Soheir M. El Nahas

Chromosome aberrations in spermatogonia and sperm abnormalities in Curacron-treated mice.

Curacron is an organophosphorus pesticide widely used in cotton fields. In order to assay its mutagenic potential in mammalian germ cells chromosomal aberrations in spermatogonial cells and sperm abnormalities were examined in mice after Curacron treatment. For studying chromosomal aberrations mice were treated both acutely (single treatment) and subacutely (for 5 consecutive days) with 3 dose levels of Curacron, 12, 36 and 72 mg/kg. Curacron was found to produce a significant increase in structural chromosomal aberrations after acute and subacute treatments. This increase was dose-dependent. A dose-dependent inhibition in mitotic activity in spermatogonia was also found. For studying sperm abnormalities mice were treated for 5 consecutive days with 20, 40 and 60 mg/kg. Morphological sperm abnormalities increased significantly after treatment with Curacron. The increase was dose-dependent. An inhibition of 40.2% in sperm count and of 74.5% in sperm motility occurred after treatment with 60 mg/kg Curacron. These results show that Curacron has a damaging effect on spermatogonial cells as well as on sperm morphology.

Specific numerical chromosomal aberrations induced by adriamycin

Fluorescence in situ hybridization with 7, 17, X, and Y chromosome‐specific DNA probe was used to investigate the ability of Adriamycin (AM) to induce aneuploidy in interphase human lymphocytes. The reliability of the probes was tested by hybridization to metaphases and interphase nuclei of untreated normal lymphocytes. Two signals were scored in over 87% of the analyzed nuclei with chromosome 7 and 17 probes, whereas one signal was recorded in over 86% of the nuclei with chromosomes X and Y. The same conditions and probe concentrations were used for hybridizing the four probes to interphase nuclei of AM‐treated and untreated lymphocytes, cultured from healthy individuals and cancer patients. AM was found to induce significant increases of trisomy 7 and 17 in lymphocytes cultured from healthy individuals and cancer patients, where the interphase nuclei showed three spots in over 70% and 72% of the cells, respectively. Only 6% of interphase nuclei of untreated cells cultured from healthy individuals and cancer patients showed three spots. No significant increase in X or Y aneuploidy was induced by exposure to AM. Environ. Mol. Mutagen. 33:161–166, 1999

Assignment of PCR markers to river buffalo chromosomes

In the process of developing a buffalo physical map using somatic cell hybrids and the cattle gene map as a template, ten PCR primers designed for four coding genes: F10, FSHB, HBB, and CYM and six DNA segments: TGLA9, TGLA227, UWCA5, CSSM6, CSSM47 and RF131 were tested on a panel of 47 buffalo-hamster somatic cell hybrids. F10-TGLA9, FSHB-HBB and UWCA5-TGLA227, respectively, were found to segregate together forming three syntenic groups. These three syntenic groups have also been reported in cattle, where they have been assigned to chromosomes BTA 12, BTA 15 and BTA 18, respectively. Comparative mapping predicts the assignment of these syntenic groups to river buffalo chromosomes BBU 13, BBU 16 and BBU 18, respectively.

Characterization of buffalo interleukin 8 (IL-8) and its expression in endometritis

River buffalo (Bubalus bubalis bubalis) with a population over 135 million heads is an important livestock. Interleukin 8 (IL-8) is a member of the chemokine family and is an important chemoattractant for neutrophils associated with a wide variety of inflammatory diseases such as endometritis. Tissue samples from the mammary gland, uterus and ovary were obtained from river buffalo (Mediterranean type) with and without endometritis. Bacteriological examination showed the presence of both gram positive and negative in all buffalo with endometritis. RNA extraction and complementary DNA (cDNA) synthesis were conducted from all tissues. Specific primer for IL8 full coding regions was designed using known cDNA sequences of Bubalus bubalis, Genbank accession number AY952930.1. IL-8 gene expression was investigated in buffalo tissues. Expression of IL-8 in buffalo with endometritis was found to increase significantly over buffalo without endometritis only in the uterus (P = 0.0159). PCR products from uterus tissues (target organs) of buffalo with and without endometritis, were purified and sequenced. No polymorphic sites were detected in the investigated samples. IL-8 cDNA nucleotide sequences of buffalo with and without endometritis were 100% identical (accession number JX413057). Buffalo IL8 cDNAs were compared with corresponding sequences of member of subfamily Bovinae (buffalo and cattle) and subfamily Caprinae (sheep and goat). IL-8 species specific differences were identified.

Genetic variation among Northern and Southern Egyptian buffaloes using polymerase chain reaction-random amplified polymorphic DNA (PCR-RAPD)

The domestic water buffalo is a species of great economic potential, especially in developing countries like Egypt. Egyptian buffalo have been classified according to minor phenotypic differences and their geographical locations. Few studies have taken place to investigate the genetic variations in Egyptian buffalo using microsatellites analysis. In the present study, 11 random primers were analyzed for the genetic diversity determination between Northern and Southern Egyptian buffaloes using polymerase chain reaction-random amplified polymorphic DNA (PCR-RAPD) analysis. 169 bands were amplified for the analyzed 11 random primers, from which 160 bands (94.67%) for North populations and 168 bands for South population (99.41%). Out of the 160 amplified bands in North populations, 152 bands were polymorphic with a percentage of 89.94% and only one specific band (0.59%). In South population, all 168 amplified bands were polymorphic, nine bands (5.33%) were specific for this population. The identity index and the genetic distance between North and South populations were measured. The results showed that the two tested populations have the same origin and belong to one breed without significant genetic difference between their animals.

AcuI identifies water buffalo CSN3 genotypes by RFLP analysis

Water buffalo population amounts to 185 million heads (http://www.fao.org). They are mainly present in Southeast Asia. Casein genes among other genes control the milk traits in water buffalo. The role of casein genotypes on composition and coagulation of milk, quality and yield traits has been investigated (Lien et al. 1995; Boettcher et al. 2004; Comin et al. 2008; Bonfatti et al. 2010, 2012; Vallas et al. 2012). The κ-casein gene has been broadly studied due to its influence on the properties of milk (Bonfatti et al. 2012). All previous reports on genotyping of water buffalo CSN3 gene (exon 4), by restriction fragment length polymorphism (RFLP) analysis used restriction enzymes HindIII, HinfI and TaqI, which successfully identify different cattle CSN3 genotypes. However in buffalo, they only resulted in BB monomorphic buffalo. This was reported in Egyptian (Othman 2005; Dayem et al. 2009; Mahmoud et al. 2010) Pakistani (Riaz et al. 2008), Indian (Gangaraj et al. 2008; Shende et al. 2009), Brazilian (Otaviano et al. 2005) buffalo and in the water buffalo genomic library (Masina et al. 2007). In a previous study on Egyptian buffalo CSN3 gene, using sequence analysis, the presence of two single-nucleotide polymorphism (SNP) in exon 4 (El Nahas et al. 2013) was reported. In the two SNP positions, C is replaced by T at codon 135 (ACC/ATC) and codon 136 (ACC/ACT) of the mature peptide. These two SNPs were also reported in Bulgarian Murrah breed (Beneduci et al. 2010), Italian buffalo (Bonfatti et al. 2012) and in Indian buffalo (Das et al. 2000). Using sequence analysis, three buffalo-CSN3 genotypes were identified (AA, AB and BB) and using in silico analysis, the restriction enzyme AcuI was suggested for buffalo-CSN3 genotyping (El Nahas et al. 2013). In this study, we show experimentally that AcuI restriction enzyme is useful in genotyping buffalo-CSN3 gene by RFLP analysis.