جستجوی مقالات مرتبط با کلیدواژه « buffalo » در نشریات گروه « پزشکی »

Sarcocystosis is a zoonotic disease worldwide caused by Sarcocystis spp., some of these species can show clinical and subclinical manifestations, resulting in financial losses. Our study was performed for identifying Sarcocystis sp., in slaughtered buffalo by PCR–RFLP based strategy with sequencing in Guilan, North of Iran.

Overall, 400 fresh muscle samples were prepared via naked-eye observation from 100 buffaloes (esophagus, diaphragm, shoulder, and thigh), followed by the digestion of samples. The PCR was done to amplify partial parts of the 18S rRNA and mitochondrial cytochrome c oxidase subunit I (Cox1) genes. Then, the PCR products were digested by endonuclease SspI, DraI, and FokI. Sequencing of all species was done to confirm the RFLP results.

Five macroscopic cysts (1.25%) were visible in the sample by naked-eye examination. Furthermore, 293 samples (73.25%) were found to be Sarcocystis sp. positive through tissue digestion and microscopic observation, whereas 376 samples (94%) were positive by PCR. In addition, the findings of PCR-RFLP and nucleotide sequence samples exhibited the infection of buffaloes with S. cruzi.

Based on the data presented herein, Bovine sarcocystosis caused by S. cruzi is very common in buffalo in the Guilan region. Regarding the high prevalence of sarcocystosis, developing disease control and prevention policies for buffaloes is necessary, and a change of attitude in traditional farming is recommended.

 Several methods have been employed to identify Coxiella burnetii isolates based on the specific Coxiella burnetii QpH1 plasmid to distinguish the acute form from the chronic form of Q fever disease in humans and animals owing to the presence of unique gene sequences in this plasmid. Therefore, the present study aimed to investigate the panel of nucleic acid fragments resulting from the enzymatic cleavage in the QpH1 plasmid isolated from cow and buffalo milk by nested polymerase chain reaction (Nested-PCR).

 A total of 86 isolates of Coxiella burnetii QpH1 plasmid, which were confirmed by the Nested-PCR method in 2018, were used to determine the RFLP panel of the QpH1 plasmid. Plasmids were first extracted with the kit and were then affected by the Hph1 restriction enzyme. Additionally, 4 nucleic acid samples were sent to Pishgam Company for sequencing with the IS1111 gene primer.

Based on the results of the PCR-RFLP test, all plasmid samples showed a similar two-fragment pattern under the influence of Hph1. The results of the nucleic acid sequencing of all 4 samples indicated that they had a Coxiella burnetii type (Nine Mile RSA493 strain).

 RFLP patterns exhibited no difference on the Coxiella burnetii QpH1 plasmid isolated from cow and buffalo milk. Hence, all isolates were genetically identical, and the infection in animals could originate from one Coxiella burnetii strain (Nine Mile RSA493 strain).

The purpose of this research was to compare the functional parameters of frozen-thawed Iranian Azaribuffalo spermatozoa with imported semen samples of Italian Mediterranean buffalo (IMB) after the thawing processand 4 hours of incubation. In this experimental study, a total of twenty-four ejaculates from four Iranian Azari buffalobulls were collected. Semen samples were diluted in AndroMed extender at a concentration of 50×106 spermatozoa/ml. The diluted samples were filled in 0.5 ml straws and were frozen in a programmable freezer. For imported semensamples, twenty-four samples of four IMB were used, which were diluted in AndroMed extender and frozen by the sameprocedure. Frozen-thawed sperm motion patterns, mitochondrial activity, membrane integrity, DNA integrity, reactiveoxygen species (ROS), and apoptosis status were evaluated immediately after thawing and 4 hours of incubation. Post-thawed sperm motility, progressive motility (PM), mitochondrial activity, membrane integrity weresignificantly higher in imported semen samples in compare with Iranian Azari buffalo. After 4 hours of incubation,sperm velocity patterns were superior in Iranian Azari semen samples. Moreover, the percentage of sperm cells withun-damaged DNA was higher in Iranian semen samples compared to imported samples at the time 0 of incubation.Following 4 hours of incubation, a significant increase in intracellular ROS level leads to reduced membrane integrity,mitochondrial activity, and DNA integrity in both buffalo breeds. At time 4, Iranian samples showed significantlylower apoptosis and higher dead spermatozoa compared to imported semen samples. Our study showed that the post-thawed quality of Iranian Azari buffalo semen was comparable with imported samples after 4 hours of incubation. Further investigations are recommended to assess the in vitro and in vivofertility rate of both buffalo breeds.

In the dairy industry, protection against species substitution or admixture is important for several reasons, including frequent human adverse reactions toward some species milk proteins, and trade and government regulations.
The objective of the present study was to assess the purity of buffalo milk and it’s products offered as “pure buffalo” in the market.
Using species-specific primers, a duplex polymerase chain reaction (PCR) assay was performed to detect the fraudulent addition of cow’s milk to buffalo’s milk and it’s products.
The limit of detection of cow’s milk in buffalo’s milk, yogurt and cheese is 1, 2 and 4%, respectively. Undeclared presence of cow’s milk was detected in 70% of the milk samples, 64% of the yogurt and 52% of the cheese samples. In 10% of the yogurt samples and 14% of the cheese samples, no apparent buffalo-related amplification product was observed, suggesting that cow’s milk was entirely substituted for buffalo’s milk in these samples.
To avoid unfair competition and to assure consumers of accurate labeling, using this method is recommended for regulatory agencies.

The seroprevalence of Toxoplasma gondii infection in buffaloes, sheep and goats in Yunnan Province, southwestern China was conducted between May 2012 and December 2013. A total of 973 (427 buffaloes, 154 sheep and 392 goats) serum samples were collected from seven administrative regions of Yunnan Province, and exam­ined for T. gondii antibodies by indirect hemagglutination (IHA) test. Some risk fac­tors related to species, age, gender and geographical origin were determined using a multinomial logistic regression. The overall seroprevalence of T. gondii in ruminant species was estimated at 11.9%. The final logistic regression model demonstrated that host species and geographical origin were the main risk factors associated with T. gondii infection (P﹤0.05). Taken together, the results of the present study revealed a high expo­sure to T. gondii in ruminant species in Yunnan Province, which has an important implication for public health.

This research studies the effects of activation and inhibition of Wnt3A signaling pathway in buffalo (Bubalus bubalis) embryonic stem (ES) cell-like cells. To carry on this experimental study, the effects of activation and inhibition of Wnt3A signaling in buffalo ES cell-like cells were examined using Bio (0.5 mM) combined with WNT3A (200 ng/ml), as an activator, and Dickkopf-1 (Dkk1, 250 ng/ml), as an inhibitor, of the pathway. ES cells were cultured up to three weeks in ES cell medium without fibroblast growth factor-2 (FGF-2) and leukemia inhibitory factor (LIF), but in the presence of Bio, WNT3A, Bio+WNT3A and Dkk1. The effects of these supplements were measured on the mean area of ES cell colonies and on the expression levels of a number of important genes related to pluripotency (Oct4, Nanog, Sox2 and c-Myc) and the Wnt pathway (β-catenin). ES cell colonies cultured in ES cell medium that contained optimized quantities of LIF and FGF-2 were used as the control. Data were collected for week-1 and week-3 treated cultures. In addition, WNT3A-transfected ES cells were compared with the respective mock-transfected colonies, either alone or in combination with Dkk1 for expression of β-catenin and the pluripotency-related genes. Data were analyzed by ANOVA, and statistical significance was accepted at P<0.05. Among various examined concentrations of Bio (0.5-5 mM), the optimum effect was observed at the 0.5 mM dose as indicated by colony area and expressions of pluripotency- related genes at both weeks-1 and -3 culture periods. At this concentration,the expressions of Nanog, Oct3/4, Sox2, c-Myc and β-catenin genes were nonsignificantly higher compared to the controls. Expressions of these genes were highest in the Bio+WNT3A treated group, followed by the WNT3A and Bio-supplemented groups, and lowest in the Dkk1-treated group. The WNT-transfected colonies showed higher expressions compared to both mock and Dkk1-treated mock transfected colonies. WNT3A functions to maintain the pluripotency of ES cell-like cells both as an exogenous growth factor as well as an endogenously expressed gene. It complements the absence of FGF-2 and LIF, otherwise propounded essential for buffalo ES cell culture. WNT3A antagonizes the inhibitory effects of Dkk1 and acts in combination with its activator, Bio, to activate the Wnt signaling pathway.

In order to retain an undifferentiated pluripotent state, embryonic stem (ES) cells have to be cultured on feeder cell layers. However, use of feeder layers limits stem cell research, since experimental data may result from a combined ES cell and feeder cell response to various stimuli. In this experimental study, a buffalo ES cell line was established from in vitro derived blastocysts and characterized by the Alkaline phosphatase (AP) and immunoflourescence staining of various pluripotency markers. We examined the effect of various factors like fibroblast growth factor 2 (FGF-2), leukemia inhibitory factor (LIF) and Y-27632 to support the growth and maintenance of bubaline ES cells on gelatin coated dishes, in order to establish feeder free culture systems. We also analyzed the effect of feeder-conditioned media on stem cell growth in gelatin based cultures both in the presence as well as in the absence of the growth factors. The results showed that Y-27632, in the presence of FGF-2 and LIF, resulted in higher colony growth and increased expression of Nanog gene. Feeder-Conditioned Medium resulted in a significant increase in growth of buffalo ES cells on gelatin coated plates, however, feeder layer based cultures produced better results than gelatin based cultures. Feeder layers from buffalo fetal fibroblast cells can support buffalo ES cells for more than two years. We developed a feeder free culture system that can maintain buffalo ES cells in the short term, as well as feeder layer based culture that can support the long term maintenance of buffalo ES cells.

The objective of the study was to investigate the effects of in vitro zinc sulphate additive to semen extender on sperm parameters (progressive motility, viability, membrane integrity and DNA stability) after cryopreservation. In this Prospective longitudinal laboratory study, semen samples of 5 buffalo bulls of 3-5 years old were collected at 5 different occasions from Iran, Urmia during summer and autumn 2011, 25 samples were used in each treatment. Sperm progressive motility, viability and abnormal morphology were measured before and at 0.5 (T0), 1(T1) and 2(T2) hours after diluting semen(1:10 v/v) in Tris-citric acid based extender (without egg yolk and glycerol) at 37˚C containing none (control group), 0.072, 0.144, 0.288, 0.576 and 1.152 mg/L zinc sulphate to investigate dose and time effects. Next, a Tris-citric acid-egg yolk-glycerol extender (20% egg yolk and 7% glycerol) containing the same amount of zinc sulphate was prepared, diluted semen (1:10 v/v) was cooled and kept into a refrigerated chamber (4˚C) for 4 hours to equilibrate. Sperm progressive motility, viability, abnormal morphology, membrane integrity and DNA damage were estimated.The equilibrated semen was loaded in 0.5 ml French straws and frozen in liquid nitrogen. Later, the frozen semen was thawed and the same parameters as well as total antioxidant capacity (TAC) of the frozen-thawed semen were determined. The results showed that zinc sulphate additive at the rate of 0.288 mg/L gave a higher protection of sperm progressive motility (53.7 ± 1.8% vs. 40.5 ± 1.7%), viability (70.8 ± 1.8% vs. 60.1 ± 1.5%), membrane integrity (67.3 ± 1.6% vs. 56.6 ± 1.7%), DNA stability (10.1 ± 0.47% vs. 11.8 ± 0.33% damaged DNA) through the process of dilution,equilibration and freeze-thawing and caused a higher TAC level (81 ± 3.3% vs. 63 ± 3.2 μmol/L) after freez-thawing compared to the control group. Adding 0.576 and 1.152 mg/L zinc sulphate, however, was deleterious to the sperm and significantly reduced the studied sperm parameters. Adding 0.288 mg/L zinc sulphate to the extender, compared to the control group, gives a better sperm preservation upon freezing processes which in turn, may results in higher semen fertility. But, addition of higher zinc sulphate concentrations (0.576 and 1.152 mg/L) are detrimental to buffalo spermatozoa.

The aim of the present study was to investigate the effects of four equilibration times (2, 4, 8 and 16 hours) and two extenders (tris or Bioxcell®) on cryopreservation of buffalo semen. In this experimental study, split pooled ejaculates (n=4), possessing more than 70% visual sperm motility were divided in two aliquots and diluted in Bioxcell ® and tris-citric egg yolk (TCE) extenders. Semen was cooled to 4˚C within 2 hours, equilibrated at 4˚C for 2, 4, 8 and 16 hours, then transferred into 0.5 ml French straws, and frozen in a programmable cell freezer before being plunged into liquid nitrogen. Postthaw motility characteristics, plasma membrane integrity, acrosome morphology and DNA integrity of the buffalo sperm were studied after thawing. There were significant interactions between equilibration times and extendersfor sperm motility and membrane integrity. Post thaw sperm motility (PMOT), progressivemotile spermatozoa (PROG), plasma membrane integrity (PMI) and normal apical ridge(NAR) measures were lower for sperm equilibrated for 2 hours in both TCE and Bioxcell®extender compared to others equilibration times. PMOT, PMI and NAR for sperm equilibrated for 4, 8 and 16 hours showed no significant differences in either extender, although PROG measures were superior in Bioxcell® compared to TCE at all equilibration times (p<0.05). Kinematic parameters such as average path velocity, curvilinear velocity and linearity in the Bioxcell® extender were superior to those in the TCE extender studied. In contrast to motility and viability, the DNA integrity of post thaw spermatozoa remained unaffected by different equilibration times. Equilibration time is necessary for preservation of the motility and integrity of buffalo sperm membranes. Equilibration times of over than 2 hours resulted in the greatest preservation of total semen parameters during cryopreservation. There were no significant interactions between equilibration times over 4 hours and type of extender which lead to greater post thaw sperm survival.

Leptin, as a 16 kDa adipokine, is a pleiotropic cytokine-like hormone that primarily secreted from adipose tissue. It also involves in the regulation of energy homeostasis, neuroendocrine function, immunity, lipid and glucose homeostasis, fatty acid oxidation, angiogenesis, puberty and reproduction. The aim of this study was to investigate the effects of in vitro addition of leptin to in vitro maturation (IVM) medium on buffalo oocyte maturation and apoptosis. In this experimental study, Ovaries from apparently normal reproductive organs of slaughtered adult buffaloes (Bubalus bubalis) with unknown breeding history were collected from Urmia Abattoir, Urmia, Iran, and were transported immediately to the laboratory in a thermos flask containing sterile normal saline with added antibiotics. Oocytes were aspirated from 2-8 mm visible follicles of the ovaries using an 18-G needle attached to a 10 ml syringe. IVM medium included tissue culture medium-199 (TCM-199), 10% fetal bovine serum (FBS), 22 μg/ml sodium pyruvate, 0.5 IU/ml ovine follicle-stimulating hormone (oFSH), 0.5 IU/ml ovine luteinizing hormone (oLH), 1 μg/ml oestradiol, 50 μg/ml gentamycin, and leptin [0 (control), 10, 50, and 100 ng/ml]. The good quality buffalo oocytes (batches of 10 oocytes) were placed in a culture plate containing six 50 μl droplets of maturation medium, covered with sterilized mineral oil, and then incubated at 38.5˚C with 5% CO2 in air for 24 hours. The maturation of oocytes was evaluated under a stereomicroscope by detecting the first polar body extrusion of oocytes. FITC-Annexin V - propidium iodide (PI) staining method was used to detect oocyte apoptosis. From a total of 115 collected ovaries, 1100 oocytes were recovered among which 283 oocyte were suitable for IVM. In the groups of leptin treated with 0 (control), 10,and 100 ng/ml, the percentage of oocytes maturation was 74.65, 83.81, 77.85, and 75.40%, while the percentage of oocytes apoptosis was 9.83, 9.54, 9.93, and 10.42%, respectively. Our results showed that addition of 10 ng/ml leptin to buffalo IVM medium increased oocyte maturation, significantly, as compared with that in control group. However, addition of leptin to IVM medium had no significant influence on buffalo oocyte apoptosis. Our findings suggested that addition of 10 ng/ml leptin to IVM medium of buffalo oocyte can improve oocyte nuclear maturation. Furthermore, we showed that there is no relation between in vitro addition of leptin to buffalo oocyte IVM medium and oocyte apoptosis.

Meat contamination has been linked to consumer health problems, as proved by outbreaks and recalls from market places. Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella spp. are considered among the most important pathogens which can be spread through meat and meat products consumption.. The aim of the present study was to determine the prevalence of E. coli O157:H7, L. monocytogenes, and Salmonella spp. in different kinds of meat marketed in Ahvaz, South-west part of Iran.. A total of 210 samples of beef, buffalo and lamb meats were collected from retail outlets and popular supermarkets. After each single pathogen and DNA extraction, multiplex PCR as a rapid and cost-effective method was carried out to determine the prevalence of the pathogens in the samples.. L. monocytogenes was detected in 2.8% of beef and buffalo samples and 4.3% of lamb samples. E. coli O157:H7 was detected in 2.8% of beef and 1.4% of buffalo samples. However, no contamination with this pathogen was found in lamb samples. The prevalence of Salmonella spp. in beef, buffalo and lamb samples was 4.3, 2.8 and 7.1%, respectively.. Due to the presence and potential hazard of E. coli O157:H7, L. monocytogenes, and Salmonella spp. in meat samples, the detection of these pathogens in different kinds of meat is crucial to safeguard public health..

The aim of the present study was to evaluate three thawing rates on the post thaw motility, viability and chromatin structure of buffalo semen frozen in 0.5-ml straws. In this experimental study semen was collected with artificial vagina (42˚C) from four buffalo bulls.Split pooled ejaculates (n=4) were extended at 37˚C with a Bioxcell® extender. Semen was cooled to 4˚C within 2 hours, equilibrated at 4˚C for 4 hours, then filled in 0.5 ml French straws, and frozen in programmable cell freezer before plunging into liquid nitrogen. Straws were thawed at water bath temperatures of 37, 50 or 70˚C for 30, 15 and 6 seconds, respectively. Semen was incubated at 37˚C for 2 hours and evaluated for post thaw motility, viability, acrosomal and DNA integrity of spermatozoa. Analysis of variance (ANOVA) was used for comparisons of means. When the ANOVA test showed statistical differences, the mean of the treatments were compared using Duncan’s multiple range tests. The initial postthaw motility (0 hour) averaged 62.7 ± 7.2%, 73.1 ± 9.77%, and 74.9 ± 8.58% for the three thaw rates, respectively. Kinematic parameters such as average path velocity, linearity and beat/cross frequency in the thaw rate of 70˚C for 6 seconds were superior to other rates studied (p<0.05). After 2 hours of incubation, proportions of progressive motility and Kinematic parameters decreased in all groups (p>0.05). A positive correlation was detected between sperm motility and thawing rate after two hours incubation times. The percentage of viable spermatozoa and spermatozoa with an intact acrosome and plasma membrane integrity were not different between the groups of samples thawed at different temperatures (p>0.05). The percentage of spermatozoa with chromatin dispersion forthe thaw rate of 70˚C for 6 seconds was significantly higher than for the to other rates studied (p< 0.05). In contrast with motility and viability, the DNA integrity of post thaw spermatozoa remained unaffected during 2 hours incubation. The post thaw motility and kinematic parameters of buffalo spermatozoa were significantly improved immediately after thawing by increasing the thawing rate from 37˚C in 30 seconds to70˚C in 6 seconds. However, this relative advantage had disappeared after incubation in a water bath at 37˚C for two hours.A thaw rate of 70˚C for 6 seconds was associated with higher chromatin dispersion than the other thaw rates studied. Sperm thawing over at 50 degrees could be safely used to improve motility recovery after sperm cryopreservation in buffalo bulls.

The sexual maturity is a major factor of fertility and reproduction activity. Since no information was availabe about microscopic developement of seminiferous tubules structure including spermatogenic cells, sertoli, PGC (Primordial Germ Cell) and laydig cells, the present detailed study on buffalo testis was undertaken. The present study was made on 30 buffaloes testes, divided into four groups Viz. 1) 3.5-6 months 2) 7-11 months 3) 12-23 months 4) 24-36 months. Tissue samples were processed for paraffin section and stained by Haematoxylin and Eosin stains. The microscopic observation showed that the seminiferous tubules had tubular structure with a define lumen at 3.5 months of age and their epithelium was consisted of abundant spermatogonia, PGC and sertoli cells. The primary and secondary spermatocyte, spermatid and spermatozoa were not seen upto 9 months of age. The coiling and thickness of seminiferous tubules were increased at one year of age. The leydig cells were seen as large clusters in interstitial connective tissue, PGC, sertoli cells, spermatogonia, primary and secondary spermatocyte, spermatid and spermatozoa also were observed in one year age. The development of seminiferous structure were observed continiued with the age. The results showed that, the perfect spermatogenesis and production of spermatozoa is observed in one year age, which it seems similar to cow (10-11 monthsage).