Detection of Campylobacter from Poultry Carcass Skin Samples at Slaughter in Southern Italy

2009 ◽  
Vol 72 (8) ◽  
pp. 1718-1721 ◽  
Author(s):  
TIZIANA PEPE ◽  
ROSARIA DE DOMINICIS ◽  
GIUSEPPINA ESPOSITO ◽  
IOLE VENTRONE ◽  
PINA M. FRATAMICO ◽  
...  
Keyword(s):  
Southern Italy ◽  
Control Strategies ◽  
Foodborne Pathogen ◽  
Raw Milk ◽  
Human Infection ◽  
Contaminated Water ◽  
Pcr Assay ◽  
Multiplex Pcr Assay ◽  
Human Infections ◽  
Poultry Carcass

Campylobacter is a major foodborne pathogen responsible for acute gastroenteritis characterized by diarrhea that is sometimes bloody, fever, cramps, and vomiting. Campylobacter species are carried in the intestinal tracts of mammals and birds, and sources of human infection include raw milk, contaminated water, direct contact with pets, and foods, particularly poultry. Campylobacter jejuni and C. coli are the species that account for the majority of human infections. The aim of this work was to determine the prevalence of Campylobacter in 190 poultry carcasses sampled at slaughter and to use a multiplex PCR assay to determine if the isolates were C. jejuni or C. coli. C. coli was not isolated, while C. jejuni was recovered from 52 (37.1%) of 140 carcasses for which pools of four sampling sites (neck, cloaca, breast, and back) were examined. In the remaining 50 carcasses, the four sites were analyzed separately, and C. jejuni was recovered from the samples in the following order: neck (n = 20), cloaca (n = 16), breast (n = 14), and back (n = 11). The results are in agreement with those of other studies, which showed that C. jejuni is more commonly associated with poultry than is C. coli. Control strategies for Campylobacter should include interventions to eliminate C. jejuni in poultry at various stages of production and processing, including at slaughter.

scholarly journals Molecular Characterization and Phylogenetic Analysis ofListeria monocytogenesIsolated from Milk and Milk Products in Kaduna, Nigeria

2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
U. B. Usman ◽  
J. K. P. Kwaga ◽  
J. Kabir ◽  
O. S. Olonitola ◽  
S. Radu ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Raw Milk ◽  
Health Concern ◽  
Sample Type ◽  
Public Health Concern ◽  
Pcr Assay ◽  
Milk Products ◽  
Multiplex Pcr Assay ◽  
Food Borne ◽  
Sporadic Cases

In this study,Listeria(L.)monocytogenesisolated from milk and milk products in Kaduna, Nigeria, were subjected to a multiplex PCR assay to identify virulence-associated genes (such asprfA,inlA,hlyA,actA, andiap). Of the 36 isolates, 9 (25%) were positive for one or two virulence-associated genes. Based on the sample type, 6 (16.9%) of the isolates that possessed virulence-associated genes were obtained from raw milk, 2 (3.2%) from “Manshanu,” and 1 (2.8%) from “Kindrimo.” Sequence and phylogenetic analysis based on the 16S rRNA revealed that NigerianL. monocytogenesisolates (NGA 34A, NGA 35A, NGA 41A, and NGA 38A), when compared with referenceL. monocytogenes, were grouped into two distinct clusters, A and B, with sequence (NGA 34A, NGA 35A, and NGA 41A) phylogenetically closer to J1776; N1-011A; R2-502; J1816; and J2-031, whereasL. monocytogenesisolate (NGA 38A) clustered with EDG; J1-220; J1926; J1817; and J2-1091. The separation of the NigerianL. monocytogenesisolates into linage A (responsible for epidemic listeriosis) and lineage B (responsible for sporadic cases of listeriosis) is of public health concern and that local isolates might have potentials for human food borne listeriosis based on the virulence factors so far identified.

Application of a multiplex PCR assay for the detection ofShigella,Escherichia coliand Shiga toxin-producingEsch. coliin milk

2009 ◽  
Vol 76 (2) ◽  
pp. 188-194 ◽  
Author(s):  
Syed Riyaz-Ul-Hassan ◽  
Saima Syed ◽  
Sarojini Johri ◽  
Vijeshwar Verma ◽  
Ghulam Nabi Qazi
Keyword(s):  
Multiplex Pcr ◽  
Shiga Toxin ◽  
Raw Milk ◽  
Pcr Assay ◽  
Milk Samples ◽  
Multiplex Pcr Assay ◽  
The Common ◽  
Different Strains ◽  
Mpcr Assay ◽  
Hygienic Conditions

A multiplex PCR (mPCR) assay using previously known genetic markers ofShigella, Escherichia coliand Shiga-toxicEsch. coliwas standardized.uidAgene was targeted for the common detection ofEsch. coliandShigella, whereasipaHandstx1genes were used as markers for the detection ofShigellaand shiga-toxin producing strains, respectively. The standardized assays detected the target organism specifically and selectively. The mPCR developed by combining all the three reactions generated specific products. The inclusivity and exclusivity tests depicted the precise specificity of the mPCR assay. Results were interpreted on the basis of the pattern of amplicons generated: amplifications of theipaHanduidAgene fragments indicated the presence ofShigellaspp., amplification ofuidAalone revealed the presence ofEsch. coliand additional presence of verotoxin gene amplicon indicated verotoxinogenic nature of the strain. Specific patterns of bands were obtained when different strains ofEsch. coliandShigellaspp. were subjected to this assay. The reactions, individually as well as in the mPCR, could detect approximately 1 cell per 20-μl PCR assay. The protocols were validated by analyzing the coded samples of full fat milk spiked with different pathogens. In naturally contaminated raw milk samples (n=100),Esch. coliwere detected in all samples and verotoxinogenicEsch. coliin 15 samples.Shigella, however, was not detected in any of the samples. When DNA purified from the samples found positive for Shiga-toxicEsch. coliwas directly used as template for the mPCR, the results showed agreement with the enrichment based detection. The mPCR assay, standardized in this study, may be used for rapid microbiological evaluation of milk samples. Further, the study emphasizes the need for better hygienic conditions in dairies.

scholarly journals Identification, Shiga toxin subtypes and prevalence of minor serogroups of Shiga toxin-producing Escherichia coli in feedlot cattle feces

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Kaylen M. Capps ◽  
Justin B. Ludwig ◽  
Pragathi B. Shridhar ◽  
Xiaorong Shi ◽  
Elisabeth Roberts ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Foodborne Pathogens ◽  
Shiga Toxin ◽  
Feedlot Cattle ◽  
Pcr Assay ◽  
Cattle Feces ◽  
Virulence Potential ◽  
Multiplex Pcr Assay ◽  
Human Infections

AbstractShiga toxin-producing Escherichia coli (STEC) are foodborne pathogens that cause illnesses in humans ranging from mild to hemorrhagic enteritis with complications of hemolytic uremic syndrome and even death. Cattle are a major reservoir of STEC, which reside in the hindgut and are shed in the feces, a major source of food and water contaminations. Seven serogroups, O26, O45, O103, O111, O121, O145 and O157, called ‘top-7’, are responsible for the majority of human STEC infections in North America. Additionally, 151 serogroups of E. coli are known to carry Shiga toxin genes (stx). Not much is known about fecal shedding and prevalence and virulence potential of STEC other than the top-7. Our primary objectives were to identify serogroups of STEC strains, other than the top-7, isolated from cattle feces and subtype stx genes to assess their virulence potential. Additional objective was to develop and validate a novel multiplex PCR assay to detect and determine prevalence of six serogroups, O2, O74, O109, O131, O168, and O171, in cattle feces. A total of 351 strains, positive for stx gene and negative for the top-7 serogroups, isolated from feedlot cattle feces were used in the study. Of the 351 strains, 291 belonged to 16 serogroups and 60 could not be serogrouped. Among the 351 strains, 63 (17.9%) carried stx1 gene and 300 (82.1%) carried stx2, including 12 strains positive for both. The majority of the stx1 and stx2 were of stx1a (47/63; 74.6%) and stx2a subtypes (234/300; 78%), respectively, which are often associated with human infections. A novel multiplex PCR assay developed and validated to detect six serogroups, O2, O74, O109, O131, O168, and O171, which accounted for 86.9% of the STEC strains identified, was utilized to determine their prevalence in fecal samples (n = 576) collected from a commercial feedlot. Four serogroups, O2, O109, O168, and O171 were identified as the dominant serogroups prevalent in cattle feces. In conclusion, cattle shed in the feces a number of STEC serogroups, other than the top-7, and the majority of the strains isolated possessed stx2, particularly of the subtype 2a, suggesting their potential risk to cause human infections.

Prevalence and Toxigenic Profiles of Bacillus cereus Isolated from Dried Red Peppers, Rice, and Sunsik in Korea

2009 ◽  
Vol 72 (3) ◽  
pp. 578-582 ◽  
Author(s):  
SUNG KI KIM ◽  
KWANG-PYO KIM ◽  
SUNG SIK JANG ◽  
EUN MI SHIN ◽  
MIN-JEONG KIM ◽  
...  
Keyword(s):  
Bacillus Cereus ◽  
Multiplex Pcr ◽  
Foodborne Pathogen ◽  
Food Poisoning ◽  
Pcr Assay ◽  
Heat Stable ◽  
Multiplex Pcr Assay ◽  
Red Peppers ◽  
Short Time

Bacillus cereus is a spore-forming foodborne pathogen responsible for diarrheal and emetic types of food poisoning. Intoxication is caused by various enterotoxins or by emetic toxin. Because of its widespread presence and the ability to form heat-stable endospores in a relatively short time, B. cereus has been difficult to control. In this study, 21 rice and 36 Sunsik (a mixture of powdered raw grains) samples were examined for the prevalence of B. cereus. A multiplex PCR assay was used to evaluate the distribution of 10 different toxigenicity-related genes among 1,082 B. cereus strains isolated from dried red peppers (919 isolates), rice (98 isolates), and Sunsik (65 isolates). The results suggest that (i) the examined foods were free of the emetic toxin but not free of enterotoxins and (ii) the distribution of enterotoxigenic genes was significantly different among the B. cereus isolates from various sources.

scholarly journals Development of a multiplex PCR assay to detect the major clonal complexes of Streptococcus suis relevant to human infection

2016 ◽  
Vol 65 (5) ◽  
pp. 392-396 ◽  
Author(s):  
Rujirat Hatrongjit ◽  
Anusak Kerdsin ◽  
Marcelo Gottschalk ◽  
Shigeyuki Hamada ◽  
Kazunori Oishi ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Streptococcus Suis ◽  
Human Infection ◽  
Pcr Assay ◽  
Multiplex Pcr Assay

Comparative Evaluation of a Phage Protein Ligand Assay with VIDAS and BAX Methodology for Detection of Escherichia coli O157:H7 Using a Standard Nonproprietary Enrichment Broth

2012 ◽  
Vol 95 (6) ◽  
pp. 1669-1671 ◽  
Author(s):  
Jingzhang Lu ◽  
Mujin Tang ◽  
Huiling Liu ◽  
Lihua Huang ◽  
Zhigang Wan ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Foodborne Pathogen ◽  
Raw Milk ◽  
Alternative Methods ◽  
Escherichia Coli O157 ◽  
Pcr Assay ◽  
Enrichment Broth ◽  
E Coli ◽  
Phage Protein ◽  
Positive Results

Abstract Escherichia coli O157:H7 is a major foodborne pathogen of concern worldwide. This study was conducted to compare the sensitivity and minimum enrichment time for detection of E. coli O157:H7 by the VIDAS ultraperformance E. coli test (VIDAS ECPT UP) with that of two other commercial detection kits, the ELISA-based VIDAS ECO system and the PCR-based BAX® system. Only VIDAS ECPT UP detected all 18 positive results of bacterial suspensions at the level of 104 CFU/mL E. coli O157:H7 and 106 CFU/mL E. coli as background flora, whereas the BAX system PCR assay detected six positive results and VIDAS ECO detected no positive results. A 6 h enrichment at 42°C is enough for detection of all 18 strains in artificial contaminated raw beef meat, raw milk, and raw chicken, and for detection of most of them in soybean sprout and fresh papaya juice with VIDAS ECPT UP, whereas enrichment of more than 8 h was required for detection of the strains with the VIDAS ECO and PCR-BAX systems. These results indicate that the VIDAS ECPT UP is superior to the other two alternative methods when a standard enrichment broth is used that is different from the broths recommended by the manufacturers.

Identification ofClostridium beijerinckii, Cl. butyricum, Cl. sporogenes, Cl. tyrobutyricumisolated from silage, raw milk and hard cheese by a multiplex PCR assay

2012 ◽  
Vol 79 (3) ◽  
pp. 318-323 ◽  
Author(s):  
Paola Cremonesi ◽  
Laura Vanoni ◽  
Tiziana Silvetti ◽  
Stefano Morandi ◽  
Milena Brasca
Keyword(s):  
16S Rrna ◽  
Multiplex Pcr ◽  
Raw Milk ◽  
Most Probable Number ◽  
Pcr Assay ◽  
Promising Alternative ◽  
Probable Number ◽  
Multiplex Pcr Assay ◽  
Microbiological Methods ◽  
Hard Cheese

Late blowing, caused by the outgrowth of clostridial spores present in raw milk and originating from silage, can create considerable product loss, especially in the production of hard and semi-hard cheeses. The conventional method for the isolation ofClostridiumspp. from cheeses with late-blowing symptoms is very complicated and the identification of isolates is problematic. The aim of this work was the development of a multiplex PCR method for the detection of the main dairy-related clostridia such as:Cl. beijerinckii, Cl. butyricum, Cl. sporogenes, Cl. tyrobutyricum.Samples derived from silage, raw milk and hard cheese were analysed by the most probable number (MPN) enumeration. Forty-four bacterial strains isolated from gas positive tubes were used to check the reliability of the multiplex PCR assay. The specificity of the primers was tested by individually analysing each primer pair and the primer pair combined in the multiplex PCR. It was interesting to note that the samples not identified by the multiplex PCR assay were amplified by V2–V3 16S rRNA primer pair and the sequencing revealed the aligned 16S rRNA sequences to bePaenibacillusandBacillusspp. This new molecular assay provides a simple promising alternative to traditional microbiological methods for a rapid, sensitive detection of clostridia in dairy products.

Usefulness of a Multiplex PCR for Detection of Diarrheagenic Escherichia coli in a Diagnostic Microbiology Laboratory Setting

2016 ◽  
Vol 1 (2) ◽  
pp. 38-42 ◽  
Author(s):  
Khairun Nessa ◽  
Dilruba Ahmed ◽  
Johirul Islam ◽  
FM Lutful Kabir ◽  
M Anowar Hossain
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Clinical Laboratory ◽  
Proteinase K ◽  
Microbiology Laboratory ◽  
Pcr Assay ◽  
E Coli ◽  
Diarrheagenic Escherichia Coli ◽  
Multiplex Pcr Assay ◽  
Diagnostic Microbiology

A multiplex PCR assay was evaluated for diagnosis of diarrheagenic Escherichia coli in stool samples of patients with diarrhoea submitted to a diagnostic microbiology laboratory. Two procedures of DNA template preparationproteinase K buffer method and the boiling method were evaluated to examine isolates of E. coli from 150 selected diarrhoeal cases. By proteinase K buffer method, 119 strains (79.3%) of E. coli were characterized to various categories by their genes that included 55.5% enteroaggregative E. coli (EAEC), 18.5% enterotoxigenic E. coli (ETEC), 1.7% enteropathogenic E. coli (EPEC), and 0.8% Shiga toxin-producing E. coli (STEC). Although boiling method was less time consuming (<24 hrs) and less costly (<8.0 US $/ per test) but was less efficient in typing E. coli compared to proteinase K method (41.3% vs. 79.3% ; p<0.001). The sensitivity and specificity of boiling method compared to proteinase K method was 48.7% and 87.1% while the positive and negative predictive value was 93.5% and 30.7%, respectively. The majority of pathogenic E. coli were detected in children (78.0%) under five years age with 53.3% under one year, and 68.7% of the children were male. Children under 5 years age were frequently infected with EAEC (71.6%) compared to ETEC (24.3%), EPEC (2.7%) and STEC (1.4%). The multiplex PCR assay could be effectively used as a rapid diagnostic tool for characterization of diarrheagenic E. coli using a single reaction tube in the clinical laboratory setting.Bangladesh J Med Microbiol 2007; 01 (02): 38-42

scholarly journals The Interplay between Campylobacter and the Caecal Microbial Community of Commercial Broiler Chickens over Time

2021 ◽  
Vol 9 (2) ◽  
pp. 221
Author(s):  
Ilaria Patuzzi ◽  
Massimiliano Orsini ◽  
Veronica Cibin ◽  
Sara Petrin ◽  
Eleonora Mastrorilli ◽  
...  
Keyword(s):  
Campylobacter Jejuni ◽  
Broiler Chickens ◽  
Control Strategies ◽  
Chicken Meat ◽  
Control Measures ◽  
Microbial Network ◽  
Gut Colonization ◽  
Commercial Broiler ◽  
Zoonotic Bacteria ◽  
Human Infections

Campylobacter is the most frequent foodborne zoonotic bacteria worldwide, with chicken meat being overwhelmingly the most important reservoir for human infections. Control measures implemented at the farm level (i.e., biosecurity or vaccination), which have been successfully applied to limit other pathogens, such as Salmonella, have not been effective in reducing Campylobacter occurrence. Thus, new approaches are needed to fully understand the ecological interactions of Campylobacter with host animals to effectively comprehend its epidemiology. The objective of this study was to analyse longitudinally the gut microbiota composition of Campylobacter-infected and non-infected farms to identify any difference that could potentially be indicative of gut colonization by Campylobacter spp. Differences in the colonization rate and timing were observed at the farms that became positive for Campylobacter jejuni over the investigated time points, even though in positive tests, the occurrence of Campylobacter jejuni gut colonization was not observed before the second week of the life of the birds. Significant differences were observed in the abundances of specific bacterial taxa between the microbiota of individuals belonging to farms that became Campylobacter positive during the study and those who remained negative with particular reference to Bacteroidales and Clostridiales, respectively. Moreover, Campylobacter colonization dramatically influenced the microbiota richness, although to a different extent depending on the infection timing. Finally, a key role of Faecalibacterium and Lactobacillus genera on the Campylobacter microbial network was observed. Understanding the ecology of the Campylobacter interaction with host microbiota during infection could support novel approaches for broiler microbial barrier restoration. Therefore, evidence obtained through this study can be used to identify options to reduce the incidence of infection at a primary production level based on the targeted influence of the intestinal microbiota, thus helping develop new control strategies in order to mitigate the risk of human exposure to Campylobacter by chicken meat consumption.

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