scholarly journals Isolation and molecular characterization of Mycobacterium tuberculosis complex isolated from raw milk in some dairy farms in Egypt

2016 ◽  
Vol 5 (2) ◽  
pp. 105
Author(s):  
Heba Hussien ◽  
Eman Mahrous
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Real Time ◽  
Real Time Pcr ◽  
Culture Method ◽  
Raw Milk ◽  
Accurate Method ◽  
Tuberculosis Complex ◽  
Milk Samples ◽  
Source Of Infection

<p>This study was conducted to detect <em>Mycobacterium tuberculosis</em> complex in milk in three Egyptian Governorates; El-Sharkia, El-Menoufia and El-Behera Governorates. 300 milk samples were collected from tuberculin positive cases, 18 (6.0%) were shedding <em>Mycobacterium tuberculosis</em> complex in their milk which detected by real time PCR. On another hand, 170 milk samples were collected from tuberculin negative cases, 5 (2.9%) were shedding <em>Mycobacterium tuberculosis</em> complex in their milk which detected by real time PCR. All milk samples were examined by three techniques including Microscopic examination, culture and real time PCR. Real time PCR is more rapid and accurate method than microscopic and culture method. The isolated colonies from culture were examined by Multiplex PCR to demonstrate the source of infection either human or animal source.</p>

scholarly journals Toxoplasma gondii and pre-treatment protocols for polymerase chain reaction analysis of milk samples: a field trial in sheep from Southern Italy

2017 ◽  
Vol 6 (1) ◽  
Author(s):  
Alice Vismarra ◽  
Elena Barilli ◽  
Maura Miceli ◽  
Carlo Mangia ◽  
Cristina Bacci ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Real Time ◽  
Dna Extraction ◽  
Real Time Pcr ◽  
Southern Italy ◽  
Raw Milk ◽  
Type I ◽  
Treatment Protocols ◽  
Milk Samples ◽  
Pre Treatment

Toxoplasmosis is a zoonotic disease caused by the protozoan <em>Toxoplasma gondii</em>. Ingestion of raw milk has been suggested as a risk for transmission to humans. Here the authors evaluated pre-treatment protocols for DNA extraction on <em>T. gondii</em> tachyzoite-spiked sheep milk with the aim of identifying the method that resulted in the most rapid and reliable PCR positivity. This protocol was then used to analyze milk samples form sheep from three different farms in southern Italy, including Real Time PCR for DNA quantification and PCR-RFLP for genotyping. The pre-treatment protocol using EDTA and Tris-HCl to remove casein gave the best results in the least amount of time compared to the others on spiked milk samples. One sample of 21 collected from sheep farms was positive on one-step PCR, Real Time PCR and resulted in a Type I genotype at one locus (SAG3). Milk usually contains a low number of tachyzoites and this could be a limiting factor for molecular identification. Our preliminary data has evaluated a rapid, cost-effective and sensitive protocol to treat milk before DNA extraction. The results of the present study also confirm the possibility of <em>T. gondii</em> transmission through consumption of raw milk and its unpasteurized derivatives.

Detection of Mycobacterium tuberculosis complex with Real Time PCR: Comparison of different primer-probe sets based on the IS6110 element

2006 ◽  
Vol 66 (1) ◽  
pp. 177-180 ◽  
Author(s):  
Paul H.M. Savelkoul ◽  
Arnold Catsburg ◽  
Sije Mulder ◽  
Ludo Oostendorp ◽  
Jurjen Schirm ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Real Time ◽  
Real Time Pcr ◽  
Mycobacterium Tuberculosis Complex ◽  
Tuberculosis Complex ◽  
Is6110 Element

scholarly journals Identification of Coxiella burnetii in Raw Milk of Livestock Animal in Iran

2021 ◽  
Vol 2021 ◽  
pp. 1-5
Author(s):  
Ashraf Mohabati Mobarez ◽  
Ehsan Mostafavi ◽  
Mohammad Khalili ◽  
Saber Esmaeili
Keyword(s):  
Real Time ◽  
Coxiella Burnetii ◽  
Real Time Pcr ◽  
Q Fever ◽  
Goat Milk ◽  
Raw Milk ◽  
Molecular Evidence ◽  
Milk Samples ◽  
Sheep Milk ◽  
Dairy Animals

Coxiella burnetii is the causative agent of Q fever in humans and animals. This study aimed to determine the frequency of C. burnetii in milk samples of dairy animals (goats, sheep, and cattle) in some selected regions in Iran, where there is no information about prevalence of C. burnetii. In this study, 162 individual milk samples were collected from 43 farms in three provinces (Tehran, Hamadan, and Mazandaran). Real-time PCR was used for the detection of IS1111a element of C. burnetii. In total, 23 of 162 samples (14.2%, 95% confidence interval (CI): 9.65–20.2%) were positive for C. burnetii by real-time PCR. C. burnetii was detected in 10.17% (95% CI: 4.74–20.46) of goat milk samples. In sheep milk samples, 18.6% (95% CI: 9.74–32.62) were positive, and C. burnetii was detected in 15% (95% CI: 8.1–26.11) of cattle milk samples. Molecular evidence of the presence of C. burnetii was seen in milk samples of dairy animals in all the studied regions. These findings demonstrated that C. burnetii infection, especially in raw milk samples, deserves more attention from the health care system and veterinary organization in Iran.

Detection of staphylococcal enterotoxin genes in raw milk samples by real-time PCR

2019 ◽  
Author(s):  
Akýn Yiðin ◽  
Mehmet Demirci ◽  
Serap Kýlýç Altun ◽  
Hikmet Dinç
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Public Health Problem ◽  
Raw Milk ◽  
Staphylococcal Enterotoxin ◽  
Donkey Milk ◽  
Milk Samples ◽  
Conventional Culture ◽  
Food Borne ◽  
Enterotoxin Genes

Presence of significant level of enterotoxigenic S. aureus in raw milk of sheep, goat and donkey may cause serious food borne disease. Many people worldwide, use raw milk in their daily life but data on presence of virulence genes in cow, sheep, goat and especially donkey milk seems to be very limited. For this reason, aim of this study was to determine the presence both S. aureus and nine staphylococcal enterotoxin genes in cow, sheep, goat and donkey milks. A total of 231 raw milk samples were collected from 48 cow, 65 goat, 65 sheep and 53 donkey were collected. To detect presence of S. aureus both conventional culture and real-time PCR were used and to detect nine staphylococcal enterotoxin genes directly in milk, real-time PCR was performed. Conventional culture and real-time PCR results were found to be similar for presence of S. aureus and it was detected in 52 (22.51%) out of 231 raw milk samples. Staphylococcal enterotoxin genes were detected in 27 out of 52 S.aureus positive samples and a total of 62 enterotoxin genes were detected in these samples. However enterotoxin genes could not be detected in two S. aureus positive donkey milk. Hence, real-time PCR proved to be reliable and faster than conventional methods to detect presence of enterotoxigenic S. aureus in milk. Raw milk samples from different animals many contain enterotoxigenic S. aureus. Therefore, one should be careful during raw milk consumption as enterotoxigenic S. aureus in milk may cause dangerous public health problem which need routine screening for this pathogen in milk

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