Main Subjects : Veterinary Microbiology

Molecular identification of allelic genotypes of pyrethroid-insecticide resistance in housefly, Iraq

Monyer Abdulameir Abd Alfatlawi; Mansour Jadaan Ali; Hassan Hachim Naser

Iraqi Journal of Veterinary Sciences, In Press
DOI: 10.33899/ijvs.2019.125538.1060

According to global-wide presence of insecticides resistance to pyrethroids, the current study identified the purpose to detect the allelic genotypes regarding this issue in house flies in Iraq. From the governorate of Al-Qadisiyah, Iraq, 60 morphologically and molecularly recognized house flies were caught from 6 different regions. Using a technique called polymerase chain reaction (PCR) amplification of specific allele (PASA), PCR was employed to reveal the presence of allele-genetic variations in the para-type sodium channel (para) gene to recognize knockdown resistance (kdr) mutation from the homozygous-wild type of complete susceptibility (sus/sus) to the mutated-homozygous type of complete resistance (kdr/kdr) or to the mutated-heterozygous type (kdr/sus). Here, these genotypes were targeted using specific primers to identify these genetic variations. The results have declared the presence of the sus/sus at 100%-frequency rate in all flies, and none of the other genotypes were detected (0%) in all flies. This valued piece of result indicates the reality of resistance persistence due to lack of insecticide-spraying programs in the governorate. This study provides high-quality information about the current status of insecticide resistance in house flies in Iraq about supporting the fact of genetic-base development of such resistance via frequent use of insecticides.

Molecular characterization of enterohemorrhagic E. coli O157 and O153 isolated from tissue camel and human stool samples in Al-Diwaniyah, Iraq

Saba Falah Klaif; Zeena Foaad Saleh; Alaa abd alkadhem Jawad

Iraqi Journal of Veterinary Sciences, Volume 33, Issue 1, Pages 81-86
DOI: 10.33899/ijvs.2019.125530.1052

The present study aimed to describe the genetic relationships of zoonotic characterization of Escherichia coli isolated from human and livestock camel clinical infection. The study includes collected (50) meat of camel and (50) stool human samples. These samples were foreword to traditional bacterial isolation and identification using enrichment culture method and biochemical tests, then confirmed by PCR technique based on Gyr B gene Escherichia coli and DNA sequencing was done on some positive isolates. The results show that Escherichia coli were isolated from animals at 42 (84%) and 39 (78%) from human infection. The PCR technique was show highly sensitive and specific confirmative detection of Escherichia coli the positive results into 40 (95%) meat sample of camel, and 35 (89.7%) stool sample of a human. To evaluate of Virulence E.coli,we used specific virulence hlyA gene from NCBI-GenBank, published sequence of E. coli hly A gene (Genbank code: X94129.1) and the results show high of presence of virulence gene hly A in camel in percentage (19) 45% than of virulence gene in human (15) 38%. DNA sequencing of a partial sequence of GyrB gene was shown highly homology sequence identity with NCBI-Blast Escherichia coli strain O157H7 isolates from human and O153H3 from the camel. The phylogenetic analysis was shown there is clear genetic similarity at between human and animal’s E. coli isolates and then the gene sequence deposited into NCBI-Genbank accession numbers (MG560867.1, MG560866.1). Also, study design for detection of some virulence gene hly A Escherichia coli. In conclusion, there prevalence E. coli in humans and camel. Therefore, it is essential to define the role of animals as an important source for the distribution of pathogen related to public health. Our study found gyrB gene sequence could be used for identification and making a phylogenetic analysis of gyrB nucleotide.

The genetic relationship for Klebsiella pneumoniae isolated from human urinary tract and beef

Saba Falah Klaif; Hassan Naser; Jenan N. Sadeq

Iraqi Journal of Veterinary Sciences, Volume 33, Issue 1, Pages 75-80
DOI: 10.33899/ijvs.2019.125531.1053

The present study aimed to describe the genetic relationships of zoonotic characterization of Klebsiella pneumoniae isolated from Human urinary tract and beef. The study includes (50) urine samples from human and (50) beef samples. The isolation and identification of Klebsiella pneumonia were done by using enrichment culture method and Vitek 2, then confirmed by PCR technique based on 16S ribosomal RNA gene which designed in this study using NCBI-GenBank (LT599801.1) and DNA sequencing was done on some positive isolates. The results show that Klebsiella pneumoniae was isolated from Beef at 38(76%) And from human at 32(64%) by vitek2. The PCR technique was show highly sensitive and specific confirmative detection of Klebsiella Pneumonia isolates at Clarify DNA sequencing of a partial sequence of 16S ribosomal RNA gene was shown homology sequence identity highly with NCBI-Blast Klebsiella pneumoniae isolates. The phylogenetic analysis was show clear genetic similarity at (0.5 genetic change) between human and beef in Klebsiella pneumoniae isolates. The gene sequence deposited into NCBI-GenBank accession numbers (MF314450.1, MF314451.1, MF314452.1, MF314453.1). In conclusion, the study presents the first report in Iraq of genetic relationship among K. pneumoniae isolates from beef and humans. Therefore, it is essential to define the role of animals as an important source for the distribution of pathogen related to public health.

Plasmid transformation and curing of nalidixic acid gene in Staphylococcus aureus isolated from buffaloes mastitis and workerʼs hands

D. A. Khaleel; R. M. Othman; B. Y. Khudaier

Iraqi Journal of Veterinary Sciences, Volume 32, Issue 2, Pages 167-174
DOI: 10.33899/ijvs.2019.153845

The present study was designed to detect resistant site of nalidixic acid through transformation and plasmid curing of S. aureus strains isolated from buffalo milk with subclinical mastitis and workers’ hands. A total of 37 S. aureus isolates including 17 isolates recovered from buffalo milk infected with subclinical mastitis, in addition to 20 isolates recovered from workers’ hands. All 37 isolates were investigated by detection of the 23S rRNA gene and various other species specific genes including coa, nuc and clfA. The antibiotic resistance of S. aureus isolates was performed by the discs diffusion method using 19 antibiotics. Plasmid transformation method was carried out by transferring the plasmid isolated from S. aureus into competent Escherichia coli HB 101 in order to detection the resistant site of nalidixic acid. Plasmid curing was accomplished by preparing different concentrations of nalidixic acid (100, 150, 200, 250 and 300 µg/ml) and cultured transformed E.coli on LB agar supported with each of the aforementioned concentrations. The molecular results showed that six isolates (five isolates from milk samples and one from workers’ hands) were identified as S. aureus by coa, nuc, and clfA species specific primers. The six S. aureus isolates were found to be resistant to at least 5 antibiotics which included the nalidixic acid. The results of plasmid transformation revealed that E. coli was able to grow on LB agar supported with 100µg/ml, 150 µg/ml, 200 µg/ml and 250 µg/mlof nalidixic acid and failed to grow on 300 µg/ml concentration.

Antimicrobial and antifungal activity of pumpkin (Cucurbita pepo) leaves extracted by four organic solvents and water

H. Mohammed; R.S. Najem; S.S.A. Altekrity

Iraqi Journal of Veterinary Sciences, Volume 32, Issue 1, Pages 33-39
DOI: 10.33899/ijvs.2018.153791

Pumpkin is a rich source of vitamin A, being high in beta-carotene, a precursor to vitamin A. It provides substantial fiber, niacin, and lutein (important antioxidant). Pumpkin seeds have many health benefits, some of which include a good source of protein, zinc, and other vitamins, and are even said to lower cholesterol, Pumpkin plant was mentioned in the holy Quran as protector to protect the prophet Yonah, peace upon him after his expulsion from the whale. The present work was design to elucidate and evaluate different organic solvents i.e. (Distilled water, Ethanol, Hexane, and Petroleum ether) extracts of pumpkin leaves against some of the pathogenic bacteria and fungi. The results showed pumpkin leaves extracts were able to inhibit bacterial (Escherichia coli, Klebsiella pneumonia, Staphylococcus aureus, Proteus mirabilis and Pseudomonas aeruginosa) and fungal (Aspergillus fumigatus, Aspergillus niger, and Candida albicans) growth, comparable with the known antibiotic Ciprofloxacin and the antifungal drug Kenazole. There were no significant differences among different solvents in their ability to produce anti- microbial activity except petroleum ether. Petroleum ether extracts did not show any bacterial growth retardation while it showed anti –fungal inhibition in higher concentrations for Aspergillus fumigates and Aspergillus niger, while Candida albicans seem to be resistant to the petroleum ether extract of pumpkin leaves.

Detection of Escherichia coli hlyA gene and Staphylococcus aureus Sea gene in raw milk of buffaloes using RT-PCR technique in AL- Qadisiyah province

J.N. Sadeq; Kh.H. Fahed; H.J. Hassan

Iraqi Journal of Veterinary Sciences, Volume 32, Issue 1, Pages 87-91
DOI: 10.33899/ijvs.2018.153815

The aim of this study was to determines the prevalence of virulence gene hemolysin A (hly A) Escherichia coli and Staphylococcal enterotoxins (sea) in Staphylococcus aureus in raw milk buffaloes. In molecular laboratory, real-time polymerase chain reaction (RT-PCR) technique has been performed for 24 samples which have been taken randomly from Buffaloes milk, using primers of high specificity for Escherichia coli hlyA gene and Staphylococcus aureus Sea genes. The results showed different degrees of the studied genes activities. Four out of 24 samples represented S. aureus Sea gene (16.6%) whereas 16 out of 24 samples represented E. coli hlyA gene (66.6%). this study concluded that buffaloes milk might be a source of contamination with pathogenic bacteria of virulent genes which may have different levels of activities.

Investigating and identifying Chlamydia psittaci in asymptomatic and symptomatic domestic dogs in middle province of Iran

M.M. Sarmeidani; P. Keihani; P. Rezaei; H. Momtaz; S.H. Heidari

Iraqi Journal of Veterinary Sciences, Volume 31, Issue 2, Pages 91-94
DOI: 10.33899/ijvs.2017.145603

C. psittaci is one of the dog’s pathogen which can cause respiratory disorders in various hosts and human beings. Chlamydiae are obligatory interacellular bacteria which belong to Chlamydiales. Conjunctival and pharyngeal swabs were taken from 50 captive dogs presented at veterinary clinics of Isfahan and Shahrekord to determine the percentage of infection and prevalence of C. Psittaci in domestic dogs. Samples were collected during 2014 from a total of 7 different breeds of dog; 1-German shepherd 2-Terrier 3- Mixed Poodle 4-Doberman pinscher 5-Persian sheepdog 6- Siberian husky 7-Pekingese breeds were sampled. The molecular PCR method was used to detect this microorganism in captive dogs and C. psittaci was detected in 9 (18%) of them.

Isolation and identification of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli from broiler in Erbil, Iraq

M.N. Al-Sharook; A.A. Hassan

Iraqi Journal of Veterinary Sciences, Volume 31, Issue 1, Pages 31-38
DOI: 10.33899/ijvs.2017.126717

Extended-spectrum beta-lactamase-producing Escherichia coli isolated from slaughtered broilers in retail market that sell live chickens in Erbil city, Iraq. Forty-one cloacal fecal samples from broiler caecum were investigated from January to April 2016. ESBLs strains were isolated using MacConkey agar supplemented with cefotaxime 1 mg/l and the isolates were identified phynotypically by biochemical tests, TBX agar and VITEK-2 compact system. A total of 34 Escherichia coli and 4 Proteus mirabilis were analysed for determination of ESBL/AmpC by disc diffusion test using antimicrobial 68DC MAST® ESβL discs group including cefpodoxime, cefpodoxime + ESBL inhibitor, cefpodoxime + AmpC inhibitor and cefpodoxime + ESBL inhibitor + AmpC inhibitor and 67DC MAST® ESβL discs group including cefpodoxime, cefpodoxime + clavulanate, ceftazidime, ceftazidime + clavulanate, cefotaxime and cefotaxime + clavulanate. The phenotypic results showed that in group 68DC discs 23.7% E. coli were resistant to cefpodoxime and in group 67DC discs 73.7% of E. coli and 7.9% of P. mirabilis were resistance to one or more of the cefpodoxime, ceftazidime and ceftazidime. Final results revealed that 78.0% of samples were ESBLs/ AmpC positive. This study is the first examination to determine phenorypically E. coli producing ESBLs/AmpC in broiler chickens in Iraq. Conclusion, the healthy broiler can be a major source of ESBLs/AmpC and the possibility that transmitted to humans through the food chain, direct contact and the surrounding environment raises the concerns about public health and safety of poultry meat and the negative consequences of drug therapy that causes the spread of antibiotic resistance.