Background With regard to the extensive production of genetically modified organisms, scientists ... more Background With regard to the extensive production of genetically modified organisms, scientists focused on the safety of antibiotic resistance marker (ARM) genes present in GM-crops produced for the consumption of human or animal. We investigated the possible transfer of these genes to the microflora present in the gastrointestinal tract and blood cells of two groups of Male Wistar Albino rats fed on a transgenic diet containing the ARM genes nptII and aadA for 90 days. The conventional PCR was employed to screen for the presence of nptII and aadA genes in the experimental diets, and animals’ samples. Results The occurrence of DNA transfer of nptII and aadA genes from GM-diet to blood and bacterial cells has been unambiguously demonstrated. Results were further confirmed by sequencing and blast analysis, indicating that both transferred segments shared significant alignment with number of Binary and Cloning Vectors with variable similarities. The frequency of transfer of DNA segmen...
Background One of the parameters required for the assessment of food and feed safety is detection... more Background One of the parameters required for the assessment of food and feed safety is detection and identification of genetically modified organisms. Legislation in some countries necessitates detection and quantification of modification in food and feed samples. Scientists have raised concern about safety of antibiotic resistance marker (ARM) genes used for transformation of crops intended for human and animal consumption. In the present work two molecular approaches have been adopted: one exploratory; for detection and quantification of ARM genes in tested plant samples and the other confirmatory; to determine the specificity/reliability of the obtained results. Results Results revealed that primers for neomycin phosphotransferase (nptII) and aminoglycoside 3″ adenyl-transferase (aadA) were amplified in the majority of the 36 DNA screened samples. Melting curve analysis using hygromycin phosphotransferase (aphIV) gene as target sequence for the fluorescent-based detection approa...
Background and Aim: Uterine lumen contamination with bacteria is ubiquitous in buffalo after part... more Background and Aim: Uterine lumen contamination with bacteria is ubiquitous in buffalo after parturition. Nearly one-third of these infected animals develop endometritis which leads to reduced fertility. The present study aimed to evaluate the expressions of IFN-Îł and NOS2 genes in uterine tissue of buffaloes with endometritis and comparing them with those in healthy animals using RT-qPCR Materials and Methods: Uterine samples were collected from 50 apparently healthy and 50 clinically infected buffaloes. RNA was extracted from the collected buffalo's uteri and cDNA was synthesized from extracted RNA. Quantitative Real Time PCR technique was performed using this synthesized cDNA. Results: Apparent up-regulation of both genes mRNA expression was recorded in endometritis-infected animals with 8.3-folds for IFN-Îł and 9.99-folds for NOS2 (P<0.001). Conclusion: The upregulation of IFN-Îł and NOS2 expression in the uterine tissue of endometritis-infected buffaloes can be used as a s...
ABSTRACT Authors&#39; contributions This work was carried out in collaboration between all au... more ABSTRACT Authors&#39; contributions This work was carried out in collaboration between all authors. Author OEO designed the study, followed up the practical work and wrote the final version of the manuscript. Author MFAS managed the analyses of the study, managed the literature searches and wrote the first draft of the manuscript. Author NAAEM performed the practical work. Author KMS followed up the steps of the search. All authors read and approved the final manuscript Article Information DOI: 10.9734/BBJ/2015/11619 Editor(s): (1) Giuliana Napolitano, ABSTRACT Aim: The hypothalamic hormone, growth hormone-releasing hormone, is the principal stimulator of pituitary growth hormone (GH) synthesis and secretion. GHRH and its receptor (GHRHR) provide important functions in the regulation of the GH axis and in the development and proliferation of pituitary somatotropic axis. This study aimed to identify the genotypes and nucleotide sequences of two multifunctional genes; growth hormone-releasing hormone (GHRH) and its receptor (GHRHR) in Egyptian buffalo. Methodology: Genomic DNA was extracted from blood samples of 100 healthy buffaloes maintained at the Mahlet Mussa and El-Gmeasa herds from 2010 to 2012. PCR was performed using primers flanking a 296-bp fragment from GHRH gene and a 425-bp fragment from GHRHR gene of Egyptian buffalo. The PCR-amplified fragments were digested with HaeIII (GHRH) and Original Research Article Othman et al.; BBJ, 5(2): 62-71, 2015; Article no.BBJ.2015.005 63 Eco57I (GHRHR), electrophoresed and analyzed on agarose gels stained with ethidium bromide. The two amplified fragments were also sequenced and aligned with published sequences. Results: Depending on the presence of the restriction site at 241^242 position (GG^CC) in 296-bp amplified fragments of GHRH, we genotyped all tested buffalo animals as AA. Due to the absence of the restriction site at position 300^301 ([CTGAAG(N) 16 ^] in the amplified fragment of GHRHR (425-bp), we genotyped the tested animals as AA. The Egyptian buffalo GHRH and GHRHR nucleotide sequences were submitted to NCBI/Bankit/GenBank and have the accession numbers JN967799 and KC295414, respectively. Conclusion: The Egyptian buffaloes are characterized by best production traits like high milk fat content as well as higher average daily gain and body weight where they are possess with fixed GHRH AA and GHRHR AA genotypes which were reported as desired genotypes for milk and growth production traits in different cattle breeds and the cattle are genetically homologous with buffaloes. To the best of our knowledge, these polymorphic sites are not identified in other buffalo populations. The identification of genotypes and nucleotide sequences of these two multifunctional genes may be useful in future marker-assisted selection (MAS) for more efficient breeding and genetic conservation programs of Egyptian buffalo.
Background With regard to the extensive production of genetically modified organisms, scientists ... more Background With regard to the extensive production of genetically modified organisms, scientists focused on the safety of antibiotic resistance marker (ARM) genes present in GM-crops produced for the consumption of human or animal. We investigated the possible transfer of these genes to the microflora present in the gastrointestinal tract and blood cells of two groups of Male Wistar Albino rats fed on a transgenic diet containing the ARM genes nptII and aadA for 90 days. The conventional PCR was employed to screen for the presence of nptII and aadA genes in the experimental diets, and animals’ samples. Results The occurrence of DNA transfer of nptII and aadA genes from GM-diet to blood and bacterial cells has been unambiguously demonstrated. Results were further confirmed by sequencing and blast analysis, indicating that both transferred segments shared significant alignment with number of Binary and Cloning Vectors with variable similarities. The frequency of transfer of DNA segmen...
Background One of the parameters required for the assessment of food and feed safety is detection... more Background One of the parameters required for the assessment of food and feed safety is detection and identification of genetically modified organisms. Legislation in some countries necessitates detection and quantification of modification in food and feed samples. Scientists have raised concern about safety of antibiotic resistance marker (ARM) genes used for transformation of crops intended for human and animal consumption. In the present work two molecular approaches have been adopted: one exploratory; for detection and quantification of ARM genes in tested plant samples and the other confirmatory; to determine the specificity/reliability of the obtained results. Results Results revealed that primers for neomycin phosphotransferase (nptII) and aminoglycoside 3″ adenyl-transferase (aadA) were amplified in the majority of the 36 DNA screened samples. Melting curve analysis using hygromycin phosphotransferase (aphIV) gene as target sequence for the fluorescent-based detection approa...
Background and Aim: Uterine lumen contamination with bacteria is ubiquitous in buffalo after part... more Background and Aim: Uterine lumen contamination with bacteria is ubiquitous in buffalo after parturition. Nearly one-third of these infected animals develop endometritis which leads to reduced fertility. The present study aimed to evaluate the expressions of IFN-Îł and NOS2 genes in uterine tissue of buffaloes with endometritis and comparing them with those in healthy animals using RT-qPCR Materials and Methods: Uterine samples were collected from 50 apparently healthy and 50 clinically infected buffaloes. RNA was extracted from the collected buffalo's uteri and cDNA was synthesized from extracted RNA. Quantitative Real Time PCR technique was performed using this synthesized cDNA. Results: Apparent up-regulation of both genes mRNA expression was recorded in endometritis-infected animals with 8.3-folds for IFN-Îł and 9.99-folds for NOS2 (P<0.001). Conclusion: The upregulation of IFN-Îł and NOS2 expression in the uterine tissue of endometritis-infected buffaloes can be used as a s...
ABSTRACT Authors&#39; contributions This work was carried out in collaboration between all au... more ABSTRACT Authors&#39; contributions This work was carried out in collaboration between all authors. Author OEO designed the study, followed up the practical work and wrote the final version of the manuscript. Author MFAS managed the analyses of the study, managed the literature searches and wrote the first draft of the manuscript. Author NAAEM performed the practical work. Author KMS followed up the steps of the search. All authors read and approved the final manuscript Article Information DOI: 10.9734/BBJ/2015/11619 Editor(s): (1) Giuliana Napolitano, ABSTRACT Aim: The hypothalamic hormone, growth hormone-releasing hormone, is the principal stimulator of pituitary growth hormone (GH) synthesis and secretion. GHRH and its receptor (GHRHR) provide important functions in the regulation of the GH axis and in the development and proliferation of pituitary somatotropic axis. This study aimed to identify the genotypes and nucleotide sequences of two multifunctional genes; growth hormone-releasing hormone (GHRH) and its receptor (GHRHR) in Egyptian buffalo. Methodology: Genomic DNA was extracted from blood samples of 100 healthy buffaloes maintained at the Mahlet Mussa and El-Gmeasa herds from 2010 to 2012. PCR was performed using primers flanking a 296-bp fragment from GHRH gene and a 425-bp fragment from GHRHR gene of Egyptian buffalo. The PCR-amplified fragments were digested with HaeIII (GHRH) and Original Research Article Othman et al.; BBJ, 5(2): 62-71, 2015; Article no.BBJ.2015.005 63 Eco57I (GHRHR), electrophoresed and analyzed on agarose gels stained with ethidium bromide. The two amplified fragments were also sequenced and aligned with published sequences. Results: Depending on the presence of the restriction site at 241^242 position (GG^CC) in 296-bp amplified fragments of GHRH, we genotyped all tested buffalo animals as AA. Due to the absence of the restriction site at position 300^301 ([CTGAAG(N) 16 ^] in the amplified fragment of GHRHR (425-bp), we genotyped the tested animals as AA. The Egyptian buffalo GHRH and GHRHR nucleotide sequences were submitted to NCBI/Bankit/GenBank and have the accession numbers JN967799 and KC295414, respectively. Conclusion: The Egyptian buffaloes are characterized by best production traits like high milk fat content as well as higher average daily gain and body weight where they are possess with fixed GHRH AA and GHRHR AA genotypes which were reported as desired genotypes for milk and growth production traits in different cattle breeds and the cattle are genetically homologous with buffaloes. To the best of our knowledge, these polymorphic sites are not identified in other buffalo populations. The identification of genotypes and nucleotide sequences of these two multifunctional genes may be useful in future marker-assisted selection (MAS) for more efficient breeding and genetic conservation programs of Egyptian buffalo.
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