Main Subjects : Molecular Biology


Molecular description of melatonin receptor 1A gene in Iraqi buffalo

hassan nima habib; Khalaf A.H. Al-Rishdy; Murthda F. AL-Hellou

Iraqi Journal of Veterinary Sciences, In Press
DOI: 10.33899/ijvs.2022.132532.2103

The water buffalo has a seasonal reproductive pattern with reduced sexual activity during the longer photoperiod. The goal of this study was to identify the single nucleotide polymorphism of melatonin receptor 1A gene in Iraqi buffalo cows and 3D structure of its protein and phylogenic with other sequences around the world. The 824 bp fragment of exon II of the MTNR1 A gene was amplified from 190 buffalo cows (4-5 years old) genomic DNA belonging to local breeders in Al-Chibayish Marshes, Southern Iraq. Amplified PCR products underwent custom sequencing at the two end. Five separate polymorphism sites, the 1st included 52 animals with 19 mutations (12 missense), the 2nd included 39 animals with 18 mutations (11 missense), the 3rd included 35 animals with 18 mutations (12 missense), the 4th included 32 animals with 18 mutations (12 missense) and the 5th included 32 animals with 14 mutations (8 missense). These polymorphic sites with accession numbers LC565046, LC565047, LC565709, LC565710 and LC565711 respectively were registered in gene bank. The phylogenetic tree reveals that in some of the Iraqi buffalo, the sequences of gene has identical to the Italianbuffalo (GU817415), and the Brazilian buffalo (JN689386). Data revealed a marked variance of the fifth polymorphism sites' 3D protein structure because of the mutations. In conclusion, as a result of mutations, the gene MTNR1A in Iraqi buffalo has polymorphisms; these polymorphisms may be linked to gene function, Therefore, further studies are needed to connect the polymorphisms of this gene with the productive and reproductive traits

Infections and molecular characterization of anisakid nematodes from two species of marine fish northwest Arabian gulf

Majid A. Bannai; Muna M. Jori

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 2, Pages 489-497
DOI: 10.33899/ijvs.2021.130613.1851

The present study provides new insight into valuable information on the diverse structure of the anisakid population, discusses the limited species richness, and also discusses the relationship with other closely diversity-related taxa in NCBI databases in the Epinephelus diacanthus and Epinephelus coioides fish. The fishing area consists of various locations in the Arabian Gulf. A total of 69 E.coioides and E. diacanthus were examined, (n= 48) were infected. Larval stages (n=1,119). Isolated larvae were encysted within the mesenteries peritoneum and viscera of fish organs, with a prevalence of 81.25% of infection and 59.459 % in the E. diacanthus and E. coioides respectively. Molecular analysis was carried out on thirty individuals of nematode parasites who have examined the morphology and showed some appearance differences, by amplifying internal transcribed spacers ITS and ITS-1 of nuclear rDNA (rDNA) by PCR using the primer sets NC5/NC2 and SS1/NC13R of DNA products. Evolutionary analyses were conducted in MEGA X. based on the identity percentage in the GenBank database showed that they belong to anisakid nematodes, in particular, they belong to nine distinct taxa within the Hysterothylacium spp. The presence of the same species individuals in one host may be the cause of these genetic variations at the species level, and that's what the current study has recorded. It has been found that there is an overlap in the order of nitrogen bases between the same species, and this occurs through the fertilization process, while the rest is clean or have only a few parasites. 

Genotyping of Salmonella enterica strains from animal and human origin using three molecular techniques

Juan S. Cruz-Méndez; Julián D. Ortiz-Muñoz; Iang S. Rondon-Barragan

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 2, Pages 531-538
DOI: 10.33899/ijvs.2021.130764.1877

This study aims to characterize different Salmonella enterica subsp molecularly. enterica strains (n=49) were isolated from human gastrointestinal cases in the Tolima region and poultry from Santander and Tolima regions using PCR-RFLP, PCR-ribotyping, and PCR-SSCP. The band patterns obtained with each technique were analyzed by building dendrograms based on the Unweighted Pair Group Method with Arithmetic mean (UPGMA) method and using the Dice coefficient. On the other hand, the discriminatory power of each technique was assessed using Simpson's discriminatory index. The genetic profiles of the gnd gene obtained with AciI restriction enzyme and the PCR-SSCP carried out with groEL gene allowed the inter-and intraserovar differentiation. Finally, the PCR-ribotyping method exhibited the highest discriminatory power (0.8571). In conclusion, we show three PCR-based genotyping methods providing an alternative for identifying similarities and differences within Salmonella enterica strains from different geographic and biological regions.

RAPD-PCR and phylogenetic analysis of E. coli isolated from human and cattle urinary tract infections

Dunya A. Mraidi; Inam J. Lafta

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue Supplement I-III, Pages 59-66
DOI: 10.33899/ijvs.2021.131100.1918

This study aimed at isolating uropathogenic Escherichia coli from urinary tract infections (UTIs) of human and cattle to examine the molecular diversity and phylogenetic relationship of the isolates. A total of 100 urine samples were collected from UTIs of human and cattle. The isolates identification was done using routine diagnostic methods and confirmed by Vitek2. Antimicrobial susceptibility was tested against 10 antimicrobials. Random amplified polymorphic DNA (RAPD)-polymerase chain reaction (PCR) was applied to identify the genetic diversity among E. coli isolates from human and animal origin by using five different octamer primers. The gelJ software for the phylogenetic analysis created Dendrograms. Out of 50 human urine samples, E. coli was isolated from 12 (24%) samples, and was positive in 5 out of 50 (10%) of cattle urine samples. Concerning the antimicrobial susceptibility test, both human and animal isolates revealed rather approximate results when tested mainly against Imipenem, Cefotaxime, and Ciprofloxacin. These antimicrobial data might indicate presence of a degree of similarity between the human and animal isolates. Using RAPD-PCR, three of the primers produced polymorphic bands; therefore, they were used for further analysis of the results. Either of P1, P3 or P4 primers showed presence of similarity between human and cow isolates. To conclude,RAPD-PCR and gelJ software might be of attractive use to identify and analyze the occurrence of genetic relationships, as this could assist in controlling the routes and sources of infection transmission between human and animal in order to prevent zoonotic infections. 

Estimation of limit of detection of Salmonella typhimurium in artificially contaminated chicken meat by cultured-based and polymerase chain reaction techniques

Yousif M. Sharif; Bizhar A. Tayeb

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue 4, Pages 621-625
DOI: 10.33899/ijvs.2020.127328.1496

The objective of this study was to develop Polymerase Chain Reaction (PCR) procedure for detection of Salmonella Typhimurium in artificially contaminated chicken meat. The experiments were conducted with various dilutions of Salmonella Typhimurium reference the American Type Culture Collection ATCC (ATCC13311TM 4.4*107) High concentration 4.4*103 Colony Forming Units (CFU)/ml, low concentration 4.4*102 CFU/ml, very low concentration 4.4*101 CFU/ml inoculated in chicken meat, in order to determine limits of detection (LOD), optimum incubation times 18 to 20 hours of pre-enrichment in Buffered Peptone Water (BPW 1%). Hence, cultural methods and DNA extraction were performed according to kits instruction. The microbiological cultural test was capable to detect 1.76 CFU/mL, whereas PCR examination was able to detect 0.18 CFU/ml of initial dilution of Salmonella Typhimurium inoculated in chicken meat. Interestingly, the results were achieved in a less time period than that of classical culture. The PCR technique is beneficial in the methodology for detection of Salmonella in chicken meat.

Bronchodilator activity of ethyl acetate extract of Nigella sativa

Ihsan Husain Mohammed Ali; Qasim Hasso Abdullah; Omer AL-Habib

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue 1, Pages 145-149
DOI: 10.33899/ijvs.2020.126455.1333

This study aims to investigate the mechanism(s) included in the bronchodilation effect exerted by Nigella sativa. Ethyl acetate extract (NS.EA) was prepared using a maceration method. Adult albino rats were recruited for thoracotomy and removal of the trachea. After cutting into pieces, the tissue was set in organ bath. The influence of cumulative concentrations of ethyl acetate extract was examined on contractile responses of isolated trachea to acetylcholine using different blockers such as Nifedipine (Ca2+channel blocker), Tetraethylammonium (Ca2+-activated K+ channel blocker), 4-aminopyridine (voltage-dependent K+ channel blocker), Glibenclamide (ATP-sensitive K+ channel blocker), BaCl2 (inward rectifier K+ channel blocker), methylene blue (soluble gaunylate cyclase inhibitor) and indomethacin (non-selective cyclooxygenase inhibitor). Significant inhibition of bronchodilation was observed when tracheal rings were pretreated with indomethacin and BaCl2 with (P<0.001), and with methylene blue and nifedipine with (P<0.05). The IC50s were (5.635, 6.9, 7.86 and 4.987 mg/ml) respectively. Conversely, 4-AP, GLIB and TEA showed no significant changes in the bronchodilation induced by the extract. Therefore, The Emax value for indomethacin significantly reduced from 101.34 to 73.28%, BaCl2 from 53.62 to 30.31%, methylene blue from 55.78 to 38.94% and nifedipine from 101.34 to 80.88%. On the other hand, the Emax for 4-AP and GLIB were non-significantly reduced from 53.62 to 40.14 and 40.13% respectively; and TEA more or less unchanged to 54.34%. In general, ethyl acetate extract of N. sativa induces bronchodilation through four mechanisms (activation of Kir channel, non-selective cyclooxygenase and to lesser extent the soluble guanylate cyclase, and blockade of Ca2+ channel).

The genotype of Entamoeba histolytica in bloody diarrhea samples of humans, cows and sheep

Hassan H. Naser

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 2, Pages 453-458
DOI: 10.33899/ijvs.2020.126135.1242

The present study was carried out to detect the genotype of E. histolytica that found in human fecal specimens and animals feces with Haemorrgic diarrhea by amplifying the SREHP gene, using RT-PCR technique, Cyber ​​green dye and by fusion curve analysis. The study also included molecular detection of amoebic parasite species using Nested-PCR technology. The study recorded presence of parasites E. histolytica; E. dispar; E. bovis with total infection rates 82.9, 26.8, 4.9%, respectively. The study revealed the presence of E. histolytica parasite in five different genotypes (I, II, III, IV, V) with rate presence 9.75, 53.65, 19.5, 9.75, 7.3%, respectively. In conclusion, there are five genotype of E. histolytica, in human and animals, most of these genotypes may be infect any host, E. bovis was recorded in sheep and cows.