| E. coli O157:H7 is considered as an important pathogen of diarrhea in adult dogs and puppies. The presented study aimed to isolate E. coli O157:H7 serotypes from dogs and puppies and to detection of the stx1, stx2 and eaeA virulence genes in these strains using conventional PCR assay. One hundred and four fecal swab samples were collected from dogs and puppies with different ages, breeds and sex. After primary cultivation of the collected samples onto MacConkey's and EMB agar media, the suspected E. coli colonies were subcultured onto CHROM and CT-SMAC agar media. The non sorbitol fermenter E. coli were serotyped using latex agglutination test for detection of O157 and H7 antigens. The results of latex test revealed 18 E. coli O157:H7 isolates. From 16 diarrheic animals, 11 (68.75%) were positive for E. coli O157:H7; 8 (72.72%) from puppies and 3 (60%) from adults. Meanwhile, 7 E. coli O157:H7 isolates were recovered from 88 non diarrheic animals (7.95 %); 4 (9.52%) from puppies and 3 (6.52%) from adults. The 18 isolates of E. coli O157:H7 were conducted to PCR assay, the results showed that 6 (33.33%) and 1(5.55%) E. coli isolates had stx1 and eaeA genes, respectively. The results also showed that three isolates (16.66%) had all the investigated genes, three isolates (16.66%) had Stx1 and eaeA genes, one isolate (5.55%) had Stx2 and eaeA genes, while four (22.22%) isolates were negative for these virulence genes. In conclusion, we may suggest that dogs and puppies can be regarded as important reservoirs for E. coli O157:H7 which is one of the main causes of diarrhea and other diseases in human.
This study is carried out to investigate the prevalence of Coxiella burnetii (C. burnetii) infections in cattle using an enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) assay targeting IS1111A transposase gene. A total of 130 lactating cows were randomly selected from different areas in Wasit province, Iraq and subjected to blood and milk sampling during the period extended between November 2018 and May 2019. ELISA and PCR tests revealed that 16.15% and 10% of the animals studied were respectively positive. Significant correlations (P<0.05) were detected between the positive results and clinical data. Two positive PCR products were analyzed phylogenetically, named as C. burnetii IQ-No.5 and C. burnetii IQ-No.6; and then recorded in the National Center for Biotechnology Information (NCBI) under an accession numbers of MN473204.1 and MN473205.1. Comparative identity of the local strains with NCBI-BLAST strains/isolates revealed 97% similarity and 0.1-0.6% of total genetic mutations/changes. NCBI-BLAST Homology Sequence reported high significant identity (P<0.05) between the local, C. burnetii IQ-No.5 and C. burnetii IQ-No.6; strains and C. burnetii 3345937 (CP014354.1) Netherlands isolate at 99.10% and 99.06%, respectively. The current study concluded that the percentage of infected cows with coxiellosis is relatively high, and Coxiella should be listed as abortive pathogen. Therefore, additional studies should be performed including different animals, samples, and regions
This study was carried out to investigate the prevalence of Escherichia coli O157:H7 serotype from diarrheic and non-diarrheic calves. The study was out in Baghdad, a province in Iraq. A total of 350 faecal samples from 35 diarrheic calves and 315 non-diarrheic calves with different ages (up to 1 year) and from both sexes. After initially enrichment, samples were streaked on sorbitol MacConkey agar plus cifixime potassium tellurite (SMA-CT) and Chrom agar™ E .coli O157:H7. Non-sorbitol fermenting (NSF) E. coli isolates were conducted to serotyping using commercial Latex agglutination test for detection of O157 and H7 antigen. E. coli Isolates were additionally tested for virulence factor eae by PCR techniques. Four isolates (11.42%) belonged to E. coli O157:H7 in 35 diarrheic calves and 28 (8.88%) in non-diarrheic calves. All four isolates from diarrheic calves were found positive for intimin (eaeA) gene (100%) and only 13 from 28 isolates (46.42%) were possessing (eaeA) gene in none diarrheic calves. In conclusion, this study revealed the importance of calves to act as a reservoir for E. coli O157:H7. Also, the eaeA genes in a high percentage in most calves suggest that they may be virulent for humans.
Q fever is an infectious disease of animals and humans, caused by globally distributed C. burnetii. In Iraq, there are no previous studies associated with the detection of the organism in cattle. An overall of 130 lactating cows were submitted to direct collection of milk samples. Initially, the samples of milk were tested using the molecular polymerase chain reaction (PCR) assay targeting three genes (16S rRNA, IS1111a transposase, and htpB). However, positive results (18.46%; 24/130) were detected only with the 16s rRNA gene. Concerning risk factors, the highest prevalence of C. burnetii was showed in the district of Badra (42.86%), whereas the lowest - in Al-Numaniyah and Al-Suwaira districts (P=0.025). There was no significant variation in positivity between the months of sampling period (P=0.082) and between age groups (P=0.076). Crossbred cows (20.69%) showed a higher positivity than local and pure breeds (P=0.043). Milk of positive samples (n=24) was used for cultivation of C. burnetii into specific pathogen free-embryonated chicken eggs (SPF-ECEs). After three passages into SPF-ECEs, contents of yolk sac were collected, subjected for DNA extraction, and re-tested by PCR assay using the primer of 16s rRNA gene only. Of 24 cultivated milk samples, 12.5% (3/24) were positive for C. burnetii. Finally, the positive local isolates were analysed phylogenetically and reported in NCBI-Genbank under the accession numbers of MN121700.1, MN121701.1, and MN121702.1. In conclusion, this is a unique study as it detected C. burnetii in Iraqi lactating cows, and confirmed that organism was shed actively through milk, suggesting that these animals can play a role as a reservoir for organism with potential risk for transmission of infection from these animals to humans as well as to other animal species.
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