ناصر شمس اسفندآبادی

This study aimed to adjust and optimize the method of separating low-motile and high-motile sperms in fresh bovine semen treated with resiquimod, an agonist of Toll-like 7/8 receptors present in X-chromosome-containing sperms and causes a decrease in ATP and energy production. This method could separate X and Y sperms in fresh cow semen. Semen samples were obtained from five bulls in three replications, treated with 0.3 µM resiquimod, and subjected to swim up in two separation tools A and B (made of polyethylene and has an angle of 60 degrees for tool A and 80 degrees for tool B). There was no significant difference in the percentage of motile sperm in the upper layers of the groups (p>0.05); however, the percentage of motile sperm was higher in the lower layer of the control group (p<0.05). The ratio of the number of counted sperm in the upper layer of the control group was significantly more than the two treated groups (p<0.05) and was higher in tool B than in tool A (p<0.05). Equating the ratio of sperms counted in the upper layer to the total number of sperms counted in the upper and lower layers showed that the number of sperms in the upper layer of tool A is almost half of the control group. It is concluded that the use of 0.3 micromol of resiquimod and swim-up in instrument A can be effectively used to separate motile sperm from immobile sperm to separate sperm containing X chromosome from Y.

It has been determined that there is a direct relationship between the severity of mastitis and the virulence factors produced by bacterial agents. Identifying bacterial virulence factors is neccecary for designing suitable vaccines against mastitis. The aim of the present study was molecular diagnosis of selected virulence factors of endemic isolates of S. aureus involved in bovine mastitis. A total of 180 milk samples were collected from cows with clinical (37 samples, 20.6%) and subclinical (143 samples, 79.4%) mastitis from 8 semi-industrial dairy farms of Chaharmahal and Bakhtiari province, Iran. After culture and purification, coagulase, catalase and oxidase tests were performed. DNA was extracted from S. aureus suspected colonies. Final confirmation was performed using PCR test on the specific 23S rRNA gene of the bacteria. Thirty one (17.22%) out of the 180 collected samples were found to be positive for S. aureus by PCR, of which 2 cases were related to clinical mastitis and 29 cases were related to subclinical mastitis. The highest frequency of virulence genes was related to the Coa gene (90.32%), followed by ClfB (87.09%), LukD, and fnbB (80.64%), LukE (77.41%), fnbA (74.19%), Hla (48.38%). The lowest frequency was related to the Hlb gene (45.16%). Based on the obtained results, the diagnosis of the virulence factors of S. aureus has the potential of being used in the development of vaccines for the prevention of mastitis.

Streptococcus uberis is a Gram-positive bacterium and the major cause of mastitis in dairy cattle. This bacterium is one of the most important causes of economic losses in dairy herds with no ordinary monitoring program for mastitis. In order to evaluate the presence of this agent in the bulk tank milk of dairy cattle farms in Isfahan province and to find out its relationship with the total bacterial and somatic cells count, 100 tank milk samples were collected. The total bacterial and somatic cells were detected in the samples and the level of infection to S. uberis was evaluated by conventional culture and RT-PCR methods. The results of RT-PCR and culture showed infectivity of 20 and 16 samples to S. uberis, respectively. All culture positive samples were also positive in RT-PCR, but 6 samples were only positive in RT-PCR, indicating that RT-PCR is more sensitive than culture. Statistical analysis showed that there is a significant relationship between the infectivity to S. uberis and the total somatic cells, but the presence of this bacterium had no significant effect on the total number of bacteria in milk samples. It can be concluded that S. uberis is usually present in dairy farms of Isfahan province and it is necessary to pay more attention to the methods of monitoring and controlling the mastitis cows.

The incidence of persistent post-mating endometritis (PPME) following natural mating or artificial insemination (AI) is always underestimated in mares that do not have abnormal clinical signs in reproductive tracts in routine examinations before mating in high-population horse breeding centers. In this study, 503 Arabian mares whose reproductive system status was confirmed before natural mating or artificial insemination in breeding soundness examination, were examined with ultrasonography at 24, 48, and 72 hours after ovulation, in terms of PPME following natural mating or AI, and the presence of uterine fluid accumulation with a diameter of 5 mm or more is recorded as PPME. The correlation between PPME and other variables such as age, type of mating, number of mattings in an estrous cycle, pre-ovulatory follicle (POF) size, volume and concentration of semen used in AI was investigated at different time intervals after ovulation. The results of this study showed that, the incidence rate of PPME following natural mating or AI in 48 and 72 hours after ovulation is 32 and 22%, respectively, in a normal population of Arab mares. Based on this rate, reproductive examinations are recommended 24 and 48 hours after ovulation in the group of sensitive Arab mares.

One of the advantages of using epididymal sperm is helping to preserve the genetic resources of elite animals and endangered species after death. This study aimed to investigate the probable protective effect of polyvinyl alcohol (PVA) and polyethylene glycol (PEG) on epididymal ram sperm under liquid storage at 4 ° C for 24 hours. Sperms were collected by making an incision on the epididymis of slaughtered rams and extended in extenders supplemented with 0 (control), 0.5, 1, 2, 5 and 10 mg/ml polyvinyl alcohol or polyethylene glycol then refrigerated for 24 hours. We found that only 0.5 mg/ml of PVA group had higher motility parameters comparing to control group (P <0.05). The lowest motility in PVA groups was observed at the concentration of 10 mg/ml. Extenders supplemented with PEG in different concentrations had no significant effect on the most kinetic indices. Both compounds were lethal to sperm at 15 mg/ml.

The endometritis form of uterine infection is considered as a common reproductive disorder deleterious to the reproductive performance of dairy herds. A wide variety of treatments with controversial results have been reported for endometritis, including local or systemic administration of antibiotics or disinfectants as well as hormone application. These treatments have side effects on endometrium, and antibiotic residues are found in milk following intrauterine and systemic antibiotic therapies. The aim of this study is to evaluate the possible effects of the mixed essential oil of Satureja bachtiarica Bunge, Artemisia Aucheri Boiss and Syzygium aromaticum (L.) Merr. & L.M.Perry on treatment of clinical endometritis in dairy cattle. One hundred and twenty cows with clinical endometritis were selected and randomly assigned to one of the following three groups: the HM group received mixed herbal essential oils, the OX group received 2.5 g oxytetracycline HCl, and the EX group received 1 g of ceftiofur sodium, all by intrauterine injection. The cleaning and first service conception rate was significantly higher in HM group than the EX, whereas the mean open days were lower in HM than the EX group. The number of service per conception was also significantly lower in HM group than OX and EX groups. In general, reproductive performance after herbal treatment was quite comparable to chemical antibiotic therapy and even better in some other reproductive indices. The mixed essential oils treatment represents an effective potential alternative to postpartum therapy for cows with clinical endometritis.

Uterine diseases are common problem that influence the reproductive output in dairy farm. Different factors include metabolic disturbances may affect the uterine defense mechanism and contribute at postpartum uterine disease occurrence. The routine metabolic profile is measurements of net values of non-esterified fatty acids (NEFA), β-hydroxybutyric acid (BHBA), and glucose (Glc) in cow serum. The present study aimed to look at the ratios of the energy metabolic products during the transition period and investigate their relationship with bovine clinical endometritis. The weekly serum samples were collected from 100 dairy cows during the transition period (2 weeks prepartum to 3 weeks postpartum). In the clean test (Day 30 postpartum), 16 cows were selected, and assigned in a case-control repeated measure design into two groups as clinical endometritis (CE: n=8) or healthy cows (n=8). The respective serum samples of the cows were assayed for NEFA, BHBA, and Glc. All the measurement units were converted to mM/L and the ratios were calculated. The trend of changes in the ratios of NEFA to BHBA was in a steady state in healthy cows during the transition period, while it raised at the time of parturition in the CE group (P<0.05). While the significant increase in NEFA to Glc ratio started after parturition in healthy cows, it was begun to increase before parturition in CE. The trend of changes in the energy metabolite ratios during the transition is different in CE compared to healthy dairy cows.

Oocyte quality is one of the most important factors in embryo production through in vitro fertilization (IVF). Developmental competence of an oocyte is highly depended on the follicular microenvironment. In this study, the effect of 2 hormonal superstimulation regimes (2 doses and 6 doses of FSH) on developmental competence of oocytes, obtained by ovum pick up (OPU) procedure, was evaluated in cattle species. A total of 120 Holstein heifers with an average age of 11 months were divided into 2 groups of 60 animals. The animals in each group received either 2 or 6 equal doses of FSH (each dose= 50 mg) at 12h interval. The FSH treatment was started 2 days after dominant follicle ablation. The ablation of dominant follicle (s) was done 6 days after synchronized estrous. After OPU, 48 h after the end of FSH therapy, the aspirated oocytes were evaluated morphologically and considered for in vitro maturation, IVF, and subsequent embryo culture. In days 3 and 7, cleavage and blastocyst rates were evaluated. The mean numbers of aspirated follicles were 6.07±0.3 and 14.9±0.9, and retrieved oocytes were 3.93 ± 0.5 and 7.27 ± 0.6, in treated animal with 2 and 6 doses of FSH, respectively, which was significantly different between two groups (P=0.0001). The difference in proportion of grade A oocytes in 2 (66.97±3.57) and 6 (73.48±3.01) doses of FSH groups was insignificant (P>0.05). The rate of cleavage in animals treated with 2 and 6 doses of FSH was 65.67 ± 3.65 and 74.27 ± 5.38, respectively, which was not significant between two groups (P>0.05). Blastocyst rate in the 6 dose FSH group (50.32±3.57) was significantly higher than that of 2 dose FSH group (38.02 ± 3.18) (P=0.011). The results of this study demonstrated that 6 doses FSH in compare to 2 doses FSH prior to OPU resulted in more aspirable follicles with higher developmental competence oocytes.

In vitro ovarian follicle culture in females at the risk of losing fertility, such premature ovarian failure (POF) and in those subjected to chemo/radio therapy as well as in animals – species at risk of extinction in a promising strategy for fertility preservation. Development of ovarian growing follicles is regulated by the coordinated action of several signaling pathways. The use of some activators or inhibitors of signaling pathways is effective in optimizing the culture conditions of immature follicles. The aim of this study was examining in vitro growth of bovine ovarian secondary stage follicles (> 80 μm) by mimicking the process of in vivo follicular development through of the Akt signaling pathway activator, PS48. Small secondary stage follicles were isolated by enzymatic digestion from slaughter ovaries by collagenase type I and Dnase I, cultured in the presence of BSA (3 mg/ml), FBS (10%), Akt (5µM PS48, first 48 h of culture) or Akt + FBS (5µM PS48, first 48 h of culture + 10% FBS) for ten days and then evaluated for growth rate. The follicular size was significantly increased in all experimental groups during 10 days culture (P<0.05). In all our groups, growth rate/day was higher in initial days of culture, after that follicular growth rate decreased. The highest growth rate was observed in FBS + PS48 group, while the lowest growth rate was detected in PS48. On the other hand, though the FBS supplemented group could increase the growth rate of follicles, its combination with PS48 was more effective . In conclusion, in our study condition the best result in follicular growth, was obtained when follicular culture medium was supplemented with FBS + PS48. In conclusion, in our study condition the best result in follicular growth, was obtained when follicular culture medium was supplemented with FBS + PS48.

Cryopreservation has led to many improvements in domestic animal breeding industry, especially in terms of spreading of superior genes. The improvement of extenders and cryoprotectants can result in a significant increase in transporting and inseminating of frozen stallion semen. In this study, we analysed the genes that would be expressed in stallion cryopreserved sperm and by providing an adequate amplification, they would be selected as reference genes in the future studies. Live sperm with above 70% motility were selected from six different stallions. Sperm samples were divided in to two parts for total mRNA extraction. The selective amplification of three candidate genes (L32, βactin and Ubiquitin) were transcribed by real-time polymerase chain reaction (RT-qPCR). Gene stabilities were calculated by GeNorm and BestKeeper softwares. Results indicated that the expression of L32 gene is less stable than that of β-actin and Ubiquitin. The expression stability of β-actin and Ubiquitin were similar. These genes can be used for normalizing of RT-qPCR reaction data. The GeNorm software introduced the best combination of genes as, β-actin and Ubiquitin with the stability value (M) of 0.083. The results of BestKeeper software showed that the most stable house-keeping gene was β-actin.

During the past years, portable ultrasonography devices with high quality pictures have routinely been used for veterinary diagnostic procedures. One of the most practical uses of this technique is reproductive management in dairy cows. The aim of this study was to designing a software for education of cattle reproductive ultrasonography. In this study, the reproductive system (uterus and ovaries) of 110 dairy cows was examined by ultrasonography during the different times postparturition. The pictures were taken and the picture legends were put under the pictures. The most practical aspects of each topic or disorder were also described. All the pictures were evaluated after gathering and only the pictures with the best quality were selected. Model View Controller (MVC) design pattern was used that is the object oriented design pattern and the graphical view was developed with JAVA. This software can be installed and used in all devices with android operating System.

The aim of the present study was to evaluate the effects of PGE2 on follicular cyst treatment in dairy cow. A total of 95 lactating dairy cows with follicular cyst were selected for a 5-month period study were performed in Zagros farm in Shahrekord.The animals were randomly divided into 4 groups; the first group received 200 µg Gonaderoline Acetate, the second received 1 mg Dinoproston, the third group received 2 mg Dinoproston and finally control group which received 2 ml normal saline. 9 days later animals in first three treatment groups were received 500 µg Cloprostenol and animals in control group received 2 ml normal saline. Cows which showed heat in 7 days period after cloprostenol injection were artificially inseminated according to am-pm rule and the remaining animals were re-examined at day 18 ± 2. To measure the concentration of serum progesterone, blood samples were collected at the first and ninth days. Submission rate was singnificantly higher in treatment groups compared to control group (p0.05). Conception rate at the first service and the percent of different types of cysts in re-test after treatments were not significantly different (p>0.05). Serum levels of progesterone in the first and third groups compared to the second and control groups were significantly increased in 9 days (p

Considering the low percentages of sperm chromatin decondensation and male pronuclear formation following intracytoplasmic sperm injection (ICSI) in ovine species which decrease the chance of normal fertilization, the current study was designed to assess the effects of some methods of sperm pretreatment on male pronuclear (MPN) formation and oocyte activation in ovine ICSI procedure.
The experimental groups of this study were comprised of: I) ICSI of untreated sperm (control), II) ICSI of pretreated sperm with GSHᷪ (GSHᷪ), III) ICSI of pretreated sperm with GSHᷪ coinjected with GSH (GSHᷪ纊.GSH), and IV) ICSI of pretreated sperm with GSHᷪ coinjected with sperm extract (GSHᷪ纊.SE). The injected oocytes were activated with IoDAMP and were examined for prenuclear formation after 16h. The MPN formation in GSHᷪ纊.GSH was higher (P

Given to the proven role of endogenous PGF2α on ovulation, the aim of the present study was to evaluate the effect of exogenous PGF2α (D-cloprostenol) administration on conception rate in a large dairy herd. Total number of 362 lactating dairy cows in Zagrous Company of Shahrekord were selected for this study in summer (From 20 August to 6 September, 2014). The estrus of all cows was synchronized by the Heatsynch protochol and following heat detection the cows were artificially inseminated according to am-pm rule. The animals were submitted to two groups; the treatment group that received 150µg D-cloprostenol intravenously at the time of AI and the control group that didn’t receive any more treatment at the time of AI. To measure the concentration of serum progesterone, blood samples were obtained from 13 cows of each group at the time of insemination and 4 and 15 days later. Pregnancy was diagnosed by rectal ultrasound examination on 35-40 days after insemination. There was no significant difference between both groups in terms of conception rate and the serum progesterone levels (P>0.05). In conclusion, this study showed no beneficial effects of intravenous administration of D-cloprostenol on conception rate of dairy cows.

The aim of this study was to improve culture conditions of bovine embryos produced by the Hand-made cloning (HMC) method with the use of serum and myoinositol. For this purpose, after oocyte in vitro maturation followed by elimination of the zona pellucida, bovine oocytes were enucleated by separation of the metaphase spindle protrusion using a very thin pulled Pasteur pipette. After attachment of adult fibroblasts as donor cell into the enucleated oocyte and subsequent fusion and activation, reconstructed embryos were cultured by Well of the Well (WOW) method in six groups, including IVCSOF, IVCSOF-CSS, IVCSOF-ci, IVCSOF-ciCSS, IVCSOFci and IVCSOFci-CSS. In these groups, the embryos were cultured in IVCSOF with or without myoinositol-sodium citrate media until the third and then CSS and/or myoinositol-sodium citrate was added. Finally, development rate was recorded and analyzed statistically. Our result showed that despite the ineffectiveness of myoinositol on embryonic development capabilities, the embryos cultured in the presence of this combination and serum significantly reached earlier to blastocyst stage (p

Despite the successful pregnancies and live births that have been reported from transferring embryos derived from cryopreserved oocytes in several mammalian species, developmental competence of frozen-thawed oocytes is low. The aim of this study was examining the effects of short term treating of the immature ovine oocyte with glycine and supplementation of vitrification and warming solution with glycine on post vitrification and warming quality. Abattoir-derived immature ovine oocytes vitrified with (GLY) or without (CON) treating with glycine and after In vitro maturation the nuclear maturation and the rate of apoptosis of them compared with their unvitrified (FRESH) counterparts. A difference at a level of P<0.05 was described significant. The survival rate of the oocytes in vitrified groups was significantly lower than of the FRESH group. Treating with glycine had no beneficial effect on post IVM survival rate. Apoptotic oocytes were significantly lower in the GLY group than CON group and were comparable to FRESH group. In conclusion, glycine can improve the quality of ovine immature oocytes after vitrification and warming.

The aim of this study was to evaluate the effect of Heatsynch and Ovsynch(two methods of ovulation synchronization) on pregnancy rate in dairy cows. This study was carried out in a large dairy herd in suburbs of Esfehan province. In total 113 dairy cows with 50-200 days in milk entered in the study. In this experiment cows were alternately assigned to three proups.1) control group (n=50) received no treatment. Cows that were detected in estrous were inseminated by on farm technician.2) Ovsynch group (n=46) received an i.m injection of 10 μg GnRH analog on days 0 and 9 and an i.m injection of 500 μg of PGF2α on day 7, then inseminated 16-20 hour after second GnRH injection. 3) Heatsynch group (n=37) received an i.m injection of 10 μg GnRH analog and 500 μg of PGF2α on days 0 and 7 respectively and then then 1mg estradiol cypiyonate 24 hour after PG injection. Cows that were detected in estrous after estradiol treatment were inseminated as a.m. / p.m.protocol. Timed artificial insemination(TAI) was performed for cows that were not detected in estrous 48 hourafter estradiol treatment.Pregnancy detection were performed on day 30 and between days 40-45 after insemination by ultrasonography and rectal palpation respectively. The differences in Conception rate between three groups were analyzed by using Chi-Square method. Although conception rate in Ovsynchgroup (%41.3) were higher than Heatsynch(%37.8) and control(%36) groups, but there was no significant differences in conception rate between three groups.

This study investigated the concentration/time dependent effects of nanosilver colloid on the motion parameters of the ejaculated ram spermatozoa in vitro. After incubation of sperm samples for 30، 60، 120 and 180 min in the presence of nanosilver colloid (0، 0. 001، 0. 01، 0. 1، 1 and 10 ppm)، the motion parameters were evaluated by computer assisted sperm analysis. There was a significant (P<0. 05) decrease in most motion of sperm parameters (the fast، slow and non-progressive motility، VCL، VSL، STR، MAD، ALH، WOB، VAP and LIN) in treated groups compared to their corresponding controls، especially after 120 min of incubation. On the other hand، there was a significant (P<0. 05) increase in sperm immotility in comparison with the control group in all times at 1 ppm nanosilver colloid. At a concentration of 10 ppm، spermatozoa were completely inactivated. It is concluded that nanosilver colloid depresses sperm functions، especially motility parameters، which can be a causative agent for sperm infertility induced by nanosilver cytotoxicity.

Heat stress causes reduced fertility and significant economic loss in dairy cattle. To override the suppressive effects of heat stress, various hormonal manipulations have been utilized. The aim of this study was to compare the effect of progesterone (in the form of CIDR) and administration of GnRH after insemination on the conception rate of heat stressed dairy cattle. All cows were inseminated at estrus and were then alternately assigned into three groups on day 5 after artificial insemination (AI): i) GnRH group (n=44) received an IM injection of 500 µg GnRH (GONAbreed, PARNEL, Australia,); ii) CIDR group (n = 44) received a CIDR (EAZI-BREED, Hamilton, NZ, containing 1/9 g progesterone) which was removed after a week; and iii) control group (n = 36), which did not receive any treatment. Conception was diagnosed on day 32-39 after AI by ultrasonography. Conception rate in GnRH, CIDR and control groups were 54.5%, 54.5% and 58.3%, respectively. The results demonstrated that there was no significant difference among the three groups (p >0.05). These treatments had had no statistically different effects on lactation, milk yield, days in milk and number of AI (p>0.05). Conception rates within GnRH and CIDR groups in <150 and >150 days in milk subgroups were 74.4%, 40.7%, 84.6% and 41.9%, respectively and differed statistically significantly (p>0.05). Conception rate within control and CIDR groups among <3 and >3 numbers of AI were 80%, 31.2%, 84.2% and 32%, respectively, which was statistically significant (p>0.05). According to the results of this study, the use of GnRH and CIDR after AI did not improve conception rates of mildly heat stressed dairy cattle

When dairy cattle are subjected to heat stress, reproductive efficiency declines.To override the suppressive effects of heat stress various environmental and hormonal manipulations have been utilized. The aim of the present study was to evaluate whether the administration of GnRH and hCG on day 5 post-insemination would improve pregnancy rate in mild heat stressed dairy cattle (summer months). All cows were inseminated at estrus and then on days 5 after AI, they were alternately assigned into three groups: GnRH group(n=44)received an IM injection (5ml) of GnRH (GONAbreed®,PARNEL, Australia,) hCG group (n=44) received an IM injection (1ml) of hCG (Pregnyl®, Daru Pakhsh, Iran) and control group (n=36) which did not received any treatment.Pregnancy was diagnosed on day 32-39 after AI by ultrasonography.. The difference in CR among three groups were analyzed by using chi-square and fisher test. CR in GnRH, hCG and control groups were 54/5%, 61% and 58/3% respectively, that weren’t significant difference among three groups (P>0.05). CR within GnRH groups in ≤150 and>150 days open subgroups were 72%, 40% that is statistically significant (P<0.05) and also CR between GnRH and hCG groups based on Open days>15o was 40%, 76% that is statistically significant (P<0.05). CR within control and hCG groups among <3 and≥3number of AI were 80%, 31%, 77% and 39%respectively, that is statistically different(P<0.05). According to the results of this study, the use of hCG after AI in repeat breeder dairy cows in summer months is recommended.