Iraqi Journal of Veterinary Sciences

Livestock-associated methicillin-resistant Staphylococcus aureus (MRSA) has become a global public health concern. The purpose of this study was to investigate the prevalence rates of MRSA infection amongst broiler chickens and broilers farm workers. The total samples used in this study were 306. Cloacal swab samples from 231 broilers and nasal swab samples from 75 broilers farm workers were collected from five farms in Duhok city, Iraq. Isolation and identification of MRSA isolates were carried out and the antibiotic susceptibility were screened. Molecular characterization of all isolates was performed by using polymerase chain reaction (PCR) technique to detect the mecA gene.S. aureus was detected among 84% (63/75) of the farms workers samples and among 84.8% (196/231) of the broiler's samples. The S. aureus isolated from farm workers and broilers appeared resistant to oxacillin 28.6% (18/63), and 32.1% (63/196), respectively. MRSA colonization in farm workers and broilers was 24% (18/75) and 27.3% (63/231) respectively. The S. aureus isolates showed the most resistant to chloramphenicol and the least resistant to vancomycin. The results of the PCR assays revealed that 85.7% (12/14) of S. aureus isolates from farm workers and 44.4% (16/36) of S. aureus isolates from broilers were positive for the mecA gene. The direct handling of broilers by farm workers plays the important role for transport the MRSA isolates from broilers to broilers farm workers.

Shigella is an intracellular bacterium can infect both human and animal. Its species especially Shigella dysenteriae cause shigellosis worldwide, with 165 million cases of severe bloody diarrhea and mucoid feces. The aim of this study was to find a rapid, sensitive and specific method for screening Shigella in raw bovine contaminated milk. For this goal, 70 samples of milk collected in sterile containers for isolating of Shigella and culturing it on selective media to identify and characterize its morphology, biochemical and molecular characteristics. This study was compared between three different DNA extraction techniques for polymerase chain reaction (direct DNA extraction using a kit, alkaline DNA extraction, and filtrated milk). Our results showed that PCR was able to detect Shigella in 15 out of 15 cases after the milk samples filtered. In other words, the filter technique can be used to detect Shigella in contaminated milk.

Consumption of food and water contaminated with heavy metals poses a huge threat to the life. Both of Lead (Pb) and Cadmium (Cd) are heavy metals and important environmental pollutants. Away from traditional treatments, the current study aims to adopt probiotic bacteria Pediococcus pentosaceu to treat heavy metal pollution. Present results indicated a good probiotic property of P. pentosaceus, where they were able to survive pH range from 3-9, during incubation periods 3 and 24 hours, and bile salt range 0.15-0.5% for the same period. The number of bacteria in gastric (pH 3) and intestinal juices (pH 8) after 24 hours of incubation was 390 and 205, respectively. Bacteria showed an inhibitory effect against pathogenic bacteria Salmonella sp. The antibiotic susceptibility test revealed them resistant to clindamycin, intermediate resistant against benzylpenicillin, ampicillin, and their sensitivity to the rest tested antibiotics. Isolated bacteria identified based on their morphology, biochemical characteristic in addition to the use of automated instrument for bacterial identification (Vitek II), and depending on the results bacteria were identified as P. pentosaceus. In bioremediation study, the lowest inhibitory concentration of lead and cadmium and (MIC) was done, followed by assay the removal capacity by P. pentosaceus, using atomic absorption spectrometry (AAS) analysis. Bacteria show high MIC (1800 and 150 ppm) for Pb and Cd respectively. With removal efficiency for Pb 62.10-68.39% in the concentrations 25 and 50 ppm, respectively, and for Cd 52.71-11.25% in the same concentrations. Depending on the present finding probiotic bacteria (P. pentosaceus) can apply in the bioremediation of heavy metals in the fish ponds when contamination occurs, in addition to their tradition used as safety additive to prevent fish disease and an enhancement agent .Finally the isolation of these bacteria from fish ponds can be considered as a good indicator for a healthy state of fish ponds in the studied area.

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.

This study was conducted to evaluate the efficacy of the fungus Metarhizium anisopliae  as a vital agent for the control of the Hyalomma anatolicum, which is infested on buffalo fields in some villages of Anbar province, Iraq. The results showed that different concentrations of the fungus 4.2*110, 4.2*310, 4.2*510 pg/ml were capable of killing the tick eggs, and the kill rate was proportional to the higher concentrations used. After 3 days of treatment, moreover causing a high proportion of phenotyping deformation in male and female ticks.

The study included the injection of Salmonella enterica serovar typhimuriumisolated from starlings bird in embryonated chicken eggs. The eggs were divided into eight groups, each group contain 6 eggs. The G1 and G2 groups were injected with the sterile normal saline solution in the choriaollantoic membrane (CAM) and yolk sac as negative control. The group G3 and G4 injected by bacterial suspension at a concentration of 10⁴ cfu/ml in the CAM and yolk sac while the G5 and G6 injected with bacterial concentration 10⁶ cfu/ml in the CAM and yolk sac respectively. Finally, the G7 and G8 groups were injected with 10⁸ cfu/ml of bacterial suspension in the CAM and yolk sac respectively. The results showed that the highest percentage of death in eggs embryos was 100% in the sixth group after 96 hours of injection. There was also a significant increase in the number of bacteria in correlation with time of incubation. The highest rate of bacterial isolate was 8,19log10, 8,26log10 after 96 and 144 hours in the sixth group, while the highest number of bacterial isolates was 7.04log10 and 6.31log10 in the third and fourth groups after 48 and 96 hours of injection respectively. The results of the statistical analysis showed a significant difference in the number of bacterial isolate after 24 hours of injection in both concentrations compared to other incubation times. A significantrelationship was also found between the amount of the dose used and the bacterial disease. this study concluded that Salmonella enterica serovar typhimurium isolated from starlings can cause pathological changes and effect on hatchery percentage in embryonated chicken eggs.

Echinococcus granulosus (E. granulosus) is a dog tapeworm cestoda; it is larval stage responsible to cystic echinococcosis, one of the most common and dangerous worldwide zoonotic parasitic disease. The aim of this study was the molecular identification of the local strain of E. granulosus isolated from sheep liver slaughtered in the principal abattoir of Aqrah city, Northern of Iraq during Jun-Nov. 2017. In this study, 37 sheep liver infected by E. granulosus, 12 of high DNA purity fertile (have protoscolices) cyst of them were considered. A molecular study conducted on the mitochondrial NADH dehydrogenase 1 (nad1) gene. Results demonstrated that E. granulosus isolates were sheep strain (G1) genotype, with fascinating highly corresponding 95% and 96% to global isolates, particularly to north African and Mediterranean countries, by employing phylogenetic tree analysis. So, the isolates of our project were deposited in Genbank (accession No. MG792129). This study findings provide that the local isolates of E. granulosus from sheep liver in Aqrah city, Northern of Iraq are loyally equivalent to global strains and isolates, in addition, nad1 gene considers a perfect biomarker in a molecular identification and phylogenetic study of this parasite.

Laboratory methods are essential for the diagnosis of Mycoplasmal infection. There are three laboratory approaches are essential for the diagnosis of Mycoplasmal infection in chicken including direct methods by culture method and polymerase chain reaction, and indirect methods by detection of Mycoplasmal antibodies by serological tests. This study aimed to detection of Mycoplasma by culture and PCR technique. Two hundred seventy-six samples were collected from infected adult boiler chicken in Salah Al-din province which suffering from respiratory signs and /or joint infection, 202 respiratory and 74 articular samples. According to the results of culture, Mycoplasma isolated in rate of 35.1% (36.6% from respiratory samples and 31.1% from articular samples). The sensitivity of culture was 100%, while the specificity of culture was 97.9% when comparing with PCR results. The current study concluded that the respiratory infection was more than articular infections, and Mycoplasma gallisepticum more distributed than Mycoplasma synoviae among chickens.

The objective of the current study was to detect the clinical signs of colibacillosis in buffalo calves, isolate E. coli O157:H7, detect its virulence gene eaeA using PCR and estimate its prevalence.The current study sampled 120 buffalo calves aged 1day to 5 months from the Al- Basra veterinary Hospital and Private veterinary clinic within the Basra province between October 2017 and July 2018. A total of 100 calves were naturally diarrheic and the other 20 calves served as controls. The clinical sings in the diarrheic subjects included a significant increase in body temperature, heart rates, respiration rates and capillary refill time as compared to control group. Other clinical signs included whitish to yellowish watery diarrhea with tincture of blood, anorexia, weakness, depression, weak suck reflex, dry oral mucous membranes, cold extremities, weak peripheral pulse, dehydration and death. Using phenotypic characterization tools like MacConky agar, EMB agar, biochemical tests and Viteck, 83 out 100 diarrhea samples confirmed E. coli. Using CT- SMACT agar, 31 out of 83 E. coli isolates were E. coli O157:H7 positive. The PCR result indicated that 47 out of the 83 isolated E. coli samples were positive for eaeA virulence gene. In conclusion, this study is a debut in the report of E. coli and E. coli O157:H7 isolation and genes identification in buffalo calves in Iraq. Therefore, proper prevention and control measures are requisite to curtail the mortality and morbidity rate caused by Colibacillosis.

This study included collect of 150 samples from brain and meat of sheep from the slaughterhouse and local butchers shop in Mosul city. 50 sample from each (brain, cutting meat, and minced meat) which used for detection of listeria monocytogenes. The International Standard Organization (ISO) methods were used for isolation. The isolated bacteria were diagnosed according to bacterial morphology, culture, and biochemical characteristics. 10 isolates were obtained, which included 2(4%) isolates from the brain of sheep, 3 (6%) isolates from cut meat and 5(10%) from minced meat. Virulence factors tests were used for bacterial isolates which include, lecithinase, lipase, protease, esterase, and hemolysin. Antibiotic sensitivity test for bacterial isolates was also used for some antibiotics. The results indicated that all isolates were sensitive to Ampicillin, Gentamycin, Chloramphenicol, and resistant to Nalidixic acid. However, they showed variant sensitivity to other antibiotics. In conclusion, this study documented that L monocytogenes can be isolated from brain and meat of sheep in Mosul city.

Flu is a highly contagious and common illness caused by influenza A, B, and C viruses. The aim of the present study was to investigate the transformation and cloning of NS1B gene with pET-32a, pET-32b and pQE-81L in Escherichia coli BL21(DE3) and DH5α. pUC57-NS1B synthetic gene was transform and clone in Escherichia coli BL21(DE3). Isolation, single digestion and ligation with pET-32b using HindIII restriction enzyme. Amplification of recombinant DNA was done with conventional PCR after transformation. Screening with IPTG of colonies. Gel electrophoresis was done for each step of cloning after isolation. Isolation, double digestion and ligation with pET-32a and pQE-81L using SacI, PstI and HindIII respectively.Recombinant DNA was attempted to be transformed into E. coli strains BL21 (DE3) and DH5α. pUC57 plasmid carrying NS1B gene was successful transformed and isolated from E. coli BL21 (DE3). Designed primers used for PCR of NS1B showed successful amplification. First screening of pET-32b-NS1B colonies using white/blue method, cloning NS1B into pET-32b using single restriction digestion with HindIII, pET-32a using double restriction digestion with SacI and HindIII and pQE-81L using double restriction digestion with PstI and HindIII gave unexpected result. This result may relate to re-ligation of digested vector for single digestion and uncompleted digestion for vectors of double restriction digestion. Current study has suggested that recombinant NS1B gene can be cloned using single digestion with other expression vectors.

Two hundred faeces sample were collected from cattle with different age and sex in Al- Diwaniyah Province. The study was conducted in the period between November 2016 and November 2017. Salmonella typhimurium bacteria identified by routine methods such as culturing on selective media, biochemical test and agglutination test using monovalent and multivalent antisera. PCR was can detection type-1 fimbriae gene coding for fimC of Salmonella typhimurium. Results showed that Salmonella isolates were 14.5% in the bovine fecal samples. Also, the serotyping of isolates by using monovalent and polyvalent antisera revealed that all Salmonella isolates in cows were S. typhimurium. The PCR technique was used for detection of type-1 fimbriae coding gene by specific primer for fimC gene. All S. typhimurium isolates in cows appeared to be contained this gene show one distinct band MW.289 bp when electrophoresed on agarose gel. The results of this score indicated that the PCR technique potentate a loud specify in the disclosing of S. typhimurium especially the serotype that encoded to fimC gene type-1 fimbriae isolated from cows in comparison to other routine diagnostic tests.

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.

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 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.

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.

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.

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.

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.

This updated review tabulates confirmed pathogens of some diseases in selected animal species in Iraq. Laboratory confirmations were done on all the reported cases.

During the 2^nd week post inoculation of thirteen rabbits with Mycobacterium bovis tuberculosis lesions appeared in the lungs, liver, spleen, kidney, mediastinal and hepatic lymph nodes and in the omentum with an equal distribution in these organs. During the 4^th week post inoculation, these tuberculosis lesions increased in size to become well developed granulomas with caseated centers. These granulomas persisted to the 6^th, 8^th and 10^th weeks post inoculation and became more encapsulated later on. Three rabbits died during the 7^th week post inoculation due to generalized tuberculosis.