(vol/vol)(Tris-egg yolk extender)or soya lecithin 2.5%(weight/vol)(Tris-soya lecithin extender)supplemented with butylated hydroxytoluene at 0.0(as the control),0.5,1.0,2.0 and 4.0 mM.Post-thawing motility(at 400×magnification),plasma(hypo-osmotic swelling test),acrosome(Trypan blue/Giemsa dual staining)membranes,DNA(comet assay),and lipid peroxidation(by malondialdehyde concentration)were assessed.Results:Spermatozoa motility was enhanced by butylated hydroxytoluene in Tris-soya lecithin extender at 0.5 mM in the two breeds,and in Tris-egg yolk extender at 1.0 mM in Boer and at 2.0 mM in Zaraibi bucks for up to 3 h post-thawing.Plasma and acrosome membranes and DNA integrity of the two breeds were maximally high with butylated hydroxytoluene at 1.0-2.0 mM in Tris-egg yolk extender and at 0.5-1.0 mM in Tris-soya lecithin extender.Lipid peroxidation was minimal with butylated hydroxytoluene at 1.0-2.0 mM in Tris-egg yolk and soya lecithin extenders in the two breeds.Butylated hydroxytoluene at 4.0 mM deteriorated spermatozoa motility,and plasma and acrosome membranes.Conclusions:The consequence of butylated hydroxytoluene on buck frozen-thawed spermatozoa varies with the levels of supplementation,buck breed,and phospholipid source in the extender.Semen parameters of Boer buck are better in their response to butylated hydroxytoluene than Zaraibi buck.Butylated hydroxytoluene at 1.0 and 2.0 mM in Tris-egg yolk extender,and at 0.5 mM in Tris-soya lecithin extender represents the best concentrations and profitably improves the semen quality of buck semen.展开更多
Background： Cytoplasmic injection of exogenous DNA into zygotes is a promising technique to generate transgenic livestock. However, it is still relatively inefficient and has not yet been demonstrated to work in buff...Background： Cytoplasmic injection of exogenous DNA into zygotes is a promising technique to generate transgenic livestock. However, it is still relatively inefficient and has not yet been demonstrated to work in buffalo. We sought to improve two key technical parameters of the procedure, namely i） how much linear DNA to inject and ii） when to inject it. For this, we introduced a constitutively expressed enhanced green fluorescent protein （EGFP） plasmid into buffalo zygotes. Results： First, we found that the proportion of EGFP-expressing blastocysts derived from zygotes injected with 20 or 50 ng/pL DNA was significantly higher than from those injected with 5 pg/mL. However, 50 ng/pL exogenous DNA compromised blastocyst development compared to non-injected IVF controls. Therefore the highest net yield of EGFP-positive blastocysts was achieved at 20 ng/pL DNA. Second, zygotes injected early （7-8 h post-insemination [hpi]） developed better than those injected at mid （12-13 hpi） or late （18-19 hpi） time points. Blastocysts derived from early injections were also more frequently EGFP-positive. As a consequence, the net yield of EGFP-expressing blastocysts was more than doubled using early vs late injections （16.4 % vs 7.7 %）. With respect to blastocyst quality, we found no significant difference in cell numbers of EGFP-positive blastocysts vs non-injected blastocysts. Following embryo transfer of six EGFP-positive blastocysts into four recipient animals, two viable buffalo calves were born. Biopsied ear tissues from both buffalo calves were analyzed for transgene presence and expression by Southern blot, PCR and confocal laser scanning microscopy, respectively. This confirmed that both calves were transgenic. Conclusions： Our cytoplasmic injection protocol improved generation of transgenic embryos and resulted in the first transgenic buffalo calves produced by this method.展开更多
Many countries in the world consider the buffalo as a priority animal for the future, since it plays a pivotal role in human food sustainability. Even though Food and Agriculture Organization has termed the buffalo as...Many countries in the world consider the buffalo as a priority animal for the future, since it plays a pivotal role in human food sustainability. Even though Food and Agriculture Organization has termed the buffalo as an important undervalued asset, this species has yet to drive the same attention as cattle. Egypt has a wealth of buffaloes dispersed in small herds all over the country, so the efforts that have been made to improve their genetic background show little return. Contrarily, other countries concerned with buffalo improvement have already used a data recording system in buffalo herds, allowing to achieve a much faster improvement progress. This review intends to survey the existing information on the application of assisted reproduction techniques to improve buffalo productivity. The strength points that may help to improve buffalo production are identified, and the obstacles hindering the genetic improvement of Egyptian buffalo are characterized. Therefore, this work will gather information related to buffalo and compile it for an audience of researchers and specialists to enforce international collaboration for the development of buffalo production. Also, it will open the way for people interested in developing a future vision for buffalo potential, which will be helpful to close or minimize the biological gaps of buffaloes' researches.展开更多
Objective:To isolate and identify microflora and fungal species at different phases during estrus synchronization of ewes and estimate their prevalence;compare the effectiveness of antimicrobial administration to intr...Objective:To isolate and identify microflora and fungal species at different phases during estrus synchronization of ewes and estimate their prevalence;compare the effectiveness of antimicrobial administration to intravaginal sponge on the changes in the vaginal micro-organisms and reproductive performance.Methods: Sixty Egyptian ewes were allocated into three equal groups (G: 1, 2 and 3). G1 was inserted with vaginal sponge containing medroxy-progesterone acetate and served as control;without antimicrobial additive. The other two groups were treated as G1, but sponges were previously injected with ciprofloxacin (G2), while sponges of G3 were injected with ciprofloxacin and clotrimazole. Vaginal swabs were collected from each treated ewe, prior sponge insertion, at sponge withdrawal and 48 h later for microbiological investigation and bacterial count. On the day of sponge removal, 300 IU/eCG was administered for each treated ewe. The identified bacterial strains before sponge insertion were tested for sensitivity with antimicrobial disks.Results:Bacterial isolates before sponge insertion were more sensitive to ciprofloxacin. Frequencies of ewes in estrus;the interval from sponge withdrawal to onset estrus and the duration of estrus were statistically similar among treated groups. The pregnancy rate in G2 (100%) was higher than G1 (66.7%) and G3 (82.4%). The total bacterial count before sponge insertion was similar between all treatments and increased significantly in all groups on the day of sponge withdraw. The prevailing bacteria on D0, D14 and 48 h after sponge removal for all treated groups wereStaphylococcus spp. followed byEscherichia coli. Regarding to fungus species, percentages of isolation increased from 5% (before sponge insertion) to 100.00% and 88.89% at sponge withdraw for G1 and G2, respectively. In G3, the fungus was declined from 10% (before sponge insertion) to 5% (at sponge removal).Conclusions:The concomitant treatments by antimicrobial to the vaginal sponge which used for estrus synchronization in ewes can improve reproductive performance.展开更多
Objective: To study the effect of adding different concentrations of soybean lecithin in Tris based extender and to compare the results of Tris soybean lecithin extender with two commercial diluents (egg yolk-based: B...Objective: To study the effect of adding different concentrations of soybean lecithin in Tris based extender and to compare the results of Tris soybean lecithin extender with two commercial diluents (egg yolk-based: BullXcell and plant-based: OptiXcell) for Damascus goat sperm cryopreservation. Methods: The ejaculates from 4 mature male Damascus goats were obtained by using an artificial vagina. Semen samples were pooled and diluted in Tris extender supplemented with soybean lecithin at different concentrations of 1.5%, 3.0%, 6.0% and 10.0% to get better concentration to be used for further experiments, with a final concentration of 240×106 spermatozoa/mL. Semen samples were packed in straws (0.25 mL), frozen by using an automated system and stored in liquid nitrogen at -196 ℃ for 48 h. After thawing (37 ℃/30 s), the samples were evaluated for sperm quality parameters, including sperm motility, membrane integrity and acrosome integrity. Malondialdehyde concentration was estimated as a marker for lipid peroxidation. Based on the previous investigations, only Tris extender supplemented with 3.0% soybean lecithin (based on its positive results) was used versus BullXcell and OptiXcell for sperm ultrastructure evaluation and artificial insemination by using electron microscope and artificial insemination of the synchronized does. Results: There was no significant difference between Tris-soybean lecithin at 3.0% and BullXcell/OptiXcell diluents in post-thaw sperm parameters and fertility following artificial insemination;meanwhile, the other concentrations of soybean lecithin (1.5%, 6.0% and 10%) showed lower sperm parameters following cryopreservation. Conclusions: Using of Tris-soybean lecithin based extender at the level of 3.0% can be an appropriate alternative to either BullXcell or OptiXcell for Damascus goat sperm cryopreservation.展开更多
Factors affecting the efficiency of nuclear transfer （NT） in rabbits were examined in the present study. When 100 V mm of pulse strength and 15 us of pulse duration were employed, 3 and 4 electronic pulses resulted ...Factors affecting the efficiency of nuclear transfer （NT） in rabbits were examined in the present study. When 100 V mm of pulse strength and 15 us of pulse duration were employed, 3 and 4 electronic pulses resulted in significantly more cytoplasts fused with donor cells compared with 2 electronic pulses （P〈 0.05）, but no significant difference was found in the cleavage rate of reconstructed embryos among the three groups （P〉0.05）. When the duration and number of electronic pulse were fixed at 15 ps and 3 times, increase of pulse intensity from 100 V mm 1 to 150 V mm^-1 and 200 V mm^-1 resulted in a significantly decrease in the cleavage rate of reconstructed embryos （P〈 0.05）, although the fusion rate did not significantly differ among the three groups （P〉 0.05）. Significantly more reconstructed embryos cleaved and developed to blastocysts when they were derived from donor embryos at the 8-16-cell stage, in comparison with the reconstructed embryos derived from donor embryos at the compact morula stage （P 〈 0.05）, although the fusion rate was similar （P 〉 0.05）. Activation of cytoplasts prior to fusion increased the cleavage rate （P〈 0.05） and blastocyst development （P〈 0.05） of reconstructed embryos, but decreased the fusion rate （P 〈 0.05） compared with cytoplasts activated post fusion. More reconstructed embryos developed to blastocysts when they were cultured in TCM ＋ 3% OCS at the first 48 h and then cultured in TCM199＋ 10% FCS, in comparison with the reconstructed embryos cultured in either TCM199＋ 10% FCS or TCM199＋3% OCS （P 〈 0.05）. When 22 NT embryos were transferred into the oviducts of one recipient rabbit, one recipient rabbit delivered a female rabbit at 34 days of gestation. In conclusion, either electrofusion parameter or developmental stage of donor embryos have a significant effect on the efficiency of NT, NT embryos require different concentration of serum at their different development stages.展开更多
[Objective] To clone ovarian-specific promoter-1 （OSP-1） fragment in rats and detect the tissue-specific expression of OSP-1 at cell level. [Method] According to the known OSP-1 sequence in rats, the specific pdmers...[Objective] To clone ovarian-specific promoter-1 （OSP-1） fragment in rats and detect the tissue-specific expression of OSP-1 at cell level. [Method] According to the known OSP-1 sequence in rats, the specific pdmers were designed. The OSP-1 fragment was amplified by PCR and compared with published OSP-1 sequence. After eukaryotic expression vector pOSP-1-EGFP-N1 was constructed by cloning OSP-1 promoter into the pEGFP-N1 vector without CMV, the recombinant plasmid was transfected into buffalo granulocytes, mammary epithelial cells and fetal fibroblast cells under mediation of liposome LipofectamineTM 2000. The green fluxorescent protein （GFP） expression was observed with the fluorescence microscope after transfection for 12, 24 and 48 h. [Result] The amplified OSP-1 fragment was 480 bp, and the homology to published rat OSP-1 sequence was 96%. The sequence analysis showed that this fragment contained a core promoter cis-element similar with TATA box and ChAT box, and multiple C/EBP beta transcription factor binding sites. The EGFP-expressing positive granulccytes were firstly observed at 12 h after transfection and they were increased. All the EGFP-expressing and non-expressing cells from granulocytes were large and round. After transfection for 48 h, the EGFP-expressing positive granulocytes were decreased. The EGFP was not expressed in buffalo mammary epithelial cell and fetal fibroblast cell. [ Conclusion] EGFP exDression controlled bv OSP-1 promoter can be found in buffao granulocvtes.展开更多
[Objective] The aim of this paper is to find the change law of reproductive hormones of female swamp buffalo in off season for estrus and to investigate the relationship between various hormones. [ Method] The concent...[Objective] The aim of this paper is to find the change law of reproductive hormones of female swamp buffalo in off season for estrus and to investigate the relationship between various hormones. [ Method] The concentrations of follicle stimulating hormone （FSH）, luteinizing hor- mone （ LH）, inhibin （ INH）, estradiol （ E2 ） and progesterone （ P4 ） were determined by the enzyme-linked immunosorbent assay （ELISA） in off season for estrue. The changes of follicles in the estrous cycle were examined using B-mode ultrasonography. [ Result] In off season for estrus of female swamp buffalo from Guangxi region, the blood concentration of E2 and P4 varied fluctuantly. The concentration of E2 had two peaks, respec- tively 3 d before ovulation and after 13 d post ovulation. The concentration of P4 increased slowly from the first day after ovulation, increased rapidly from 5 to 13 d, peaked on Day 16 and decreased to the baseline 3 d before ovulation. The concentrations of FSH, LH and INH roached a peak re- spectively after 10 d post ovulation, 2 d before ovulation, and after 4 d post ovulation. The changes of the hormones had correlations. The concen- trations of FSH, LH and INH had highly significant correlations with each other, while the concentration of INH had highly significant correlations with that of E2 or P4- The concentration of E2 also had a highly significant correlation with that of P4- [ Conclusionl The study can guide the breeding of swamp buffalo from Guanqxi reqion and also help to improve their roproduction performance.展开更多
Objective: To study the genetic and non-genetic factors and their interactions affecting growth rate and body weights at birth, weaning and at 6 months of age in Saudi Ardi, Damascus goats and their crosses. Methods: ...Objective: To study the genetic and non-genetic factors and their interactions affecting growth rate and body weights at birth, weaning and at 6 months of age in Saudi Ardi, Damascus goats and their crosses. Methods: Crossbreeding program between Saudi Ardi(A) goats with Damascus(D) was carried out to improve the meat productivity of Ardi goats through crossbreeding. The pedigree records of the body weights were obtained from 754 kids (397 males and 357 females) produced from 46 Sires and 279 Dams. Birth weight, weaning weigh and 6 months weight as well as average daily gain during different growth stages from birth to weaning (D1), weaning to 6 months (D2) and from birth to 6 months of age (D3) were recorded during winter/autumn and summer/spring. Data were classified according to breed, generation, sex, season, year, and type of birth. Data were analyzed using GLM procedure for the least-squares means of the fixed factors. Heritability and genetic parameters were estimated with derivative-free restricted maximum likelihood procedures using the MTDFREML program. Results: The percentages of variations were moderate for body weights and high for daily gains. Genetic groups had a highly significant (P<0.01) effect on the body weights traits. Damascus goats had higher (P<0.01) birth and weaning weights, but ?D?A group kids had a higher (P<0.01) body weight at 6 months. The genetic groups had a significant effects on the daily weight gains for D1 (P<0.01) and D3 (P<0.05) periods, whereas, it had no effects on D2 period. The fixed effects of sex, season, year and type of birth were significant differences for body weights. Male kids were heavier (P<0.01) than females for different growth stages. Body weights and daily gains during winter/autumn were significantly higher (P<0.01) than summer/spring. Kids born and raised as singles were significantly (P<0.01) heavier than those were born as twins or triplets. The genetic and phenotypic correlations between birth and weaning weights were positive for both Damascus and Ardi goats. Conclusions: Genetic program for Ardi goats through upgrading with Damascus is possible to improve meat production.展开更多
The human and bovine lactoferrin have been studied extensively, but very few reports have been published concerning camel lactoferrin (cLf). The present study aimed to isolate cLf and evaluate its efficiency including...The human and bovine lactoferrin have been studied extensively, but very few reports have been published concerning camel lactoferrin (cLf). The present study aimed to isolate cLf and evaluate its efficiency including antimicrobial activity and immunomodulator effects. cLf isolation was attempted from camel milk whey using a cation exchange chromatography by SP-Sepharose. The antimicrobial activity of the isolated cLf was investigated against Staphylococcus aureus (S. aureus), Streptococcus agalactiae (S. agalactiae), Escherichia coli (E. coli) and Pseudomonas aeruginosa (P. aerogenosa) strains. The immune effect of cLf was studied by lymphocyte transformation test. It was found that cLf was separated around molecular weight of 80 kDa and showed significant inhibitory effect against E. coli followed by P. aeruginosa, S. agalactiae and S. aureus. cLf increased lymphocyte transformations mean values in a dose dependant manner. The highest transformations mean value was determined at 50 μg/mL. In conclusion, these results suggest that cLf is a potent natural antimicrobial and novel immunomodulator agent.展开更多
Vitrification of immature oocytes at the germinal vesicle (GV) stage is important to preserve female gametes. The standard formula for vitrification solutions has long been a debate. Herein, we investigated the effect...Vitrification of immature oocytes at the germinal vesicle (GV) stage is important to preserve female gametes. The standard formula for vitrification solutions has long been a debate. Herein, we investigated the effect of the presence or absence of trehalose in vitrification solution on viability, in vitro maturation (IVM) rates, and development of vitrified/warmed immature dromedary camel oocytes. Cumulus oocyte complexes (COCs) obtained at slaughter from the ovaries of mature she-camels were randomly allocated into three groups;namely, control group, oocytes were directly subjected to IVM without vitrification, vitrification solution 1 (VS1) group, oocytes were vitrified in a solution composed of 25% ethylene glycol (EG) plus 25% dimethyl sulfoxide (DMSO) + 0.5 M trehalose;and vitrification solution 2 (VS2) group, oocytes were vitrified in a solution composed of 25% EG plus 25% DMSO. Vitrification of COCs was conducted by open pulled straws (OPS) method. Following vitrification and warming, morphologically viable oocytes were matured in vitro for 36 h. COCs were then fertilized and cultured in vitro for 7 days. The percentage of viable oocytes was significantly higher (P 0.05) in VS2 than VS1 group (80.0% vs. 63.3%, respectively). Nuclear maturation, cleavage (48 h post-insemination;pi), and blastocyst rates (7 days pi) were significantly higher (P < 0.05) in VS2 than in VS1 groups. No significant differences were observed in oocyte maturation and development rates between VS2 and control groups. In conclusion, vitrification of immature dromedary camel oocytes in trehalose-free solution (VS2) was more advantageous than that in trehalose supplemented media since it did not reduce viability and development.展开更多
文摘Objective: To evaluate the effects of reduced glutathione on the quality of cryopreserved Boer buck spermatozoa. Methods: The current study was carried out on five Boer bucks from which semen samples were collected by artificial vagina. After microscopical evaluation at 37 ℃, semen samples that fulfill the ideal requirements for extension were diluted in a tris–based extender including different concentrations of reduced glutathione (2, 5, 7 and 10 mM) and those without glutathione served as a control. Sperm motility, viability, acrosome integrity, DNA integrity, total antioxidant capacity and lipid peroxidation were assessed post-thawing. Results: The current results revealed that post-thawing motility, viability and acrosomal integrity were significantly improved [(66.67±5.50)%, 168.30±18.59 and (12.75±2.45)%, respectively] when 5 mM glutathione was added to semen extender;especially as compared with the control ((40.00 ±2.88)%, 95.00±8.90 and (25.75±3.46)%, respectively)Similarly, at this concentration (5 mM) sperm DNA damage, tail length and tail moment of cryopreserved semen were significantly (P<0.05) reduced [(2.32±0.27)%, (1.64±0.49) μm and 3.55±0.63, respectively] compared with the control extender ((6.66±0.84)%, (4.09±0.47) μm and 26.47±0.51, respectively)Moreover, addition of 5 mM glutathione to buck semen extender significantly (P<0.05) increased total antioxidant capacity [0.51±0.07) mμ/mL] and decreased lipid peroxidation of cryopreserved spermatozoa [(8.68±2.72) nmol/mL] compared with the control ((0.18±0.02) mμ/mL and (24.92±5.80) nmol/mL, respectively)Conclusions: The addition of 5 mM glutathione to semen diluent improve freezability of Boer buck spermatozoa through DNA protection from deterioration and oxidative stress reduction. Moreover, 10 mM of glutathione exerts cytotoxic effects on Boer buck semen.
文摘Mycoplasma species(spp.)are bacteria that are difficult to detect.Currently,the polymerase chain reaction(PCR)is considered the most effective diagnostic tool to detect these microorganisms in both human and veterinary medicine.There are 13 known species of human Mycoplasma and 15 species of canine Mycoplasma.Owing to the difficulties in identifying the individual species of Mycoplasma,there is a lack of information regarding which species are saprophytic and which are pathogenic.The prevalence of the individual species is also unknown.In addition,in both humans and dogs,the results of some studies on the impact of Mycoplasma are conflicting.The presence of Mycoplasma spp.on the epithelium of reproductive tract is often associated with infertility,although they are also detected in healthy individuals.The occurrence of Mycoplasma spp.is more common in dogs(even 89%)than in humans(1.3%–4%).This is probably because the pH of a dog's genital is more conducive to the growth of Mycoplasma spp.than that of humans.Phylogenetically,human and canine Mycoplasma are related,and majority of them belong to the same taxonomic group.Furthermore,40%of canine Mycoplasma spp.are placed in common clusters with those of human.This suggests that species from the same cluster can play a similar role in the canine and human reproductive tracts.This review summarizes the current state of knowledge about the impact of Mycoplasma on canine and human male fertility as well as the prospects of further development in this field.
文摘Objective:To evaluate the cryoprotective effect of butylated hydroxytoluene on buck frozen semen.Methods:Semen was collected from Boer(n=6)and Zaraibi(n=6)bucks by electroejaculator for 5 weeks.Semen aliquots were diluted at 38℃in Tris-buffer with egg yolk 15.0%(vol/vol)(Tris-egg yolk extender)or soya lecithin 2.5%(weight/vol)(Tris-soya lecithin extender)supplemented with butylated hydroxytoluene at 0.0(as the control),0.5,1.0,2.0 and 4.0 mM.Post-thawing motility(at 400×magnification),plasma(hypo-osmotic swelling test),acrosome(Trypan blue/Giemsa dual staining)membranes,DNA(comet assay),and lipid peroxidation(by malondialdehyde concentration)were assessed.Results:Spermatozoa motility was enhanced by butylated hydroxytoluene in Tris-soya lecithin extender at 0.5 mM in the two breeds,and in Tris-egg yolk extender at 1.0 mM in Boer and at 2.0 mM in Zaraibi bucks for up to 3 h post-thawing.Plasma and acrosome membranes and DNA integrity of the two breeds were maximally high with butylated hydroxytoluene at 1.0-2.0 mM in Tris-egg yolk extender and at 0.5-1.0 mM in Tris-soya lecithin extender.Lipid peroxidation was minimal with butylated hydroxytoluene at 1.0-2.0 mM in Tris-egg yolk and soya lecithin extenders in the two breeds.Butylated hydroxytoluene at 4.0 mM deteriorated spermatozoa motility,and plasma and acrosome membranes.Conclusions:The consequence of butylated hydroxytoluene on buck frozen-thawed spermatozoa varies with the levels of supplementation,buck breed,and phospholipid source in the extender.Semen parameters of Boer buck are better in their response to butylated hydroxytoluene than Zaraibi buck.Butylated hydroxytoluene at 1.0 and 2.0 mM in Tris-egg yolk extender,and at 0.5 mM in Tris-soya lecithin extender represents the best concentrations and profitably improves the semen quality of buck semen.
基金supported by the National Transgenic Project(2009ZX08007-009B, 2011ZX08007-003)Guangxi natural science funding(2012GXNSFCB053002)+1 种基金funding of State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources(KSL-CUSAb-2012-02)supported by AgResearch core funding
文摘Background： Cytoplasmic injection of exogenous DNA into zygotes is a promising technique to generate transgenic livestock. However, it is still relatively inefficient and has not yet been demonstrated to work in buffalo. We sought to improve two key technical parameters of the procedure, namely i） how much linear DNA to inject and ii） when to inject it. For this, we introduced a constitutively expressed enhanced green fluorescent protein （EGFP） plasmid into buffalo zygotes. Results： First, we found that the proportion of EGFP-expressing blastocysts derived from zygotes injected with 20 or 50 ng/pL DNA was significantly higher than from those injected with 5 pg/mL. However, 50 ng/pL exogenous DNA compromised blastocyst development compared to non-injected IVF controls. Therefore the highest net yield of EGFP-positive blastocysts was achieved at 20 ng/pL DNA. Second, zygotes injected early （7-8 h post-insemination [hpi]） developed better than those injected at mid （12-13 hpi） or late （18-19 hpi） time points. Blastocysts derived from early injections were also more frequently EGFP-positive. As a consequence, the net yield of EGFP-expressing blastocysts was more than doubled using early vs late injections （16.4 % vs 7.7 %）. With respect to blastocyst quality, we found no significant difference in cell numbers of EGFP-positive blastocysts vs non-injected blastocysts. Following embryo transfer of six EGFP-positive blastocysts into four recipient animals, two viable buffalo calves were born. Biopsied ear tissues from both buffalo calves were analyzed for transgene presence and expression by Southern blot, PCR and confocal laser scanning microscopy, respectively. This confirmed that both calves were transgenic. Conclusions： Our cytoplasmic injection protocol improved generation of transgenic embryos and resulted in the first transgenic buffalo calves produced by this method.
文摘Many countries in the world consider the buffalo as a priority animal for the future, since it plays a pivotal role in human food sustainability. Even though Food and Agriculture Organization has termed the buffalo as an important undervalued asset, this species has yet to drive the same attention as cattle. Egypt has a wealth of buffaloes dispersed in small herds all over the country, so the efforts that have been made to improve their genetic background show little return. Contrarily, other countries concerned with buffalo improvement have already used a data recording system in buffalo herds, allowing to achieve a much faster improvement progress. This review intends to survey the existing information on the application of assisted reproduction techniques to improve buffalo productivity. The strength points that may help to improve buffalo production are identified, and the obstacles hindering the genetic improvement of Egyptian buffalo are characterized. Therefore, this work will gather information related to buffalo and compile it for an audience of researchers and specialists to enforce international collaboration for the development of buffalo production. Also, it will open the way for people interested in developing a future vision for buffalo potential, which will be helpful to close or minimize the biological gaps of buffaloes' researches.
文摘Objective:To isolate and identify microflora and fungal species at different phases during estrus synchronization of ewes and estimate their prevalence;compare the effectiveness of antimicrobial administration to intravaginal sponge on the changes in the vaginal micro-organisms and reproductive performance.Methods: Sixty Egyptian ewes were allocated into three equal groups (G: 1, 2 and 3). G1 was inserted with vaginal sponge containing medroxy-progesterone acetate and served as control;without antimicrobial additive. The other two groups were treated as G1, but sponges were previously injected with ciprofloxacin (G2), while sponges of G3 were injected with ciprofloxacin and clotrimazole. Vaginal swabs were collected from each treated ewe, prior sponge insertion, at sponge withdrawal and 48 h later for microbiological investigation and bacterial count. On the day of sponge removal, 300 IU/eCG was administered for each treated ewe. The identified bacterial strains before sponge insertion were tested for sensitivity with antimicrobial disks.Results:Bacterial isolates before sponge insertion were more sensitive to ciprofloxacin. Frequencies of ewes in estrus;the interval from sponge withdrawal to onset estrus and the duration of estrus were statistically similar among treated groups. The pregnancy rate in G2 (100%) was higher than G1 (66.7%) and G3 (82.4%). The total bacterial count before sponge insertion was similar between all treatments and increased significantly in all groups on the day of sponge withdraw. The prevailing bacteria on D0, D14 and 48 h after sponge removal for all treated groups wereStaphylococcus spp. followed byEscherichia coli. Regarding to fungus species, percentages of isolation increased from 5% (before sponge insertion) to 100.00% and 88.89% at sponge withdraw for G1 and G2, respectively. In G3, the fungus was declined from 10% (before sponge insertion) to 5% (at sponge removal).Conclusions:The concomitant treatments by antimicrobial to the vaginal sponge which used for estrus synchronization in ewes can improve reproductive performance.
文摘Objective: To study the effect of adding different concentrations of soybean lecithin in Tris based extender and to compare the results of Tris soybean lecithin extender with two commercial diluents (egg yolk-based: BullXcell and plant-based: OptiXcell) for Damascus goat sperm cryopreservation. Methods: The ejaculates from 4 mature male Damascus goats were obtained by using an artificial vagina. Semen samples were pooled and diluted in Tris extender supplemented with soybean lecithin at different concentrations of 1.5%, 3.0%, 6.0% and 10.0% to get better concentration to be used for further experiments, with a final concentration of 240×106 spermatozoa/mL. Semen samples were packed in straws (0.25 mL), frozen by using an automated system and stored in liquid nitrogen at -196 ℃ for 48 h. After thawing (37 ℃/30 s), the samples were evaluated for sperm quality parameters, including sperm motility, membrane integrity and acrosome integrity. Malondialdehyde concentration was estimated as a marker for lipid peroxidation. Based on the previous investigations, only Tris extender supplemented with 3.0% soybean lecithin (based on its positive results) was used versus BullXcell and OptiXcell for sperm ultrastructure evaluation and artificial insemination by using electron microscope and artificial insemination of the synchronized does. Results: There was no significant difference between Tris-soybean lecithin at 3.0% and BullXcell/OptiXcell diluents in post-thaw sperm parameters and fertility following artificial insemination;meanwhile, the other concentrations of soybean lecithin (1.5%, 6.0% and 10%) showed lower sperm parameters following cryopreservation. Conclusions: Using of Tris-soybean lecithin based extender at the level of 3.0% can be an appropriate alternative to either BullXcell or OptiXcell for Damascus goat sperm cryopreservation.
文摘Factors affecting the efficiency of nuclear transfer （NT） in rabbits were examined in the present study. When 100 V mm of pulse strength and 15 us of pulse duration were employed, 3 and 4 electronic pulses resulted in significantly more cytoplasts fused with donor cells compared with 2 electronic pulses （P〈 0.05）, but no significant difference was found in the cleavage rate of reconstructed embryos among the three groups （P〉0.05）. When the duration and number of electronic pulse were fixed at 15 ps and 3 times, increase of pulse intensity from 100 V mm 1 to 150 V mm^-1 and 200 V mm^-1 resulted in a significantly decrease in the cleavage rate of reconstructed embryos （P〈 0.05）, although the fusion rate did not significantly differ among the three groups （P〉 0.05）. Significantly more reconstructed embryos cleaved and developed to blastocysts when they were derived from donor embryos at the 8-16-cell stage, in comparison with the reconstructed embryos derived from donor embryos at the compact morula stage （P 〈 0.05）, although the fusion rate was similar （P 〉 0.05）. Activation of cytoplasts prior to fusion increased the cleavage rate （P〈 0.05） and blastocyst development （P〈 0.05） of reconstructed embryos, but decreased the fusion rate （P 〈 0.05） compared with cytoplasts activated post fusion. More reconstructed embryos developed to blastocysts when they were cultured in TCM ＋ 3% OCS at the first 48 h and then cultured in TCM199＋ 10% FCS, in comparison with the reconstructed embryos cultured in either TCM199＋ 10% FCS or TCM199＋3% OCS （P 〈 0.05）. When 22 NT embryos were transferred into the oviducts of one recipient rabbit, one recipient rabbit delivered a female rabbit at 34 days of gestation. In conclusion, either electrofusion parameter or developmental stage of donor embryos have a significant effect on the efficiency of NT, NT embryos require different concentration of serum at their different development stages.
基金supported by the Major Projects of Genetically Modified Organism (GMO) New Varieties Breeding of Ministry of Agriculture ( 2008ZX08007-003)
文摘[Objective] To clone ovarian-specific promoter-1 （OSP-1） fragment in rats and detect the tissue-specific expression of OSP-1 at cell level. [Method] According to the known OSP-1 sequence in rats, the specific pdmers were designed. The OSP-1 fragment was amplified by PCR and compared with published OSP-1 sequence. After eukaryotic expression vector pOSP-1-EGFP-N1 was constructed by cloning OSP-1 promoter into the pEGFP-N1 vector without CMV, the recombinant plasmid was transfected into buffalo granulocytes, mammary epithelial cells and fetal fibroblast cells under mediation of liposome LipofectamineTM 2000. The green fluxorescent protein （GFP） expression was observed with the fluorescence microscope after transfection for 12, 24 and 48 h. [Result] The amplified OSP-1 fragment was 480 bp, and the homology to published rat OSP-1 sequence was 96%. The sequence analysis showed that this fragment contained a core promoter cis-element similar with TATA box and ChAT box, and multiple C/EBP beta transcription factor binding sites. The EGFP-expressing positive granulccytes were firstly observed at 12 h after transfection and they were increased. All the EGFP-expressing and non-expressing cells from granulocytes were large and round. After transfection for 48 h, the EGFP-expressing positive granulocytes were decreased. The EGFP was not expressed in buffalo mammary epithelial cell and fetal fibroblast cell. [ Conclusion] EGFP exDression controlled bv OSP-1 promoter can be found in buffao granulocvtes.
基金funded by the China Natural Science Foundation(30760171)
文摘[Objective] The aim of this paper is to find the change law of reproductive hormones of female swamp buffalo in off season for estrus and to investigate the relationship between various hormones. [ Method] The concentrations of follicle stimulating hormone （FSH）, luteinizing hor- mone （ LH）, inhibin （ INH）, estradiol （ E2 ） and progesterone （ P4 ） were determined by the enzyme-linked immunosorbent assay （ELISA） in off season for estrue. The changes of follicles in the estrous cycle were examined using B-mode ultrasonography. [ Result] In off season for estrus of female swamp buffalo from Guangxi region, the blood concentration of E2 and P4 varied fluctuantly. The concentration of E2 had two peaks, respec- tively 3 d before ovulation and after 13 d post ovulation. The concentration of P4 increased slowly from the first day after ovulation, increased rapidly from 5 to 13 d, peaked on Day 16 and decreased to the baseline 3 d before ovulation. The concentrations of FSH, LH and INH roached a peak re- spectively after 10 d post ovulation, 2 d before ovulation, and after 4 d post ovulation. The changes of the hormones had correlations. The concen- trations of FSH, LH and INH had highly significant correlations with each other, while the concentration of INH had highly significant correlations with that of E2 or P4- The concentration of E2 also had a highly significant correlation with that of P4- [ Conclusionl The study can guide the breeding of swamp buffalo from Guanqxi reqion and also help to improve their roproduction performance.
文摘Objective: To study the genetic and non-genetic factors and their interactions affecting growth rate and body weights at birth, weaning and at 6 months of age in Saudi Ardi, Damascus goats and their crosses. Methods: Crossbreeding program between Saudi Ardi(A) goats with Damascus(D) was carried out to improve the meat productivity of Ardi goats through crossbreeding. The pedigree records of the body weights were obtained from 754 kids (397 males and 357 females) produced from 46 Sires and 279 Dams. Birth weight, weaning weigh and 6 months weight as well as average daily gain during different growth stages from birth to weaning (D1), weaning to 6 months (D2) and from birth to 6 months of age (D3) were recorded during winter/autumn and summer/spring. Data were classified according to breed, generation, sex, season, year, and type of birth. Data were analyzed using GLM procedure for the least-squares means of the fixed factors. Heritability and genetic parameters were estimated with derivative-free restricted maximum likelihood procedures using the MTDFREML program. Results: The percentages of variations were moderate for body weights and high for daily gains. Genetic groups had a highly significant (P<0.01) effect on the body weights traits. Damascus goats had higher (P<0.01) birth and weaning weights, but ?D?A group kids had a higher (P<0.01) body weight at 6 months. The genetic groups had a significant effects on the daily weight gains for D1 (P<0.01) and D3 (P<0.05) periods, whereas, it had no effects on D2 period. The fixed effects of sex, season, year and type of birth were significant differences for body weights. Male kids were heavier (P<0.01) than females for different growth stages. Body weights and daily gains during winter/autumn were significantly higher (P<0.01) than summer/spring. Kids born and raised as singles were significantly (P<0.01) heavier than those were born as twins or triplets. The genetic and phenotypic correlations between birth and weaning weights were positive for both Damascus and Ardi goats. Conclusions: Genetic program for Ardi goats through upgrading with Damascus is possible to improve meat production.
文摘The human and bovine lactoferrin have been studied extensively, but very few reports have been published concerning camel lactoferrin (cLf). The present study aimed to isolate cLf and evaluate its efficiency including antimicrobial activity and immunomodulator effects. cLf isolation was attempted from camel milk whey using a cation exchange chromatography by SP-Sepharose. The antimicrobial activity of the isolated cLf was investigated against Staphylococcus aureus (S. aureus), Streptococcus agalactiae (S. agalactiae), Escherichia coli (E. coli) and Pseudomonas aeruginosa (P. aerogenosa) strains. The immune effect of cLf was studied by lymphocyte transformation test. It was found that cLf was separated around molecular weight of 80 kDa and showed significant inhibitory effect against E. coli followed by P. aeruginosa, S. agalactiae and S. aureus. cLf increased lymphocyte transformations mean values in a dose dependant manner. The highest transformations mean value was determined at 50 μg/mL. In conclusion, these results suggest that cLf is a potent natural antimicrobial and novel immunomodulator agent.
文摘Vitrification of immature oocytes at the germinal vesicle (GV) stage is important to preserve female gametes. The standard formula for vitrification solutions has long been a debate. Herein, we investigated the effect of the presence or absence of trehalose in vitrification solution on viability, in vitro maturation (IVM) rates, and development of vitrified/warmed immature dromedary camel oocytes. Cumulus oocyte complexes (COCs) obtained at slaughter from the ovaries of mature she-camels were randomly allocated into three groups;namely, control group, oocytes were directly subjected to IVM without vitrification, vitrification solution 1 (VS1) group, oocytes were vitrified in a solution composed of 25% ethylene glycol (EG) plus 25% dimethyl sulfoxide (DMSO) + 0.5 M trehalose;and vitrification solution 2 (VS2) group, oocytes were vitrified in a solution composed of 25% EG plus 25% DMSO. Vitrification of COCs was conducted by open pulled straws (OPS) method. Following vitrification and warming, morphologically viable oocytes were matured in vitro for 36 h. COCs were then fertilized and cultured in vitro for 7 days. The percentage of viable oocytes was significantly higher (P 0.05) in VS2 than VS1 group (80.0% vs. 63.3%, respectively). Nuclear maturation, cleavage (48 h post-insemination;pi), and blastocyst rates (7 days pi) were significantly higher (P < 0.05) in VS2 than in VS1 groups. No significant differences were observed in oocyte maturation and development rates between VS2 and control groups. In conclusion, vitrification of immature dromedary camel oocytes in trehalose-free solution (VS2) was more advantageous than that in trehalose supplemented media since it did not reduce viability and development.
