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.

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.

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>

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

Application of an F57 Sequence-Based Real-Time PCR Assay for Mycobacterium paratuberculosis Detection in Bulk Tank Raw Milk and Slaughtered Healthy Dairy Cows

2006 ◽  
Vol 69 (7) ◽  
pp. 1662-1667 ◽  
Author(s):  
C. BOSSHARD ◽  
R. STEPHAN ◽  
T. TASARA
Keyword(s):  
Detection Limit ◽  
Real Time ◽  
Real Time Pcr ◽  
Raw Milk ◽  
Mesenteric Lymph Nodes ◽  
Pcr Assay ◽  
Milk Samples ◽  
Light Cycler ◽  
Bulk Tank ◽  
Mycobacterium Avium Subsp Paratuberculosis

A light cycler–based real-time PCR assay that targets the F57 sequence was used to collect data on the prevalence of Mycobacterium avium subsp. paratuberculosis (MAP) in 100 bulk tank raw milk samples and in a population of 101 slaughtered dairy cattle. The assay's reproducible detection limit in total genomic DNA templates isolated from 10-ml samples of MAP-spiked raw milk was 100 cells per ml. Similarly, the evaluation of MAP-spiked bovine feces also demonstrated that the assay had a reproducible detection limit of 100 cells if they were contained within 200 mg of fecal sample material. Among the 100 bulk tank milk samples that were tested, we found 3 samples (3%) to be positive for MAP. In the slaughterhouse part of the study, 8.9% (9 of 101) of the cows were positive for MAP DNA in fecal samples, 4.9% (5 of 101) in mesenteric lymph nodes, 0.9% (1 of 101) in ileum tissue, and 3.6% (3 of 84) in milk. Meanwhile, for 2.9% (3 of 101) of the culled cows, MAP DNA was detected in samples of diaphragmatic muscles.

scholarly journals PREVALENCE OF COXIELLA BURNETII INFECTION IN CATTLE, BLACK BENGAL GOATS AND TICKS IN BANGLADESH

2016 ◽  
Vol 14 (1) ◽  
pp. 65-68 ◽  
Author(s):  
A. Chakrabartty ◽  
P. K. Bhattacharjee ◽  
R. R. Sarker ◽  
A. K. M. A. Rahman ◽  
K. Henning ◽  
...  
Keyword(s):  
Real Time ◽  
Coxiella Burnetii ◽  
Real Time Pcr ◽  
Q Fever ◽  
Elisa Test ◽  
Quantitative Real Time Pcr ◽  
Serum Samples ◽  
Test Kit ◽  
History Of ◽  
Cattle Serum

The objectives of this study were to determine the prevalence of Coxiella burnetii infection in domestic ruminants and to detect Coxiella burnetii DNA from ticks and serum samples. A total of 24 ticks, 91 goats and 81 cattle serum samples with the history of abortion and reproductive disorders were collected from the different areas in Bangladesh. The serum samples were tested by CHEKIT Q-Fever Antibody ELISA Test Kit and Coxiella burnetii DNA was detected by multiplex quantitative real- time PCR. The overall prevalence was 7.6% and 6.1% in goats and cattle, respectively. However, none of seropositive samples and tick samples was positive in quantitative real-time PCR.

scholarly journals Real-time PCR Biochip for On-Site Detection of Coxiella Burnetii in Ticks

2021 ◽  
Author(s):  
A-Tai Truong ◽  
Bo-Ram Yun ◽  
Jiyeon Lim ◽  
Subin Min ◽  
Mi-Sun Yoo ◽  
...  
Keyword(s):  
Real Time ◽  
Coxiella Burnetii ◽  
Real Time Pcr ◽  
Q Fever ◽  
Pcr Amplification ◽  
Pcr Detection ◽  
Economic Damage ◽  
Haemaphysalis Longicornis ◽  
Perfect Agreement ◽  
Close Relationship

Abstract Background: Q fever, a zoonosis caused by Coxiella burnetii, has adverse effects on public health. Ticks are the natural reservoirs of C. burnetii and they contribute to the transmission of the pathogen. A tool for rapid, sensitive, and accurate detection of C. burnetii from ticks is important for the prevention of Q fever. Methods: Ultra-rapid real-time PCR (UR-qPCR) as a chip-based real-time PCR system was developed for the detection of C. burnetii from ticks. The UR-qPCR system was established and evaluated for the rapidity, sensitivity, and specificity of C. burnetii detection. Results: C. burnetii was detected using UR-qPCR from 5,644 larval, nymphal, and adult ticks from 408 pools collected from livestock and epidemiologically linked environments in two provinces, Gangwon and Jeju, in Korea. Ticks from three species were identified; Haemaphysalis longicornis accounted for the highest number, present in 333 of 408 pools (81.62%), followed by Haemaphysalis flava in 62 pools (15.19%) and Ixodes nipponensis in 13 pools (3.19%). The rapidity and sensitivity of PCR detection was demonstrated with the sufficient amplification and detection of approximately 56 copies of C. burnetii DNA with only 20 min of PCR amplification. The kappa value for the diagnostic agreement between UR-qPCR and stationary qPCR was in perfect agreement (p = 1). PCR detection and sequencing indicated that C. burnetii was present in 5 of the 408 pools (1.23%), in which four pools contained H. longicornis and one pool contained H. flava. The infection rates of C. burnetii in the tick pools collected from Gangwon and Jeju Provinces were 1.70% and 0.58%, respectively. Phylogenetic analysis indicated a close relationship between the detected C. burnetii and those originated from goats, humans, and ticks in different countries, such as USA, France, Germany, and Serbia. Conclusions: The results of this study could be important for the prevention and control of Q fever in the two provinces. The UR-qPCR with its features of mobility, sensitivity, and rapidity is helpful for constructing early alert systems in the field for C. burnetii in ticks and for alleviating the transmission and economic damage due to Q fever.

scholarly journals Evaluation of the Effectiveness of Q Fever Treatment with Oxytetracycline

2012 ◽  
Vol 56 (4) ◽  
pp. 513-517 ◽  
Keyword(s):  
Real Time ◽  
Coxiella Burnetii ◽  
Real Time Pcr ◽  
Q Fever ◽  
The Third ◽  
Molecular Studies ◽  
Fever Treatment ◽  
Specific Sequences

Abstract The aim of the study was to evaluate the effectiveness of Q fever treatment with oxytetracycline based on the level of Coxiella burnetii antibodies in the sera of infected goats and cows, and excretion of Coxiella burnetii in milk. The study was performed in naturally infected goats and cattle. Forty-six goat sera and 35 cows’ sera were investigated three times before, and twice after treatment with oxytetracycline. The percentage of seronegative goats after treatment (the third examination) was 86.96% while the percentage of seronegative cows after treatment was 52.77%. Moreover, the molecular studies (real-time PCR) of cheese from milk of these animals showed that the specific sequences of DNA for Coxiella burnetii were present despite treatment with oxytetracycline.

scholarly journals The Sensitivity of the Real-time PCR and Nested-PCR for Detection of Coxiella burnetii in Milk Samples

2017 ◽  
Vol 4 (2) ◽  
pp. 11
Author(s):  
Mojtaba Bonyadian ◽  
Hamdollah Moshtaghi ◽  
Hamidreza Kazemeini
Keyword(s):  
Real Time ◽  
Coxiella Burnetii ◽  
Real Time Pcr ◽  
Nested Pcr ◽  
Pcr Assay ◽  
Milk Samples ◽  
Bulk Milk ◽  
Significant Difference ◽  
Pcr Assays ◽  
Nested Pcr Assays

Coxiella burnetii is the causative agent of Q-fever, a widespread zoonosis. In domestic animals infection remains either asymptomatic or presents as infertility or abortion. Clinical presentation in humans can range from mild flu-like illness to acute pneumonia and hepatitis. In humans serology is the gold standard for diagnosis but is inadequate for early case detection, so real-time PCR and nested-PCR assays were developed in this study to measure amounts of C.burnetii shed in milk. Our study was to assess the sensitivity of the realtime PCR and nested-PCR for detection of Coxiella burnetii in bovine bulk milk samples from dairy herds in 3 provinces (Chaharmahal and Bakhtiari , Isfahan and Yazd) of Iran. In the present study, 300 bulk milk samples from 89 dairy cattle herds were tested for C. burnetii using real-time PCR and nested-PCR assays. The animals which their milk samples collected for this study were clinically healthy. In total, 74 of 300 (24.7%) cow milk samples were positive in real-time PCR assay and 26 of 300 (8.7%) samples were positive in nested-PCR assay. McNemar test shows a significant difference in detection of C. burnetii between real-time PCR and nested-PCR. Also the results of this study indicate those clinically healthy dairy cows are important sources of C. burnetii infection in Iran.

Sign in / Sign up

Export Citation Format

Share Document