pcr

The aim of the present study was to detect Hepatozoon canis infection in dogs in Urmia municipality, northwestern Iran. The effects of age, sex, lifestyle, and tick infestation as risk factors in the incidence of the disease were studied. During years 2018 and 2019, a total of 246 whole blood samples were taken from the jugular vein of each examined dog (103 stray dogs, 99 shelter dogs, and 44 pets) and subjected to microscopic and molecular examinations. Ixodid ticks were also collected from the body surface and identified. Microscopically, infected neutrophils with Hepatozoon spp. were detected in 5 of 246 (2/03%) thin stained blood smears. Molecularly, 23 out of 246 (9.34%) blood samples were found to be infected with H. canis. There was no significant difference in different age groups and the sex of sampled dogs. However, stray dogs had a higher significant infection rate than pets and shelter ones. In body inspection, all ticks were belonging to the specie of Rhipicephalus sanguineus. The obtained sequence was transferred to GenBank/NCBI (samples accession numbers MT001887). BLAST analysis of obtaining sequences isolated from dogs indicated a 100% similarity with H. canis 18S rRNA gene sequences in GenBank. Based on our results canine hepatozoonosis was common in dogs in the study area.

In this research in order to identify the part of the gene polymorphism transforming growth factor beta-3 (TGFβ3) in broiler chickens Ross, polymerase chain reaction technique and single-strand conformation polymorphism (PCR-SSCP) were performed. For this purpose randomly selection was done between 200 chicks and selected chicks were bled, and genomic DNA was extracted. To determine the quality of the DNA extracted 1% agarose gel was used. Then, by using a pair of specific primers through polymerase chain reaction (PCR) gene fragment with the size of 294 bp from parts of fourth introns and entire fifth exons of the gene was amplified, and finally to determine the bands acrylamide gel and stained with silver nitrate were used. The analysis of band patterns lead to identify three band patterns AA, AB and BB, with 0.52, 0.42 and 0.06 frequency, respectively. Two alleles A and B, with a frequency of 0.73 and 0.27 in the study population was also identified. Shannon index and effective number of alleles for the TGFβ3 gene were 0.5833 and 1.6507, respectively. To confirm the results of PCR-SSCP some samples of each genotype were sequenced directly. Compare of least squares mean different genotypes showed that the gene polymorphism TGFβ3 has significant relation with leg weight and carcass traits.

A number of bacillus species have been recognized as effective and beneficial probiotics. However, some of the bacillus species are known to carry virulence genes which are the main cause of disease, hence, it is essential to evaluate their safety before considering them as probiotic. To date, in our country no studies have been performed on the genotypic virulence determinants of Bacillus species recognized as probiotics. In this study we aimed to assess the safety of indigenous bacillus species isolated from broiler chicken (Bacillus subtilis and B.leichniformis) and honey bee (B.subtilis and B.megaterium) samples. The selected bacillus species were identified based on biochemical and molecular, and evaluated for their probiotic properties such as acid and bile resistance, antimicrobial activity against Streptococcus agalactiae, Enterococcus faecalis, Staphylococcus aureus, Pseudomonas aeruginosa, Salmonella typhimurium, Listeria monocytogenes and Escherichia coli, and antibiotic susceptibility pattern. The hemolytic activity, DNase, lecithinase, lipase, gelatinase, and coagulase activity were also studied. The presence of virulence genes, including hbl, Nhe, cytK, bceT, lipA, ces, was analyzed by molecular methods in a PCR reaction. According to results, all 11 Bacillus species showed considerable resistance to acid and bile, and appeared sensitive to all tested antibiotics. Seven isolates were beta hemolytic and lecitinase positive, one isolate was DNase positive, four isolates were cytK positive, while TA0044 and TA0046 were LipA positive. Only two isolates of poultry origin lacked all tested virulence genes and were classified as safe probiotic.

The aim of this research was to utilize a simple, rapid, cheap and safe method for sex determination of newly born canaries through feather to help accurately breeding this bird. The external morphology of males and females of many birds such as canary are fairly similar to each other, Therefore, determining their sexes is very difficult. The different length of CHD (chromo-helicase-DNA-binding) gene in Z and W chromosomes can recignized by using molecular technique based and PCR method, for the sex determination of young birds. In this study, blood and feathers samples of 140 canaries from 10 different breeds were used and their DNA were extracted. It should be mentioned that for PCR, at first 5 primers designed and finally 1 specific primer were confirmed and used. The results showed that the amplification of a fragment with the length of 252 bp for males (ZZ) and two fragments with the lengths of 252 and 309 bp for female (ZW). The bands had a length difference of 57 bp. The DNA sequencing of these two fragments was done for the first time, the sequence of these two fragments of the CHD gene in canary species was submitted and accepted in the NCBI site under the names of CHDZ-1 and CHDW-1 with the accession numbers of MG679825.1 and MG679826.1.

Introduction: Mycobacterium avium subsp. aratuberculosis (Map) is a very slow growing bacterium that solely infect the digestive tract and leads to Johne’s disease (JD) that is characterized by immedicable diarrhea. Considering innate resistance and different transmission modes of Map and its possible role in development or progression of some human diseases (mainly Crohn’s disease), strong zoonotic potential has been proposed for Map. Diagnosis of JD depends on isolation of viable Map (culture), tracking of its consequences on host immune system (ELISA) or amplification of Map genome (PCR). Fecal culture considered as gold standard for Map detection but obstacles like very slow growth rate of Map, decontamination issues and its high expenses led to development of alternatives like PCR based approaches to gain PCR advantages like high sensitivity, specificity and rapidity for diagnosis of Map. Since no published report was found on Map infection status in Kerman, the current study aimed to investigate the presence of Map in bovine feces samples using 3 different detection methods including culture, IS900 PCR and IS900 nested PCR. Also IS1311 PCR-REA was used for strain typing of Map isolates. Materials and Methods: 212 fecal samples were taken from 4 dairy cattle farms located in Kerman province, southeast Iran. The fecal fractions for culture and PCR ere prepared separately to inhibit potential crosscontaminations. Per each sample, 300 μl of prepared inoculum was cultured on slope of Herrold’s egg yolk medium (HEYM) alone, and two HEYM + 2.0 mg/L of mycobactin J slopes. Inoculated slopes were incubated at 37 °C for 4 months and monitored at biweekly intervals. After extraction of DNA from fecal samples, PCR reaction performed in 25 μl volume and products were analyzed by electrophoresis on 2% agarose gels. The samples were considered as positive if 413 bp amplified band was present. To avoid false positive results, a REA approach using AlwI restriction enzyme was adopted. For IS900 nested PCR reaction, 3 ml from IS900 PCR products were used as template. Resultant products were screened as before. To evaluate the limit of detection of PCR reactions, the DNA extracted from confirmed MAP field strain culture was 10 fold serially diluted (1 μg – 1 fg) and the minimum detection level of PCR and nested PCR were observed. Strain typing of Map isolates was based on IS1311 PCR-REA. Results and Discussion: It takes 3-4 months of incubation for appearance of MAP positive colonies. Map was detected in 32/212 (15.1%) of cultured samples. All colonies were verified as MAP using IS900 PCR and REA with AlwI. 44 positive samples were detected by IS900 PCR (20.7%) while nested PCR was able to find 52 infected samples (24.5%). All isolates detected by PCR based methods were also verified as MAP by IS900 PCR-REA. According to expectations, nPCR offered higher sensitivity than conventional PCR. However, comparing the methods used in the present study showed that as a simple and cheap assay, nested PCR is inherently successful in amplification even with rare amount of starting template and has the better ability to detect the Map infection in fecal samples and can be used as a routine method in diagnostic processes. Although this study was the first attempt to access the Map infection status in Kerman province, as summarized in table 3, many studies were conducted all around Iran that can be compared with. Altogether, Map incidence in other central Iran provinces (Isfahan, Fars and Chaharmahal and Bakhtiari) were between 14.1 to 31.8 percent of tested populations based on examination of fecal samples. Considering that most of reported values came from clinically infected or suspected animals, the estimates obtained in this study showed the possibility that the Map incidence in Kerman province may be higher than other parts of central Iran, but it more investigations needed to create a realistic scheme of Map infection status in Kerman province. Remembering that a copy of genome was considered as equivalent to 5 fg of DNA, sensitivity analysis using serially diluted DNA preparations revealed that the minimum detection level were 1 pg (200 genomes) and 10 fg (2 genomes) for PCR and nested PCR, respectively. According to results, compared to nested PCR, conventional PCR is not sensitive enough for diagnostic tests on field samples. Analysis of fragments produced after IS1311 REA showed the presence of explicit patterns of four bands for all obtained isolates that is the indication of MAP C type. Obtained DNA sequences from the amplified IS1311 locus was exactly consistent with previously published sequence for MAP C type. S type was not detected in current study that was consistent with results of researches performed in Razavi Khorasan province. Failure to find S Map strain was not surprising, since its global prevalence in cow population is very low and hard to find. Also, there are few papers dealing with Map strain typing in Iran, so it is conceivable to find S Map type in Iran’s cattle population if more research conducted in this respect. Although paratuberculosis is a global neglected disease, the situation is worse in developing countries like Iran. Contrary to bovine tuberculosis, due to absence of a national program to control paratuberculosis infection in Iran’s dairy herds, no reliable data is available for arrangement of tools for efficient restriction of disease which leads to inadequate attention to JD and eventually receive the least priority to control. Unfortunately, paratuberculosis as an overlooked disease, is a severe hazard for cows’ health and also for economic activities related to dairy cattle industry. On the other hand, Map is still considered as an important suspicious zoonotic agent which cause serious health problems for humans. Meanwhile, considering aforementioned warnings, enactment of crucial health standards is vital to minimize the chance of infection transmission to non-infected populations and alleviating health problems and economic losses due to Map infection.

Transcription factor 7 gene (TCF7) is located on genomic candidate region for fat traits on sheep chromosome no. 5 and it is excepted to affect fat traits. In the present study, blood samples were collected from 164 Lori-Bakhtiari sheep in Shouli breeding station located at Shahr-e-Kord city, and 113 Zel sheep in Shirang breeding station located at Gorgan city. DNA was extracted from blood samples using modified salting out method and polymerase chain reactions were performed for amplification of 643 bp fragment of exon 2 of locous TCF7 gene. Determination of genotype patterns of sheep were obtained according to result of vertical electrophoresis and AA, AB and BB genotypes were revealed. In Zel breed, AA and AB genotype patterns were observed with frequency of 71 and 29 percent respectively and in Lori-bakhtiari breed, AB and BB genotype patterns were observed with frequency of 10 and 90 percent. There was no significant effect of genotype patterns on fat traits In Zel breed but it was significant effect in Lori-Bakhtiari breed. AB genotype had the greatest impact on fat-tail weight and blood cholesterol and BB genotype had considerable effect on blood triglycerides.

This research was carried out in order to detect the polymorphisms in four sites of PEPCK-C gene by PCR-RFLP method in a commercial layer and broiler strains and breeder hens of Mazandaran native fowls breeding station. Blood samples were collected randomly from 150 birds from a three populations and DNA was extracted using modified salting out method. Using polymerase chain reaction (PCR) and specific primers pairs a region with the length of 3792 bp from position -1723 to exon 4 from PEPCK-C gene was amplified. In RsaI marker site two alleles of A (33%) and B (67%) were detected in Mazandaran native fowls, A (34%), B (66%) in commercial broiler and A (24%) and B (76%) in commercial layer population. Two genotypes of AB and BB were detected with the frequency of 66 and 34% in Mazandaran native fowls 68 and 32% in commercial broiler and 48 and 52% in commercial layer hens. In BstEII marker site two alleles of A (28%) and B (72%) were detected in Mazandaran native fowls population, A (19%), B (81%) in commercial broiler and A (59%) and B (41%) in commercial layer lines. Three genotypes of AA, AB and BB were detected with the frequency of 2, 52 and 46 in Mazandaran native fowls, 2, 34 and 64% in commercial broiler and 42, 34 and 24% commercial layer. In PstI marker site two alleles of A (43%) and B (57%) were detected in Mazandaran native fowls population, A (52%), B (48%) in commercial broiler and A and B (50%) in commercial layer samples. Three genotypes of AA, AB and BB were detected with the frequency of 16, 54 and 30% in Mazandaran native fowls, 18, 68 and 14% in commercial broiler and 6, 88 and 6% in commercial layer population. In BstEII marker site (position to 흍) the results showed that all samples from different strains had the same banding pattern with monomorphic BB genotype. Among 3 observed SNPs 2 were in the promoter site and 1 in intron 2 region. As a result of finding polymorphism in this marker site and also its major effect on production traits such as egg for production, feed conversion ratio and growth rate it can be considered as a candidate gene in detection of QTL related to these traits in poultry industry.

To determine of MEG9 gene polymorphism and its association with birth weight in Sistani cattle, the blood sample of 130 calves (male and female) were randomly collected belonging to Sistani cattle Research Center. The DNA extraction from whole blood was performed using phenol-chloroform procedure. Polymerase chain reaction was performed for amplification of 210-bp segments of exon 3 of MEG9 gene using specific primers. Detection of mutations were carried out by electrophoresis of poly-acrylamide gel 8% and silver staining using SSCP method. Association study of allele patterns with birth weight was done with fixed linear model and comparison of means was conducted by Tukey-kramer method. Results showed that three allele patterns including q1, q2 and q3 in PCR product of MEG9 gene in Sistani cattle with frequency of 37, 34 and 29 % respectively. The genetic diversity in MEG9 gene had significant correlation with birth weight (P

The most important traits of animal are affected by quantitative trait loci and environmental effects. Peroxisome proliferator’s active receptor gamma co activator 1 alpha (PPARGC1A) gene is a member of transcription co activators family, which plays a central role in cellular energy hemostasis regulating. In this study, blood samples were collected from hundred Brown Swiss cow and DNA was extracted from whole blood by modified salting out procedure. Two SNP positions, one located in the 3́-untranslated region (A/C) and the other one in iron 9 of PPARGC1A-T19C were genotyped by PCR-RFLP. Breeding values were individually predicted for milk production, fat and protein milk percent traits by DFREML model one. Association between polymorphisms and breeding values were investigated by SAS at 5% significant level. Genotype frequencies of AA, AC and CC genotypes at PPARGC1a-A968C were 0.593, 0.385 and 0.022, as well as, the allele frequencies of A and C were 0.786 and 0.214, respectively. Genotype frequencies of TT, TC and CC for PPARGC1a-T19C were 0.165, 0.428 and 0.407 and the allele frequencies of T and C were 0.379 and 0.621, respectively. Chi-square test of the both SNPs indicated Hardy-Weinberg expectations (P

Introduction The accumulation of improperly folded forms of host-encoded cellular prion protein (PrPc) in the central nervous system (CNS) lead to a fatal neurodegenerative disease in sheep and goats, namely Scrapie. The application of genetic breeding programs to eradicate transmissible spongiform encephalopathies in goats is an important aim for reasons of animal welfare as well as human food safety and food security. Scrapie and scrapie-like diseases are associated with polymorphisms and mutations of the gene coding for PrP, a host neuronal membrane glycoprotein which is found in an aggregated form (scrapie-associated fibrils) in extracts of brain tissues from all mammals affected by these diseases. The different allelic forms of PrP gene have been shown to make animals variably susceptible to this disease. It has been established that presence of abnormal forms of the prion protein (PrP) is associated with scrapie. Several amino acid polymorphisms caprine PrP encoding genes have been reported to be associated with scrapie susceptibility. Sheep exposed to Scrapie have been shown to gain highest scrapie resistance in the presence of Q171R polymorphism and maximum scrapie susceptibility in the presence of A136V polymorphism. Based on A136V, R154H and Q171R/H polymorphisms and their five alleles (ARQ, VRQ, AHQ, ARR and ARH) sheep and goats can be classified into five groups (R1-R5). The most resistant genotype (R1) is ARR/ARR and the most susceptible genotypes (R5) are VRQ/VRQ, VRQ/ARQ, VRQ/ARH and VRQ/AHQ. Additionally, other caprine PrP polymorphisms I142M, H143R, N146S/D, R154H, R211Q and Q222K have been shown to be associated with low scrapie risk. Although scrapie is an animal health issue, its presence has not been investigated in Iranian goat breeds, where sheep and goats are major livestock species. Based on the positive impact of PrP genetics on sheep scrapie in Europe in the past decade, we have established caprine PrP gene variation in 120 Naeinian goats from the Isfahan, Iran to evaluation of genetic variation in this important region of PrP gene.
Material and Methods The DNA was extracted from 120 blood samples of Naeinian goats from Isfahan province using modified salting out method. After amplification of the desired fragment by polymerase chain reaction (PCR), genotyping of samples was carried out by PCR-SSCP Analysis (Single-Strand conformation Polymorphism). In the following, direct sequencing method was used to confirm the genotyping results. Obtained sequences were analyzed by Chromas Pro and BioEdit. In order to evaluate the amino acid polymorphisms caprine PrP encoding gene was used from Expasy server site. In addition, to evaluate the Hardy-Weinberg equilibrium, we used the chi square test in SAS 9.1 software. All statistical tests were considered significant with a level of P≤0.05.
Results and Discussion The results of the present study showed that nine binding patterns were observed at PrP locus in studied goat population. The results of direct sequencing were confirmed the PCR-SSCP analysis results. Genetic analysis on protein sequences revealed an amino acid polymorphism in codon 186 (T- M or K) and two silent polymorphisms in codons 138 and 143. Based on codons 136, 154 and 171, all goats showed ARQ haplotype and there is no variation in these three codons. Additionally, the results of Hardy-Weinberg test confirmed that this population was not compatible with the HWE (P Material and The DNA was extracted from 120 blood samples of Naeinian goats from Esfahan province using modified salting out method. After amplification of the desired fragment by polymerase chain reaction (PCR), genotyping of samples was carried out by PCR-SSCP Analysis. In the following, direct sequencing method was used to confirm the genotyping results. Then, obtained sequences were analyzed by bioinformatics softwars, for example, Chromas Pro, BioEdit. In order to evaluation of amino acid polymorphisms caprine PrP encoding gene was used from Expasy server site.
The results of the present study showed that nine binding patterns were observed at PrP locus in studied goat population. The results of direct sequencing was confirmed the PCR-SSCP analysis results. Genetic analysis revealed an amino acid polymorphism in codon 186 (T- M or K) and two silent polymorphism in codons 138 and 143.
it is noticed that all of polymorphisms in exon 3 of PrP gene are important and can be used to improve the breeding programs. According to three codon system (codons 136, 154 and 171), all of the studied goats had shown ARQ haplotype which offer Naeinian goats were classified in low resistance group (R3).

Insulin-like growth factor-1 gene (IGF1) is a biological candidate gene for investigation traits such as growth, body weight and muscles growth in different species. In this research eight pairs of white and wild quails were crossed reciprocally as a base population. A total of 34 quails were produced in first generation (F1) and 422 quails were generated by intercrossing the F1 population. Body weights at the time of hatching and different weeks were recorded in the second generation. The association between IGF1 promoter region polymorphism and body weight in different ages was investigated in F2 generation. Using PCR-SSCP assay and sequencing, a novel single nucleotide polymorphism (SNP) was identified in IGF1 promoter region in 472 birds from three Generations. Genotypic frequencies of AA, AG and GG genotypes were 0.05, 0.62 and 0.33, respectively for all generations. The frequency of A and G alleles were 0.36 and 0.64, respectively. The AA genotype was lower than AG and GG genotypes for body weights in ages of one to five weeks and slaughter time and the average of daily gain was also lower from week first to four but these differences weren’t significant. The average of daily gain was higher in females compared to males for AG genotype (P

For decreasing the number of breeding ewes on pastures and prevention of demolition pastures, it seems that animal breeding programs are benefit for identifying effective candidate genes on litter size in Iranian sheep breeds and using those. The TGFβ family genes are one of the most important effective factors on litter size in sheep. The GDF9 gene is one of the most important members from this family. The aim of the present study was to identify available mutations in exon 2 of GDF9 gene in crossbred sheep (Romanov rams × Lori-Bakhtiari ewes) using PCR-SSCP. In this study, after extraction genomic DNA from blood samples of 36 crossbred animals with 3-6 months old (17 males and 19 females), the first region (from 5’ end) of exon 2 (634bp segments) of GDF9 gene was amplified using PCR. The single stranded conformation polymorphism (SSCP) patterns of PCR products were studied using acrylamide gel electrophoresis and silver staining method and then from any pattern one sample sequenced. In the studied population, 3 banding patterns 1, 2, and 3 obtained with frequencies of 0.305, 0.584 and 0.111, respectively. The sequencing results showed presence of 4 mutations (471, 477, 520 and 721 situations) in the studied population.

Introduction The compounds known as β-adrenergic agonists (β-AA) are organic molecules that have the ability to bind to β-adrenergic receptors (β-AR) and start biochemical reactions that will result in increase of accretion of skeletal muscle and reduction in accretion of fat. The anabolic responses to β-AA are temporary, with a peak time occurring during the first 14 days, after which there is a linear decline in growth response, due to either down-regulation or desensitization of the β-AR. Based on down-regulation of β-AR research, the hypothesis that intermittent feeding of β-AA could enhance response on growth performance was created. The objective of this study was therefore to ascertain the effects of continuous and intermittent use of ZH for a period of 42 d on β-AR gene expression in Lori-Bakhtiari feedlot lambs.
Materials and Methods The continuous feeding of ZH (daily regimen), intermittent 1 d feeding ZH followed by 1 d of withdrawal (1 on 1 off regimen), and intermittent 2 d feeding ZH followed by 2 d of withdrawal (2 on 2 off regimen) were employed as the different feeding methods. Thirty two Lori-Bakhtiari male lambs (initial BW=44±4.7 kg) were assigned to one of four treatments (8 lambs/treatment) based on initial BW and were fed with a diet content of 14% protein and supplemented with 0.2 mg/kg of live weight d-1ZH. The basal diet without ZH was the added control group. For evaluating gene expression, biopsy samples of the semimembranosus muscle were collected from 3 lambs per treatment before ZH supplementation on d 0, and subsequently at d 21, and d 42. Samples were rapidly frozen in liquid N2. In laboratory after total RNA isolation from muscle, the RNA was then reverse-transcribed into complementary DNA (cDNA). Real time PCR for cDNA samples was performed using an iQ5 BioRad instrument.
Results and Discussion The results of this study showed that the main effects and period × regimen interactions effect was not significant on β1 and β2-adrenergic receptors gene expression. Also there were no significant differences between treatments and control group.
Most of the physiological effects of catecholamines are due to specific interactions with β-adrenergic receptors (β¬-AR). The growth response to β-AA administration commonly is often reduced over time which is possibly a result of desensitization of the β-AR after chronic continuous exposure to the β-AA. The modification in the dosage of β-AA during the feeding period was previously proven to be a useful approach for compensating the reduced response in rats and lambs. Likewise, Hossner (2005) suggested that the intermittent treatment, e.g. treating for 2 days and resting for 2 days is one of the solutions for avoiding the desensitization and down-regulation of receptors. In studies evaluating mRNA responses to β-AA, no change occurred in β1-AR mRNA abundance due to β-AA treatment. Baxa et al. (2010) explained that no detectable changes of β1-AR could be attributed to generally low abundance, which results in differences hidden by sample-to-sample variation. We were unable to detect differences in the mRNA expression of β2-AR mRNA in muscle samples. The decrease in β2-AR protein expression due to ZH treatment may be in response to a posttranscriptional event.
Conclusion According to our results, it can be concluded that we could use ZH intermittently (1 on 1 off-42 d) in feedlot lambs to decrease fattening costs versus of daily administration of ZH. Further investigation is suggested to detect the alteration of β-adrenergic receptors in continuous and intermittent feeding regimens for clarifying possibly down-regulation or desensitization of them.

European foul brood, is a honey bee larvae disease. EFB is a dangerous disease of the bee colonies that considered to be caused by Gram-positive Melissococcus plutonius. The disease is widely distributed in the world and has created a growing problem in some areas. Because of recent advances in molecular technology, identification of bacteria is very easy and nucleic acid detection technology is rapidly replacing traditional methods of microbiology. European foul brood, is located in the list of dangerous diseases for the health of animals. The disease is endemic in many parts of areas and provide seasonal epidemic. The scenario of this disease varies in different countries.
In this study with standard strain, PCR was developed by specific primers. PCR products electrophpresed on 1 % agarose gel.Larval samples using culture and PCR methode, were tested simultaneously.Eightsamples werepositive by PCRand2 sampleswere positiveby culture.

This research was conducted to study polymorphism of intron 44 of ACACA gene and its association with milk traits in Mahabadi goats. Blood samples were taken from 81 Mahabadi goats. DNA was extracted from blood using salting out method. 897 base pair fragment of intron 44 ACACA gene was amplified using polymerase chain reaction. SSCP method was used to investigate the amplified DNA fragment polymorphism. Amplified products were sequenced in both directions. Finally, seven different patterns were obtained and sequences compared with each other also with Capra hircus ACACA gene sequence on NCBI (NC_022311). Seven novel SNPs were identified which were NC_022311: nucleotide 154503(T/G), 154518(A/G), 154577(A/G), 154640(C/T), 154712(A/C), 154928(C/T), 154956(C/T). Then a mixed animal model was used by SAS9.1 analysis system to evaluate the association of these SNPs with milk traits. In this model, variables of age of dam, genotypes and mount of recording were considered as fixed effects. There were significant association of SNP2 (P=0.05), SNP3 (P=0.05) and SNP5 (P<0.01) with milk yield. SNPs 1 (P=0.017), 6 (P=0.017) and 7 (P<0.01) had also significant effect on milk fat-percentage.

In this study, blood samples were collected from the jugular vein of 109 Kermani sheep from Kermani sheep breeding station. Genomic DNA was extracted from blood sample using modified salting out method and polymerase chain reactions were performed for amplification of 214 bp fragment containing a part of exon 4 of GH gene. Single Strand Conformation Polymorphism (SSCP) was used for genotyping. For this purpose, vertical electrophoresis of PCR products was performed on 12% acrylamide gel, at 300 V, for 17 h at 4 C˚. Silver-staining of gels, resulted three genotypic patterns of 1, 2 and 3 with frequencies of 20.18%, 35.77% and 44.03%, respectively. Analysis of variance was performed using SAS software. The results showed no­ significant association between the different patterns of GH gene and growth traits.

The objective of this study was detection of polymorphism in β-Lctoglobulin gene and its association with milk production traits in Mahabadi goats using PCR-SSCP method. For this purpose, blood samples were taken from 89 Mahabadi goat that reared in the farm of animal science department at Tehran university (Karaj). DNA was extracted from whole blood using optimized salting out method. Specific primers used for amplification of 177 bp fragment from then 7 β-Lactoglobulin gene. For identifying the polymorphism, PCR products were electrophoresed on polyacrylamide gel (SSCP method) and stained with silver nitrate method. Results indicated the different patterns, may be due to polymorphism in this fragment. The frequency of the six pattern were 7.14%, 10.71%, 13.09%, 33.33%, 9.52% and 26.19% respectively. The statistical analysis indicated that the effect of different patterns were significant on milk production and somatic cell count (p<0.01), but on fat and protein percentage had no significant effect.