Main Subjects : 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

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.

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. 

Molecular characterization and microbial resistance of different bacterial isolates in some dairy products

Waleed K. Alkhafaje; Zakia A. Olama; Safaa M. Ali

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 2, Pages 333-339
DOI: 10.33899/ijvs.2021.130206.1764

Bacterial contamination of milk and dairy products is a common problem. In the last two years, the foodborne diseases caused by the intake of milk and dairy products have been mostly disturbed with Salmonella entertica, Listeria monocytogenes Escherichia coli 0157:H7 and Campylobacter jejune. The study aims to isolate multidrug resistance (MDR) bacteria in dairy products and study of the molecular characterization of that isolates. MDR bacteria were found in 30 out of the 131 bacterial isolates. The incidence of MDR bacterial isolates revealed the abundance of Staphylococcus sp. with 43.3%, Bacillus sp 16.7%, Salmonella 13.3%, E. coli 10 %, Enterococcus 6.7 % Psedoumonas 3.3 %, Shegella 3.3 % and Proteous 3.3 %. Molecular studies of genes presence or absence for class A contain TEM, CTX and BSHV, class B contain VIM, IMP, KPC and NDM, class C contain FOX and class D contain OXA-10, OXA-24 and OXA-58 were tested. NDM, TEM, CITM and OXA -10 genes were the most abundant the selected bacterial isolates. The results of this study indicate that cheese made from unpasteurized milk can pose a significant risk to consumers. Product manufacturing processes should be subject to health control-to-control pathogens. The novelty in this work depend on screening of gene responsible of the resistance from the bacteria isolated from dairy product using the molecular technique.

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.

Antimicrobial activity of peptides extracted from camels' blood neutrophils against some pathogenic bacteria

M.A. Mahmood; M.A. Essa

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue 1, Pages 33-37
DOI: 10.33899/ijvs.2020.126239.1270

The aim of the current research was to isolate and purify antimicrobial cationic peptides (Amps) taken from camel blood and to study its antibacterial activity against some multidrug-resistant pathogenic bacteria Salmonella typhimurium, Bacillus subtilis. The results showed the possibility of obtaining these peptides from camel's white blood cells, and was separated by using Reversed-Phased-High Performance Liquid Chromatography (RP-HPLC) technical equipment. The results showed the presence of 21 absorption peaks of the isolated peptide extract. The two-stage-radial diffusions method was used to study the activity of the crude peptide against the studied bacteria. The results showed that these peptides were effective without any resistance. The best effect was on the Bacillus subtilis with a diameter of 31 mm inhibition, while the less effect was on the Salmonella typhimurium with a diameter of 20 mm.

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.

Application of ultrasound contra infection with secondary hydatidosis in mice

Baraa B. Ramdan; Asmaa A. Ali

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 2, Pages 397-403
DOI: 10.33899/ijvs.2019.126132.1240

The present research explained the impact of ultrasound on the viability of Echinococcus granulosus protoscolices in vitro application of fixed frequency waves at 20000 pulse/sec and 1.8 w/cm2, with different exposing periods, thirty, twenty, fifteen, twelve, ten and five seconds respectively, subsequently prevention of mice against interjected with minor hydatid cysts, in comparison with the control animals infected with protoscolices without exposure to ultrasound waves, depending on many criteria including the numbers, weights and diameters of developed hydatid cysts. The results displayed an obvious impact of ultrasound waves on Echinococcus granulosus protoscolices viability number by increase of exposure period in vitro, in addition to a decrease of 100% of hydatid cysts numbers in the dealt ones with 17% fertility exposed for 20 seconds, the reduction rate was 99.23% in the group of fertility 35% which exposed for 15 seconds, after four and five months of infection.

Detection of similarity and genetic distance between Iraqi chicken varieties and different standard strains

Thamer A. Ezzulddin; Dhyaa M. Jwher; Sufian A. Dabdoub

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 2, Pages 333-337
DOI: 10.33899/ijvs.2019.126109.1235

Forty-eight wing vein blood samples were collected from different locations of poultry rearing farms and back yard chickens of Nineveh governorate from the of local and exotic chicken. The chicken divided into twelve groups four birds each according to colors and phenotype for the local and exotic chicken respectively. Blood DNA was extracted and amplified by thermocycler apparatus and the electrophoresis was done using 1.2% agarose gel for DNA bands exhibiting. The results showed high genetic similarity within the local chickens ranged between 0.78- 0.96 at an average of 0.88, while it ranged between 0.73- 0.86 at an average of 0.78 in exotic breeds. The degree of similarity between Iraqi and exotic breeds was 0.74-0.88 at average of 0.80. The calculated average of differences among each of Iraqi and exotic chickens and in between were 0.12, 0.22 and 0.20, respectively. However, the genetic distance within the local chicken, exotic breed and in between them was 0.128, 0.24 and 0.21 respectively. The study concluded that the genetic similarity was higher within local chicken groups than those of exotic breeds.