Main Subjects : Animal Bacteria

Multi-drug resistant phenotypes of extended-spectrum β-lactamase (ESBL)-producing E. coli from layer chickens

Adewale Olopade; Asinamai A. Bitrus; Asabe H. Halimat Momoh-Zekeri; Pwaveno H. Bamaiyi

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 4, Pages 945-951
DOI: 10.33899/ijvs.2022.132655.2117

Antimicrobial resistance (AMR) is a growing and emerging public health problem worldwide. This research determines the occurrence of ESBL E. coli and antimicrobial resistance profiles of E. coli on eggshells from selected layer chickens. The shells of 270 egg samples were swabbed to detect the presence of E. coli. E. coli isolates were recovered from 73(23%) of the 270 samples collected. The isolates were subjected to antimicrobial susceptibility testing using six panel antibiotics (ampicillin, tetracycline, sulphamethoxazole-trimethoprim, gentamicin, imipenem, and ciprofloxacin) using the disk diffusion method. The isolates showed the highest resistance to Ampicillin 95.9%, closely followed by tetracycline 89%, sulphamethoxazole-trimethoprim 72%, gentamicin 41.1%, and imipenem 1.4%. Also, 78% of the isolates were multi-drug resistant. A 56/73 (76.7%) out of seventy-three isolates were screened as presumptive ESBL-E. coli by culture on ESBL CHROM agar and 42/56 (75%) of the isolates yielded ESBL-producing E. coli based on the production of ESBL by double disc diffusion method. The questionnaire survey results showed that all farms used antimicrobial agents for therapeutic or prophylactic purposes. Also, not all the farms had suitable biosecurity measures. The findings of this study indicated that eggshells are potential reservoirs for multi-drug resistant E. coliand ESBL-Producing E. coli.

Detection of Campylobacter fetus in aborted ewes in Sulaimani province by PCR

Eman D. Arif

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 3, Pages 647-651
DOI: 10.33899/ijvs.2021.131225.1931

Abortion is one of the most critical factors affecting lambing rates and, as a result, sheep farm profitability. It is also significant from a zoonotic viewpoint, in addition to financial losses. In sheep flocks, Campylobacter fetus causes infectious infertility, embryonic death, and miscarriages. The study investigated C. fetus from aborted fetuses and vaginal swab samples collected from sheep flocks in the Sulaimani province by the polymerase chain reaction. Thirty-eight aborted fetuses and 70 vaginal swabs were collected from sheep flocks in three districts of Sulaimani province (Kalar, Said Sadiq, and Chamchamal) from March 2018 to June 2019. The pathogen was identified in clinical specimens using conventional PCR. C. fetus was isolated in 16 of 38 aborted fetuses (42.1%) and 13 of 70 vaginal swabs from aborted ewes (18.6 %). The C. fetus gene 16S rRNAwas sequenced and received the accession number MW694741 in NCBI GenBank. Phylogenetic analysis of 16S rRNA gene sequences designated that the C. fetus isolates formed a separate branch displayed the highest similarity and clustered with MN203686.1 and EU773268.1 accessions in a specific clade. A lower degree of affinity of C. fetus was revealed with Campylobacter coli and Campylobacter jejuni.

Molecular detection of ESBL/AmpC ß-Lactamase Escherichia coli isolated from sheep in Mosul city

Fatma R. Rafee Mahmood; Ihsan M. Ahmed

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 2, Pages 387-392
DOI: 10.33899/ijvs.2021.130380.1810

Globally, extended-spectrum ß-lactamase (ESBL)/Ampicillin ß-lactamase (AmpC) producing Escherichia coli has become the greatest threat for distributing antibiotic resistance. Accordingly, this study was designed to detect and screen the genes that confer resistance in E. coli isolated from sheep as main livestock in Mosul city. Forty E. coli isolates previously recovered from milk and fecal samples were included in this study. These isolates were obtained from healthy ewes, their lambs, and also from ewes with clinical mastitis. Polymerase chain reaction (PCR) was used to confirm the E. coli isolates targeting the 16sRNA gene. Furthermore, screening of different genotypes of ESBL/AmpC was conducted using specific primers. The results showed that the CTX-M gene was predominant among ESBL genotypes and recorded 40/40 (100%). While, SHV and TEM genes recorded 7/40 (17.5%) and 5/40 (12.5%), respectively. Moreover, fecal carriage of resistance genes was more than that obtained from milk in both healthy and diseased animals. However, none of the 40 isolates showed positive results for AmpC genes. The presence of different genotypes of ESBL E. coli isolated from feces or milk origin may act as a potential source for transferring antibiotic resistance to humans, other animals, and the environment.

Molecular characterization of extended spectrum beta-lactamase producing Klebsiella pneumoniae isolated from cows in Mosul city, Iraq

Sumaya Y. Aldabbagh

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 2, Pages 375-380
DOI: 10.33899/ijvs.2021.130341.1803

Cows are considered as reservoir hosts for many pathogenic bacteria that are resistant to broad-spectrum beta lactamase(ESBL). Presence of ESBL resistant K. pneumoniae in nasal of cows and beef meat constitutes a risk for public health due to transfer of antibiotic resistance gene from cows to environment, humans and farm animals. Therefore, the current study was concluded the detection of molecular characteristics of ESBL producing Klebsiella pneumoniae that was isolated from both cow's nostrils and from local beef samples. Fifty nasal swabs were collected from farms cows in Mosul city, and 50 samples of beef from local butcher shops for the period from February to August 2020. Bacterial isolation and identification tests were conducted for ESBL resistant Klebsiella pneumoniae using MacConkey agar medium with-1 μg/ml cefotaxime. PCR was carried out to confirm the results using special primers (SSKP 1 F and SSKP 1 R) for Klebsiella (the target gene 16srRNA). Then, a molecular examination was performed using the precursors CTX-M, TEM and SHV. Through bacterial isolation, 36 (72%) and 28 (56%) isolates were belonging to ESBL resistant Klebsiella pneumoniae from nasal samples and beef meat respectively. The CTX-M, TEM and SHV genes formed (100, 89.2, 85.7) % and (100, 72.2, 71.4) % for each the meat and nasal samples respectively. This study showed that cows play a major role in transferring ESBL producing Klebsiella pneumoniae from cows to humans as a result of environmental handling or consumption contaminated meat. 

Molecular characterization and phylogenetic analysis of Escherichia coli isolated from milk of cattle affected by mastitis

Azhar A. Neamah; Khilood H. Fahad; Jenan N. Sadeq; Monyer A. Al-Fatlawi

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 1, Pages 251-254
DOI: 10.33899/ijvs.2021.129934.1702

This evolution-based study aimed to reliably identify the epidemiological prevalence of Escherichia coli that wasrecovered from affected milk of cattle by mastitis, study the evolution of this bacterium, and describe some isolates using polymerase chain reaction (PCR) technique and DNA sequencing. Here, we collected 50 cattle milk samples and submitted them to conventional bacterial isolation and identification using enrichment culture method and biochemical tests. Then, we confirmed the results by PCR technique based on 16S ribosomal RNA gene. The results showed that E. coli was isolated from cattle at (36%), and this was confirmed by PCR that showed highly specific detection of E. coli isolates at (100%). DNA sequencing of partial 16S ribosomal RNA gene showed (99%) homological identity with NCBI-Blast E. coli isolates and the phylogenetic analysis showed genetic similarity (0.5 genetic changes). In conclusion, this was the first study in Iraq to report genetic relationship between E. coli isolated from milk of mastitis-infected cattle. Therefore, it is essential to define the role of animals as an important source in the distribution of some pathogens that are related to public health.

Major-surface-protein-4-gene-based detection of Anaplasma marginale isolated from sheep in Al-Diwaniyah province, Iraq

Saba F. Klaif; Asaad J. Abid; Monyer A. Al-Fatlawi; Mansoor J. Ali

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 1, Pages 85-88
DOI: 10.33899/ijvs.2021.129230.1635

This study was purposed for confirming detection and typing of Anaplasma spp in infected sheep from Al-Diwaniyah province, Iraq. Sampling of 50 blood specimens was performed from clinically-identified infection of anaplasmosis. The samples of the blood were subjected to DNA extraction followed by polymerase-chain-reaction (PCR)-based detection of the Anaplasma marginale using major surface protein (MSP4) gene. The results have shown that 8 blood samples were infected with A. marginale. The PCR-based identification has revealed a confirmative identification of the Anaplasma marginale in the infected sheep. The study identifies Anaplasma marginale as a member of infectious agents that affect sheep in the study city. 

Immunotherapeutic effect of chitosan and listeriolysin O on Listeria monocytogenes infection in mice

Hero I. Ali; Shaimaa N. Yasssein

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue Supplement I-III, Pages 149-155
DOI: 10.33899/ijvs.2021.132332.2082

This study aimed to evaluate the effect of dietary chitosan and listeriolysin O (LLO) on the immune response against Listeria monocytogenes infection in mice. L. monocytogenes isolate was obtained from the unit of Zoonotic Disease, College of Veterinary Medicine, University of Baghdad. It was reactivated and cultured on PALCAM agar and exposed to a confirmatory diagnostic test. listeriolysin O (LLO) was extracted and purified. Sixty mice were used and divided into four groups each one involving 15 mice; the first group was fed on diet supplemented with a chitosan for 4 weeks. At the end of the 2nd week, normal saline was injected S/C; the second group was fed on a normal diet without any addition. At the end of 2nd week, 0.3 ml of LLO was injected S/C, then the booster dose of LLO was given after 14 days of 1st injection; the third group was fed on a diet supplemented with chitosan as in G1, and treated as in G2 and the fourth group fed on a normal diet without any addition and treated as in G1. A skin test was performed on 5 mice of each group while a Challenge test by injection of 0.2ml of 1x109 CFU/ml of viable L. monocytogenes intraperitoneally was performed on other mice. The concentration of IgG titer and IL6 were measured. The results revealed that the third group recorded significantly higher values in the skin thickness, IgG, and IL6 concentrations when compared with other groups at different times which indicate that LLO and chitosan may support each other to provide the most beneficial effect by eliciting of good cellular and humeral immunity against listeriosis.

Genetic diversity and clonal relatedness of Aeromonas hydrophila strains isolated from hemorrhagic septicemia’s cases in common Carp (Cyprinus carpio) farms

Zanan M. Taha

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue 4, Pages 643-648
DOI: 10.33899/ijvs.2020.127566.1511

The objective of this study was to determine molecular typing and comparison analysis of 24 Aeromonas hydrophila isolated from the diseased fish with hemorrhagic septicemia in freshwater ponds and cage in Mosul and Duhok cities, Iraq. A total of 24 A. hydrophila isolates that were collected from various fish ponds and cage, were used in this study. Identification of isolates was made by the standard microbiological and molecular methods. ERIC-PCR was done with different primers to establish the genetic relationship between strains. ERIC-PCR typing showed that 24 strains of A. hydrophila were classified into 11 ERIC types (genotypes). Genotypes 9 and 7 represented the most prevalent clone. All A. hydrophila strains that were isolated from the same fish were genetically diverse. There was minimal genetic similarity between some strains which were retrieved from the same geographical source area. Also, some isolates from different geographic source area were showed a 100% genetically similar. Aeromonas hydrophila was genotypically heterogeneous and clonally dispersed among different fish ponds and cage in Mosul and Duhok cities, Iraq. Besides, one fish can be infected with more than one strains of A. hydrophila.

Detection of CTX-M gene in extended spectrum β-lactamases producing Enterobacteriaceae isolated from bovine milk

Ihsan M. Ahmed

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue 2, Pages 397-402
DOI: 10.33899/ijvs.2020.126909.1412

Extended spectrum β-lactamases producing Enterobacteriaceae (ESBL-E) have emerged recently as the main cause that facilitates the spreading of antibiotic resistance worldwide. Due to its composition and nutritive values, raw cow milk is vulnerable to bacterial contamination from different sources, especially ESBL-E. Accordingly, present study aimed to detect the ESBL-E in the raw milk of healthy cows. 80 raw cow milk samples were collected from unorganized farms and cows belong to individual owners and investigated for the presence of ESBL-E with the main focusing on CTX-M type. The bacterial isolation was performed using selective MacConkey agar plus cefotaxime (MC+), while PCR was used to confirm the species of the isolated bacteria and presence of CTX-M gene. The results showed that 28.75%(23/80) samples were ESBL-E positive and distributed as following, 82.61%(19/23) were pure E. coli isolates, 4.35%(1/23) was pure K. pneumoniae isolate and finally, 13.04%(3/23) were mixed of both E. coli and K. pneumoniae isolates. Moreover, the total number of positive ESBL-E was 26 isolates with the majority of them were belong to E. coli and recorded 84.61%(22/26), while K. pneumoniae was recorded less 15.39%(4/26). Additionally, the CTX-M gene was successfully identified in all ESBL-E positive isolates by using PCR, including E. coli and K. pneumoniae isolates. The results of this study assert the importance of raw cow milk as a potential source of ESBL-E that might be transmitted to humans.

Molecular analysis of ompA gene Pasteurella multocida Indonesia local isolates

A. T. Soelih Estoepangestie; Arini Rahmi Dewi; Suwarno Suwarno; Didik Handijatno; Rahaju Ernawati; Wiwiek Tyasningsih

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue 2, Pages 211-216
DOI: 10.33899/ijvs.2019.125934.1191

The aim of this research was to analyze ompA molecular gene of Pasteurella multocida buffalo isolate and bovine isolate from Nusa Tenggara Timur, Indonesia and Katha strain isolate from hemorrhagic septicemia vaccine. Determinant of P. multocida local isolates ompA gene amplification sequencing PCR then conducted to see the sequence of nucleotide sequences of ompA gene. The results of PCR amplification showed an amplicon of 559 bp of all isolates. The homology analysis result of the isolates ranged from 93 - 100% with 13 P. multocida isolates from GenBank, and phylogenetic tree analysis shows that buffalo isolate was closely related to Katha strain, Iran, India and China isolate. Whereas bovine isolate far enough with buffalo and Katha strain isolate. Nucleotide sequences were compared to amino acids then by the method of Kolaskar and Tongaonkar antigenicity predicted antigens in P. multocida. B cell epitope predictions from local isolates and Katha strain were found in five peptides QVSPVFAG, IPELALRVEYQ, GQSVYVPEVVSKT, LKSASVAVAG, and ANYLVAKG.

Evaluation the safety and synergistic effect of NiFe2O4 nanoparticles with antibiotic against Pseudomonas aeruginosa

Hala M. Majeed; Heba Younis Khalef; Halah Abdulkhaleq Awadh; Bashar Sadeq Noomi; Nihad Abdul-Hussain jafar; Khaild Ahmad Hadi

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue 1, Pages 71-77
DOI: 10.33899/ijvs.2020.126298.1294

The antimicrobial resistance currently impedes and threatens the future of effective prevention and treatment of the continually expanding range of infections caused by bacteria. This study aimed to identify the bacterial causes the wound infection among animals and using the antibiotic/nanoparticles mixture as a new attempt for the treatment the wound infection induced in rats. For this purpose, 112 swabs wound infection cases in the different animal types (36 sheep, 21 goats, 12 cows, 4 horses, 8 dogs, 9 rabbits, 7 genies pigs and 15 rats) were studied in the for bacterial isolation. The Pseudomonas aeruginosa was tested for its sensitivity to the antibiotics and the nanoparticles (CoFe2O4 and NiFe2O4) in vitro by using the MIC method. Also the wound infection was induced in the rats and the effect of nanoparticles/antibiotics mixture were tested in vivo. The results showed that P. aeruginosa was the predominant bacterial type that the caused wound infection. The minimum inhibitor concentration of NiFe2O4 and CoFe2O4 nanoparticles were 32 µg /ml and 16 µg /ml respectively. A clear synergistic effect of antibiotic/ nanoparticles as antibacterial were noticed which appear as a decrease in MIC and increase of the inhibitory diameter zone. According to the result of Random Amplification of Polymorphic DNA test, the nanoparticles effects on genetic material of P. aeruginosa observed as an appearance/disappearance of bands, increase in thickness and clarity of the bands.

Molecular fingerprinting of methicillin resistant Staphylococcus aureus strains isolated from human and poultry in Duhok, Iraq

Hishiyar A Hado; Mahde S Assafi

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue 1, Pages 99-103
DOI: 10.33899/ijvs.2020.126375.1310

Methicillin-resistant Staphylococcus aureus (MRSA) has been recently identified in poultry and farm workers. The aim of this work was to investigate the epidemiological relatedness of MRSA among chickens and farmworker. MRSA isolates (n=50) from human (n=14) and from chikens (n=36) were tested for molecular epidemiological relatedness between human and poultry. RAPD-PCR was carried out for fingerprinting of MRSA isolates genome. Seven genotypes group (A-G) have been identified. All human MRSA were belonging to genotype A. Whereas, chickens MRSA isolates was belonging to different genotype patterns groups (A-G). To conclude, human MRSA was belonging to one genotype pattern but the chickens MRSA strains were belonging to seven genotypes. The genotype pattern A was the most dominant among all MRSA isolates. It is possible that the chickens play an important role for the human exposure to MRSA by direct contact. Further studies are required to address the relatedness between human and chicken MRSA.

Genetic detection to Aeromonas hydrophila proteolytic activity in milk samples (cows, buffaloes and goats) in Basra governorate

Rawa B Banay

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 2, Pages 253-258
DOI: 10.33899/ijvs.2019.125888.1174

This study aim is to determine the incidence and the virulence of Aeromonas hydrophila in raw milk, randomly collected from Basra governorate by using of polymerase chain reaction (PCR) technique. In this study, the total number of raw milk samples collected from cows, buffaloes and goats that kept from different the regions of Basra governorate were 90 samples. The PCR technique is modern method which regarded as a reliable tool to detect virulent gene of the A. hydrophila isolates. The PCR assays using the primers sets SerAh-F and SerAh-R resulted in the amplification of 650-bp bands from the targeted proteases gene of the A. hydrophil. The result of the present study showed that the results of PCR concerning the proteolytic activity of A. hydrophila in the tested raw milk samples according to animals' source. The higher percentage of the proteolytic activity was found in the cow's raw milk samples 40% and in the buffalo's milk samples was 26.7% while, the proteolytic activity did not find in the goat's milk samples. The association between the source of the milk sample and proteolytic A. hydrophila positive results was considered to be statistically highly significant. The higher percentage of the A. hydrophila isolates found in the raw cow milk was 40%, and the A. hydrophila isolates found in the raw buffalo milk was 26.7%, while, the A. hydrophila isolates did not find in the goat milk.

Bioremediation of lead and cadmium and the strive role of Pediococcus pentosaceus probiotic

Raghad Jaafar

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 1, Pages 51-57
DOI: 10.33899/ijvs.2019.125581.1092

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.

Detection of Mycobacterium paratuberculosis in raw cow’s milk using polymerase chain reaction (PCR) technique

Ihsan M. Ahmed; Raad A. Al-Sanjary; Haiffa H. Alkazaly

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 1, Pages 83-86
DOI: 10.33899/ijvs.2019.125556.1075

Paratuberculosis or Johne’s disease is a chronic debilitating disease mainly infects ruminants and caused by Mycobacterium paratuberculosis. Previous serological studies in Mosul city confirm the presence of positive reactants for paratuberculosis in cattle. However, culture methods to confirm the disease need a long incubation period and also special media. Raw cow’s milk is considered as potential source for transmission of M. paratuberculosis in cows’ herds. Accordingly, this study aimed to detect the presence of M. paratuberculosis specifically in the raw cow’s milk using polymerase chain reaction (PCR) technique as a rapid, sensitive and reliable method. A total of 50 samples of raw cow’s milk were collected from cows suffering from emaciation and unresponsive to antibiotic treatment. All the samples were subjected to DNA extraction and direct amplification PCR. The results showed that 3 (6%) out of 50 milk samples were positive for M. paratuberculosis. This is the first study in Mosul city that confirms the presence of M. paratuberculosis in raw cow’s milk using PCR technique. In conclusion, raw cow’s milk could be an important source for M. paratuberculosis infection in dairy cows, and also PCR technique could be helpful in rapid diagnosis of paratuberculosis.