Molecular characterization of alpha subunit 1 of sodium pump (ATP1A1) gene in Camelus dromedarius: its differential tissue expression potentially interprets the role in osmoregulation.

BACKGROUND: Dromedary or one-humped camel (Camelus dromedarius) is distinctively acclimatized to survive the arid conditions of the desert environment. It has an excellent ability to compete dehydration with substantial tolerance for rapid dehydration. Therefore, it offers an excellent model for studying osmoregulation. Molecular characterization of Na+/K+ ATPase as a central regulator of electrolyte normohemostasis affords a better understanding of this mechanism in camel. Here is the first to resolve the full-length of alpha-1 subunit of sodium pump (ATP1A1) gene with its differential expression in dromedary tissues. RESULTS: The nucleotide sequence for the recovered full cDNA of ATP1A1was submitted to the GenBank (NCBI GenBank accession #MW628635) and bioinformatically analyzed. The cDNA sequence was of 3760 bp length with an open reading frame (ORF) of 3066 bp encoding a putative 1021 amino acids polypeptide with a molecular mass of 112696 Da. Blast search analysis revealed the shared high similarity of dromedary ATP1A1gene with other known ATP1A1genes in different species. The comparative analysis of its protein sequence confirmed the high identity with other mammalian ATP1A1 proteins. Further transcriptomic investigation for different organs was performed by real-time PCR to compare its level of expression among different organs. The results confirm a direct function between the ATP1A1 gene expression and the order of vital performance of these organs. The expression of ATP1A1 mRNA in the adrenal gland and brain was significantly higher than that in the other organs. The noticed down expression in camel kidney concomitant with overexpression in the adrenal cortex might interpret how dromedary expels access sodium without water loss with relative high ability to restrain mineralocorticoid-induced sodium retention on drinking salty water. CONCLUSION: The results reflect the importance of sodium pump in these organs. Na+/K+ ATPase in the adrenal gland and brain than other organs.

Prevalence of Clostridium perfringens type A isolates in commercial broiler chickens and parent broiler breeder hens in Egypt.

The aim of this study was to determine the presence of genes coding for alpha (cpalpha), beta (cpbeta), epsilon (epsilontx), iota (iotaA), enterotoxin (cpepsilon) and beta2 (cpbeta2) toxins in Clostridium perfringens isolates from broiler chickens and parent broiler breeder hens, using multiplex polymerase chain reaction (PCR) assay. The prevalence of C. perfringens in the intestinal segments and the effects of age were also investigated. The highest isolation rate was from the duodenum, at 41.7% in broiler chickens and 58.4% in parent broiler breeder hens; the lowest isolation rates came from the ileum, at 15.6% and 27.1%, respectively. Chickens harboured C. perfringens in the intestine and this increased with age. Clostridium perfringens was detected in 35.4% (17/48) of asymptomatic broiler chickens and 22.1% (17/77) of asymptomatic parent broiler breeder hens. The bacterium was detected in 100% of the broiler chickens and parent broiler breeder hens with clinical signs (31/31 and 60/60, respectively). The multiplex PCR assay indicated that in 99 (79.2%) of the 125 samples that tested positive for C. perfringens the strains isolated were type A and were shown to carry the cpalpha gene (99/99, or 100%). The gene encoding cpbeta2-toxin was present in 62.6% (62/99) of the isolates. A significant association was found between C. perfringens possessing the beta2-toxin gene and necrotic enteritis in broiler chickens and parent broiler breeder hens, suggesting that this gene might play a key role in the pathogenesis of the disease in Egypt. The authors suggest that the presence of the cpbeta2-toxin gene in C. perfringens isolates found in broiler chickens and parent broiler breeder hens during this study poses a risk of transmission to humans through the food chain.

Body composition and peak aerobic power in male international level Hungarian athletes.

Body size, physique, body composition and physiological performance of elite athletes are independent aspects, have aroused the interest of exercise scientists, but studies that combine these aspects in elite athletes are scarcely available. The aim of the present study was to describe the selected anthropometric and exercise physiological characteristics of some Hungarian top athletes. The investigated subjects were qualified Hungarian water polo players (n=25), paddlers (n=24) and modem pentathlonists (n=20), all of whom had been medalists at several continental and intercontinental competitions. The athletes' body composition was estimated by the Drinkwater-Ross (45) body mass fractionation technique. Peak physiological performance was estimated by graded exhausting spiroergometric treadmill exercise. Intergroup differences in mean height, body mass and body composition characteristics were significant at the 5% level of random error. By the results of spiroergometry, all the three groups compared could be qualified as physically excellently trained. The greatest oxygen uptake relative to body mass was found in the modern pentathlonists (73.22 ml x kg(-1) x min(-1)) and the lowest one (59.79) in the water polo players. The authors do not disregard the favourable effects of regular and adequate trainings in the development of the studied characteristics, but in their opinion the process of proper selection has been the most important factor that explains the observed significant intergroup differences.

Cloning and expression of recombinant MPB70 protein antigen from Mycobacterium bovis BCG for diagnosis of tuberculosis.

In a search for developing new skin test reagents, MPB70 protein antigen was a candidate antigen for the Diagnosis of bovine tuberculosis. First M. bovis BCG genomic DNA was extracted purified and the mpb70 gene was amplified by PCR. The gene was then ligated to an expression vector, PQE. After transformation of the expression E. coil M15 host strain with the PQE plasmid, the expression was induced using 10 mM of IPTG. Two bands were seen in the SDS-PAGE analysis the 44 and 50 KDa represents the dimmers of the nonglycosylated and glycosylated form of the reMPB70 antigen. The His-tagged reMPB70 antigen was then purified by metal affinity chromatography using Ni-NTA agarose. Protein refolding was done by the use of the co solvent Polyethylene glycol MW 3000. The diagnostic potential of the re-MPB70 was evaluated using sera from experimentally sensitized guinea pigs with different strains of mycobacteria (M. bovis BCG, M. tuberculosis, M. kansasii and M. intracellular) using ELISA test. The results indicated the efficiency of MPB70 but not bovine PPD to discriminate between M. bovis sensitized guinea pigs and those sensitized with other mycobacterial strains at serum dilution of 1150. In a field trials to using reMPB70 antigen for the serodiagnosis of bovine tuberculosis using ELISA test. Fifty serum samples from tuberculin +ve and 6 from tuberculin -ve cattle were used as well as 10 tuberculin +ve buffaloes. All +ve animals were confirmed to be M. bovis infected by P/M analysis, bacteriological examination. ELISA results revealed that reMPB70 could recognize the tuberculin +ve infected animals at serum dilution of 1/50 and that it could diagnose tuberculosis in cattle as well as buffaloes.

Molecular detection of Babesia equi in infected and carrier horses by polymerase chain reaction.

Twenty-three blood samples were used in this study; five were from five naturally infected horses with Babesia equi (B. equi), while eighteen were from asymptomatic horses with equine babesiasis from different localities in Egypt. All samples were subjected to microscopic examination, indirect fluorescent antibody test (IFA) and polymerase chain reaction (PCR). The carrier animals were microscopically detected in 7 out of 18 samples (38.8%) and in 9 of 18 by using IFA (50%), whereas PCR revealed that 14 samples were positive (78%). Two synthetic oligonucleotide primers, based on the B. equi merozoite antigen gene (EMA-1) were used. A 819 bps DNA fragment is specifically amplified from the gene encoding EMA-1 of B. equi. Our results demonstrate that PCR is a valuable technique for routine detection of B. equi in chronically infected horses, even at low parasitaemia levels.