Detection and pathogenicity of Listeria monocytogenes in common carp (Cyprinus carpio) fish in Baghdad, Iraq

Al-Gburi, Nagham M.

doi:10.33899/ijvs.2019.125980.1205

Cited by 10 publications

(5 citation statements)

References 24 publications

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“…The PCR tube included l0 µL of master mix, 5 µL of DNA, and l µL (10 pmol) of each forward and reversed primer. The ultimate volume of 20 µL was achieved by adding 3 µL of DEPC-treated water (25,26). The thermal cycler method began with an initial denaturation at 95 °C for five minutes.…”

Section: Oligonucleotides and Pcr Amplificationmentioning

confidence: 99%

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

Arif

2022

IJVS

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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 rRNA was 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.

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Section: Oligonucleotides and Pcr Amplificationmentioning

confidence: 99%

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

Arif

2022

IJVS

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“…During this work, a positive control was used in order to optimize RT-PCR technique and to match with the PCR primers and hybridization probe. This control was represented by a DNA extract of L. monocytogenes (accession numbers MH092995.1) confirmed previously by PCR and DNA sequence obtained from the BLAST-N program (National Center for Biotechnology Information) and recorded by a previous study on carp fish sample [30 ]. Forward primer -TCGCAAACAGATCTAGACCAAGTT-3` inlA-R 5`…”

Section: The Isolation Of L Monocytogenesmentioning

confidence: 99%

Detection of Listeria monocytogenes in Several Types of Frozen Meat in Baghdad city

Yassin

Nadhom

Al-Gburi

2021

eijs

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Detection of pathogenic bacteria, such as Listeria monocytogenes, in food is crucial for safeguarding public health in Iraq. Forty five samples of frozen meat (15 samples of each of minced red meat, chicken, and fish) were collected from different markets in Baghdad city. Molecular (RT-PCR) and culturing (conventional microbiological examination) methods were used to determine the level of contamination of L. monocytogenes in these types of meat. For the culturing method, TSYEB broth was used as an enrichment medium, whereas BALCAM medium (HiMedia) with the listeria selective supplement FD061 was used as a selective medium, for the isolation and identification of this bacterium. The isolates were confirmed microscopically and biochemically. The results of the culturing method showed that the total number of the isolates of L. monocytogenes was 14/45 (31.1%). The incidence of this bacterium was high in fish (11/15, 73.3%), while it was low in the other two types of meat. 2/15 (13.3%) in red meat and 1/15 (6.7 %) in chicken. Molecular detection of each sample of the bacteria was performed using RT-PCR technique after preparing the Genomic DNA extraction of these samples according to the protocol provided by ReliaPrep™ Blood gDNA Miniprep System kit (Promega, USA). The PCR primers and the hybridization probe ((Macrogen, Korea) were used to target the inlA gene sequence (specific for L. monocytogenes). The results of the RT-PCR assay showed that 10/45 (22.2%) of the samples were positive for L. monocytogenes, which was detected only in fish samples ((10/15, 66.7%), while not found in minced red meat and chicken. However, our results showed differences when compared to other previous works because there were many studies found that the highest contamination rate was in red meat products. We conclude that the PCR kit used for the detection of L monocytogenes appears to give accurate results in the diagnoses of this bacterium in meat products and in comparison with the other routine diagnosis methods in the laboratory, which included culturing and doing biochemical tests which last for approximately 7 days, the RT-PCR technique was able to confirm the findings within 48 hours.

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“…The bacterial sample was achieved from Media Diagnostic Center in Erbil, Iraq. virulent Listeria monocytogenes were commonly cultivated at 37°C on Brain heart infusion agar for stimulation of bacteria and after activation on it transported to Listeria Identification Agar Base (PALCAM) (HiMedia, India) M1064 with Listeria Selective Supplement (PALCAM) (HiMedia, India) to avoid mutation of bacteria also as selective identification of L. monocytogenes by providing a grey-green with a black center and a black halo (12).…”

Section: Bacteriamentioning

confidence: 99%

Immunohistochemical and pathological changes in BALB/c mice immunized with whole sonicated Listeria monocytogenes antigens and the effect of probiotics

Shihab¹,

Ibrahim²

2021

IJVS

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The current study was undertaken to investigate the role of macrophages as a cellular immune function against immunization with whole sonicated Listeria monocytogenes antigens (WSLMAgs) and the effect of probiotics. A preparation of WSLMAgs containing whole L. monocytogenes cell, after two subcutaneous immunization of BALB/c mice with 0.5ml WSLMAgs 0.5 mg/ml at an interval of two weeks. The bacterial identification was conducted by a conventional culture method using Listeria selective media PALCAM and confirmed by Polymerase Chain Reaction (PCR), As well as, the immunohistochemical and pathological change of it was studied in vivo by inoculating mice with pre-challenge WSLMAgs and post-challenge with virulent L. monocytogenes 1×10 8 CFU/mL. The results revealed the cellular immune function against pre-and post-immunization in spleen organ via lymphocytic hyperplasia in white pulp and coalescence of lymphoid follicles and marker F4/80 + show the immune-positive cells in aggregation adjacent to lymphoid follicle or focal aggregation of macrophages between follicles. In conclusion, the effectiveness of sonicated L. monocytogenes pre and post-immunization then challenge with virulent L. monocytogenes in the induction of cellular immune response, might serve as an immunization platform for applicants.

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Detection and pathogenicity of Listeria monocytogenes in common carp (Cyprinus carpio) fish in Baghdad, Iraq

Cited by 10 publications

References 24 publications

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

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

Detection of Listeria monocytogenes in Several Types of Frozen Meat in Baghdad city

Immunohistochemical and pathological changes in BALB/c mice immunized with whole sonicated Listeria monocytogenes antigens and the effect of probiotics

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