The mitochondrial cyt b gene plays a serious role in investigating untruthful meat species. This study aimed to authenticate the species of poultry products (Escallop, Nugget, Steak, and Sausage) depending on cyt b gene by using universal cyt b primer. DNA was isolated, and then a band of 359 bp of a mitochondrial cytochrome b gene was produced during the PCR amplification. The PCR products were exposed to Hinf1 and Rsa I restriction enzymes. The restricted fragments produced by restriction fragment length polymorphism technique (RFLP), were run by agarose gel electrophoresis. Results showed that all products had a similar band except sausage product does not follow the rule and showed mislabeling product by the REs, Two bands were yielded by HinfI RE for all products (114 and 245) bp with the differentiated sausage among other products based on the fake product (63 and 296) bp, while digestion by Rsa I produced three bands for escallop, nugget, and steak, (63, 100, 196), but only two bands for sausage was generated (148 and 211). As result, the study offered that analyzing meat products to detect the origin species via a PCR-RFLP technique by using these restriction enzymes can give reliable results. In short sausage is considered as fraud products because the results showed different bands as compared with poultry meat.
The study aimed to identify the species origin of imported buffalo meat from three countries Ukraine, Brazil and India to Erbil using PCR-RFLP technique. The pair of universal cyt b primer was designed to amplify a 359 bp DNA fragment in PCR amplification. Then the amplified fragments were digested with Hinf1 and Rsa I restriction enzymes, achieving a characteristics banding pattern in a 2% agarose which produced evidence to identify origin meat species. The results presented that digestion of samples with the Hinf1 RE, produced two bands in each (Ukraine and Brazil), (58 and 301) bp while it was showed different bands in Indian buffalo meat (85, 274) bp. On the other hand the outcomes in the Rsa I RE were two bands in Ukraine and Brazil (156, 203) bp and two bands were obtained in Indian buffalo meat (106, 253) bp. The results realized that the Indian buffalo meat species was not acceptable and showed mislabeling products. Thus, the obtained results recommend that the PCR-RFLP technique with HinfI and Rsa I restriction enzymes play an important role to detect the origin meat species, since it is a fast, simple and easily handle technique for detection of meat species.
The current study was carried out to evaluate and identify the genetic variation between Local Black chicken lines with two commercial Layer (Isa Brown) and Broiler (Rose) breeds using RAPD markers and sequencing approach of 18s rRNA gene. From the result of the RAPD marker, all primers used were produced 152 scorable bands ranging from 2 to 9 with the size ranging from 320 to 2990 bp with percentage polymorphic loci 64.86% among chicken populations. The highest amplified fragments by primer OPC-11 and lowest by OPAA-03 were 24 and 8, respectively. The mean of the observed number of alleles (na), effective number of alleles (ne), gene diversity (h), Shannon's information index (I) for all loci found to be 1.6486, 1.5189, 0.2883 and 0.4129, respectively. The existence of a high level of polymorphisms and targeted (74) loci throughout all chicken populations/primers indicate sufficient genetic distance and more genetic variability among chicken populations using RAPD-PCR techniques. Result of blasted sequences of 18srRNA gene of local chicken has GenBank accession number MT808178 and MT808179 by BLAST tool against Gallus gallus, it showed the highest identity 95.74% and 94.88% for data of first and second part, respectively. The overall dendrograms clustered showed that the local chicken was closer to the commercial layer than to the broiler chicken lines. Therefore, it suggests that improving the local Black chicken line according to the layer breeding program to collect genetically invaluable genetic resources.
The present study investigated and identified genetic diversity between native White chicken lines and two commercial Broiler (Rose) and Layer (Isa Brown) chicken breeds using RAPD markers and a sequencing technique. All primers applied produced 151 scorable bands with percentage polymorphic loci of 54.93% within chicken populations, as per the results of the RAPD marker. The maximum amplified fragment by primer OPC-11 was 22 and the fewest by primer OPAA-03 was 7. For all loci analyzed, the effective number of alleles (ne), means the observed number of alleles (na), Shannon's information index (I), and gene diversity (h) was 1.4394, 1.5493, 0.3496, and 0.2441, respectively. The presence of a high number of polymorphisms and targeted (71) loci across all chicken populations indicates that RAPD-PCR techniques provide sufficient genetic distance and higher genetic variation among chicken populations. The highest identity of the blasted sequences of the 18srRNA gene of local white chicken is 90.41% and 84.23%. Likewise, a total of 46 and 27 nucleotides are altered with 27 and 10 gaps in both sequences for the first and second regions, respectively. According to both phylogenetic trees, the local white chicken had a stronger sense of individuality and was slightly closer to the commercial broiler breeds than the layer chicken breeds. As a result, it suggests that enhancing the local chicken line requires a broiler breeding program, as well as cross-breeding with other native chicken lines to obtain hereditarily significant new strains.
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