buffalo

Since, feeding in animal husbandry is one of the most critical and costly issues, therefore should focus on cheap resources like agricultural wastes which provide adequate nutrients for livestock. Olive pomace is a residue of the oil extraction of olive fruit, which after drying can be used as a by-product in animal feed. Olive pomace contains high lignin (32% DM), cellulose and hemicellulose (Sadeghi et al., 2009). Raw olive pomace contains high amounts of 16 and 18 fatty acids; and about 96% of olive pomace fatty acids include linoleic acid and oleic acid (Nazari et al., 2008). Olive pomace is containing anti-nutritional components such as phenolic compounds and tannins (0.58 to 1.1 g/kg DM), and it may have adverse effects on livestock (Molina et al, 2003). Feeding dairy ruminants with non-core olive cake containing tocopherol, retinol and phenols can cause positive changes on milk quality, and improved oxidative stability of milk (Pauselli et al, 2007). Most of buffalo breeders use wheat flour in buffalo diet. Using wheat flour, can destroy the rumen microbial population and as an important part in human foods, increases costs. However, there is no research on wheat flour replaced with olive pomace in buffalo nutrition. Therefore, this study was conducted to explore the effect of olive cake on digestibility, blood biochemical parameters and production performance of Khuzestani dairy Buffalo.

Fifteen dairy buffaloes average 4 years, average body weight 570 kg and 2 parities were used in a completely randomized design with 3 treatments and 5 repetitions. Experimental treatments were included 0%, 7.5%, and 15% olive pomace substituted with wheat flour, which fed two times daily for 45 days in a completely randomized design. Diets were formulated according to the NRC. The olive pomace used in this study dried by air drying. The pomaces were stirred and mixed several times daily. The crude protein, metabolizable energy, neutral detergent fiber, acid detergent fibers, and fat of olive pomace were 6.5%, 2.65 Mk / kg DM, 56, 45, and 9 %, respectively.At the end of the experiment, feed intake, nutrient digestibility, production performance and blood parameters were measured. The diets were prepared completely mixed and given daily at 7:00 and 17:00. Fresh water was also available to them. The combination of control diet consisted of corn silage, wheat straw, olive cake, wheat flour, wheat bran, rice bran, mineral supplement and vitamins, limestone. Milking was done every day in the morning and evening and milk production was recorded. Milk scanner was used to measure milk protein, fat, and lactose. The milk pH determined by using pH papers or electrical pH meters; and milk solids and non-fat solids (SNF), by lactometer.Also, blood samples were taken from all buffaloes at the end of the period, 3 to 4 hours after morning intake and blood metabolites determined. For apparent digestibility of the diets, during three consecutive days, buffalo feces were collected. Also, sampling of feed residuals in the last three days of the period was weighed. Then, the digestibility of organic matter, dry matter, crude protein, crude fat, acid detergent fibers and neutral detergent fibers were measured.The obtained data were analyzed using the General Linear Model procedure of the SAS. A comparison of the mean was made with Duncan. The model of this design was a completely randomized design with three treatments, and each treatment includes five replications based on the statistical model: Yij= µ + Ti+ eij. Also, repeated data per time were analyzed based on the mixed statistical model. Yci = µ + Tc + Ɛic + (T*P)ik.

Milk fat and SNF increased in experimental treatments at the end of the period (P<0.05). However, milk production, protein, acidity, density and pH of milk were not affected by the experimental treatments (P>0.05). Milk production increased by using dried olive kernels in dairy buffaloes (Terramoccia et al., 2013). Inclusion of unsaturated fatty acids often reduces the saturated fatty acids of milk. Also, high fat in olive pomace can be a major reason for high percentage of milk fat in the olive pomace treatment compared to control (Chilofalo et al. 2003). Also, the concentration of blood parameters was not affected by experimental diets. But, blood cholesterol in 7.5% olive cake increased and the concentration of alkaline phosphatase enzyme increased by 15% olive cake compared to the other treatments (P<0.05). Obeidat (2017) reported that adding olive pomace to the diet of Awasi lambs had no effect on blood parameters (glucose, blood urea nitrogen, aspartate aminotransferase and alanine aminotransferase). But Hashish and Al-Sameh (2012) declared that saturated fatty acids in olive pomace can increase the concentration of saturated fatty acids and blood cholesterol. Other researchers also concluded that with the increase in dietary fat concentration, blood cholesterol increased (Bhatta et al., 2011). Nutrient digestibility was not affected by experimental treatments (P>0.05). Using olive pomace up to 20% instead of barley in Lori lambs increased microbial protein production in the rumen. Also, olive pomace in the Dalagh sheep diet increased feeding efficiency and carcass weight; and reduced production costs up to 66% (Samadi, 2009). The use of olive pomace in Awasi lambs, did not influence dry matter intake, digestibility of crude protein, neutral detergent fiber and acidic detergent fiber (Obeidat, 2017).

According to the recent study, replacing wheat flour with olive cake had no negative effect on production, blood parameters and digestibility of nutrients in dairy buffaloes, and due to the reasonable price of olive cake; replacing olive cake up to 15% with wheat flour in the diet of dairy buffaloes is recommended.

The rapid development of sequencing technologies and bioinformatic tools has made it possible to sequence the entire genome of many species, providing a starting point for exploring the genome-wide genetic diversity of organisms. The buffalo is one of the most important domesticated animals in the world and has great economic value for humans, being used for milk, meat, leather and most importantly as labor. Khuzestani buffalo are among the most important buffalo in Iran. These animals are scattered in western and southwestern Iran. The largest population of this breed is in the Khuzestan province and they are fully adapted to the geographical features of this region

In this study, whole genome sequencing of the Khuzestan buffalo was performed in paired-end format using Illumina technology. Quality control of the readings obtained met all quality parameters and processing of the readings showed that all samples were of high quality. The BWA-MEM software package was then used to match the data to the reference genome. Genome variants were obtained with the Freebayes software. The final filtering of the variant file then took place. This filtering serves to increase the quality of the analysis and to sort out inferior variants.

The alignment percentage for all samples was over 97.44% and also the coverage in the sequenced samples ranged from x 4.3 to x 9.12, indicating reasonable quality of the reads. Also, in this study we identified 76,676,529 million variants fairly evenly distributed on the chromosomes of the Khuzestan buffalo. The highest and the lowest number of variants were observed on chromosome 1 chromosome 28, repectively. In this study, the total number of translocation and inversion mutations in the genome level was 298,193,017 and 135,694,466, respectively. In addition, the ratio of transitions to transversion was estimated at 2.1989 (Ts/Tv).

The presence of genetic diversity in the native population of Iranian buffalos are valuable parameters that can be studied to evaluate the genetic potential of economic traits in these populations in order to design suitable breeding programs.

Due to the importance of Sarcocystis as a common zoonotic disease in public health, in this study, the infection rate of Sarcocystis infection and molecular identification of its species by using COX1 and 18srRNA genes in slaughtered cattle and buffaloes of the abattoir of Urmia was investigated. For this purpose, samples were taken from the intercostal muscles, diaphragm, esophagus, and thighs of 100 cows and 100 buffaloes slaughtered in the slaughterhouse of Urmia city in 2021. At first, macroscopic and microscopic evaluation were performed on samples. Then, DNA was extracted from homogenized tissues and molecular investigations were done using COX1 and 18srRNA genes. In the macroscopic examination, none of the samples obtained from cows were infected with sarcocystis, while this protozoan was isolated from 11 (%11) samples obtained from buffaloes. The infection rate in the impression smear method in cow and buffaloes samples were 27 (%27) and 16 (%16) and in peptic digestion method was 37 (%37) and 23 (%23), respectively. In parallel, molecular evaluation confirmed the presence of sarcocystis infection in 39 cases (39%) out of 100 cow carcasses and 26 cases (26%) out of 100 studied buffalo carcasses. The findings of this study indicate relatively high infection rate in cattle and buffalo populations of the province. The findings of this study indicate relatively high infection rate in cattle and buffalo populations of the province. So, it is suggested to avoid eating raw and undercooked meat.

Metabolites of medicinal plants are used to improve energy efficiency in the rumen and animal productivity. Malva sylverstris is a plant whose cultivation yield in a quarterly period is estimated at 10 tons per hectare. Malva sylverstris leaves and flowers contain tannins, calcium oxalate, pectic substances and mucilage, anthocyanins, rich in vitamins, and due to the presence of vitamin C and flavonoids, many uses have been mentioned for this plant. In some studies, the use of herbs had an increasing effect on milk production performance. This experiment was conducted to investigate the effect of Malva sylverstris nutrition on milk production and composition, liver enzymes, nutrient digestibility, and blood biochemical parameters of dairy buffaloes.

Eight khuzestani dairy buffaloes, 4-5 parities, average weight 600±12 kg in the early lactation (forth months), average milk production 6 liter, were used. Buffaloes randomly fed with a diet without Malva sylverstris and diet containing 500 g Malva sylverstri/ dry matter plant for one month. Animals were kept in individual stables and adaption to the diet was performed for one week. Sufficient and clean water was provided to the buffaloes.At the end of the experiment, feed intake, nutrient digestibility, milk production, quality and microbial load of milk were measured. Buffalo blood parameters such as glucose, cholesterol and triglyceride, urea and liver enzymes were determined. The data were analyzed in a completely randomized design and the means were compared with the Tukey test at significance level of 5%.

The results showed that milk fat, protein, and lactose of milk did not change in the experimental treatments (P >0.05), but dry matter intake and milk production of buffaloes increased. According to the results, the milk microbial load of buffaloes fed Malva sylverstris decreased (P <0.05). Blood glucose, cholesterol, and triglyceride levels in the Malva sylverstris treatment were lower than the control treatment (P <0.05). Blood urea and liver enzymes were not affected by the treatments (P >0.05). Nutrient digestibility like dry matter, crud protein, and nautral detergent fiber in buffaloes fed Malva sylverstris increased (P <0.05).

According to the result of the present study, the use of Malva sylverstris plant in the diet of dairy buffaloes not only did not have negative effects on the production performance of dairy buffaloes but also increased milk production and reduced the microbial load of milk, therefore it can be used as an appropriate supplement in the dairy buffaloes.

In this study, in order to genetic and economic evaluation of the progeny testing selection program and the usual selection program of buffaloes in Khuzestan, 3000 buffalo cows were simulated using R program. In each progeny test programs, different levels of milk recording (100, 80, 50 and 20%) and artificial insemination (100, 80, 50 and 20%) were used and in usual program, the selection was based on pedigree information of male calf and its mother milk production. In the selection index, two traits of milk production and fat percentage were considered. The studied programs were compared based on genetic improvement, production changes, total genotypic value and costs and revenues of programs. In study of selection programs, it was observed that with increasing the proportion of herds covered by artificial insemination, genetic progress and changes in total genotypic value were increased in progeny testing programs. This increasing was also observed for the cost and revenue of each of considered programs. In addition to artificial insemination, the milk recording system was also effective in increasing the two parameters of cost and revenue. However, the additional costs of using milk recording system and artificial insemination were offset by increased revenue from performance of better buffaloes. In general, based on changes in total genotypic value, genetic improvement of milk production and minimum cost and maximum income of the programs, the progeny testing selection program with 20% of milk recording and 100% of artificial insemination was selected for buffalo herds as the suitable progeny testing program.

In this study, production, population and economic parameters of 30 buffalo herds of Khuzestan province in two sale conditions during 2016-2017 were used to estimate the economic value for production, reproductive and survival traits. Economic value of the traits were estimated by one unit increase for each trait using the maximum profit trend. The average annual revenue, cost and profit per head of buffalo for milk and local cream condition were 60480817, 59270944 and 1209873 Rials, respectively, and for just milk sale condition were 59220881, 59270944, and -50062.7 Rials, respectively. In the milk and local cream sale condition, as a proper sale, the economic value of production traits including milk yield, fat percentage, fattening male calf weight and weaning weight were 16774, 178011, 16051 and -46070 Rials, respectively, the economic value of reproductive traits including conception rate, live birth rate, calving interval, buffalo heifer weight and age at first calving were 476142, 527296, 9504, -36668 and -76743 Rials, respectively, and the economic value of survival traits including survival rate of buffalo cow, survival rate of calf from birth to 4 months of age, survival rate of buffalo heifer from 4 months to parturition and longevity were 364946, 216406, 143588 and 72030 Rials, respectively, per head of buffalo in a year. According to the results, selection for increasing the average milk yield, fat percentage and fattening male calf weight would improve the economic condition of buffalo breeder. Considering the negative economic value of calving interval and the age at first calving, breeding in order to shorten them can increase the economic efficiency of the herd. Also, due to the superiority of milk and local cream sale condition compared to milk sale as well as taking into account that due to the lower milk production in buffalo raw milk selling was unprofitable, dairy products selling along with milk can be regarded to improve the economic efficiency.

Total of 43 milking buffaloes were considered in this experiment to investigate the effect of body condition score of calving (BCSc) on reproductive performance at North-west Buffalo Rearing and Breeding Center, West Azerbaijan, Iran. The experiment was carried out in the form of completely randomized design with 2 factors of BCS and Parity. The BCS had 4 of 2.5-2.99, 3.0-3.49, 3.5-3.99, and 4.0-4.49, and Parity had 3 levels of 1: first calving, 2: 2nd- 5th parity, and 3: 5+ parity. The BCSc of individual animals was determined by measuring animals’ backfat thickness using an ultrasound machine at the 12–13th rib (mm) at the start of the trial. The results showed a linear relationship between BCSc and parity with calf birth weight (P<0.05). The lowest age at first estrus after calving was in animals within group of BCS with 2.5 to 2.99 (P<0.05), but the effect of parity on age at first estrus after calving was not significant (P<0.05). The main effect of BCSc and parity were significant on conception rate in the first artificial insemination such that buffaloes within BCSc group of 3-3.49 and within parity group 2 had higher conception rate. In this study the open days after calving and calving intervals increased with increasing BCS (P<0.05). The effect of BCS and parity group was not significant (P>0.05) on the number of artificial insemination resulted in pregnancy. Breeding efficiency was linearly associated with number of parity (P>0.05), but it did not affected by BCS (P>0.05).

The aim of this study was to compare the Wood, Dijkstra and Sineus models for fitting the lactation curve of the first abdomen of Khuzestan Buffalo. The effect of fix factor such as year, season and herd on the curve of milk parameters were studied. In this study, 73169 test day records of milk were used. Records were collected in the animal breeding center. According to the test day record and using different initial values, using SAS software and NLIN procedure were estimated the parameters of each lactation models. R2adj, AIC, MSE, RMSE and Iterations were used for comparison of models. The results of the model's comparison were showed that all three models of Wood, Dijkstra and Sinus were able to fit the buffalo curve. But the Wood model, having the lowest AIC, MSE and RMSE was goodness than the other two models. The mean of Wood parameters such as a, b and c were 2.36, 0.656 and 0.0009 were respectively. Average the peak of lactation was 84 days. The most and least lactation continuous was in summer and spring, respectively (p<0.05). All Wood parameters and lactation indexes were affected year, herd and their interaction (p<0.05). By using the lactation curve and determining the components of lactation, it is possible to plan for feeding and management of buffaloes.

The aim of this study was to determine genomewide linkage disequilibrium (LD), Haplotype block and effective population size using the information obtained from 243 Azarbaijani breed buffalo using a high density SNP panel (Axiom® Buffalo Genotyping 90K). After quality control of SNP markers data, 62,141 SNP markers remained for identification of linkage disequiliberum, haplotype blocks and effective population size. LD was measured by the square of the correlation coefficient (r2) between alleles. The maximum LD measured by r2 varied from 0.25 to 0.29 at a distance of < 2.5 kb, and the minimum average values of r2 varied from 0.012 to 0.014 at distances ranging from 900 to 1000 kb, clearly showing that the average r2 reduced with the increase in SNP pair distances. Overall, 1693 blocks were observed through the genome. Eleven percent of all SNPs were clustered into haplotype blocks, covering 202 ±3.4 Mb of the total autosomal genome size. Effective population size (Ne) was estimated based on expected linkage disequilibrium. Ne was estimated to be 422 in our population.

In order to investigate the milk and its components curve of Khuzestani buffaloes a total of 9061 data of test-day milk, fat and protein yield of the first lactation period buffaloes collected during 1993-2013 were used. The buffaloes having less than 4 test-day records were removed. The lactation days was considered from the 5th to 210th days of lactation. Five different functions of Wood, Wilmink, Parabolic, Mixed Logarithmic and Reverse Polynomial were tested for fitting the lactation curve of buffaloes using the NLIN procedure of SAS 9.1 software. Comparing the studied functions, the Wood function was the best predictor of the lactation curve. The most suitable models for the fat and protein percentage, fat and protein content were Parabolic and Inverse Polynomial with the same results, Parabolic, Parabolic and Inverse Polynomial with Wood functions, respectively. Based on the suitable models, the expected daily milk yield, fat percentage, fat and protein content and protein percentage at the beginning of lactation (the 5th day of lactation) were 6.10 kg with a peak yield of 8.46 kg on the 89th day of lactation, 5.51% with a peak of 6.35% on the 210th day of lactation, 427.75 g with a peak of 513.73 g on the 153th day of lactation, 4.20% with a peak of 4.23% on the 210th day of lactation, 317.70 g with a peak of 344.43 g on the 82th day of lactation, respectively. The results showed that Khuzestani buffaloes have the same production as other breeds in the world in terms of milk production and peak yield or close to them. Considering the rural breeding situation of buffaloes in Khuzestan province, it is expected that improvement of environmental conditions such as nutrition and management provide the condition for incidence of the maximum genetic ability of the buffaloes.

This experiment was conducted to investigate effect of replacing alfalfa with subabul pod (0, 9 and 18 % of DM) on digestibility, in vitro fermentation and ruminal degradability in fistulated cow and buffalo. Fermentation of experimental diets was determined by gas production, digestibility by tilly and terry and degradability by in situ (4 replicates per diet) and the data were analyzed by split plot design. Digestibility of dry mater, NDF and ADF of diets containing different levels of subabul pod were not different in cow and buffalo (P>0.05). But potential of gas production in diets containing subabul pod was lower than control diet (P0.05). Cell wall degradability was greatest for control treatment (in buffalo 23.9 % and cow 35.9 %, respectively) and the lowest value was for diet containing 18 % subabul pod (in buffalo 64.1% and cow 64.3%, respectively) (P0.05). According the result, replacement of alfalfa hay with subabul pod in cow and buffalo diet, didn’t affect in vitro digestibility and gas production parameters, but improved in situ ruminal degradability. Therefore, it seems 50 % subabul pod can be used instead of alfalfa in cow and buffalo nutrition.

Understanding of population genetic structure is valuable for better implementation of breeding programs and most importantly, preservation of genetic resources. Genomic data provide an opportunity to consider complex evolutionary history of populations and reconstruct rare historical events. In this research, the structure of Iranian buffalo populations was studied by using principal component analysis and discriminant analysis principal component methods. For this purpose, the number of 404 buffalos from three breeds including North, Azari and Khozestani were sampled and genotyped by SNPChip 90k from Padano Company in Italy. The results of principal component analysis and discriminant analysis principal component showed a clear picture of the genetic structure of the studied populations. Assessing the optimal number of clusters with criteria BIC, K = 3 by the DAPC method showed the best results. The result of cross-validation for retaining principal components was optimized to 50 first components that showed the lowest MSE. In this study, DAPC predicted assignment of individuals to clusters and membership probabilities with 100% accuracy. PCA method was not able to provide a group assessment and DAPC method outperformed than PCA in achieving a clear variance difference between populations. DAPC method can be applied in quality control and stratification population correction of GWAS as an alternative to the PCA because of summarizing the genetic differentiation between groups and overlooking within-group variation and providing better population structure.

Introduction Siris is a tropical legume. It has common names such as silk tree. Compositional studies indicated carbohydrates as major components and potassium was found in the highest amount and copper in the lowest. Siris leaf has high content of N (16 to 23 percent crude protein) and Ca, a low content of tannins and phenolic compounds. Levels of total tannins in the leaf of siris is 4%. The amino acids profile indicated that arginine and lysine are present in large amounts in seeds while glutamic acid and aspartic acid are higher in pods. Siris is an economically important plant for industrial and medicinal uses. This experiment was conducted to investigate the different levels (0, 50 and 100 %) of siris leave or silk tree instead of alfalfa on degradability parameters, fermentative and in vitro digestibility in cows and buffalo Khouzestan.
Materials and methods The leave of siris were collected from shushtar and milled. Levels of 0, 50 and 100% leave replaced alfalfa in the diet. Rumen fluid was collected from fistulated cow and buffalo before the morning feeding. About 500 ± 10 mg experimental samples (1.0 mm screen) incubated with 35 ml buffered rumen fluid under continuous CO2 reflux in 100 ml vials for 2, 4, 6, 8, 10, 12, 16, 24, 48, 72 and 96 h, in a water bath maintained at 39°C. Cumulative gas production data were fitted to the exponential equation Y=B (1−e−Ct). Partitioning factor, microbial biomass, actually degradable organic matter was calculated. Digestibility of dry matter and NDF of the samples were determined using an in vitro procedure. Dry matter and crude protein degradability were measured by in situ technique using fistulated animals (fed 60% forage% concentrate). Samples put in the polyester bags and incubated in the rumen for 2, 4, 6, 8, 16, 24, 48, 72 and 96 hours. After the specific incubation periods, the bags immediately were hand-rinsed under cold tap water until clear, and dried in a forced-air oven (60 °C, 48 hour). Disappearance of DM and CP of samples from bags with incubation time were calculated using the equation of, P = a b (1- e -ct), P= fraction degraded in the time t, a= soluble fraction, b= potentially degradable fraction, c= degradation rate and t= incubation time. The effective degradability was calculated using the equation of ED = a (bc/(c). Data were subjected to analysis as a split plot design using the General Linear Model (GLM) procedure of SAS.
Results and Discussion Dry matter digestibility of the diet containing 50% siris leave in Buffalo was 85.70 % and in cows, 82/94 %. Regardless of the type of treatment digestibility of dry matter and NDF was not significant between cow and buffalo. Regardless of the type of animal, the potential of gas production in the diets containing 0, 50 and 100 % siris leave was 175.05, 108.44 and 103.14 ml, respectively. The effect of diets containing leave on pf, microbial biomass, microbial biomass efficiency and organic matter digested was not significant. The efficiency of the microbial biomass in Buffalo (0.7281%) was higher than the cow (0.6308%). Slowly degradable fraction (b), constant degradable rate (c), potential of degradability (PD) and effective degradability (ED) of dry matter at 50 % leaf was the highest. Fraction a rapidly degradable (a) and effective degradability (ED) protein at 50 and 100 % the leave was the highest amount in compared to the control diet. In agreement with results, the researchers reported tropical legumes because of high quantity of protein and carbohydrate and lower fiber can be used as a suitable substrate for rumen microbial growth. Also legumes ability to provide required nitrogen, energy and vitamins for microorganisms. But presence of anti-nutritional factors such as tannin, saponin and oxalate in siris might be reduce fibrolytic enzyme activity and consequently fiber digestibility and fermentation.
According to the results it ccan be concluded siris leaves or silk tree improved fermentation condition in the rumen, therefore can be used instead of alfalfa in the diet of cow and buffalo in vitro.

The purpose of this research was to classify buffaloes from different areas of the two Azari (West and East Azarbayjan and Ardabil provinces) and North (Guilan province) ecotypes using support vector machine method. A total of 258 buffalo were sampled and genotyped using the Axiom Buffalo 90K Genotyping Array at the Parco Technologic Padano lab in Italy. Two metric methods of cross validation and the area under the receiver operating characteristic (AUC) were used to determine the predictive performance of support vector machine (SVM) to classify individuals. The results of cross validation and methods for classifying different regions of the two ecotypes (4 provinces) were 92% and 96%, respectively that showed despite the difficulty of identifying individuals from provinces close to each other, support vector machine (SVM) method shows higher accuracy in assigning animals to their herds. Result of two ecotypes showed accuracy about 96% and 98% which represents the better ability to separate the two ecotypes. Machine learning method provides predictions for classification of each individual which can be efficient in quality control and genetic studies.

Milk production and its components are belong to quantitative and polygenic traits and affected by many genes. This study was conducted to determine the polymorphism in β-lactoglobulin, prolactin and DGAT1 genes of buffaloes in Khuzestan province using PCR-RFLP technique. Therefore, blood samples were collected from 200 buffaloes of Khuzestan province and then DNA samples were extracted using optimal and modified salting-out method. After DNA extraction, the genotype of the animals was determined by PCR-RFLP. For genotyping of animals, the 262 bp fragment of exon and intron 4 of the beta-lactoglobulin, the 411 bp fragment of exon 8 of the DGAT1 and the 156 bp fragment of exon 3 of the prolactin genes were amplified and then digested using restriction enzyme HeaIII, CfrI and RsaI, respectively. Then restricted PCR products electrophoresed on 2% agaros gel and stained in etediumbromide. Results of this study showed that the allele of B, the allele of A and the allele of K has been fixed in beta-lactoglobulin, prolactin and DGAT1 loci in buffalo population of Khuzestan, respectively and polymorphism was not seen for any of these loci.

In the present study karyotype of Mazani buffalo was studied in comparison with those of Azeri buffalo populations from Iran. Karyotyping analysisis a powerful instrument to determine the karyotype of farm animal and to turn up more about fundamental basis for chromosomal abnormalities.Blood samples were taken from ten (5 males and 5 females) Mazani buffaloes and thirty (15 males and 15 females) Azeri buffaloes. The Mazani buffaloes belong to state of Mazendaran and Azeri buffaloes belong to states of west and east Azerbaijan and Ardebil. Blood lymphocytes cultured at 37ºC for 72 hours in the presence of phytohemagglutinin and the metaphase spreads were performed on microscopic slide. Giemsa was used to stain chromosomes. The Mazani and Azeri Buffalo exhibited the same karyotype with diploid number of 2n = 50. The fundamental numbers (NF) were 60 in male and female. Stebbins classifications show both Mazani and Azeri buffaloes belong to 3B group. Both of Mazani and Azeri buffaloes are riverine. Based on the karyotype symmetry parameters, Mazani buffalo populations showed more symmetry that Azeri buffalo populations. The chromosomal evidences and other detailed karyotype analysis allows us to detect interrelationships of spices from a chromosomal point of view, to group the different populations and postulate relationships among them.

The aim of the present study is to investigate the effects of in vitro supplementation of Tempol on spermatozoa of buffalo bulls before and after freeze-thawing. A total of 25 samples from five healthy buffalo bulls (5 ejaculates from each bull on 5 different occasions) were used. Each ejaculate was diluted at 37°C in tris-citric acid based extender (without egg yolk) containing 0 (control), 0.5, 1, 4, 8 and 12 mM Tempol. Sperm progressive motility and viability were evaluated in diluted semen at 0 (T0) (immediatelyafter dilution), 60 (T1) and 120 (T2) minutes after semen dilution. In the next step, the same doses of Tempol, egg yolk and glycerol were added to the above extender and semen samples diluted, cooled to 4°C within 2 h, equilibrated for 4 hours and sperm parameters (progressive motility, viability, membrane integrity and DNA damage) estimated. The equilibrated semen was packed into 0.5 mL French straws and frozen in liquid nitrogen. Later, the semen was thawed and analyzed for the same parameters, as well as total antioxidant capacity (TAC). Results showed that adding 0.5 and 1 mM Tempol to the semen extender preserved the sperm progressive motility in diluted and equilibrated semen compared to the control without affecting other parameters. However, in frozen-thawed semen, extenders containing 0.5 and 1 mM Tempol significantly preserved sperm progressive motility, viability, membrane integrity and semen total antioxidant capacity and also resulted in lower DNA damaged sperm counts. It was concluded that addition of 0.5 or 1mM tempol to the extender would preserve semen quality during freeze-thawing.