This evolution-based study aimed to reliably identify the epidemiological prevalence of Escherichia coli that was recovered 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.
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.
Hemolysin protein is exotoxin produce by organisms that cause lysis of blood cells. This study was conducted to screen the presence of hemolysin gene from 20 isolates of Klebsiella pneumonia based 16S rRNA genes by using specific primer. This gene potent the pathogenesis of Klebsiella pneumonia. The primer was designed in this study by NCBI-GenBank and primer3 plus. (Bioneer Company provided the primers. Korea). Molecular detection of isolates, which give away specific PCR products of 505bp for hly gene, hemolysin gene, was detected in 70% (14/20).
Iraqi's local markets contains a significant variety of imported meats from different sources, beef burger is one of them, the current study aimed to knowledge occurrence and prevalence of bacteria in meat of beef hamburger by using vitek 2 technique in Al-Diwaniyah city. Forty Samples of beef burger were chosen randomly from local markets, all the isolates submitted to culture on many of media agar like MacConkey, blood base agar, SS agar, manitol salt agar, Eosin-methylene blue agar, Chromagar Orientation, , and salmonella Chromagar, then the isolates tested by vitek 2 technique to confirm final diagnosis, the study was shown there are many of bacteria reside in beef hamburger at different percentage, E.coli (55%), Klebsiella pneumonia (38%), Klebsiella oxytoca (2%), Staphylococcus aureus (44%), Proteus mirabilis (13%), Salmonella typhimurium (18%), Enterococcus feacalis (8%) and serratia marcescens (3%). It could consideration meat of beef burger is an eligible nutrient to the multiplication of many of bacteria types that may be very dangerous if it transmitted to human.
Real-time polymerase chain reaction assay was used for detection of Clostridium perfringens toxins genes alpha (cpa )directly from the fecal samples of cattle and sheep.Fecal samples from 20 clinically healthy cattle and 20 clinically healthy sheep were collected randomly from different farms located in AL-Diwanyia city ,All the strains of Clostridium perfringens were analyzed by Real-Time PCR using specific primers for alpha (cpa)toxin genes ,its reveald that (cpa)toxin gene of Clostridium perfringens ,were detected in cattles in 8(40%) and 14 (70%) in sheep Real time positive respectively samples ,This results indicate that Clostridium perfringens alpha toxin producing strains were prevalent in sheep and cattles and its possibly play an important role in the diarrheal disease caused by Clostridium perfringens .
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