scholarly journals Detection ofYersinia enterocoliticain Retail Chicken Meat, Mashhad, Iran

2018 ◽  
Vol 2018 ◽  
pp. 1-4 ◽  
Author(s):  
Khadigeh Sirghani ◽  
Tayebeh Zeinali ◽  
Abdollah Jamshidi
Keyword(s):  
Yersinia Enterocolitica ◽  
Health Hazard ◽  
Culture Method ◽  
Good Alternative ◽  
Food Samples ◽  
Selective Medium ◽  
Chicken Meat ◽  
Poultry Meat ◽  
Rrna Gene ◽  
Pcr Assay

Poultry meat is one of the most important sources of infection ofYersiniaspp. for humans. The aim of the present study was to evaluate the incidence ofYersinia enterocoliticain chicken meat by using culture method on selective medium and confirmation by PCR assay. Also, biochemical methods were used for biotyping. A total of 100 chicken thigh meat samples were collected randomly from retail outlets in Mashhad, Iran. Samples were enriched in Peptone-Sorbitol-Bile (PSB) broth and then cultured on Cefsulodin-Irgasan-Novobiocin (CIN) agar containing antibiotics supplement. The DNA was extracted from suspected colonies ofYersiniaspp. and then PCR test using specific primers for 16S rRNA gene ofYersinia enterocoliticawas performed. In this study, 30% of chicken meat was contaminated withYersiniaspp. by culture method and 25% of chicken meat was contaminated withYersinia enterocolitica. Biotyping of isolated colonies showed that all of the isolates belonged to biotype 1A. Culture and detection ofYersiniaspp. from food samples traditionally take 4 days. Due to high accuracy and speed of PCR assay, it is a good alternative method for microbiological techniques. In conclusion, poultry meat can act as a source ofY. enterocoliticaand could be considered as a public health hazard.

An Improved PCR Primer Pair Based on 16S rDNA for the Specific Detection of Salmonella Serovars in Food Samples

2004 ◽  
Vol 67 (7) ◽  
pp. 1335-1343 ◽  
Author(s):  
CHIEN-KU LIN ◽  
CHO-LIEN HUNG ◽  
SHU-CHEN HSU ◽  
CHENG-CHIH TSAI ◽  
HAU-YANG TSEN
Keyword(s):  
Sequence Data ◽  
Complete Sequence ◽  
Food Samples ◽  
Chicken Meat ◽  
Rrna Gene ◽  
Whole Milk ◽  
Sporadic Cases ◽  
Salmonella Serovars ◽  
Primer Annealing ◽  
Pcr Primer

Salmonella serovars are some of the major bacterial pathogens that can cause sporadic cases and outbreaks of foodborne illness. Based on the sequence data in the V3 region of the 16S rRNA gene, two PCR primer pairs have been designed for the detection of all serovars of Salmonella. However, none of these primers were specific for Salmonella because complete sequence homology with certain non-Salmonella strains has been found within each of them. Thus, the specificities of these two primer pairs could not rely on only one of the two primers. In this study, we modified our previous 16SFI primer by extending one base at the 5′ end and three bases at the 3′ end. The modified primer, 16S-Sal, was designed with one or more mismatched bases near the 3′ end of the primer annealing to the corresponding sequences of non-Salmonella strains. Such modification eliminates interference from Citrobacter freundii and Enterobacter cloacae as occurs with the 16SFI primer. When 16S-Sal and a degenerate primer, 16S-CCR, were used as a primer pair, detection specificity of Salmonella serovars was achieved. Because this primer pair was used for PCR detection of the salmonellae in food samples, such as whole milk and chicken meat, as low as 1 to 9 CFU/g (ml) of the food sample could be detected when a 8-h preculture step was performed prior to the PCR. For chicken meat, the endogenous microflora did not interfere with the PCR results.

Specific PCR Detection of Arcobacter butzleri, Arcobacter cryaerophilus, Arcobacter skirrowii, and Arcobacter cibarius in Chicken Meat

2009 ◽  
Vol 72 (7) ◽  
pp. 1491-1495 ◽  
Author(s):  
DANIELA PENTIMALLI ◽  
NICOLETTE PEGELS ◽  
TERESA GARCÍA ◽  
ROSARIO MARTÍN ◽  
ISABEL GONZÁLEZ
Keyword(s):  
Pcr Amplification ◽  
Chicken Meat ◽  
Pcr Analysis ◽  
Rrna Gene ◽  
Pcr Assay ◽  
Gyra Gene ◽  
Specific Primers ◽  
Specific Pcr ◽  
Arcobacter Butzleri ◽  
Species Specific

An enrichment PCR assay using species-specific primers was developed for the detection of Arcobacter butzleri, Arcobacter cryaerophilus, Arcobacter skirrowii, and Arcobacter cibarius in chicken meat. Primers for A. cryaerophilus, A. skirrowii, and A. cibarius were designed based on the gyrA gene to amplify nucleic acid fragments of 212, 257, and 145 bp, respectively. The A. butzleri–specific primers were designed flanking a 203-bp DNA fragment in the 16S rRNA gene. The specificity of the four primer pairs was assessed by PCR analysis of DNA from a panel of Arcobacter species, related Campylobacter, Helicobacter species, and other food bacteria. The applicability of the method was then validated by testing 42 fresh retail-purchased chicken samples in the PCR assay. An 18-h selective preenrichment step followed by PCR amplification with the four Arcobacter primer sets revealed the presence of Arcobacter spp. in 85.7% of the retail chicken samples analyzed. A. butzleri was the only species present in 50% of the samples, and 35.7% of the samples were positive for both A. butzleri and A. cryaerophilus. A. skirrowii and A. cibarius were not detected in any of the chicken samples analyzed. The enrichment PCR assay developed is a specific and rapid alternative for the survey of Arcobacter contamination in meat.

scholarly journals Virulence genotyping and antimicrobial resistance profiles of Yersinia enterocolitica isolated from meat and meat products in Egypt

2020 ◽  
Author(s):  
G. A. Younis ◽  
R. M. Elkenany ◽  
H. A. Dowidar
Keyword(s):  
Antimicrobial Resistance ◽  
Yersinia Enterocolitica ◽  
Genetic Relatedness ◽  
Meat Product ◽  
Meat Products ◽  
High Sensitivity ◽  
Resistance Rate ◽  
Chicken Meat ◽  
Nucleotide Sequence Analysis ◽  
Rrna Gene

Abstract Pathogenic Yersinia enterocolitica (Y. enterocolitica) is one of the food-borne entero-pathogen responsible for yersiniosis in humans. The purpose of this research was to survey the prevalence, virulence-associated genes, and antimicrobial resistance of Y. enterocolitica isolated from meat and meat product samples in Egypt. Forty-one (5.9%) out of 700- samples of chicken meat, beef, ground beef, and sausage were positive Y. enterocolitica with a high prevalence in chicken meat (12%). Five virulence genes (ail, inv, ystA, ystB, and yadA) were characterized among 41 Y. enterocolitica isolates with variable frequencies. Among the strains tested, the ystB gene was detected with a high percentage (78.1%), followed by inv gene (70.7%), ail gene (14.6%), ystA gene (12.2%), and yadA gene (2.4%). A high resistance rate was estimated to amoxicillin-clavulanic acid (100%), followed by cefazolin (95%), ampicillin (65.9%), and doxycycline (51.2%), whilst a high sensitivity rate was observed to gentamicin and ciprofloxacin (97.6% each). Interestingly, the multidrug resistance was specified in the 70.7% of strains and showing 13 resistance patterns. Based on nucleotide sequence analysis of the 16s rRNA gene, the phylogenetic tree showed the genetic relatedness amongst Y. enterocolitica isolates. These findings highlighted the emergence of virulent and multidrug-resistant pathogenic Y. entrocolitica in retailed meat and meat products in Egypt.

scholarly journals DETECTION OF MULTI–DRUG RESISTANT SALMONELLA FROM MILK AND MEAT IN BANGLADESH

2018 ◽  
Vol 16 (1) ◽  
pp. 115-120 ◽  
Author(s):  
M. A. Rahman ◽  
A. K. M. A. Rahman ◽  
M. A. Islam ◽  
M. M. Alam
Keyword(s):  
Health Hazard ◽  
Dairy Farm ◽  
Food Samples ◽  
Chicken Meat ◽  
Multi Drug Resistance ◽  
Drug Resistant ◽  
Salmonella Spp ◽  
Gram Staining ◽  
Hygienic Measures ◽  
Agar Media

This study was conducted to investigate the prevalence of Salmonella spp. in milk, chicken meat and beef and to determine the multi-drug resistance (MDR) profile of Salmonella spp. in Mymensingh and Gazipur districts, Bangladesh. A total of 169 samples of milk (n=108), chicken meat (n=51) and beef (n=10) were collected from Bangladesh Agricultural University (BAU) dairy farm, American dairy farm, Gazipur and different  small dairy farms of municipal area during July 2016 to June 2017. Salmonella spp. were isolated on various selective agar media such as: Salmonella-Shigella (SS) agar, Xylose-Lysine Deoxycholate (XLD) agar, Eosine-Methylene Blue (EMB) agar. Identification of Salmonella spp. was done by colony characteristics, Gram staining, biochemical test and Polymerase Chain Reaction (PCR). Multi-drug resistant Salmonella spp. was detected by disc diffusion test using 10 commonly used antibiotics. The overall prevalence of Salmonella spp. in all food samples was 21.89%. A total of 29 (56.86%) chicken meat, 02 (1.85%) milk, and 06 (60%) beef samples were Salmonella spp. positive. Antibiogram study showed that an overall 89.19% of Salmonella spp. was found multi-drug resistant. Specifically 100%, 66.67% and 93.10% of the Salmonella spp. isolates originated from milk, beef and chicken meat respectively were multi-drug resistant. The result of this study suggests that MDR Salmonella spp. is prevalent in the milk and meat which might cause public health hazard if proper hygienic measures are not undertaken at farm and marketing level.

Simple, Rapid, and Reliable Detection of Escherichia coli O26 Using Immunochromatography

2013 ◽  
Vol 76 (5) ◽  
pp. 748-754 ◽  
Author(s):  
TARO YONEKITA ◽  
TATSUYA FUJIMURA ◽  
NAOKI MORISHITA ◽  
TAKASHI MATSUMOTO ◽  
FUMIKI MORIMATSU
Keyword(s):  
Escherichia Coli ◽  
Diarrheal Disease ◽  
Culture Method ◽  
Food Samples ◽  
Pcr Assay ◽  
Beef Liver ◽  
E Coli ◽  
Pure Cultures ◽  
Alfalfa Sprout ◽  
Meat Samples

Shiga toxin–producing Escherichia coli (STEC) O26 has been increasingly associated with diarrheal disease all over the world. We developed an immunochromatographic (IC) strip for the rapid detection of E. coli O26 in food samples. To determine the specificity of the IC strip, pure cultures of 67 E. coli and 22 non–E. coli strains were tested with the IC strip. The IC strip could detect all (18 of 18) E. coli O26 strains tested and did not react with strains of any other E. coli serogroup or non–E. coli strains tested (0 of 71). The minimum detection limits for E. coli O26 were 2.2 ×103 to 1.0 ×105 CFU/ml. To evaluate the ability of the IC strip to detect E. coli O26 in food, 25-g food samples (ground beef, beef liver, ground chicken, alfalfa sprout, radish sprout, spinach, natural cheese, and apple juice) were spiked with E. coli O26. The IC strip was able to detect E. coli O26 at very low levels (approximately 1 CFU/25 g of food samples) after an 18-h enrichment, and the IC strip results were in 100% agreement with the results of the culture method and PCR assay. When 115 meat samples purchased from supermarkets were tested, 5 were positive for E. coli O26 with the IC strip; these results were confirmed with a PCR assay. These results suggest that the IC strip is a useful tool for detecting E. coli O26 in food samples.

Development of a Quantitative Real-Time PCR Method To Enumerate Total Bacterial Counts in Ready-to-Eat Fruits and Vegetables

2006 ◽  
Vol 69 (10) ◽  
pp. 2504-2508 ◽  
Author(s):  
HAJIME TAKAHASHI ◽  
HIROTAKA KONUMA ◽  
YUKIKO HARA-KUDO
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Fruits And Vegetables ◽  
Bacterial Species ◽  
Culture Method ◽  
Plate Count ◽  
Rrna Gene ◽  
Pcr Assay ◽  
Cycle Number ◽  
Wide Range

A newly developed real-time PCR assay rapidly quantifies the total bacterial numbers in contaminated ready-to-eat vegetables and fruits compared with the standard plate count method. Primers targeting the rpoB gene, which encodes for the β subunit of the bacterial RNA polymerase and which is common to most bacterial species, was used instead of the 16S rRNA gene, which has multiple copies and varies among bacterial species. A primer pair specific for rpoB was confirmed to amplify rpoB in a wide range of bacterial species after we assessed 49 strains isolated from five kinds of fruits and vegetables. We purchased fruits and vegetables from retail shops and enumerated the bacteria associated with them by use of real-time PCR and compared this to the number found by the culture method. We found a high correlation between the threshold PCR cycle number when compared with the plate count culture number. The real-time PCR assay developed in this study can enumerate the dominant bacterial species in ready-to-eat fruits and vegetables.

scholarly journals Isolation and Identification of Listeria Monocytogenes in Bangalore City from Various Food Samples

2021 ◽  
Vol 14 (4) ◽  
pp. 2271-2276
Author(s):  
Vedavati Goudar ◽  
Kanthesh B M ◽  
Nagalambika Prasad
Keyword(s):  
Listeria Monocytogenes ◽  
Ice Cream ◽  
Raw Milk ◽  
Food Samples ◽  
Selective Medium ◽  
Chicken Meat ◽  
Isolation And Identification ◽  
Fresh Milk ◽  
Biochemical Identification ◽  
Raw Fish

The current research emphasis on the isolation and differentiation of Listeria monocytogenes from different food samples most frequently infected with Listeriosis outbreaks. Crude chicken meat, raw milk, pasteurized cheese, ice cream and raw fish are samples from the city of Bangalore. The selective medium mainly used for the isolation of Listeria is oxford agar. Using isolated L. monocytogenes from food samples, morphologic and biochemical identification was carried out. 2 samples (fresh milk and Ice cream) were positive out of 5 samples; 3 samples (raw chicken meat, raw fish, and pasteurized cheese) were negative. The results conferred during this study indicate the contamination of Ice- cream and Raw Milk samples with L. monocytogenes.

scholarly journals Development of IMBs-qPCR Detection Method for Yersinia enterocolitica Based on the foxA Gene

2021 ◽  
Author(s):  
Jingxuan Shi ◽  
Heng Chi ◽  
Aiping Cao ◽  
Yinna Song ◽  
Min Zhu ◽  
...  
Keyword(s):  
Yersinia Enterocolitica ◽  
Culture Method ◽  
Food Samples ◽  
Pcr Detection ◽  
Immunomagnetic Beads ◽  
Foxa Gene ◽  
Qpcr Method ◽  
Virulence Factor Gene ◽  
Food Safety Risk ◽  
Pork Samples

Abstract Yersinia enterocolitica is an important zoonotic pathogen, which seriously endangers food safety risk. In this study, the recombinant outer membrane protein OmpF and its antibody were prepared and coupled with immunomagnetic beads (IMBs) to capture Y. enterocolitica in food samples, combining the quantitative PCR detection with primers of virulence factor gene fox A for Yersinia enterocolitica contamination. The results showed that the capture efficiency of approximately 80% using anti-OmpF antibody-immunomagnetic beads and linearly dependent capture under 10 1 -10 5 CFU/mL Y. enterocolitica . compare with less than 10% capture of other bacteria. The detection limit of 64 CFU/mL was obtained by based on fox A gene PCR detection combined with capture of the anti-OmpF antibody-immunomagnetic beads to detect Yersinia enterocolitica in artificially contaminated milk and pork samples. Comparing with the culture method, the developed IMBs-qPCR method has higher consistency, less time consuming, which providing an effective alternative method for rapid detection of Y. enterocolitica in food.

Evaluation of a 5′ -Nuclease (TaqMan) Assay with the Thin Agar Layer Oxyrase Method for the Detection of Yersinia enterocolitica in Ground Pork Samples†

2004 ◽  
Vol 67 (2) ◽  
pp. 271-277 ◽  
Author(s):  
V. C. H. WU ◽  
D. Y. C. FUNG ◽  
R. D. OBERST
Keyword(s):  
Detection Limit ◽  
Yersinia Enterocolitica ◽  
Food Samples ◽  
Selective Medium ◽  
Taqman Assay ◽  
Ground Pork ◽  
Sensitivity Studies ◽  
Agar Layer ◽  
Pork Samples ◽  
Nuclease Assay

A 5′-nuclease (TaqMan) assay was evaluated for its capability to recover and detect stressed Yersinia enterocolitica. Sensitivity studies of a 5′-nuclease assay for detecting Y. enterocolitica O:8 in a pure culture system and spiked ground pork samples demonstrated that the assay has reliable sensitivity with a detection limit of 3 to 4 log CFU/ml or CFU/g. The PCR 5′-nuclease (TaqMan) assay was evaluated with the Thin Agar Layer Oxyrase method (TALO, overlaying 14 ml of Trypticase soy agar with a 1:30 dilution of “Oxyrase® for Agar” onto a prepoured pathogen-specific, selective medium), and it was compared against the selective medium cefsulodin-irgasan-novobiocin (CIN) for recovering and detecting Y. enterocolitica from inoculated nonfrozen and frozen (−15°C, 2 days) ground pork samples. The TALO method showed more sensitivity (detection limit, 2 log CFU/ml), and it has greater recovery capability (0.5 to 1 log CFU/ml) than CIN (P < 0.05). The 5′-nuclease assay provided rapid detection processing (5 versus 24 h after an 18-h enrichment). The sensitivity per PCR was calculated to as low as 0 to 1 log CFU per PCR reaction; however, in the method's current developmental stage, target pathogens should be enriched to 3 to 4 log CFU/ml or CFU/g to show consistent results. In a survey of 100 ground pork samples using TALO, CIN, and PCR methods, no Y. enterocolitica was recovered. A combined cultivation and an automated PCR TaqMan could be used as a presumptive screening test for detecting Y. enterocolitica in food samples.

scholarly journals Rapid Identification of Yersinia enterocolitica in Blood by the 5′ Nuclease PCR Assay

2000 ◽  
Vol 38 (5) ◽  
pp. 1953-1958 ◽  
Author(s):  
Keya Sen
Keyword(s):  
Yersinia Enterocolitica ◽  
Rapid Identification ◽  
Taqman Assay ◽  
Rrna Gene ◽  
Chromosomal Dna ◽  
Pcr Assay ◽  
Clinical Sepsis ◽  
Taqman Pcr ◽  
Stored Blood ◽  
The 16S Rrna Gene

Yersinia enterocolitica accounts for 50% of the clinical sepsis episodes caused by the transfusion of contaminated red blood cells. A 5′ nuclease TaqMan PCR assay was developed to detectY. enterocolitica in blood. Primers and a probe based on the nucleotide sequence of the 16S rRNA gene from Y. enterocolitica were designed. Whole-blood samples were spiked with various numbers of Y. enterocolitica cells, and total chromosomal DNA was extracted. When the TaqMan PCR assay was performed, as few as six bacteria spiked in 200 μl of blood could be detected. The assay was specific and did not detect other Yersiniaspecies. The TaqMan assay is easy to perform, takes 2 h, and has the potential for use in the rapid detection of Y. enterocolitica contamination in stored blood units.

Sign in / Sign up

Export Citation Format

Share Document