Molecular detection assay of five Salmonella serotypes of public interest: Typhimurium, Enteritidis, Newport, Heidelberg, and Hadar.

PubMed

Bugarel, M; Tudor, A; Loneragan, G H; Nightingale, K K

2017-03-01

Foodborne illnesses due to Salmonella represent an important public-health concern worldwide. In the United States, a majority of Salmonella infections are associated with a small number of serotypes. Furthermore, some serotypes that are overrepresented among human disease are also associated with multi-drug resistance phenotypes. Rapid detection of serotypes of public-health concern might help reduce the burden of salmonellosis cases and limit exposure to multi-drug resistant Salmonella. We developed a two-step real-time PCR-based rapid method for the identification and detection of five Salmonella serotypes that are either overrepresented in human disease or frequently associated with multi-drug resistance, including serotypes Enteritidis, Typhimurium, Newport, Hadar, and Heidelberg. Two sets of four markers were developed to detect and differentiate the five serotypes. The first set of markers was developed as a screening step to detect the five serotypes; whereas, the second set was used to further distinguish serotypes Heidelberg, Newport and Hadar. The utilization of these markers on a two-step investigation strategy provides a diagnostic specificity of 97% for the detection of Typhimurium, Enteritidis, Heidelberg, Infantis, Newport and Hadar. The diagnostic sensitivity of the detection makers is >96%. The availability of this two-step rapid method will facilitate specific detection of Salmonella serotypes that contribute to a significant proportion of human disease and carry antimicrobial resistance. Published by Elsevier B.V.

Comparative evaluation of two rapid Salmonella-IgM tests and blood culture in the diagnosis of enteric fever.

PubMed

Prasad, K J; Oberoi, J K; Goel, N; Wattal, C

2015-01-01

Enteric fever is a major public health problem in developing countries like India. An early and accurate diagnosis is necessary for a prompt and effective treatment. We have evaluated the diagnostic accuracy of two Rapid Salmonella-IgM tests (Typhidot-IgM and Enteroscreen-IgM) as compared to blood culture in rapid and early diagnosis of enteric fever. A total of 2,699 patients' serum samples were tested by Rapid Salmonella-IgM tests and blood culture. Patients were divided into two groups. Test group - patients with enteric fever and blood culture positives for Salmonella Typhi; and three types of Controls, i.e. patients with non-enteric fever illnesses, normal healthy controls and patients positive for S. Paratyphi- A. In addition to this we have also evaluated the significance of positive Salmonella-IgM tests among blood culture-negative cases. The overall sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of the Typhidot-IgM test and Enteroscreen-IgM test considering blood culture as gold standard were 97.29% and 88.13%, 97.40% and 87.83%, 98.18% and 92.03%, 96.15% and 82.27%, respectively. Typhidot-IgM test was found to be significantly more sensitive and specific as compared to Enteroscreen-IgM. Among blood culture-negative patients, Rapid Salmonella-IgM tests detected 72.25% additional cases of enteric fever. Although the Rapid Salmonella-IgM tests are meant to diagnose S. Typhi only, but these tests detect S. Paratyphi- A also. Thirty-eight patients who were blood culture-positive for S. Paratyphi- A were also positive by Rapid Salmonella-IgM tests. Rapid Salmonella-IgM tests offer an advantage of increased sensitivity, rapidity, early diagnosis and simplicity over blood culture.

Electrochemical characterization of an immunosensor for Salmonella spp. detection

USDA-ARS?s Scientific Manuscript database

Immunosensors represent a rapid alternative method for diagnosing Salmonella contamination. The objective of this study was to develop and evaluate the performance of an electrochemical immunosensor for the detection of Salmonella spp., the most common foodborne pathogen worldwide. In the immunosens...

Salmonella rarely detected in Mississippi coastal waters and sediment.

PubMed

Carr, M R; Wang, S Y; McLean, T I; Flood, C J; Ellender, R D

2010-12-01

Standards for the rapid detection of individual pathogens from environmental samples have not been developed, but in their absence, the use of molecular-based detection methods coupled with traditional microbiology techniques allows for rapid and accurate pathogen detection from environmental waters and sediment. The aim of this research was to combine the use of enrichment with PCR for detection of Salmonella in Mississippi coastal waters and sediment and observe if that presence correlated with levels of enterococci and climatological variables. Salmonella were primarily found in samples that underwent nutrient enrichment and were present more frequently in freshwater than marine waters. Salmonella were detected infrequently in marine and freshwater sediments. There was a significant positive correlation between the presence of detectable Salmonella and the average enterococcal count. An inverse relationship, however, was observed between the frequency of detection and the levels of salinity, turbidity and sunlight exposure. Results from this study indicated the presence of Salmonella in Mississippi coastal waters, and sediments are very low with significant differences between freshwater and marine environments. Using pathogenic and novel nonpathogenic molecular markers, Salmonella do not appear to be a significant pathogenic genus along the Mississippi Coast. © 2010 The Authors. Journal of Applied Microbiology © 2010 The Society for Applied Microbiology.

Surface enhanced raman spectroscopy technique in rapid detection of live and dead salmonella cells

USDA-ARS?s Scientific Manuscript database

Many research proved that Surface Enhanced Raman Spectroscopy (SERS) can detect pathogens rapidly and accurately. In this study, a silver metal substrate was used for the selected common food pathogen Salmonella typhimurium bacteria. Nano silver rods were deposited on a thin titanium coating over t...

Interdigitated microelectrode based impedance biosensor for detection of salmonella enteritidis in food samples

NASA Astrophysics Data System (ADS)

Kim, G.; Morgan, M.; Hahm, B. K.; Bhunia, A.; Mun, J. H.; Om, A. S.

2008-03-01

Salmonella enteritidis outbreaks continue to occur, and S. enteritidis-related outbreaks from various food sources have increased public awareness of this pathogen. Conventional methods for pathogens detection and identification are labor-intensive and take days to complete. Some immunological rapid assays are developed, but these assays still require prolonged enrichment steps. Recently developed biosensors have shown great potential for the rapid detection of foodborne pathogens. To develop the biosensor, an interdigitated microelectrode (IME) was fabricated by using semiconductor fabrication process. Anti-Salmonella antibodies were immobilized based on avidin-biotin binding on the surface of the IME to form an active sensing layer. To increase the sensitivity of the sensor, three types of sensors that have different electrode gap sizes (2 μm, 5 μm, 10 μm) were fabricated and tested. The impedimetric biosensor could detect 103 CFU/mL of Salmonella in pork meat extract with an incubation time of 5 minutes. This method may provide a simple, rapid and sensitive method to detect foodborne pathogens.

Immunochromatographic strip assay for the rapid and sensitive detection of Salmonella Typhimurium in artificially contaminated tomato samples.

PubMed

Shukla, Shruti; Leem, Hyerim; Lee, Jong-Suk; Kim, Myunghee

2014-06-01

This study was designed to confirm the applicability of a liposome-based immunochromatographic assay for the rapid detection of Salmonella enterica subsp. enterica serovar Typhimurium (Salmonella Typhimurium) in artificially contaminated tomato samples. To determine the detection limit and pre-enrichment incubation time (10, 12, and 18 h pre-enrichment in 1% buffered peptone water), the tests were performed with different cell numbers of Salmonella Typhimurium (3 × 10(0), 3 × 10(1), 3 × 10(2), and 3 × 10(3) CFU·mL(-1)) inoculated into 25 g of crushed tomato samples. The assay was able to detect as few as 30 Salmonella Typhimurium cells per 25 g of tomato samples (1.2 cells·g(-1)) after 12 h pre-enrichment incubation. Moreover, when the developed assay was compared with traditional morphological and biochemical culture-based methods as well as colloidal gold nanoparticle-based commercial test strips, the developed assay yielded positive results for the detection of Salmonella Typhimurium within a shorter period time. These findings confirm that the developed assay may have practical application for the sensitive detection of Salmonella Typhimurium in various food samples, including raw vegetables, with a relatively low detection limit and shorter analysis time.

Classification of Salmonella Enterica serotypes with selective bands using visible/NIR hyperspectral imaging

USDA-ARS?s Scientific Manuscript database

Optical detection of foodborne bacteria such as Salmonella classifies bacteria by analyzing spectral data, and has potential for rapid detection. In this experiment hyperspectral microscopy is explored as a means for classifying five Salmonella serotypes. Initially, the microscope collects 89 spect...

Rapid and early detection of salmonella serotypes with hyperspectral microscope and multivariate data analysis

USDA-ARS?s Scientific Manuscript database

This study was designed to evaluate hyperspectral microscope images for early and rapid detection of Salmonella serotypes: S. Enteritidis, S. Heidelberg, S. Infantis, S. Kentucky, and S. Typhimurium at incubation times of 6, 8, 10, 12, and 24 hours. Images were collected by an acousto-optical tunab...

Label-Free Detection and Serotyping of Salmonellae by Surface Enhanced Raman Spectroscopy with Immunomagnetic Separation

USDA-ARS?s Scientific Manuscript database

Salmonella spp. are one of the leading causes of foodborne outbreaks in the United States and globally. Current detection and characterization techniques for Salmonella are time consuming and rapid methods could greatly benefit outbreak investigation, new case prevention and disease treatment. In th...

Simultaneous detection and serotyping of Salmonellae by immunomagnetic separation and label-free surface enhanced Raman spectroscopy

USDA-ARS?s Scientific Manuscript database

Salmonella spp. are one of the leading causes of foodborne outbreaks in the United States and globally. Current detection and characterization techniques for Salmonellae are time consuming and costly, and rapid methods could greatly benefit outbreak investigation, new case prevention and disease tre...

The development of methods for the detection of Salmonella in chickens by a combination of immunomagnetic separation and PCRs.

PubMed

Dai, Fengying; Zhang, Miao; Xu, Dixin; Yang, Yin; Wang, Jiaxiao; Li, Mingzhen; Du, Meihong

2017-11-01

Micro- and nanoimmunomagnetic beads (MIMBs and NIMBs) used for immunomagnetic separation (IMS) with PCR were studied for the rapid detection of Salmonella. The capture efficiency of the two different IMBs was evaluated by a conventional plate counting method, and the binding pattern was studied using scanning electron microscopy. The specificity of the IMBs was tested with Salmonella, Shigella flexneri, enterohemorrhagic Escherichia coli O157:H7, and Listeria monocytogenes. By comparing the pre-enrichment IMS and the IMS enrichment steps with a 5.5-H enrichment time, this study developed a rapid and sensitive method for the detection of Salmonella in chicken. The method was implemented by IMS enrichment and PCR with MIMBs and NIMBs, with a total analysis time of 8 H. We showed that the method was sensitive based on NIMBs with a detection limit of 10° CFU for Salmonella in 25 g of chicken. © 2016 International Union of Biochemistry and Molecular Biology, Inc.

Relative effectiveness of the Bacteriological Analytical Manual method for the recovery of Salmonella from whole cantaloupes and cantaloupe rinses with selected preenrichment media and rapid methods.

PubMed

Hammack, Thomas S; Valentin-Bon, Iris E; Jacobson, Andrew P; Andrews, Wallace H

2004-05-01

Soak and rinse methods were compared for the recovery of Salmonella from whole cantaloupes. Cantaloupes were surface inoculated with Salmonella cell suspensions and stored for 4 days at 2 to 6 degrees C. Cantaloupes were placed in sterile plastic bags with a nonselective preenrichment broth at a 1:1.5 cantaloupe weight-to-broth volume ratio. The cantaloupe broths were shaken for 5 min at 100 rpm after which 25-ml aliquots (rinse) were removed from the bags. The 25-ml rinses were preenriched in 225-ml portions of the same uninoculated broth type at 35 degrees C for 24 h (rinse method). The remaining cantaloupe broths were incubated at 35 degrees C for 24 h (soak method). The preenrichment broths used were buffered peptone water (BPW), modified BPW, lactose (LAC) broth, and Universal Preenrichment (UP) broth. The Bacteriological Analytical Manual Salmonella culture method was compared with the following rapid methods: the TECRA Unique Salmonella method, the VIDAS ICS/SLM method, and the VIDAS SLM method. The soak method detected significantly more Salmonella-positive cantaloupes (P < 0.05) than did the rinse method: 367 Salmonella-positive cantaloupes of 540 test cantaloupes by the soak method and 24 Salmonella-positive cantaloupes of 540 test cantaloupes by the rinse method. Overall, BPW, LAC, and UP broths were equivalent for the recovery of Salmonella from cantaloupes. Both the VIDAS ICS/SLM and TECRA Unique Salmonella methods detected significantly fewer Salmonella-positive cantaloupes than did the culture method: the VIDAS ICS/SLM method detected 23 of 50 Salmonella-positive cantaloupes (60 tested) and the TECRA Unique Salmonella method detected 16 of 29 Salmonella-positive cantaloupes (60 tested). The VIDAS SLM and culture methods were equivalent: both methods detected 37 of 37 Salmonella-positive cantaloupes (60 tested).

Rapid multiplex PCR and Real-Time TaqMan PCR assays for detection of Salmonella enterica and the highly virulent serovars Choleraesuis and Paratyphi C

USDA-ARS?s Scientific Manuscript database

Salmonella enterica is a human pathogen with over 2,500 serovars characterized. S. enterica serovars Choleraesuis (Cs) and Paratyphi C (Pc) are two globally distributed serovars. We have developed a rapid molecular typing method to detect Cs and Pc in food samples by using a comparative genomics ap...

Rapid detection of Salmonella spp. in food by use of the ISO-GRID hydrophobic grid membrane filter.

PubMed Central

Entis, P; Brodsky, M H; Sharpe, A N; Jarvis, G A

1982-01-01

A rapid hydrophobic grid-membrane filter (HGMF) method was developed and compared with the Health Protection Branch cultural method for the detection of Salmonella spp. in 798 spiked samples and 265 naturally contaminated samples of food. With the HGMF method, Salmonella spp. were isolated from 618 of the spiked samples and 190 of the naturally contaminated samples. The conventional method recovered Salmonella spp. from 622 spiked samples and 204 unspiked samples. The isolation rates from Salmonella-positive samples for the two methods were not significantly different (94.6% overall for the HGMF method and 96.7% for the conventional approach), but the HGMF results were available in only 2 to 3 days after sample receipt compared with 3 to 4 days by the conventional method. Images PMID:7059168

Rapid detection of Salmonella in pet food: design and evaluation of integrated methods based on real-time PCR detection.

PubMed

Balachandran, Priya; Friberg, Maria; Vanlandingham, V; Kozak, K; Manolis, Amanda; Brevnov, Maxim; Crowley, Erin; Bird, Patrick; Goins, David; Furtado, Manohar R; Petrauskene, Olga V; Tebbs, Robert S; Charbonneau, Duane

2012-02-01

Reducing the risk of Salmonella contamination in pet food is critical for both companion animals and humans, and its importance is reflected by the substantial increase in the demand for pathogen testing. Accurate and rapid detection of foodborne pathogens improves food safety, protects the public health, and benefits food producers by assuring product quality while facilitating product release in a timely manner. Traditional culture-based methods for Salmonella screening are laborious and can take 5 to 7 days to obtain definitive results. In this study, we developed two methods for the detection of low levels of Salmonella in pet food using real-time PCR: (i) detection of Salmonella in 25 g of dried pet food in less than 14 h with an automated magnetic bead-based nucleic acid extraction method and (ii) detection of Salmonella in 375 g of composite dry pet food matrix in less than 24 h with a manual centrifugation-based nucleic acid preparation method. Both methods included a preclarification step using a novel protocol that removes food matrix-associated debris and PCR inhibitors and improves the sensitivity of detection. Validation studies revealed no significant differences between the two real-time PCR methods and the standard U.S. Food and Drug Administration Bacteriological Analytical Manual (chapter 5) culture confirmation method.

Direct rapid detection of beef lymph nodes containing high levels of Salmonella

USDA-ARS?s Scientific Manuscript database

Category: Post-harvest research Published: Unpublished to date Objective: The objective of this work was to determine if beef lymph nodes containing Salmonella could be rapidly identified through direct testing without enrichment. Experimental Design & Analysis: Beef lymph nodes (1,038) were co...

Evaluation of immunomagnetic separation for the detection of Salmonella in surface waters by polymerase chain reaction.

PubMed

Hsu, Chao-Yu; Hsu, Bing-Mu; Chang, Tien-Yu; Hsu, Tsui-Kang; Shen, Shu-Min; Chiu, Yi-Chou; Wang, Hung-Jen; Ji, Wen-Tsai; Fan, Cheng-Wei; Chen, Jyh-Larng

2014-09-19

Salmonella spp. is associated with fecal pollution and capable of surviving for long periods in aquatic environments. Instead of the traditional, time-consuming biochemical detection, polymerase chain reaction (PCR) allows rapid identification of Salmonella directly concentrated from water samples. However, prevalence of Salmonella may be underestimated because of the vulnerability of PCR to various environmental chemicals like humic acid, compounded by the fact that various DNA polymerases have different susceptibility to humic acid. Because immunomagnetic separation (IMS) theoretically could isolate Salmonella from other microbes and facilitate removal of aquatic PCR inhibitors of different sizes, this study aims to compare the efficiency of conventional PCR combined with immunomagnetic separation (IMS) for Salmonella detection within a moderately polluted watershed. In our study, the positive rate was increased from 17.6% to 47% with nearly ten-fold improvement in the detection limit. These results suggest the sensitivity of Salmonella detection could be enhanced by IMS, particularly in low quality surface waters. Due to its effects on clearance of aquatic pollutants, IMS may be suitable for most DNA polymerases for Salmonella detection.

Gold Nanoparticle Labeling Based ICP-MS Detection/Measurement of Bacteria, and Their Quantitative Photothermal Destruction

PubMed Central

Lin, Yunfeng

2015-01-01

Bacteria such as Salmonella and E. coli present a great challenge in public health care in today’s society. Protection of public safety against bacterial contamination and rapid diagnosis of infection require simple and fast assays for the detection and elimination of bacterial pathogens. After utilizing Salmonella DT104 as an example bacterial strain for our investigation, we report a rapid and sensitive assay for the qualitative and quantitative detection of bacteria by using antibody affinity binding, popcorn shaped gold nanoparticle (GNPOPs) labeling, surfance enchanced Raman spectroscopy (SERS), and inductively coupled plasma mass spectrometry (ICP-MS) detection. For qualitative analysis, our assay can detect Salmonella within 10 min by Raman spectroscopy; for quantitative analysis, our assay has the ability to measure as few as 100 Salmonella DT104 in a 1 mL sample (100 CFU/mL) within 40 min. Based on the quantitative detection, we investigated the quantitative destruction of Salmonella DT104, and the assay’s photothermal efficiency in order to reduce the amount of GNPOPs in the assay to ultimately to eliminate any potential side effects/toxicity to the surrounding cells in vivo. Results suggest that our assay may serve as a promising candidate for qualitative and quantitative detection and elimination of a variety of bacterial pathogens. PMID:26417447

Detection of Salmonella enterica Serovar Typhimurium by Using a Rapid, Array-Based Immunosensor

PubMed Central

Taitt, Chris Rowe; Shubin, Yura S.; Angel, Roselina; Ligler, Frances S.

2004-01-01

The multianalyte array biosensor (MAAB) is a rapid analysis instrument capable of detecting multiple analytes simultaneously. Rapid (15-min), single-analyte sandwich immunoassays were developed for the detection of Salmonella enterica serovar Typhimurium, with a detection limit of 8 × 104 CFU/ml; the limit of detection was improved 10-fold by lengthening the assay protocol to 1 h. S. enterica serovar Typhimurium was also detected in the following spiked foodstuffs, with minimal sample preparation: sausage, cantaloupe, whole liquid egg, alfalfa sprouts, and chicken carcass rinse. Cross-reactivity tests were performed with Escherichia coli and Campylobacter jejuni. To determine whether the MAAB has potential as a screening tool for the diagnosis of asymptomatic Salmonella infection of poultry, chicken excretal samples from a private, noncommercial farm and from university poultry facilities were tested. While the private farm excreta gave rise to signals significantly above the buffer blanks, none of the university samples tested positive for S. enterica serovar Typhimurium without spiking; dose-response curves of spiked excretal samples from university-raised poultry gave limits of detection of 8 × 103 CFU/g. PMID:14711637

Detection of Salmonella enterica in Meat in Less than 5 Hours by a Low-Cost and Noncomplex Sample Preparation Method

PubMed Central

Hoorfar, J.; Hansen, F.; Christensen, J.; Mansdal, S.; Josefsen, M. H.

2016-01-01

ABSTRACT Salmonella is recognized as one of the most important foodborne bacteria and has wide health and socioeconomic impacts worldwide. Fresh pork meat is one of the main sources of Salmonella, and efficient and fast methods for detection are therefore necessary. Current methods for Salmonella detection in fresh meat usually include >16 h of culture enrichment, in a few cases <12 h, thus requiring at least two working shifts. Here, we report a rapid (<5 h) and high-throughput method for screening of Salmonella in samples from fresh pork meat, consisting of a 3-h enrichment in standard buffered peptone water and a real-time PCR-compatible sample preparation method based on filtration, centrifugation, and enzymatic digestion, followed by fast-cycling real-time PCR detection. The method was validated in an unpaired comparative study against the Nordic Committee on Food Analysis (NMKL) reference culture method 187. Pork meat samples (n = 140) were either artificially contaminated with Salmonella at 0, 1 to 10, or 10 to 100 CFU/25 g of meat or naturally contaminated. Cohen's kappa for the degree of agreement between the rapid method and the reference was 0.64, and the relative accuracy, sensitivity, and specificity for the rapid method were 81.4, 95.1, and 97.9%, respectively. The 50% limit of detections (LOD50s) were 8.8 CFU/25 g for the rapid method and 7.7 CFU/25 g for the reference method. Implementation of this method will enable faster release of Salmonella low-risk meat, providing savings for meat producers, and it will help contribute to improved food safety. IMPORTANCE While the cost of analysis and hands-on time of the presented rapid method were comparable to those of reference culture methods, the fast product release by this method can provide the meat industry with a competitive advantage. Not only will the abattoirs save costs for work hours and cold storage, but consumers and retailers will also benefit from fresher meat with a longer shelf life. Furthermore, the presented sample preparation might be adjusted for application in the detection of other pathogenic bacteria in different sample types. PMID:27986726

Detection of Salmonella enterica in Meat in Less than 5 Hours by a Low-Cost and Noncomplex Sample Preparation Method.

PubMed

Fachmann, M S R; Löfström, C; Hoorfar, J; Hansen, F; Christensen, J; Mansdal, S; Josefsen, M H

2017-03-01

Salmonella is recognized as one of the most important foodborne bacteria and has wide health and socioeconomic impacts worldwide. Fresh pork meat is one of the main sources of Salmonella , and efficient and fast methods for detection are therefore necessary. Current methods for Salmonella detection in fresh meat usually include >16 h of culture enrichment, in a few cases <12 h, thus requiring at least two working shifts. Here, we report a rapid (<5 h) and high-throughput method for screening of Salmonella in samples from fresh pork meat, consisting of a 3-h enrichment in standard buffered peptone water and a real-time PCR-compatible sample preparation method based on filtration, centrifugation, and enzymatic digestion, followed by fast-cycling real-time PCR detection. The method was validated in an unpaired comparative study against the Nordic Committee on Food Analysis (NMKL) reference culture method 187. Pork meat samples ( n = 140) were either artificially contaminated with Salmonella at 0, 1 to 10, or 10 to 100 CFU/25 g of meat or naturally contaminated. Cohen's kappa for the degree of agreement between the rapid method and the reference was 0.64, and the relative accuracy, sensitivity, and specificity for the rapid method were 81.4, 95.1, and 97.9%, respectively. The 50% limit of detections (LOD 50 s) were 8.8 CFU/25 g for the rapid method and 7.7 CFU/25 g for the reference method. Implementation of this method will enable faster release of Salmonella low-risk meat, providing savings for meat producers, and it will help contribute to improved food safety. IMPORTANCE While the cost of analysis and hands-on time of the presented rapid method were comparable to those of reference culture methods, the fast product release by this method can provide the meat industry with a competitive advantage. Not only will the abattoirs save costs for work hours and cold storage, but consumers and retailers will also benefit from fresher meat with a longer shelf life. Furthermore, the presented sample preparation might be adjusted for application in the detection of other pathogenic bacteria in different sample types. Copyright © 2017 American Society for Microbiology.

Amperometric biosensor for Salmonella typhimurium detection in milk

USDA-ARS?s Scientific Manuscript database

This paper reports an amperometric biosensor for rapid and sensitive Salmonella Typhimurium detection in milk. The biosensor was assembled from the self-assembled monolayers technique on a gold surface. In this device, polyclonal antibodies were oriented by protein A. The biosensor structure was cha...

Development of a quantitative fluorescence single primer isothermal amplification-based method for the detection of Salmonella.

PubMed

Wang, Jianchang; Li, Rui; Hu, Lianxia; Sun, Xiaoxia; Wang, Jinfeng; Li, Jing

2016-02-16

Food-borne disease caused by Salmonella has long been, and continues to be, an important global public health problem, necessitating rapid and accurate detection of Salmonella in food. Real time PCR is the most recently developed approach for Salmonella detection. Single primer isothermal amplification (SPIA), a novel gene amplification technique, has emerged as an attractive microbiological testing method. SPIA is performed under a constant temperature, eliminating the need for an expensive thermo-cycler. In addition, SPIA reactions can be accomplished in 30 min, faster than real time PCR that usually takes over 2h. We developed a quantitative fluorescence SPIA-based method for the detection of Salmonella. Using Salmonella Typhimurium genomic DNA as template and a primer targeting Salmonella invA gene, we showed the detection limit of SPIA was 2.0 × 10(1)fg DNA. Its successful amplification of different serotypic Salmonella genomic DNA but not non-Salmonella bacterial DNA demonstrated the specificity of SPIA. Furthermore, this method was validated with artificially contaminated beef. In conclusion, we showed high sensitivity and specificity of SPIA in the detection of Salmonella, comparable to real time PCR. In addition, SPIA is faster and more cost-effective (non-use of expensive cyclers), making it a potential alternative for field detection of Salmonella in resource-limited settings that are commonly encountered in developing countries. Copyright © 2015 Elsevier B.V. All rights reserved.

Electrochemical Biosensors for Rapid Detection of Foodborne Salmonella: A Critical Overview

PubMed Central

Cinti, Stefano; Volpe, Giulia; Piermarini, Silvia; Delibato, Elisabetta; Palleschi, Giuseppe

2017-01-01

Salmonella has represented the most common and primary cause of food poisoning in many countries for at least over 100 years. Its detection is still primarily based on traditional microbiological culture methods which are labor-intensive, extremely time consuming, and not suitable for testing a large number of samples. Accordingly, great efforts to develop rapid, sensitive and specific methods, easy to use, and suitable for multi-sample analysis, have been made and continue. Biosensor-based technology has all the potentialities to meet these requirements. In this paper, we review the features of the electrochemical immunosensors, genosensors, aptasensors and phagosensors developed in the last five years for Salmonella detection, focusing on the critical aspects of their application in food analysis. PMID:28820458

TECRA Unique test for rapid detection of Salmonella in food: collaborative study.

PubMed

Hughes, D; Dailianis, A E; Hill, L; McIntyre, D A; Anderson, A

2001-01-01

The TECRA Unique Salmonella test uses the principle of immunoenrichment to allow rapid detection of Salmonellae in food. A collaborative study was conducted to compare the TECRA Salmonella Unique test with the reference culture method given in the U.S. Food and Drug Administration's Bacteriological Analytical Manual. Three food types (milk powder, pepper, and soy flour) were analyzed in Australia and 2 food types (milk chocolate and dried egg) were analyzed in the United States. Forty-one collaborators participated in the study. For each of the 5 foods at each of the 3 levels, a comparison showed no significant differences (p > or = 0.05) in the proportion of positive test samples for Unique and that for the reference method using the Chi-square test for independence with continuity correction.

Rapid combined assay for Salmonella detection in food samples.

PubMed

Gadó, I; Major, P; Király, M; Pláveczky, M G

2000-01-01

A rapid method was developed to detect salmonellae in food samples. The method gave a possibility to obtain results after 28 h 30 min. The preenrichment in buffered peptone water lasted for 6 h, the enrichment in Rappaport-Vassiliadis medium was applied for 18 h followed by PCR with INVA1-INVA2 primer pair, adapting Chiu and Ou's method. This procedure was suitable to demonstrate salmonella contamination at min. 10 cfu/25 g sample. Out of 18 samples there was a good agreement between the results of the conventional and rapid methods in case of 17 samples. PCR with SPVC1-SPVC2 primer pair informing about the presence of virulence plasmid was performed in separate tubes, because decreased sensitivity was observed in case of multiplex PCR.

Comparison of two commercially available rapid detection methods and a conventional culture method to detect naturally occurring salmonellae on broiler carcasses

USDA-ARS?s Scientific Manuscript database

Many different screening devices and sampling methods have been used to detect the presence of naturally occurring Salmonella on commercially processed broiler carcasses. The objective of this study was to compare two commercial screening systems (BAX® and Roka®) to a standard cultural procedure use...

Microfluidic Chip-Based Detection and Intraspecies Strain Discrimination of Salmonella Serovars Derived from Whole Blood of Septic Mice

PubMed Central

Patterson, Adriana S.; Heithoff, Douglas M.; Ferguson, Brian S.; Soh, H. Tom; Mahan, Michael J.

2013-01-01

Salmonella is a zoonotic pathogen that poses a considerable public health and economic burden in the United States and worldwide. Resultant human diseases range from enterocolitis to bacteremia to sepsis and are acutely dependent on the particular serovar of Salmonella enterica subsp. enterica, which comprises over 99% of human-pathogenic S. enterica isolates. Point-of-care methods for detection and strain discrimination of Salmonella serovars would thus have considerable benefit to medical, veterinary, and field applications that safeguard public health and reduce industry-associated losses. Here we describe a single, disposable microfluidic chip that supports isothermal amplification and sequence-specific detection and discrimination of Salmonella serovars derived from whole blood of septic mice. The integrated microfluidic electrochemical DNA (IMED) chip consists of an amplification chamber that supports loop-mediated isothermal amplification (LAMP), a rapid, single-temperature amplification method as an alternative to PCR that offers advantages in terms of sensitivity, reaction speed, and amplicon yield. The amplification chamber is connected via a microchannel to a detection chamber containing a reagentless, multiplexed (here biplex) sensing array for sequence-specific electrochemical DNA (E-DNA) detection of the LAMP products. Validation of the IMED device was assessed by the detection and discrimination of S. enterica subsp. enterica serovars Typhimurium and Choleraesuis, the causative agents of enterocolitis and sepsis in humans, respectively. IMED chips conferred rapid (under 2 h) detection and discrimination of these strains at clinically relevant levels (<1,000 CFU/ml) from whole, unprocessed blood collected from septic animals. The IMED-based chip assay shows considerable promise as a rapid, inexpensive, and portable point-of-care diagnostic platform for the detection and strain-specific discrimination of microbial pathogens. PMID:23354710

Rapid and specific detection of Salmonella in water samples using real-time PCR and High Resolution Melt (HRM) curve analysis.

PubMed

van Blerk, G N; Leibach, L; Mabunda, A; Chapman, A; Louw, D

2011-01-01

A real-time PCR assay combined with a pre-enrichment step for the specific and rapid detection of Salmonella in water samples is described. Following amplification of the invA gene target, High Resolution Melt (HRM) curve analysis was used to discriminate between products formed and to positively identify invA amplification. The real-time PCR assay was evaluated for specificity and sensitivity. The assay displayed 100% specificity for Salmonella and combined with a 16-18 h non-selective pre-enrichment step, the assay proved to be highly sensitive with a detection limit of 1.0 CFU/ml for surface water samples. The detection assay also demonstrated a high intra-run and inter-run repeatability with very little variation in invA amplicon melting temperature. When applied to water samples received routinely by the laboratory, the assay showed the presence of Salmonella in particularly surface water and treated effluent samples. Using the HRM based assay, the time required for Salmonella detection was drastically shortened to less than 24 h compared to several days when using standard culturing methods. This assay provides a useful tool for routine water quality monitoring as well as for quick screening during disease outbreaks.

Using a surface plasmon resonance biosensor for rapid detection of salmonella typhimurium in chicken carcass

USDA-ARS?s Scientific Manuscript database

Chicken is one of the most popular meat products in the world. Salmonella Typhimurium is a common foodborne pathogens associated with the processing of poultry. An optical Surface Plasmon Resonance (SPR) biosensor was sensitive to the presence of Salmonella Typhimurium in chicken carcass. The Spr...

Rapid and Sensitive Salmonella Typhi Detection in Blood and Fecal Samples Using Reverse Transcription Loop-Mediated Isothermal Amplification.

PubMed

Fan, Fenxia; Yan, Meiying; Du, Pengcheng; Chen, Chen; Kan, Biao

2015-09-01

Typhoid fever caused by Salmonella enterica serovar Typhi remains a significant public health problem in developing countries. Although the main method for diagnosing typhoid fever is blood culture, the test is time consuming and not always able to detect infections. Thus, it is very difficult to distinguish typhoid from other infections in patients with nonspecific symptoms. A simple and sensitive laboratory detection method remains necessary. The purpose of this study is to establish and evaluate a rapid and sensitive reverse transcription-based loop-mediated isothermal amplification (RT-LAMP) method to detect Salmonella Typhi infection. In this study, a new specific gene marker, STY1607, was selected to develop a STY1607-RT-LAMP assay; this is the first report of specific RT-LAMP detection assay for typhoid. Human-simulated and clinical blood/stool samples were used to evaluate the performance of STY1607-RT-LAMP for RNA detection; this method was compared with STY1607-LAMP, reverse transcription real-time polymerase chain reaction (rRT-PCR), and bacterial culture methods for Salmonella Typhi detection. Using mRNA as the template, STY1607-RT-LAMP exhibited 50-fold greater sensitivity than STY1607-LAMP for DNA detection. The STY1607-RT-LAMP detection limit is 3 colony-forming units (CFU)/mL for both the pure Salmonella Typhi samples and Salmonella Typhi-simulated blood samples and was 30 CFU/g for the simulated stool samples, all of which were 10-fold more sensitive than the rRT-PCR method. RT-LAMP exhibited improved Salmonella Typhi detection sensitivity compared to culture methods and to rRT-PCR of clinical blood and stool specimens from suspected typhoid fever patients. Because it can be performed without sophisticated equipment or skilled personnel, RT-LAMP is a valuable tool for clinical laboratories in developing countries. This method can be applied in the clinical diagnosis and care of typhoid fever patients as well as for a quick public health response.

Applications of immunomagnetic capture and time-resolved fluorescence detection for Salmonella enteriditis in liquid eggs

NASA Astrophysics Data System (ADS)

Tu, Shu-I.; Gehring, Andrew; Paoli, George

2008-04-01

An immuno sandwich method was evaluated for the detection of Salmonella in liquid eggs. Liquid eggs spiked with different out-break strains of Salmonella were mixed with proper enrichment media and incubated at 37 C for 4 to 20 h. After enrichment, immunomagnetic beads (IMB) coated with anti Salmonella antibodies were used to capture the bacteria. Samarium (Sm) labeled anti Salmonella antibodies were then used to form sandwiched complexes with IMB captured bacteria. Sandwiched Salmonella were then treated with Sm-chelator to allow the measurement of the released Sm by time-resolved fluorescence (TRF). The processes ranging from IMB capture to Sm chelation were performed using an automated KingFisher apparatus. With this approach, the presence of ~ 1 CFU of outbreak strains of Salmonella Enteritidis per egg (~50 g of liquid eggs) could be detected after enrichment for 20 h at 37 C. For higher levels of Salmonella Enteritidis contamination, e.g., 10 CFU per 50 g of liquid eggs, the enrichment time could be reduced to 5 h at 37 C. The results demonstrated that a combination of IMB capture and TRF measurement could be a rapid and sensitive method for Salmonella Enteritidis detection in liquid eggs.

Molecular identification of common Salmonella serovars using multiplex DNA sensor-based suspension array.

PubMed

Aydin, Muhsin; Carter-Conger, Jacqueline; Gao, Ning; Gilmore, David F; Ricke, Steven C; Ahn, Soohyoun

2018-04-01

Salmonella is one of major foodborne pathogens and the leading cause of foodborne illness-related hospitalizations and deaths. It is critical to develop a sensitive and rapid detection assay that can identify Salmonella to ensure food safety. In this study, a DNA sensor-based suspension array system of high multiplexing ability was developed to identify eight Salmonella serovars commonly associated with foodborne outbreaks to the serotype level. Each DNA sensor was prepared by activating pre-encoded microspheres with oligonucleotide probes that are targeting virulence genes and serovar-specific regions. The mixture of 12 different types of DNA sensors were loaded into a 96-well microplate and used as a 12-plex DNA sensor array platform. DNA isolated from Salmonella was amplified by multiplex polymerase chain reaction (mPCR), and the presence of Salmonella was determined by reading fluorescent signals from hybridization between probes on DNA sensors and fluorescently labeled target DNA using the Bio-Plex® system. The developed multiplex array was able to detect synthetic DNA at the concentration as low as 100 fM and various Salmonella serovars as low as 100 CFU/mL within 1 h post-PCR. Sensitivity of this assay was further improved to 1 CFU/mL with 6-h enrichment. The array system also correctly and specifically identified serotype of tested Salmonella strains without any cross-reactivity with other common foodborne pathogens. Our results indicate the developed DNA sensor suspension array can be a rapid and reliable high-throughput method for simultaneous detection and molecular identification of common Salmonella serotypes.

Production of recombinant flagellin to develop ELISA-based detection of Salmonella Enteritidis.

PubMed

Mirhosseini, Seyed Ali; Fooladi, Abbas Ali Imani; Amani, Jafar; Sedighian, Hamid

Food-borne diseases, caused by the pathogenic bacteria, are highly prevalent in the world. Salmonella is one of the most important bacterial genera responsible for this. Salmonella Enteritidis (SE) is one of the non-typhoid Salmonellae that can be transmitted to human from poultry products, water, and contaminated food. In recent years, new and rapid detection methods such as enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) have been developed. In this study, recombinant FliC (rFliC) was produced to be used as an antigen. The immunization was conducted in mice with the purified recombinant FliC (rFliC). The mice were subcutaneously immunized with rFliC and elicited significant rFliC specific serum IgG antibodies. An indirect ELISA system was established for the detection of Salmonella Enteritidis. Our results confirmed that the recombinant flagellin can be one of the excellent indicators for the detection of Salmonella Enteritidis. Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

Detection of Salmonella enterica serovar Enteritidis (SE) Antibodies in Serum Using A Polystyrene Bead/SE Flagella Agglutination Assay

USDA-ARS?s Scientific Manuscript database

Serologic screening of flocks can be an important method to detect Salmonella enteritidis (SE) infections but can be labor intensive or lack specificity. Our goal was to develop a rapid agglutination assay using SE flagella adsorbed to polystyrene beads as a simple, relatively specific test to dete...

Loop-Mediated Isothermal Amplification for Salmonella Detection in Food and Feed: Current Applications and Future Directions

PubMed Central

Yang, Qianru; Domesle, Kelly J.

2018-01-01

Abstract Loop-mediated isothermal amplification (LAMP) has become a powerful alternative to polymerase chain reaction (PCR) for pathogen detection in clinical specimens and food matrices. Nontyphoidal Salmonella is a zoonotic pathogen of significant food and feed safety concern worldwide. The first study employing LAMP for the rapid detection of Salmonella was reported in 2005, 5 years after the invention of the LAMP technology in Japan. This review provides an overview of international efforts in the past decade on the development and application of Salmonella LAMP assays in a wide array of food and feed matrices. Recent progress in assay design, platform development, commercial application, and method validation is reviewed. Future perspectives toward more practical and wider applications of Salmonella LAMP assays in food and feed testing are discussed. PMID:29902082

Rapid detection of Salmonella Typhimurium in chicken carcass using a SPR biosensor

NASA Astrophysics Data System (ADS)

Wang, Shizhou; Lan, Yubin; Yin, Yongguang; Dasari, Thirumala R.

2005-11-01

The SPR biosensor was sensitive to the presence of Salmonella Typhimurium in chicken carcass. The selectivity of the SPR biosensor was assayed using a series of antibody concentrations and dilution series of the organism. The SPR biosensor was specific to Salmonella Typhimurium at concentrations of 106 CFU/ml. Initial results show potential for its application for pathogenic bacteria monitoring.

Identification of Salmonella Typhimurium-specific DNA aptamers developed using whole-cell SELEX and FACS analysis.

PubMed

Moon, Jihea; Kim, Giyoung; Lee, Sangdae; Park, Saetbyeol

2013-11-01

Conventional methods for detection of infective organisms, such as Salmonella, are complicated and require multiple steps, and the need for rapid detection has increased. Biosensors show great potential for rapid detection of pathogens. In turn, aptamers have great potential for biosensor assay development, given their small size, ease of synthesis and labeling, lack of immunogenicity, a lower cost of production than antibodies, and high target specificity. In this study, ssDNA aptamers specific to Salmonella Typhimurium were obtained by a whole bacterium-based systematic evolution of ligands by exponential enrichment (SELEX) procedure and applied to probing S. Typhimurium. After 10 rounds of selection with S. Typhimurium as the target and Salmonella Enteritidis, Escherichia coli and Staphylococcus aureus as counter targets, the highly enriched oligonucleic acid pool was sorted using flow cytometry. In total, 12 aptamer candidates from different families were sequenced and grouped. Fluorescent analysis demonstrated that aptamer C4 had particularly high binding affinity and selectivity; this aptamer was then further characterized. © 2013 Elsevier B.V. All rights reserved.

Multiplex PCR for simultaneous identification of E. coli O157:H7, Salmonella spp. and L. monocytogenes in food.

PubMed

Nguyen, Thuy Trang; Van Giau, Vo; Vo, Tuong Kha

2016-12-01

The rapid detection of pathogens in food is becoming increasingly critical for ensuring the safety of consumers, since the majority of food-borne illnesses and deaths are caused by pathogenic bacteria. Hence, rapid, sensitive, inexpensive and convenient approaches to detect food-borne pathogenic bacteria is essential in controlling food safety. In this study, a multiplex PCR assay for the rapid and simultaneous detection of Escherichia coli O157:H7, Salmonella spp. and Listeria monocytogenes was established. The invA, stx and hlyA genes specifically amplified DNA fragments of 284, 404 and 510 bp from Salmonella spp., L. monocytogenes and E. coli O157:H7, respectively. The 16S rRNA gene was targeted as an internal control gene in the presence of bacterial DNA. The specificity and sensitivity of the multiplex PCR were performed by testing different strains. The multiplex PCR assay was able to specifically simultaneously detect ten colony-forming unit/mL of each pathogen in artificially inoculated samples after enrichment for 12 h. The whole process took less than 24 h to complete, indicating that the assay is suitable for reliable and rapid identification of these three food-borne pathogens, which could be suitable in microbial epidemiology investigation.

Nano-particle enhanced impedimetric biosensor for detedtion of foodborne pathogens

NASA Astrophysics Data System (ADS)

Kim, G.; Om, A. S.; Mun, J. H.

2007-03-01

Recent outbreaks of foodborne illness have been increased the need for rapid and sensitive methods for detection of these pathogens. Conventional methods for pathogens detection and identification involve prolonged multiple enrichment steps. Even though some immunological rapid assays are available, these assays still need enrichment steps result in delayed detection. Biosensors have shown great potential for rapid detection of foodborne pathogens. They are capable of direct monitoring the antigen-antibody reactions in real time. Among the biosensors, impedimetric biosensors have been widely adapted as an analysis tool for the study of various biological binding reactions because of their high sensitivity and reagentless operation. In this study a nanoparticle-enhanced impedimetric biosensor for Salmonella enteritidis detection was developed which detected impedance changes caused by the attachment of the cells to the anti-Salmonella antibodies immobilized on interdigitated gold electrodes. Successive immobilization of neutravidin followed by anti-Salmonella antibodies was performed to the sensing area to create a biological detection surface. To enhance the impedance responses generated by antigen-antibody reactions, anti-Salmonella antibody conjugated nanoparticles were introduced on the sensing area. Using a portable impedance analyzer, the impedance across the interdigital electrodes was measured after the series of antigen-antibody bindings. Bacteria cells present in solution attached to capture antibodies and became tethered to the sensor surface. Attached bacteria cells changed the dielectric constant of the media between the electrodes thereby causing a change in measured impedance. Optimum input frequency was determined by analyzing frequency characteristics of the biosensor over ranges of applied frequencies from 10 Hz to 400 Hz. At 100 Hz of input frequency, the biosensor was most sensitive to the changes of the bacteria concentration and this frequency was used for the detection experiments. The biosensor was able to detect 106 CFU/mL in phosphate buffered saline (PBS) with a detection time of 3 minutes. Additional use of nanoparticles significantly enhanced the detection performance. By using the nanoparticles the biosensor could detect 104 CFU/mL of Salmonella enteritidis in PBS and 105 CFU/mL of cells in milk.

A multiplex single nucleotide polymorphism typing assay for detecting mutations that result in decreased fluoroquinolone susceptibility in Salmonella enterica serovars Typhi and Paratyphi A

PubMed Central

Song, Yajun; Roumagnac, Philippe; Weill, François-Xavier; Wain, John; Dolecek, Christiane; Mazzoni, Camila J.; Holt, Kathryn E.; Achtman, Mark

2010-01-01

Objectives Decreased susceptibility to fluoroquinolones has become a major problem for the successful therapy of human infections caused by Salmonella enterica, especially the life-threatening typhoid and paratyphoid fevers. Methods By using Luminex xTAG beads, we developed a rapid, reliable and cost-effective multiplexed genotyping assay for simultaneously detecting 11 mutations in gyrA, gyrB and parE of S. enterica serovars Typhi and Paratyphi A that result in nalidixic acid resistance (NalR) and/or decreased susceptibility to fluoroquinolones. Results This assay yielded unambiguous single nucleotide polymorphism calls on extracted DNA from 292 isolates of Salmonella Typhi (NalR = 223 and NalS = 69) and 106 isolates of Salmonella Paratyphi A (NalR = 24 and NalS = 82). All of the 247 NalR Salmonella Typhi and Salmonella Paratyphi A isolates were found to harbour at least one of the target mutations, with GyrA Phe-83 as the most common one (143/223 for Salmonella Typhi and 18/24 for Salmonella Paratyphi A). We also identified three GyrB mutations in eight NalS Salmonella Typhi isolates (six for GyrB Phe-464, one for GyrB Leu-465 and one for GyrB Asp-466), and mutations GyrB Phe-464 and GyrB Asp-466 seem to be related to the decreased ciprofloxacin susceptibility phenotype in Salmonella Typhi. This assay can also be used directly on boiled single colonies. Conclusions The assay presented here would be useful for clinical and reference laboratories to rapidly screen quinolone-resistant isolates of Salmonella Typhi and Salmonella Paratyphi A, and decipher the underlying genetic changes for epidemiological purposes. PMID:20511368

Nano-materials for use in sensing of salmonella infections: Recent advances.

PubMed

Pashazadeh, Paria; Mokhtarzadeh, Ahad; Hasanzadeh, Mohammad; Hejazi, Maryam; Hashemi, Maryam; de la Guardia, Miguel

2017-01-15

Salmonella infectious diseases spreading every day through food have become a life-threatening problem for millions of people and growing menace to society. Health expert's estimate that the yearly cost of all the food borne diseases is approximately $5-6 billion. Traditional methodologies for salmonella analysis provide high reliability and very low limits of detection. Among them immunoassays and Nucleic acid-based assays provide results within 24h, but they are expensive, tedious and time consuming. So, there is an urgent need for development of rapid, robust and cost-effective alternative technologies for real-time monitoring of salmonella. Several biosensors have been designed and commercialized for detection of this pathogen in food and water. In this overview, we have updated the literature concerning novel biosensing methods such as various optical and electrochemical biosensors and newly developed nano- and micro-scaled and aptamers based biosensors for detection of salmonella pathogen. Furthermore, attention has been focused on the principal concepts, applications, and examples that have been achieved up to diagnose salmonella. In addition, commercial biosensors and foreseeable future trends for onsite detecting salmonella have been summarized. Copyright © 2016 Elsevier B.V. All rights reserved.

Evaluation of the 3M™ Molecular Detection Assay (MDA) 2 - Salmonella for the Detection of Salmonella spp. in Select Foods and Environmental Surfaces: Collaborative Study, First Action 2016.01.

PubMed

Bird, Patrick; Flannery, Jonathan; Crowley, Erin; Agin, James R; Goins, David; Monteroso, Lisa

2016-07-01

The 3M™ Molecular Detection Assay (MDA) 2 - Salmonella uses real-time isothermal technology for the rapid and accurate detection of Salmonella spp. from enriched select food, feed, and food-process environmental samples. The 3M MDA 2 - Salmonella was evaluated in a multilaboratory collaborative study using an unpaired study design. The 3M MDA 2 - Salmonella was compared to the U.S. Food and Drug Administration Bacteriological Analytical Manual Chapter 5 reference method for the detection of Salmonella in creamy peanut butter, and to the U.S. Department of Agriculture, Food Safety and Inspection Service Microbiology Laboratory Guidebook Chapter 4.08 reference method "Isolation and Identification of Salmonella from Meat, Poultry, Pasteurized Egg and Catfish Products and Carcass and Environmental Samples" for the detection of Salmonella in raw ground beef (73% lean). Technicians from 16 laboratories located within the continental United States participated. Each matrix was evaluated at three levels of contamination: an uninoculated control level (0 CFU/test portion), a low inoculum level (0.2-2 CFU/test portion), and a high inoculum level (2-5 CFU/test portion). Statistical analysis was conducted according to the probability of detection (POD) statistical model. Results obtained for the low inoculum level test portions produced difference in collaborator POD values of 0.03 (95% confidence interval, -0.10 to 0.16) for raw ground beef and 0.06 (95% confidence interval, -0.06 to 0.18) for creamy peanut butter, indicating no statistically significant difference between the candidate and reference methods.

Aptasensors for quantitative detection of Salmonella Typhimurium.

PubMed

Ansari, Najmeh; Yazdian-Robati, Rezvan; Shahdordizadeh, Mahin; Wang, Zhouping; Ghazvini, Kiarash

2017-09-15

Salmonella is one of the most frequent causes of food borne infectious disease. Among nearly 2500 documented serotypes are reported, Salmonella Typhimurium is the number one serotype associated with salmonellosis worldwide. Many different methods have been developed for the detection and quantification of S. typhimurium. Most of these assays are usually expensive, time consuming and require difficult sample preparation steps. Therefore, it is necessary to develop rapid, robust, cost-effective and sensitive alternative detection methods. In the last years, aptasensors, used for detection of S. typhimurium in different samples. In this review, recent advances and applications of aptasensors for the detection and quantification of S. typhimurium in details have been summarized. Copyright © 2017 Elsevier Inc. All rights reserved.

Real-time PCR method combined with immunomagnetic separation for detecting healthy and heat-injured Salmonella Typhimurium on raw duck wings.

PubMed

Zheng, Qianwang; Mikš-Krajnik, Marta; Yang, Yishan; Xu, Wang; Yuk, Hyun-Gyun

2014-09-01

Conventional culture detection methods are time consuming and labor-intensive. For this reason, an alternative rapid method combining real-time PCR and immunomagnetic separation (IMS) was investigated in this study to detect both healthy and heat-injured Salmonella Typhimurium on raw duck wings. Firstly, the IMS method was optimized by determining the capture efficiency of Dynabeads(®) on Salmonella cells on raw duck wings with different bead incubation (10, 30 and 60 min) and magnetic separation (3, 10 and 30 min) times. Secondly, three Taqman primer sets, Sal, invA and ttr, were evaluated to optimize the real-time PCR protocol by comparing five parameters: inclusivity, exclusivity, PCR efficiency, detection probability and limit of detection (LOD). Thirdly, the optimized real-time PCR, in combination with IMS (PCR-IMS) assay, was compared with a standard ISO and a real-time PCR (PCR) method by analyzing artificially inoculated raw duck wings with healthy and heat-injured Salmonella cells at 10(1) and 10(0) CFU/25 g. Finally, the optimized PCR-IMS assay was validated for Salmonella detection in naturally contaminated raw duck wing samples. Under optimal IMS conditions (30 min bead incubation and 3 min magnetic separation times), approximately 85 and 64% of S. Typhimurium cells were captured by Dynabeads® from pure culture and inoculated raw duck wings, respectively. Although Sal and ttr primers exhibited 100% inclusivity and exclusivity for 16 Salmonella spp. and 36 non-Salmonella strains, the Sal primer showed lower LOD (10(3) CFU/ml) and higher PCR efficiency (94.1%) than the invA and ttr primers. Moreover, for Sal and invA primers, 100% detection probability on raw duck wings suspension was observed at 10(3) and 10(4) CFU/ml with and without IMS, respectively. Thus, the Sal primer was chosen for further experiments. The optimized PCR-IMS method was significantly (P=0.0011) better at detecting healthy Salmonella cells after 7-h enrichment than traditional PCR method. However there was no significant difference between the two methods with longer enrichment time (14 h). The diagnostic accuracy of PCR-IMS was shown to be 98.3% through the validation study. These results indicate that the optimized PCR-IMS method in this study could provide a sensitive, specific and rapid detection method for Salmonella on raw duck wings, enabling 10-h detection. However, a longer enrichment time could be needed for resuscitation and reliable detection of heat-injured cells. Copyright © 2014 Elsevier B.V. All rights reserved.

Evaluation of PCR and high-resolution melt curve analysis for differentiation of Salmonella isolates.

PubMed

Saeidabadi, Mohammad Sadegh; Nili, Hassan; Dadras, Habibollah; Sharifiyazdi, Hassan; Connolly, Joanne; Valcanis, Mary; Raidal, Shane; Ghorashi, Seyed Ali

2017-06-01

Consumption of poultry products contaminated with Salmonella is one of the major causes of foodborne diseases worldwide and therefore detection and differentiation of Salmonella spp. in poultry is important. In this study, oligonucleotide primers were designed from hemD gene and a PCR followed by high-resolution melt (HRM) curve analysis was developed for rapid differentiation of Salmonella isolates. Amplicons of 228 bp were generated from 16 different Salmonella reference strains and from 65 clinical field isolates mainly from poultry farms. HRM curve analysis of the amplicons differentiated Salmonella isolates and analysis of the nucleotide sequence of the amplicons from selected isolates revealed that each melting curve profile was related to a unique DNA sequence. The relationship between reference strains and tested specimens was also evaluated using a mathematical model without visual interpretation of HRM curves. In addition, the potential of the PCR-HRM curve analysis was evaluated for genotyping of additional Salmonella isolates from different avian species. The findings indicate that PCR followed by HRM curve analysis provides a rapid and robust technique for genotyping of Salmonella isolates to determine the serovar/serotype.

Quasi-metagenomics and realtime sequencing aided detection and subtyping of Salmonella enterica from food samples.

PubMed

Hyeon, Ji-Yeon; Li, Shaoting; Mann, David A; Zhang, Shaokang; Li, Zhen; Chen, Yi; Deng, Xiangyu

2017-12-01

Metagenomics analysis of food samples promises isolation-independent detection and subtyping of foodborne bacterial pathogens in a single workflow. Selective concentration of Salmonella genomic DNA through immunomagnetic separation (IMS) and multiple displacement amplification (MDA) were shown to shorten culture enrichment of Salmonella -spiked raw chicken breast samples by over 12 hours while permitting serotyping and high-fidelity single nucleotide polymorphisms (SNP) typing of the pathogen using short shotgun sequencing reads. The herein termed quasi-metagenomics approach was evaluated on Salmonella -spiked lettuce and black peppercorn samples as well as retail chicken parts naturally contaminated with different serotypes of Salmonella. Between 8 and 24 h culture enrichment was required for detecting and subtyping naturally occurring Salmonella from unspiked chicken parts compared with 4 to 12 h culture enrichment when Salmonella -spiked food samples were analyzed, indicating the likely need for longer culture enrichment to revive low levels of stressed or injured Salmonella cells in food. Further acceleration of the workflow was achieved by real-time nanopore sequencing. After 1.5 hours of analysis on a potable sequencer, sufficient data were generated from sequencing IMS-MDA product of a cultured-enriched lettuce sample to allow serotyping and robust phylogenetic placement of the inoculated isolate. Importance Both culture enrichment and next-generation sequencing remain to be time-consuming processes for food testing where rapid methods for pathogen detection are widely available. Our study demonstrated substantial acceleration of the respective process through IMS-MDA and real-time nanopore sequencing. In one example, the combined use of the two methods delivered a less than 24 h turnaround time from a Salmonella -contaminated lettuce sample to phylogenetic identification of the pathogen. Improved efficiency like this is important for further expanding the use of whole genome and metagenomics sequencing in microbial analysis of food. Our results suggest the potential of the quasi-metagenomics approach in areas where rapid detection and subtyping of foodborne pathogens is important, such as foodborne outbreak response and precision tracking and monitoring of foodborne pathogens in production environments and supply chains. Copyright © 2017 American Society for Microbiology.

Samsung Salmonella Detection Kit. AOAC Performance Tested Method(SM) 021203.

PubMed

Li, Jun; Cheung, Win Den; Opdyke, Jason; Harvey, John; Chong, Songchun; Moon, Cheol Gon

2012-01-01

Salmonella, one of the most common causes of foodborne illness, is a significant public health concern worldwide. There is a need in the food industry for methods that are simple, rapid, and sensitive for the detection of foodborne pathogens. In this study, the Samsung Salmonella Detection Kit, a real-time PCR assay for the detection of Salmonella, was evaluated according to the current AOAC guidelines. The validation consisted of lot-to-lot consistency, stability, robustness, and inclusivity/exclusivity studies, as well as a method comparison of 10 different food matrixes. In the validation, the Samsung Salmonella Detection Kit was used in conjunction with the Applied Biosystems StepOnePlus PCR system and the Samsung Food Testing Software for the detection of Salmonella species. The performance of the assays was compared to the U.S. Department of Agriculture/Food Safety and Inspection Service-Microbiology Laboratory Guidebook (USDA/FSIS-MLG) 4.05: Isolation and Identification of Salmonella from Meat, Poultry, Pasteurized Egg, and Catfish and the and U.S. Food and Drug Administration/Bacteriological Analytical Manual (FDA/BAM) Chapter 5 Salmonella reference methods. The validation was conducted using an unpaired study design for detection of Salmonella spp. in raw ground beef, raw pork, raw ground pork, raw chicken wings, raw salmon, alfalfa sprouts, pasteurized orange juice, peanut butter, pasteurized whole milk, and shell eggs. The Samsung Salmonella Detection Kit demonstrated lot-to-lot consistency among three independent lots as well as ruggedness with minor modifications to changes in enrichment incubation time, enrichment incubation temperature, and DNA sample volume for PCR reaction. Stability was observed for 13 months at -20 degrees C and 3 months at 5 degrees C. For the inclusivity/exclusivity study, the Samsung Salmonella Detection Kit correctly identified 147 Salmonella species isolates out of 147 isolates tested from each of three different enrichment broths (a total of 441 isolates detected), and correctly excluded all 31 nontarget strains analyzed. For the method comparison, statistical analysis was conducted according to the Mantel-Haenszel Chi-square formula for unpaired test portions, and there was no significant difference in the number of positive samples detected between the Samsung Salmonella Detection Kit and the USDA/FSIS-MLG and FDA/BAM reference methods for all 10 food matrixes.

Rapid Detection of Pathogenic Bacteria from Fresh Produce by Filtration and Surface-Enhanced Raman Spectroscopy

NASA Astrophysics Data System (ADS)

Wu, Xiaomeng; Han, Caiqin; Chen, Jing; Huang, Yao-Wen; Zhao, Yiping

2016-04-01

The detection of Salmonella Poona from cantaloupe cubes and E. coli O157:H7 from lettuce has been explored by using a filtration method and surface-enhanced Raman spectroscopy (SERS) based on vancomycin-functionalized silver nanorod array substrates. It is found that with a two-step filtration process, the limit of detection (LOD) of Salmonella Poona from cantaloupe cubes can be as low as 100 CFU/mL in less than 4 h, whereas the chlorophyll in the lettuce causes severe SERS spectral interference. To improve the LOD of lettuce, a three-step filtration method with a hydrophobic filter is proposed. The hydrophobic filter can effectively eliminate the interferences from chlorophyll and achieve a LOD of 1000 CFU/mL detection of E. coli O157:H7 from lettuce samples within 5 h. With the low LODs and rapid detection time, the SERS biosensing platform has demonstrated its potential as a rapid, simple, and inexpensive means for pathogenic bacteria detection from fresh produce.

Aptasensors for rapid detection of Escherichia coli O157:H7 and Salmonella typhimurium

NASA Astrophysics Data System (ADS)

Wu, Wen-he; Li, Min; Wang, Yue; Ouyang, Hou-xian; Wang, Lin; Li, Ci-xiu; Cao, Yu-chen; Meng, Qing-he; Lu, Jian-xin

2012-11-01

Herein we reported the development of aptamer-based biosensors (aptasensors) based on label-free aptamers and gold nanoparticles (AuNPs) for detection of Escherichia coli ( E. coli) O157:H7 and Salmonella typhimurium. Target bacteria binding aptamers are adsorbed on the surface of unmodified AuNPs to capture target bacteria, and the detection was accomplished by target bacteria-induced aggregation of the aptasensor which is associated as red-to-purple color change upon high-salt conditions. By employing anti- E. coli O157:H7 aptamer and anti- S. typhimurium aptamer, we developed a convenient and rapid approach that could selectively detect bacteria without specialized instrumentation and pretreatment steps such as cell lysis. The aptasensor could detect as low as 105colony-forming units (CFU)/ml target bacteria within 20 min or less and its specificity was 100%. This novel method has a great potential application in rapid detection of bacteria in the near future.

High prevalence of Salmonella spp. in wastewater reused for irrigation assessed by molecular methods.

PubMed

Santiago, Paula; Jiménez-Belenguer, Ana; García-Hernández, Jorge; Estellés, Rosa Montes; Hernández Pérez, Manuel; Castillo López, M Angeles; Ferrús, María Antonia; Moreno, Yolanda

2018-01-01

Salmonella spp. is one of the most important causal agents of food-borne illness in developed countries and its presence in irrigation water poses a risk to public health. Its detection in environmental samples is not easy when culture methods are used, and molecular techniques such as PCR or ribosomal rRNA probe hybridization (Fluorescent in situ Hybridization, FISH) are outstanding alternatives. The aim of this work was to determine the environmental risk due to the presence of Salmonella spp. in wastewater by culture, PCR and FISH. A new specific rDNA probe for Salmonella was designed and its efficiency was compared with the rest of methods Serotype and antibiotic resistance of isolated strains were determined. Forty-five wastewater samples (collected from two secondary wastewater treatment plants) were analysed. Salmonella strains were isolated in 24 wastewater samples (53%), two of them after disinfection treatment. Twenty-three Salmonella strains exhibited resistance to one or more antimicrobial agent. Analysis of wastewater samples yielded PCR positive results for Salmonella in 28 out of the 45 wastewater samples (62%). FISH analysis allowed for the detection of Salmonella in 27 (60%) samples. By using molecular methods, Salmonella was detected in four samples after disinfection treatment. These results show the prevalence of Salmonella in reclaimed wastewater even after U.V. disinfection, what is a matter of public health concern, the high rates of resistance to antibiotics and the adequacy of molecular methods for its rapid detection. FISH method, with SA23 probe developed and assayed in this work provides a tool for detecting Salmonella in water within few hours, with a high rate of effectiveness. Copyright © 2017 Elsevier GmbH. All rights reserved.

Real-Time PCR Method for Detection of Salmonella spp. in Environmental Samples.

PubMed

Kasturi, Kuppuswamy N; Drgon, Tomas

2017-07-15

The methods currently used for detecting Salmonella in environmental samples require 2 days to produce results and have limited sensitivity. Here, we describe the development and validation of a real-time PCR Salmonella screening method that produces results in 18 to 24 h. Primers and probes specific to the gene invA , group D, and Salmonella enterica serovar Enteritidis organisms were designed and evaluated for inclusivity and exclusivity using a panel of 329 Salmonella isolates representing 126 serovars and 22 non- Salmonella organisms. The invA - and group D-specific sets identified all the isolates accurately. The PCR method had 100% inclusivity and detected 1 to 2 copies of Salmonella DNA per reaction. Primers specific for Salmonella -differentiating fragment 1 (Sdf-1) in conjunction with the group D set had 100% inclusivity for 32 S Enteritidis isolates and 100% exclusivity for the 297 non-Enteritidis Salmonella isolates. Single-laboratory validation performed on 1,741 environmental samples demonstrated that the PCR method detected 55% more positives than the V itek i mmuno d iagnostic a ssay s ystem (VIDAS) method. The PCR results correlated well with the culture results, and the method did not report any false-negative results. The receiver operating characteristic (ROC) analysis documented excellent agreement between the results from the culture and PCR methods (area under the curve, 0.90; 95% confidence interval of 0.76 to 1.0) confirming the validity of the PCR method. IMPORTANCE This validated PCR method detects 55% more positives for Salmonella in half the time required for the reference method, VIDAS. The validated PCR method will help to strengthen public health efforts through rapid screening of Salmonella spp. in environmental samples.

Real-Time PCR Method for Detection of Salmonella spp. in Environmental Samples

PubMed Central

Drgon, Tomas

2017-01-01

ABSTRACT The methods currently used for detecting Salmonella in environmental samples require 2 days to produce results and have limited sensitivity. Here, we describe the development and validation of a real-time PCR Salmonella screening method that produces results in 18 to 24 h. Primers and probes specific to the gene invA, group D, and Salmonella enterica serovar Enteritidis organisms were designed and evaluated for inclusivity and exclusivity using a panel of 329 Salmonella isolates representing 126 serovars and 22 non-Salmonella organisms. The invA- and group D-specific sets identified all the isolates accurately. The PCR method had 100% inclusivity and detected 1 to 2 copies of Salmonella DNA per reaction. Primers specific for Salmonella-differentiating fragment 1 (Sdf-1) in conjunction with the group D set had 100% inclusivity for 32 S. Enteritidis isolates and 100% exclusivity for the 297 non-Enteritidis Salmonella isolates. Single-laboratory validation performed on 1,741 environmental samples demonstrated that the PCR method detected 55% more positives than the Vitek immunodiagnostic assay system (VIDAS) method. The PCR results correlated well with the culture results, and the method did not report any false-negative results. The receiver operating characteristic (ROC) analysis documented excellent agreement between the results from the culture and PCR methods (area under the curve, 0.90; 95% confidence interval of 0.76 to 1.0) confirming the validity of the PCR method. IMPORTANCE This validated PCR method detects 55% more positives for Salmonella in half the time required for the reference method, VIDAS. The validated PCR method will help to strengthen public health efforts through rapid screening of Salmonella spp. in environmental samples. PMID:28500041

Visual detection technique for efficient screening and isolation of Salmonella based on a novel enrichment assay using chromatography membrane.

PubMed

Tang, F; Xiong, Y; Zhang, H; Wu, K; Xiang, Y; Shao, J-B; Ai, H-W; Xiang, Y-P; Zheng, X-L; Lv, J-R; Sun, H; Bao, L-S; Zhang, Z; Hu, H-B; Zhang, J-Y; Chen, L; Lu, J; Liu, W-Y; Mei, H; Ma, Y; Xu, C-F; Fang, A-Y; Gu, M; Xu, C-Y; Chen, Y; Chen, Z; Sun, Z-Y

2016-03-01

To detect Salmonella more efficiently and isolate strains more easily, a novel and simple detection method that uses an enrichment assay and two chromogenic reactions on a chromatography membrane was developed. Grade 3 chromatography paper is used as functionalized solid phase support (SPS), which contains specially optimized medium. One reaction for screening is based on the sulfate-reducing capacity of Salmonella. Hydrogen sulfide (H2S) generated by Salmonella reacts with ammonium ferric citrate to produce black colored ferrous sulfide. Another reaction is based on Salmonella C8 esterase that is unique for Enterobacteriaceae except Serratia and interacts with 4-methylumbelliferyl caprylate (MUCAP) to produce fluorescent umbelliferone, which is visible under ultraviolet light. A very low detection limit (10(1) CFU ml(-1)) for Salmonella was achieved on the background of 10(5) CFU ml(-1) Escherichia coli. More importantly, testing with more than 1,000 anal samples indicated that our method has a high positive detection rate and is relatively low cost, compared with the traditional culture-based method. It took only 1 day for the preliminary screening and 2 days to efficiently isolate the Salmonella cells, indicating that the new assay is specific, rapid, and simple for Salmonella detection. In contrast to the traditional culture-based method, this method can be easily used to screen and isolate targeted strains with the naked eye. The results of quantitative and comparative experiments showed that the visual detection technique is an efficient alternative method for the screening of Salmonella spp. in many applications of large-sized samples related to public health surveillance.

Label-free SERS detection of Salmonella Typhimurium on DNA aptamer modified AgNR substrates

USDA-ARS?s Scientific Manuscript database

Salmonella Typhimurium is an important foodborne pathogen which causes gastroenteritis in both humans and animals. Currently available rapid methods have relied on antibodies to offer specific recognition of the pathogen from the background. As a substitute of antibodies, nucleic acid aptamers offer...

MicroSEQ® Salmonella spp. Detection Kit Using the Pathatrix® 10-Pooling Salmonella spp. Kit Linked Protocol Method Modification.

PubMed

Wall, Jason; Conrad, Rick; Latham, Kathy; Liu, Eric

2014-03-01

Real-time PCR methods for detecting foodborne pathogens offer the advantages of simplicity and quick time to results compared to traditional culture methods. The addition of a recirculating pooled immunomagnetic separation method prior to real-time PCR analysis increases processing output while reducing both cost and labor. This AOAC Research Institute method modification study validates the MicroSEQ® Salmonella spp. Detection Kit [AOAC Performance Tested Method (PTM) 031001] linked with the Pathatrix® 10-Pooling Salmonella spp. Kit (AOAC PTM 090203C) in diced tomatoes, chocolate, and deli ham. The Pathatrix 10-Pooling protocol represents a method modification of the enrichment portion of the MicroSEQ Salmonella spp. The results of the method modification were compared to standard cultural reference methods for diced tomatoes, chocolate, and deli ham. All three matrixes were analyzed in a paired study design. An additional set of chocolate test portions was analyzed using an alternative enrichment medium in an unpaired study design. For all matrixes tested, there were no statistically significant differences in the number of positive test portions detected by the modified candidate method compared to the appropriate reference method. The MicroSEQ Salmonella spp. protocol linked with the Pathatrix individual or 10-Pooling procedure demonstrated reliability as a rapid, simplified, method for the preparation of samples and subsequent detection of Salmonella in diced tomatoes, chocolate, and deli ham.

Rapid detection of Salmonella in milk by electrochemical magneto-immunosensing.

PubMed

Liébana, Susana; Lermo, Anabel; Campoy, Susana; Cortés, María Pilar; Alegret, Salvador; Pividori, María Isabel

2009-10-15

A very simple and rapid method for the detection of Salmonella in milk is reported. In this approach, the bacteria are captured and preconcentrated from milk samples with magnetic beads through an immunological reaction. A second polyclonal antibody labeled with peroxidase is used as serological confirmation with electrochemical detection based on a magneto-electrode. The 'IMS/m-GEC electrochemical immunosensing' approach shows a limit of detection of 5 x 10(3) and 7.5 x 10(3)CFU mL(-1) in LB and in milk diluted 1/10 in LB broth, respectively, in 50 min without any pretreatment. If the skimmed-milk is preenriched for 6h, the method is able to detect as low as 1.4 CFU mL(-1), while if it is preenriched for 8h, as low as 0.108 x CFU mL(-1) (2.7 x CFU in 25 g of milk, in 5 samples of 5 mL) are detected accordingly with the legislation. Moreover, the method is able to clearly distinguish between food pathogenic bacteria such as Salmonella and Escherichia coli. The features of this approach are discussed and compared with classical culture methods.

Rapid identification of salmonella serotypes with stereo and hyperspectral microscope imaging Methods

USDA-ARS?s Scientific Manuscript database

The hyperspectral microscope imaging (HMI) method can reduce detection time within 8 hours including incubation process. The early and rapid detection with this method in conjunction with the high throughput capabilities makes HMI method a prime candidate for implementation for the food industry. Th...

Rapid Identification of Salmonella Serotypes with Stereo and Hyperspectral Microscope Imaging Methods

USDA-ARS?s Scientific Manuscript database

The hyperspectral microscope imaging (HMI) method can reduce detection time within 8 hours including incubation process. The early and rapid detection with this method in conjunction with the high throughput capabilities makes HMI method a prime candidate for implementation for the food industry. Th...

Multiplex surface plasmon resonance imaging platform for label-free detection of foodborne pathogens

USDA-ARS?s Scientific Manuscript database

Salmonellae are among the leading causes of foodborne outbreaks in the United States, and more rapid and efficient detection methods are needed. Surface plasmon resonance imaging (SPRi) is an emerging optical technique, which allows for rapid and label-free screening of multiple targets simultaneous...

Rectal swab sampling followed by an enrichment culture-based real-time PCR assay to detect Salmonella enterocolitis in children.

PubMed

Lin, L-H; Tsai, C-Y; Hung, M-H; Fang, Y-T; Ling, Q-D

2011-09-01

Although routine bacterial culture is the traditional reference standard method for the detection of Salmonella infection in children with diarrhoea, it is a time-consuming procedure that usually only gives results after 3-4 days. Some molecular detection methods can improve the turn-around time to within 24 h, but these methods are not applied directly from stool or rectal swab specimens as routine diagnostic methods for the detection of gastrointestinal pathogens. In this study, we tested the feasibility of a bacterial enrichment culture-based real-time PCR assay method for detecting and screening for diarrhoea in children caused by Salmonella. Our results showed that the minimum real-time PCR assay time required to detect enriched bacterial culture from a swab was 3 h. In all children with suspected Salmonella diarrhoea, the enrichment culture-based real-time PCR achieved 85.4% sensitivity and 98.1% specificity, as compared with the 53.7% sensitivity and 100% specificity of detection with the routine bacterial culture method. We suggest that rectal swab sampling followed by enrichment culture-based real-time PCR is suitable as a rapid method for detecting and screening for Salmonella in paediatric patients. © 2011 The Authors. Clinical Microbiology and Infection © 2011 European Society of Clinical Microbiology and Infectious Diseases.

Detection of viable Salmonella in ice cream by TaqMan real-time polymerase chain reaction assay combining propidium monoazide.

PubMed

Wang, Yuexia; Yang, Ming; Liu, Shuchun; Chen, Wanyi; Suo, Biao

2015-09-01

Real-time polymerase chain reaction (PCR) allows rapid detection of Salmonella in frozen dairy products, but it might cause a false positive detection result because it might amplify DNA from dead target cells as well. In this study, Salmonella-free frozen ice cream was initially inoculated with heat-killed Salmonella Typhimurium cells and stored at -18°C. Bacterial DNA extracted from the sample was amplified using TaqMan probe-based real-time PCR targeting the invA gene. Our results indicated that DNA from the dead cells remained stable in frozen ice cream for at least 20 days, and could produce fluorescence signal for real-time PCR as well. To overcome this limitation, propidium monoazide (PMA) was combined with real-time PCR. PMA treatment can effectively prevent PCR amplification from heat-killed Salmonella cells in frozen ice cream. The PMA real-time PCR assay can selectively detect viable Salmonella at as low as 10 3  CFU/mL. Combining 18 hours of pre-enrichment with the assay allows for the detection of viable Salmonella at 10 0  CFU/mL and avoiding the false-positive result of dead cells. The PMA real-time PCR assay provides an alternative specifically for detection of viable Salmonella in ice cream. However, when the PMA real-time PCR assay was evaluated in ice cream subjected to frozen storage, it obviously underestimated the contamination situation of viable Salmonella, which might lead to a false negative result. According to this result, the use of enrichment prior to PMA real-time PCR analysis remains as the more appropriate approach. Copyright © 2015. Published by Elsevier B.V.

Development and evaluation of probe based real time loop mediated isothermal amplification for Salmonella: A new tool for DNA quantification.

PubMed

Mashooq, Mohmad; Kumar, Deepak; Niranjan, Ankush Kiran; Agarwal, Rajesh Kumar; Rathore, Rajesh

2016-07-01

A one step, single tube, accelerated probe based real time loop mediated isothermal amplification (RT LAMP) assay was developed for detecting the invasion gene (InvA) of Salmonella. The probe based RT LAMP is a novel method of gene amplification that amplifies nucleic acid with high specificity and rapidity under isothermal conditions with a set of six primers. The whole procedure is very simple and rapid, and amplification can be obtained in 20min. Detection of gene amplification was accomplished by amplification curve, turbidity and addition of DNA binding dye at the end of the reaction results in colour difference and can be visualized under normal day light and in UV. The sensitivity of developed assay was found 10 fold higher than taqman based qPCR. The specificity of the RT LAMP assay was validated by the absence of any cross reaction with other members of enterobacteriaceae family and other gram negative bacteria. These results indicate that the probe based RT LAMP assay is extremely rapid, cost effective, highly specific and sensitivity and has potential usefulness for rapid Salmonella surveillance. Crown Copyright © 2016. Published by Elsevier B.V. All rights reserved.

Isolation and identification of Salmonella spp. in environmental water by molecular technology in Taiwan

NASA Astrophysics Data System (ADS)

Kuo, Chun Wei; Hao Huang, Kuan; Hsu, Bing Mu; Tsai, Hsien Lung; Tseng, Shao Feng; Shen, Tsung Yu; Kao, Po Min; Shen, Shu Min; Chen, Jung Sheng

2013-04-01

Salmonella spp. is one of the most important causal agents of waterborne diseases. The taxonomy of Salmonella is very complicated and its genus comprises more than 2,500 serotypes. The detection of Salmonella in environmental water samples by routines culture methods using selective media and characterization of suspicious colonies based on biochemical tests and serological assay are generally time consuming. To overcome this drawback, it is desirable to use effective method which provides a higher discrimination and more rapid identification about Salmonella in environmental water. The aim of this study is to investigate the occurrence of Salmonella using molecular technology and to identify the serovars of Salmonella isolates from 70 environmental water samples in Taiwan. The analytical procedures include membrane filtration, non-selective pre-enrichment, selective enrichment of Salmonella. After that, we isolated Salmonella strains by selective culture plates. Both selective enrichment and culture plates were detected by Polymerase Chain Reaction (PCR). Finally, the serovars of Salmonella were confirmed by using biochemical tests and serological assay. In this study, 15 water samples (21.4%) were identified as Salmonella by PCR. The positive water samples will further identify their serotypes by culture method. The presence of Salmonella in environmental water indicates the possibility of waterborne transmission in drinking watershed. Consequently, the authorities need to provide sufficient source protection and to maintain the system for disease prevention. Keywords: Salmonella spp., serological assay, PCR

Evaluation of 3M molecular detection assay (MDA) Salmonella for the detection of Salmonella in selected foods: collaborative study.

PubMed

Bird, Patrick; Fisher, Kiel; Boyle, Megan; Huffman, Travis; Benzinger, M Joseph; Bedinghaus, Paige; Flannery, Jonathan; Crowley, Erin; Agin, James; Goins, David; Benesh, DeAnn; David, John

2013-01-01

The 3M Molecular Detection Assay (MDA) Salmonella is used with the 3M Molecular Detection System for the detection of Salmonella spp. in food, food-related, and environmental samples after enrichment. The assay utilizes loop-mediated isothermal amplification to rapidly amplify Salmonella target DNA with high specificity and sensitivity, combined with bioluminescence to detect the amplification. The 3M MDA Salmonella method was compared using an unpaired study design in a multilaboratory collaborative study to the U.S. Department of Agriculture/Food Safety and Inspection Service-Microbiology Laboratory Guidebook (USDA/FSIS-MLG 4.05), Isolation and Identification of Salmonella from Meat, Poultry, Pasteurized Egg and Catfish Products for raw ground beef and the U.S. Food and Drug Administration/Bacteriological Analytical Manual (FDA/BAM) Chapter 5 Salmonella reference method for wet dog food following the current AOAC guidelines. A total of 20 laboratories participated. For the 3M MDA Salmonella method, raw ground beef was analyzed using 25 g test portions, and wet dog food was analyzed using 375 g test portions. For the reference methods, 25 g test portions of each matrix were analyzed. Each matrix was artificially contaminated with Salmonella at three inoculation levels: an uninoculated control level (0 CFU/test portion), a low inoculum level (0.2-2 CFU/test portion), and a high inoculum level (2-5 CFU/test portion). In this study, 1512 unpaired replicate samples were analyzed. Statistical analysis was conducted according to the probability of detection (POD). For the low-level raw ground beef test portions, the following dLPOD (difference between the POD of the reference and candidate method) values with 95% confidence intervals were obtained: -0.01 (-0.14, +0.12). For the low-level wet dog food test portions, the following dLPOD with 95% confidence intervals were obtained: -0.04 (-0.16, +0.09). No significant differences were observed in the number of positive samples detected by the 3M MDA Salmonella method versus either the USDA/FSIS-MLG or FDA/BAM methods.

Enzymatic Digestion for Improved Bacteria Separation from Leafy Green Vegetables.

PubMed

Wang, Danhui; Wang, Ziyuan; He, Fei; Kinchla, Amanda J; Nugen, Sam R

2016-08-01

An effective and rapid method for the separation of bacteria from food matrix remains a bottleneck for rapid bacteria detection for food safety. Bacteria can strongly attach to a food surface or internalize within the matrix, making their isolation extremely difficult. Traditional methods of separating bacteria from food routinely involve stomaching, blending, and shaking. However, these methods may not be efficient at removing all the bacteria from complex matrices. Here, we investigate the benefits of using enzyme digestion followed by immunomagnetic separation to isolate Salmonella from spinach and lettuce. Enzymatic digestion using pectinase and cellulase was able to break down the structure of the leafy green vegetables, resulting in the detachment and release of Salmonella from the leaves. Immunomagnetic separation of Salmonella from the liquefied sample allowed an additional separation step to achieve a more pure sample without leaf debris that may benefit additional downstream applications. We have investigated the optimal combination of pectinase and cellulase for the digestion of spinach and lettuce to improve sample detection yields. The concentrations of enzymes used to digest the leaves were confirmed to have no significant effect on the viability of the inoculated Salmonella. Results reported that the recovery of the Salmonella from the produce after enzyme digestion of the leaves was significantly higher (P < 0.05) than traditional sample preparation methods to separate bacteria (stomaching and manually shaking). The results demonstrate the potential for use of enzyme digestion prior to separation can improve the efficiency of bacteria separation and increase the likelihood of detecting pathogens in the final detection assay.

Next day Salmonella spp. detection method based on real-time PCR for meat, dairy and vegetable food products.

PubMed

Rodriguez-Lazaro, David; Gonzalez-García, Patricia; Delibato, Elisabetta; De Medici, Dario; García-Gimeno, Rosa Maria; Valero, Antonio; Hernandez, Marta

2014-08-01

The microbiological standard for detection of Salmonella relies on several cultural steps and requires more than 5 days for final confirmation, and as consequence there is a need for an alternative rapid methodology for its detection. The aim of this study was to compare different detection strategies based on real-time PCR for a rapid and sensitive detection in an ample range of food products: raw pork and poultry meat, ready to eat lettuce salad and raw sheep milk cured cheese. Three main parameters were evaluated to reduce the time and cost for final results: the initial sample size (25 and 50 g), the incubation times (6, 10 and 18 h) and the bacterial DNA extraction (simple boiling of the culture after washing the bacterial pellet, the use of the Chelex resin, and a commercial silica column). The results obtained demonstrate that a combination of an incubation in buffered peptone water for 18 h of a 25 g-sample coupled to a DNA extraction by boiling and a real-time PCR assay detected down to 2-4 Salmonella spp.CFU per sample in less than 21 h in different types of food products. This RTi-PCR-based method is fully compatible with the ISO standard, providing results more rapidly and cost-effectively. The results were confirmed in a large number of naturally contaminated food samples with at least the same analytical performance as the reference method. Copyright © 2014 Elsevier B.V. All rights reserved.

Specificity tests of an oligonucleotide probe against food-outbreak salmonella for biosensor detection

NASA Astrophysics Data System (ADS)

Chen, I.-H.; Horikawa, S.; Xi, J.; Wikle, H. C.; Barbaree, J. M.; Chin, B. A.

2017-05-01

Phage based magneto-elastic (ME) biosensors have been shown to be able to rapidly detect Salmonella in various food systems to serve food pathogen monitoring purposes. In this ME biosensor platform, the free-standing strip-shaped magneto-elastic sensor is the transducer and the phage probe that recognizes Salmonella in food serves as the bio-recognition element. According to Sorokulova et al. at 2005, a developed oligonucleotide probe E2 was reported to have high specificity to Salmonella enterica Typhimurium. In the report, the specificity tests were focused in most of Enterobacterace groups outside of Salmonella family. Here, to understand the specificity of phage E2 to different Salmonella enterica serotypes within Salmonella Family, we further tested the specificity of the phage probe to thirty-two Salmonella serotypes that were present in the major foodborne outbreaks during the past ten years (according to Centers for Disease Control and Prevention). The tests were conducted through an Enzyme linked Immunosorbent Assay (ELISA) format. This assay can mimic probe immobilized conditions on the magnetoelastic biosensor platform and also enable to study the binding specificity of oligonucleotide probes toward different Salmonella while avoiding phage/ sensor lot variations. Test results confirmed that this oligonucleotide probe E2 was high specific to Salmonella Typhimurium cells but showed cross reactivity to Salmonella Tennessee and four other serotypes among the thirty-two tested Salmonella serotypes.

Recombinase polymerase amplification combined with lateral flow dipstick for equipment-free detection of Salmonella in shellfish.

PubMed

Gao, Weifang; Huang, Hailong; Zhu, Peng; Yan, Xiaojun; Fan, Jianzhong; Jiang, Jinpo; Xu, Jilin

2018-05-01

Salmonella is a major pathogen that causes acute foodborne outbreaks worldwide. Seafood, particularly shellfish, is a proven source of Salmonella spp. infection because many people prefer to eat it raw or lightly cooked. However, traditional identification methods are too time-consuming and complex to detect contamination of bacteria in the food chain in a timely manner, and few studies have aimed to identify Salmonella in shellfish early in the supply chain. We herein developed a method for rapid detection of Salmonella in shellfish based on the method of recombinase polymerase amplification (RPA) combined with lateral flow dipstick (LFD), which targets the invasion gene A (invA). The RPA-LFD was able to function at 30-45 °C, and at the temperature of 40 °C, it only took 8 min of amplification to reach the test threshold of amplicons. The established method had both a good specificity and a sensitivity of 100 fg DNA per reaction (20 µL). Regarding practical performance, RPA-LFD performed better than real-time PCR. Another advantage of RPA-LFD is that it was capable of being performed without expensive equipments. Thus, RPA-LFD has potential for further development as a detection kit for Salmonella in shellfish and other foods under field conditions.

Recombinant plasmid-based quantitative Real-Time PCR analysis of Salmonella enterica serotypes and its application to milk samples.

PubMed

Gokduman, Kurtulus; Avsaroglu, M Dilek; Cakiris, Aris; Ustek, Duran; Gurakan, G Candan

2016-03-01

The aim of the current study was to develop, a new, rapid, sensitive and quantitative Salmonella detection method using a Real-Time PCR technique based on an inexpensive, easy to produce, convenient and standardized recombinant plasmid positive control. To achieve this, two recombinant plasmids were constructed as reference molecules by cloning the two most commonly used Salmonella-specific target gene regions, invA and ttrRSBC. The more rapid detection enabled by the developed method (21 h) compared to the traditional culture method (90 h) allows the quantitative evaluation of Salmonella (quantification limits of 10(1)CFU/ml and 10(0)CFU/ml for the invA target and the ttrRSBC target, respectively), as illustrated using milk samples. Three advantages illustrated by the current study demonstrate the potential of the newly developed method to be used in routine analyses in the medical, veterinary, food and water/environmental sectors: I--The method provides fast analyses including the simultaneous detection and determination of correct pathogen counts; II--The method is applicable to challenging samples, such as milk; III--The method's positive controls (recombinant plasmids) are reproducible in large quantities without the need to construct new calibration curves. Copyright © 2016 Elsevier B.V. All rights reserved.

Comparative Evaluation of Veriflow® Salmonella Species to USDA and FDA Culture-Based Methods for the Detection of Salmonella spp. in Food and Environmental Samples.

PubMed

Puri, Amrita; Joelsson, Adam C; Terkhorn, Shawn P; Brown, Ashley S; Gaudioso, Zara E; Siciliano, Nicholas A

2017-09-01

Veriflow® Salmonella species (Veriflow SS) is a molecular-based assay for the presumptive detection of Salmonella spp. from environmental surfaces (stainless steel, sealed concrete, plastic, and ceramic tile), dairy (2% milk), raw meat (20% fat ground beef), chicken carcasses, and ready-to-eat (RTE) food (hot dogs). The assay utilizes a PCR detection method coupled with a rapid, visual, flow-based assay that develops in 3 min post-PCR amplification and requires only an 18 h enrichment for maximum sensitivity. The Veriflow SS system eliminates the need for sample purification, gel electrophoresis, or fluorophore-based detection of target amplification and does not require complex data analysis. This Performance Tested MethodSM validation study demonstrated the ability of the Veriflow SS method to detect low levels of artificially inoculated or naturally occurring Salmonella spp. in eight distinct environmental and food matrixes. In each reference comparison study, probability of detection analysis indicated that there was no significant difference between the Veriflow SS method and the U.S. Department of Agriculture Food Safety and Inspection Service Microbiology Laboratory Guidebook Chapter 4.06 and the U.S. Food and Drug Administration Bacteriological Analytical Manual Chapter 5 reference methods. A total of 104 Salmonella strains were detected in the inclusivity study, and 35 nonspecific organisms went undetected in the exclusivity study. The study results show that the Veriflow SS method is a sensitive, selective, and robust assay for the presumptive detection of Salmonella spp. sampled from environmental surfaces (stainless steel, sealed concrete, plastic, and ceramic tile), dairy (2% milk), raw meat (20% fat ground beef), chicken carcasses, and RTE food (hot dogs).

Detection and Identification of Salmonella spp. in Surface Water by Molecular Technology in Taiwan

NASA Astrophysics Data System (ADS)

Tseng, S. F.; Hsu, B. M.; Huang, K. H.; Hsiao, H. Y.; Kao, P. M.; Shen, S. M.; Tsai, H. F.; Chen, J. S.

2012-04-01

Salmonella spp. is classified to gram-negative bacterium and is one of the most important causal agents of waterborne diseases. The genus of Salmonella comprises more than 2,500 serotypes and its taxonomy is also very complicated. In tradition, the detection of Salmonella in environmental water samples by routines culture methods using selective media and characterization of suspicious colonies based on biochemical tests and serological assay are generally time and labor consuming. To overcome this disadvantage, it is desirable to use effective method which provides a higher discrimination and more rapid identification about Salmonella in environmental water. The aim of this study is to investigate the occurrence of Salmonella using novel procedures of detection method and to identify the serovars of Salmonella isolates from 157 surface water samples in Taiwan. The procedures include membrane filtration, non-selective pre-enrichment, selective enrichment of Salmonella, and then isolation of Salmonella strains by selective culture plates. The selective enrichment and culture plates were both detected by PCR. Finally, we used biochemical tests and serological assay to confirm the serovars of Salmonella and also used Pulsed-field gel electrophoresis (PFGE) to identify their sarovar catagories by the genetic pattern. In this study, 44 water samples (28%) were indentified as Salmonella. The 44 positive water samples by culture method were further identified as S. Agona(1/44), S. Albany (10/44), S. Bareilly (13/44),S. Choleraesuis (2/44),S. Derby (4/44),S. Isangi (3/44),S.Kedougou(3/44),S. Mbandaka(1/44),S.Newport (3/44), S. Oranienburg(1/44), S. Potsdam (1/44),S. Typhimurium (1/44), andS. Weltevreden(1/44) by PFGE. The presence of Salmonella in surface water indicates the possibility of waterborne transmission in drinking watershed if water is not adequately treated. Therefore, the authorities need to have operating systems that currently provide adequate source protection and maintaining the system to prevent disease. Keywords: Salmonella spp.; biochemical tests; Serological assay; PCR; PFGE

Rapid biochemical screening for Salmonella, Shigella, Yersinia, and Aeromonas isolates from stool specimens.

PubMed Central

De Ryck, R; Struelens, M J; Serruys, E

1994-01-01

Four screens for the rapid (4 to 6 h) biochemical detection of pathogens from enteric isolation media are described. The Salmonella screen consisted of Kligler iron agar (KIA), motility-indole-urea-tryptophan-deamination semisolid medium (MIU-TDA), and the o-nitrophenyl-beta-D-galactopyranoside (ONPG) test; the Shigella screen consisted of KIA, MIU-TDA, the ONPG test, and the lysine decarboxylation-indole test; the Yersinia screen consisted of a rhamnose broth; the Aeromonas screen consisted of a xylose agar plate. When tested on 2,102 fresh isolates and 71 stock strains, the screens correctly detected 212 enteric pathogens (sensitivity, 100%), with a specificity of 98.1%. PMID:8077408

Biomagnetic separation of Salmonella Typhimurium with high affine and specific ligand peptides isolated by phage display technique

NASA Astrophysics Data System (ADS)

Steingroewer, Juliane; Bley, Thomas; Bergemann, Christian; Boschke, Elke

2007-04-01

Analyses of food-borne pathogens are of great importance in order to minimize the health risk for customers. Thus, very sensitive and rapid detection methods are required. Current conventional culture techniques are very time consuming. Modern immunoassays and biochemical analysis also require pre-enrichment steps resulting in a turnaround time of at least 24 h. Biomagnetic separation (BMS) is a promising more rapid method. In this study we describe the isolation of high affine and specific peptides from a phage-peptide library, which combined with BMS allows the detection of Salmonella spp. with a similar sensitivity as that of immunomagnetic separation using antibodies.

Detection of Salmonella spp. in veterinary samples by combining selective enrichment and real-time PCR.

PubMed

Goodman, Laura B; McDonough, Patrick L; Anderson, Renee R; Franklin-Guild, Rebecca J; Ryan, James R; Perkins, Gillian A; Thachil, Anil J; Glaser, Amy L; Thompson, Belinda S

2017-11-01

Rapid screening for enteric bacterial pathogens in clinical environments is essential for biosecurity. Salmonella found in veterinary hospitals, particularly Salmonella enterica serovar Dublin, can pose unique challenges for culture and testing because of its poor growth. Multiple Salmonella serovars including Dublin are emerging threats to public health given increasing prevalence and antimicrobial resistance. We adapted an automated food testing method to veterinary samples and evaluated the performance of the method in a variety of matrices including environmental samples ( n = 81), tissues ( n = 52), feces ( n = 148), and feed ( n = 29). A commercial kit was chosen as the basis for this approach in view of extensive performance characterizations published by multiple independent organizations. A workflow was established for efficiently and accurately testing veterinary matrices and environmental samples by use of real-time PCR after selective enrichment in Rappaport-Vassiliadis soya (RVS) medium. Using this method, the detection limit for S. Dublin improved by 100-fold over subculture on selective agars (eosin-methylene blue, brilliant green, and xylose-lysine-deoxycholate). Overall, the procedure was effective in detecting Salmonella spp. and provided next-day results.

Real-time and rapid detection of Salmonella Typhimurium using an inexpensive lab-built surface plasmon resonance setup

NASA Astrophysics Data System (ADS)

Lukose, Jijo; Shetty, Vignesh; Ballal, Mamatha; Chidangil, Santhosh; Sinha, Rajeev K.

2018-07-01

Cost-effective diagnostic platforms for rapid pathogen detection are always incumbent in both developing and developed worlds. However, exorbitant diagnostic expenses and the inability to detect pathogens early are a matter of concern for the sustainability and affordability of healthcare devices, which are crucial for deciding how to provide healthcare solutions to the masses, especially in developing countries. Herein, we present the rapid and real-time detection of Salmonella Typhimurium using an inexpensive lab-built surface plasmon resonance (SPR) imaging set up. Pathogen detection is accomplished with the aid of a monoclonal antibody immobilized on a 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide): N-hydroxysuccinimide-modified self-assembled monolayer covalently bonded to a Au thin film. Successful pathogen detection is performed at two concentrations, ~1.5  ×  108 and ~1  ×  106 cfu ml‑1, in phosphate-buffered saline solution. The developed system is capable of detecting bacterial cells within 6–7 min after their injection into the SPR sensor surface. The present study reveals a cost-effective device having high potential for pathogen detection without any labelling tags.

Rapid detection of bacterial pathogens using flourescence spectroscopy and chemometrics

USDA-ARS?s Scientific Manuscript database

This work presents the development of a method for rapid bacterial identification based on the fluorescence spectroscopy combined with multivariate analysis. Fluorescence spectra of pure three different genera of bacteria (Escherichia coli, Salmonella, and Campylobacter) were collected from 200...

Rapid and Sensitive Detection of Shigella spp. and Salmonella spp. by Multiple Endonuclease Restriction Real-Time Loop-Mediated Isothermal Amplification Technique

PubMed Central

Wang, Yi; Wang, Yan; Luo, Lijuan; Liu, Dongxin; Luo, Xia; Xu, Yanmei; Hu, Shoukui; Niu, Lina; Xu, Jianguo; Ye, Changyun

2015-01-01

Shigella and Salmonella are frequently isolated from various food samples and can cause human gastroenteritis. Here, a novel multiple endonuclease restriction real-time loop-mediated isothermal amplification technology (MERT-LAMP) were successfully established and validated for simultaneous detection of Shigella strains and Salmonella strains in only a single reaction. Two sets of MERT-LAMP primers for 2 kinds of pathogens were designed from ipaH gene of Shigella spp. and invA gene of Salmonella spp., respectively. Under the constant condition at 63°C, the positive results were yielded in as short as 12 min with the genomic DNA extracted from the 19 Shigella strains and 14 Salmonella strains, and the target pathogens present in a sample could be simultaneously identified based on distinct fluorescence curves in real-time format. Accordingly, the multiplex detection assay significantly reduced effort, materials and reagents used, and amplification and differentiation were conducted at the same time, obviating the use of postdetection procedures. The analytical sensitivity of MERT-LAMP was found to be 62.5 and 125 fg DNA/reaction with genomic templates of Shigella strains and Salmonella strains, which was consist with normal LAMP assay, and at least 10- and 100-fold more sensitive than that of qPCR and conventional PCR approaches. The limit of detection of MERT-LAMP for Shigella strains and Salmonella strains detection in artificially contaminated milk samples was 5.8 and 6.4 CFU per vessel. In conclusion, the MERT-LAMP methodology described here demonstrated a potential and valuable means for simultaneous screening of Shigella and Salmonella in a wide variety of samples. PMID:26697000

Development of Multiplex Real-time PCR with Internal Amplification Control for Simultaneous Detection of Salmonella and Cronobacter sakazakii in Powdered Infant Formula.

USDA-ARS?s Scientific Manuscript database

Contamination of powdered infant formula (PIF) by the bacteria Cronobacter sakazakii and Salmonella enterica was deemed a matter of great concern by the World Health Organization and the Food and Agriculture Organization of the United Nations in 2004. Therefore, we developed a rapid and sensitive m...

Rapid detection of salmonella using SERS with silver nano-substrate

NASA Astrophysics Data System (ADS)

Sundaram, J.; Park, B.; Hinton, A., Jr.; Windham, W. R.; Yoon, S. C.; Lawrence, K. C.

2011-06-01

Surface Enhanced Raman Scattering (SERS) can detect the pathogen in rapid and accurate. In SERS weak Raman scattering signals are enhanced by many orders of magnitude. In this study silver metal with biopolymer was used. Silver encapsulated biopolymer polyvinyl alcohol nano-colloid was prepared and deposited on stainless steel plate. This was used as metal substrate for SERS. Salmonella typhimurium a common food pathogen was selected for this study. Salmonella typhimurium bacteria cells were prepared in different concentrations in cfu/mL. Small amount of these cells were loaded on the metal substrate individually, scanned and spectra were recorded using confocal Raman microscope. The cells were exposed to laser diode at 785 nm excitation and object 50x was used to focus the laser light on the sample. Raman shifts were obtained from 400 to 2400 cm-1. Multivariate data analysis was carried to predict the concentration of unknown sample using its spectra. Concentration prediction gave an R2 of 0.93 and standard error of prediction of 0.21. The results showed that it could be possible to find out the Salmonella cells present in a low concentration in food samples using SERS.

Comparison of reverse transcriptase PCR, reverse transcriptase loop-mediated isothermal amplification, and culture-based assays for Salmonella detection from pork processing environments.

PubMed

Techathuvanan, Chayapa; Draughon, Frances Ann; D'Souza, Doris Helen

2011-02-01

Novel rapid Salmonella detection assays without the need for sophisticated equipment or labor remain in high demand. Real-time reverse transcriptase PCR (RT-PCR) assays, though rapid and sensitive, require expensive thermocyclers, while a novel RT loop-mediated isothermal amplification (RT-LAMP) method requires only a simple water bath. Our objective was to compare the detection sensitivity of Salmonella Typhimurium from the pork processing environment by RT-LAMP, RT-PCR, and culture-based assays. Carcass and surface swabs and carcass rinses were obtained from a local processing plant. Autoclaved carcass rinses (500 ml) were spiked with Salmonella Typhimurium and filtered. Filters were placed in stomacher bags containing tetrathionate broth (TTB) and analyzed with or without 10-h enrichment at 37 °C. Natural swabs were stomached with buffered peptone water, and natural carcass rinses were filtered, preenriched, and further enriched in TTB. Serially-diluted enriched samples were enumerated by spread plating on xylose lysine Tergitol 4 agar. RNA was extracted from 5 ml of enriched TTB with TRIzol. RT-LAMP assay using previously described invA primers was conducted at 62 °C for 90 min in a water bath with visual detection and by gel electrophoresis. SYBR Green I-based-real-time RT-PCR was carried out with invA primers followed by melt temperature analysis. The results of RT-LAMP detection for spiked carcass rinses were comparable to those of RT-PCR and cultural plating, with detection limits of 1 log CFU/ml, although they were obtained significantly faster, within 24 h including preenrichment and enrichment. RT-LAMP showed 4 of 12 rinse samples positive, while RT-PCR showed 1 of 12 rinse samples positive. For swabs, 6 of 27 samples positive by RT-LAMP and 5 of 27 by RT-PCR were obtained. This 1-day RT-LAMP assay shows promise for routine Salmonella screening by the pork industry. Copyright ©, International Association for Food Protection

In vitro selection of RNA aptamer specific to Salmonella typhimurium.

PubMed

Han, Seung Ryul; Lee, Seong-Wook

2013-06-28

Salmonella is a major foodborne pathogen that causes a variety of human diseases. Development of ligands directly and specifically binding to the Salmonella will be crucial for the rapid detection of, and thus for efficient protection from, the virulent bacteria. In this study, we identified a RNA aptamer-based ligand that can specifically recognize Salmonella Typhimurium through SELEX technology. To this end, we isolated and characterized an RNase-resistant RNA aptamer that bound to the OmpC protein of Salmonella Typhimurium with high specificity and affinity (Kd ~ 20 nM). Of note, the selected aptamer was found to specifically bind to Salmonella Typhimurium, but neither to Gram-positive bacteria (Staphylococcus aureus) nor to other Gram-negative bacteria (Escherichia coli O157:H7). This was evinced by aptamer-immobilized ELISA and aptamer-linked precipitation experiments. This Salmonella species-specific aptamer could be useful as a diagnostic ligand against pathogen-caused foodborne sickness.

Detection and characterization of foodborne pathogenic bacteria with hyperspectral microscope imaging

USDA-ARS?s Scientific Manuscript database

Rapid detection and identification of pathogenic microorganisms naturally occurring during food processing are important in developing intervention and verification strategies. In the poultry industry, contamination of poultry meat with foodborne pathogens (especially, Salmonella and Campylobacter) ...

Magnetic focusing immunosensor for the detection of Salmonella typhimurium in foods

NASA Astrophysics Data System (ADS)

Pivarnik, Philip E.; Cao, He; Letcher, Stephen V.; Pierson, Arthur H.; Rand, Arthur G.

1999-01-01

From 1988 through 1992 Salmonellosis accounted for 27% of the total reported foodborne disease outbreaks and 57% of the outbreaks in which the pathogen was identified. The prevalence of Salmonellosis and the new requirements to monitor the organism as a marker in pathogen reduction programs will drive the need for rapid, on-site testing. A compact fiber optic fluorometer using a red diode laser as an excitation source and fiber probes for analyte detection has been constructed and used to measure Salmonella. The organisms were isolated with anti-Salmonella magnetic beads and were labeled with a secondary antibody conjugated to a red fluorescent dye. The response of the system was proportional to the concentration of Salmonella typhimurium from 3.2 X 105 colony forming units (CFU)/ml to 1.6 X 107 CFU/ml. The system was developed to utilize a fiber-optic magnetic focusing problem that attracted the magnetic microspheres to the surface of a sample chamber directly in front of the excitation and emission fibers. The signal obtained from a homogenous suspension of fluorescent magnetic microspheres was 9 to 10 picowatts. After focusing, the signal from the fluorescent labeled magnetic microspheres increased to 200 picowatts, approximately 20 times greater than the homogeneous suspension. The magnetic focusing assay detected 1.59 X 105 colony forming units/ml of Salmonella typhimurium cultured in growth media. The process of magnetic focusing in front of the fibers has the potential to reduce the background fluorescence from unbound secondary antibodies, eliminating several rinsing steps, resulting in a simple rapid assay.

A rapid method to identify Salmonella enterica serovar Gallinarum biovar Pullorum using a specific target gene ipaJ.

PubMed

Xu, Lijuan; Liu, Zijian; Li, Yang; Yin, Chao; Hu, Yachen; Xie, Xiaolei; Li, Qiuchun; Jiao, Xinan

2018-06-01

Salmonella enterica serovar Gallinarum biovar Pullorum (S. Pullorum) is the pathogen of pullorum disease, which leads to severe economic losses in many developing countries. Traditional methods to identify S. enterica have relied on biochemical reactions and serotyping, which are time-consuming with accurate identification if properly carried out. In this study, we developed a rapid polymerase chain reaction (PCR) method targeting the specific gene ipaJ to detect S. Pullorum. Among the 650 S. Pullorum strains isolated from 1962 to 2016 all over China, 644 strains were identified to harbour ipaJ gene in the plasmid pSPI12, accounting for a detection rate of 99.08%. Six strains were ipaJ negative because pSPI12 was not found in these strains according to whole genome sequencing results. There was no cross-reaction with other Salmonella serotypes, including Salmonella enterica serovar Gallinarum biovar Gallinarum (S. Gallinarum), which show a close genetic relationship with S. Pullorum. This shows that the PCR method could distinguish S. Gallinarum from S. Pullorum in one-step PCR without complicated biochemical identification. The limit of detection of this PCR method was as low as 90 fg/μl or 10 2 CFU, which shows a high sensitivity. Moreover, this method was applied to identify Salmonella isolated from the chicken farm and the results were consistent with what we obtained from biochemical reactions and serotyping. Together, all the results demonstrated that this one-step PCR method is simple and feasible to efficiently identify S. Pullorum.

Evaluation of Modification of the 3M™ Molecular Detection Assay (MDA) Salmonella Method (2013.09) for the Detection of Salmonella in Selected Foods: Collaborative Study.

PubMed

Bird, Patrick; Fisher, Kiel; Boyle, Megan; Huffman, Travis; Benzinger, M Joseph; Bedinghaus, Paige; Flannery, Jonathon; Crowley, Erin; Agin, James; Goins, David; Benesh, DeAnn; David, John

2014-01-01

The 3M(™) Molecular Detection Assay (MDA) Salmonella utilizes isothermal amplification of nucleic acid sequences with high specificity, efficiency, rapidity and bioluminescence to detect amplification of Salmonella spp. in food, food-related, and environmental samples after enrichment. A method modification and matrix extension study of the previously approved AOAC Official Method(SM) 2013.09 was conducted, and approval of the modification was received on March 20, 2014. Using an unpaired study design in a multilaboratory collaborative study, the 3M MDA Salmonella method was compared to the U.S. Department of Agriculture/Food Safety and Inspection Service (USDA/FSIS) Microbiology Laboratory Guidebook (MLG) 4.05 (2011), Isolation and Identification of Salmonella from Meat, Poultry, Pasteurized Egg, and Catfish Products for raw ground beef and the U.S. Food and Drug Administration (FDA)/Bacteriological Analytical Manual (BAM) Chapter 5, Salmonella reference method for wet dog food following the current AOAC guidelines. A total of 20 laboratories participated. For the 3M MDA Salmonella method, raw ground beef was analyzed using 25 g test portions, and wet dog food was analyzed using 375 g test portions. For the reference methods, 25 g test portions of each matrix were analyzed. Each matrix was artificially contaminated with Salmonella at three inoculation levels: an uninoculated control level (0 CFU/test portion), a low inoculum level (0.2-2 CFU/test portion), and a high inoculum level (2-5 CFU/test portion). In this study, 1512 unpaired replicate samples were analyzed. Statistical analysis was conducted according to the probability of detection (POD). For the low-level raw ground beef test portions, the following dLPOD (difference between the LPODs of the reference and candidate method) values with 95% confidence intervals were obtained: -0.01 (-0.14, +0.12). For the low-level wet dog food test portions, the following dLPOD with 95% confidence intervals were obtained: -0.04 (-0.16, +0.09). No significant differences were observed in the number of positive samples detected by the 3M MDA Salmonella method versus either the USDA/FSIS-MLG or FDA/BAM methods.

Rapid, sensitive, and simultaneous detection of three foodborne pathogens using magnetic nanobead-based immunoseparation and quantum dot-based multiplex immunoassay.

PubMed

Wang, Hong; Li, Yanbin; Wang, Andrew; Slavik, Michael

2011-12-01

Losses caused by foodborne diseases are enormous in terms of human life, illness, medical costs, and food product recalls. Rapid detection of multiple bacterial pathogens in foods is extremely important to ensure food safety. The objective of this research was to develop a multiplex immunoassay by integrating magnetic nanobeads (MNBs) for immunoseparation with quantum dots (QDs) as fluorescent labels for rapid, sensitive, and simultaneous detection of three major pathogenic bacteria, Salmonella Typhimurium, Escherichia coli O157:H7, and Listeria monocytogenes, in food products. In this research, both streptavidin-conjugated MNBs (30- and 150-nm diameter) and QDs (530-, 580-, and 620-nm emission wavelength) were separately coated with biotinylated anti-Salmonella, anti-E. coli, and anti-Listeria antibodies. The immuno-MNBs were mixed with a food sample to capture the three target bacteria. After being magnetically separated from the sample, the MNB-cell conjugates were mixed with the immuno-QDs to form the MNB-cell-QD complexes, and unattached QDs were removed. The fluorescence intensity of the MNB-cell-QD complexes was measured at wavelengths of 530, 580, and 620 nm to determine the populations of Salmonella Typhimurium, E. coli O157:H7, and L. monocytogenes, respectively. This multiplex immunoassay simultaneously detected Salmonella Typhimurium, E. coli O157:H7, and L. monocytogenes at levels as low as 20 to 50 CFU/ml in food samples in less than 2 h without enrichment. The change in fluorescence intensity was linearly correlated (R(2) > 0.96) with the logarithmic value of bacterial level in the range of 10 to 10(3) CFU/ml. More than 85% of the three target pathogens could be simultaneously separated from food samples. The multiplex immunoassay could be expanded to detect more target pathogens, depending on the availability of specific antibodies and QDs with different emission wavelengths.

Characteristics of Clusters of Salmonella and Escherichia coli O157 Detected by Pulsed-Field Gel Electrophoresis that Predict Identification of Outbreaks.

PubMed

Jones, Timothy F; Sashti, Nupur; Ingram, Amanda; Phan, Quyen; Booth, Hillary; Rounds, Joshua; Nicholson, Cyndy S; Cosgrove, Shaun; Crocker, Kia; Gould, L Hannah

2016-12-01

Molecular subtyping of pathogens is critical for foodborne disease outbreak detection and investigation. Many clusters initially identified by pulsed-field gel electrophoresis (PFGE) are not confirmed as point-source outbreaks. We evaluated characteristics of clusters that can help prioritize investigations to maximize effective use of limited resources. A multiagency collaboration (FoodNet) collected data on Salmonella and Escherichia coli O157 clusters for 3 years. Cluster size, timing, extent, and nature of epidemiologic investigations were analyzed to determine associations with whether the cluster was identified as a confirmed outbreak. During the 3-year study period, 948 PFGE clusters were identified; 849 (90%) were Salmonella and 99 (10%) were E. coli O157. Of those, 192 (20%) were ultimately identified as outbreaks (154 [18%] of Salmonella and 38 [38%] of E. coli O157 clusters). Successful investigation was significantly associated with larger cluster size, more rapid submission of isolates (e.g., for Salmonella, 6 days for outbreaks vs. 8 days for nonoutbreaks) and PFGE result reporting to investigators (16 days vs. 29 days, respectively), and performance of analytic studies (completed in 33% of Salmonella outbreaks vs. 1% of nonoutbreaks) and environmental investigations (40% and 1%, respectively). Intervals between first and second cases in a cluster did not differ significantly between outbreaks and nonoutbreaks. Molecular subtyping of pathogens is a rapidly advancing technology, and successfully identifying outbreaks will vary by pathogen and methods used. Understanding criteria for successfully investigating outbreaks is critical for efficiently using limited resources.

Evaluation of 3M molecular detection system and ANSR pathogen detection system for rapid detection of salmonella from egg products

USDA-ARS?s Scientific Manuscript database

Loop-mediated isothermal amplification (LAMP) is a novel simple detection technology that amplifies DNA with high speed, efficiency, and specificity under isothermal conditions. The objective of this study was to evaluate the effectiveness of 3M Molecular Detection System (MDS) and ANSR Pathogen Det...

Inclusivity, exclusivity and limit of detection of commercially available real-time PCR assays for the detection of Salmonella.

PubMed

Margot, H; Stephan, R; Guarino, S; Jagadeesan, B; Chilton, D; O'Mahony, E; Iversen, C

2013-08-01

The traditional cultural detection of Salmonella spp. is both time- and labour-intensive. Salmonella is often a release criterion for the food industry and time to result is therefore an important factor. Storage of finished products and raw materials can be costly and may adversely impact available shelf-life. The application of real-time PCR for the detection of Salmonella spp. in food samples enables a potential time-saving of up to four days. The advancement of real-time PCR coupled with the development of commercially available systems in different formats has made this technology accessible for laboratories in an industrial environment. Ideally these systems are reliable and rapid as well as easy to use. The current study represents a comparative evaluation of seven commercial real-time PCR systems for the detection of Salmonella. Forty-nine target and twenty-nine non-target strains were included in the study to assess inclusivity and exclusivity. The limit of detection for each of the method was determined in four different food products. All systems evaluated were able to correctly identify the 49 Salmonella strains. Nevertheless, false positive results (Citrobacter spp.) were obtained with four of the seven systems. In milk powder and bouillon powder, the limit of detection was similar for all systems, suggesting a minimal matrix effect with these samples. Conversely, for black tea and cocoa powder some systems were prone to inhibition from matrix components. Up to 100% of the samples were inhibited using the proprietary extracts but inhibition could be reduced considerably by application of a DNA clean-up kit. Copyright © 2013 Elsevier B.V. All rights reserved.

Evaluation of detection methods for screening meat and poultry products for the presence of foodborne pathogens.

PubMed

Bohaychuk, Valerie M; Gensler, Gary E; King, Robin K; Wu, John T; McMullen, Lynn M

2005-12-01

Rapid and molecular technologies such as enzyme-linked immunosorbent assay (ELISA), PCR, and lateral flow immunoprecipitation can reduce the time and labor involved in screening food products for the presence of pathogens. These technologies were compared with conventional culture methodology for the detection of Salmonella, Campylobacter, Listeria, and Escherichia coli O157:H7 inoculated in raw and processed meat and poultry products. Recommended protocols were modified so that the same enrichment broths used in the culture methods were also used in the ELISA, PCR, and lateral flow immunoprecipitation assays. The percent agreement between the rapid technologies and culture methods ranged from 80 to 100% depending on the pathogen detected and the method used. ELISA, PCR, and lateral flow immunoprecipitation all performed well, with no statistical difference, compared with the culture method for the detection of E. coli O157:H7. ELISA performed better for the detection of Salmonella, with sensitivity and specificity rates of 100%. PCR performed better for the detection of Campylobacter jejuni, with 100% agreement to the culture method. PCR was highly sensitive for the detection of all the foodborne pathogens tested except Listeria monocytogenes. Although the lateral flow immunoprecipitation tests were statistically different from the culture methods for Salmonella and Listeria because of false-positive results, the tests did not produce any false negatives, indicating that this method would be suitable for screening meat and poultry products for these pathogens.

A magneto-DNA nanoparticle system for the rapid and sensitive diagnosis of enteric fever

PubMed Central

Park, Ki Soo; Chung, Hyun Jung; Khanam, Farhana; Lee, Hakho; Rashu, Rasheduzzaman; Bhuiyan, Md. Taufiqur; Berger, Amanda; Harris, Jason B.; Calderwood, Stephen B.; Ryan, Edward T.; Qadri, Firdausi; Weissleder, Ralph; Charles, Richelle C.

2016-01-01

There is currently no widely available optimal assay for diagnosing patients with enteric fever. Here we present a novel assay designed to detect amplified Salmonella nucleic acid (mRNA) using magneto-DNA probes and a miniaturized nuclear magnetic resonance device. We designed primers for genes specific to S. Typhi, S. Paratyphi A, and genes conserved among Salmonella enterica spp. and utilized strongly magnetized nanoparticles to enhance the detection signal. Blood samples spiked with in vitro grown S. Typhi, S. Paratyphi A, S. Typhimurium, and E. coli were used to confirm the specificity of each probe-set, and serial 10-fold dilutions were used to determine the limit of the detection of the assay, 0.01–1.0 CFU/ml. For proof of principle, we applied our assay to 0.5 mL blood samples from 5 patients with culture-confirmed enteric fever from Bangladesh in comparison to 3 healthy controls. We were able to detect amplified target cDNA in all 5 cases of enteric fever; no detectable signal was seen in the healthy controls. Our results suggest that a magneto-DNA nanoparticle system, with an assay time from blood collection of 3.5 hours, may be a promising platform for the rapid and culture-free diagnosis of enteric fever and non-typhoidal Salmonella bacteremia. PMID:27605393

[Sensitivity of three inmunocromathographic tests in faeces samples for Campylobacter and Salmonella detection in comparison to culture].

PubMed

Liébana-Martos, Ma del Carmen; Gutierrez, José; Riazzo, Cristina; Navarro, José Ma

2014-06-01

Introduction: Campylobacter sp. and Salmonella enterica are two of the main organisms causing gastroenteritis in our environment. Immunochromatographic tests for antigen detection performed directly on stool samples for its simplicity and rapid results may make them useful diagnostic elements in the context of primary care. During October 2012 we selected all feces in which enteropathogenic bacteria are isolated from those received for stool culture in the laboratory of Microbiology of the University Hospital Virgen de las Nieves of Granada. After standard management of faeces samples and isolation of any enteropathogen, the commercial kits: Campy Leti, Ridaquick Campylobacterscreen and Salmonella Leti were tested for simultaneous research of Campylobacter and Salmonella antigens. Sensitivity and specificity were determined. Two hundred and thirty five stool samples were received in which 8 Salmonella enterica (7 B serogroup and 1 D serogroup), 7 Campylobacter jejuni, 4 Aeromonas hydrophila and 1 Yersinia enterocolitica were isolated. Campy Leti, Ridaquick Campylobacterscreen and Salmonella Leti presented a sensitivity of 100%, 100% and 75%, respectively. Specificities corresponded to 46%, 69% and 100%, respectively. Immunocromatographic tests can be useful for a first screening of enteropathogen in primary care.

Evaluation of a rapid method to exclude the presence of certain enteric pathogens in stool specimens.

PubMed Central

Teti, G; Burdash, N M; Zamboni, C; Fava, C; Tomasello, F; Mastroeni, P

1984-01-01

A new commercial method intended to exclude the presence of Salmonella spp., Shigella spp., and Yersinia enterocolitica and to presumptively identify Salmonella isolates within 2 h after primary isolation from stool specimens was evaluated. This system is marketed in Europe as API Z and in the United States as Rapid SST. The strip consists of five pairs of cupules for the screening of five lactose-negative colonies. The first cupule of each pair detects the presence of five enzymatic activities, whereas the second serves to maintain the strain for additional testing if necessary. A total of 197 fresh isolates from stool specimens and 217 stock cultures of Salmonella spp., Shigella spp., and Yersinia enterocolitica were tested, with the API 20E system as a reference method. In the stool specimens, 77.3% of the bacteria could be excluded from further workup for the presence of these organisms within 2 h. Over 97% of the stock strains and each of three fresh Salmonella isolates tested produced a reaction pattern corresponding to a correct presumptive identification. This reaction pattern was not produced by any isolate other than the Salmonella isolates. The API Z system can be used as a screen for the presence of Salmonella and Shigella spp. and can provide an accurate presumptive identification of Salmonella isolates within 2 h after primary isolation. PMID:6394610

Rapid Salmonella detection in experimentally inoculated equine faecal and veterinary hospital environmental samples using commercially available lateral flow immunoassays.

PubMed

Burgess, B A; Noyes, N R; Bolte, D S; Hyatt, D R; van Metre, D C; Morley, P S

2015-01-01

Salmonella enterica is the most commonly reported cause of outbreaks of nosocomial infections in large animal veterinary teaching hospitals and the closure of equine hospitals. Rapid detection may facilitate effective control practices in equine populations. Shipping and laboratory testing typically require ≥48 h to obtain results. Lateral flow immunoassays developed for use in food-safety microbiology provide an alternative that has not been evaluated for use with faeces or environmental samples. We aimed to identify enrichment methods that would allow commercially available rapid Salmonella detection systems (lateral flow immunoassays) to be used in clinical practice with equine faecal and environmental samples, providing test results in 18-24 h. In vitro experiment. Equine faecal and environmental samples were inoculated with known quantities of S. enterica serotype Typhimurium and cultured using 2 different enrichment techniques for faeces and 4 enrichment techniques for environmental samples. Samples were tested blindly using 2 different lateral flow immunoassays and plated on agar media for confirmatory testing. In general, commercial lateral flow immunoassays resulted in fewer false-negative test results with enrichment of 1 g faecal samples in tetrathionate for 18 h, while all environmental sample enrichment techniques resulted in similar detection rates. The limit of detection from spiked samples, ∼4 colony-forming units/g, was similar for all methods evaluated. The lateral flow immunoassays evaluated could reliably detect S. enterica within 18 h, indicating that they may be useful for rapid point-of-care testing in equine practice applications. Additional evaluation is needed using samples from naturally infected cases and the environment to gain an accurate estimate of test sensitivity and specificity and to substantiate further the true value of these tests in clinical practice. © 2014 EVJ Ltd.

High resolution melting (HRM) analysis as a new tool for rapid identification of Salmonella enterica serovar Gallinarum biovars Pullorum and Gallinarum.

PubMed

Ren, Xingxing; Fu, Ying; Xu, Chenggang; Feng, Zhou; Li, Miao; Zhang, Lina; Zhang, Jianmin; Liao, Ming

2017-05-01

Salmonella enterica serovar Gallinarum biovars Pullorum and Gallinarum represent the most common causative agents of chicken salmonellosis, which result in high mortality and morbidity throughout the world. It is difficult and laborious to discriminate these diseases based on biochemical or phenotypic methods. Herein, we report the development of a single nucleotide polymorphism (SNP) PCR-high resolution melt (PCR-HRM) assay for the detection and discrimination of both S. Pullorum and S. Gallinarun. The gene rfbS, which encodes a factor involved in the biosynthesis of ADP paratose in serogroup D of Salmonella, has been identified as a robust genetic marker for the identification of S. Pullorum and S. Gallinarun based on polymorphisms at positions 237 and 598. Therefore, PCR-HRM analyses were used to characterize this gene. A total of 15 reference and 33 clinical isolates of Salmonella and related Gram-negative bacteria were detected using 2 sets of primers. Our PCR-HRM assay could distinguish S. Pullorum from S. Gallinarun and other strains using the primer pair SP-237F/237R. Similarly, S. Gallinarun could be distinguished from S. Pullorum and other strains using primer set SG-598F/598R. These 2 assays showed high specificity (100%) for both S. Pullorum and S. Gallinarun; the sensitivity of these 2 assays was at least 100-fold greater than that of the allele-specific PCR assay. This present study demonstrated that HRM analysis represents a potent, simple, and economic tool for the rapid, specific, and sensitive detection of S. Pullorum and S. Gallinarun. Our approach also may aid efforts for purification of Avian Salmonella disease. © 2016 Poultry Science Association Inc.

Specific Discrimination of Three Pathogenic Salmonella enterica subsp. enterica Serotypes by carB-Based Oligonucleotide Microarray

PubMed Central

Shin, Hwa Hui; Hwang, Byeong Hee; Seo, Jeong Hyun

2014-01-01

It is important to rapidly and selectively detect and analyze pathogenic Salmonella enterica subsp. enterica in contaminated food to reduce the morbidity and mortality of Salmonella infection and to guarantee food safety. In the present work, we developed an oligonucleotide microarray containing duplicate specific capture probes based on the carB gene, which encodes the carbamoyl phosphate synthetase large subunit, as a competent biomarker evaluated by genetic analysis to selectively and efficiently detect and discriminate three S. enterica subsp. enterica serotypes: Choleraesuis, Enteritidis, and Typhimurium. Using the developed microarray system, three serotype targets were successfully analyzed in a range as low as 1.6 to 3.1 nM and were specifically discriminated from each other without nonspecific signals. In addition, the constructed microarray did not have cross-reactivity with other common pathogenic bacteria and even enabled the clear discrimination of the target Salmonella serotype from a bacterial mixture. Therefore, these results demonstrated that our novel carB-based oligonucleotide microarray can be used as an effective and specific detection system for S. enterica subsp. enterica serotypes. PMID:24185846

Specific discrimination of three pathogenic Salmonella enterica subsp. enterica serotypes by carB-based oligonucleotide microarray.

PubMed

Shin, Hwa Hui; Hwang, Byeong Hee; Seo, Jeong Hyun; Cha, Hyung Joon

2014-01-01

It is important to rapidly and selectively detect and analyze pathogenic Salmonella enterica subsp. enterica in contaminated food to reduce the morbidity and mortality of Salmonella infection and to guarantee food safety. In the present work, we developed an oligonucleotide microarray containing duplicate specific capture probes based on the carB gene, which encodes the carbamoyl phosphate synthetase large subunit, as a competent biomarker evaluated by genetic analysis to selectively and efficiently detect and discriminate three S. enterica subsp. enterica serotypes: Choleraesuis, Enteritidis, and Typhimurium. Using the developed microarray system, three serotype targets were successfully analyzed in a range as low as 1.6 to 3.1 nM and were specifically discriminated from each other without nonspecific signals. In addition, the constructed microarray did not have cross-reactivity with other common pathogenic bacteria and even enabled the clear discrimination of the target Salmonella serotype from a bacterial mixture. Therefore, these results demonstrated that our novel carB-based oligonucleotide microarray can be used as an effective and specific detection system for S. enterica subsp. enterica serotypes.

High Resolution Melting Analysis for Rapid Mutation Screening in Gyrase and Topoisomerase IV Genes in Quinolone-Resistant Salmonella enterica

PubMed Central

Thong, Kwai Lin

2014-01-01

The increased Salmonella resistance to quinolones and fluoroquinolones is a public health concern in the Southeast Asian region. The objective of this study is to develop a high resolution melt curve (HRM) assay to rapidly screen for mutations in quinolone-resistant determining region (QRDR) of gyrase and topoisomerase IV genes. DNA sequencing was performed on 62 Salmonella strains to identify mutations in the QRDR of gyrA, gyrB, parC, and parE genes. Mutations were detected in QRDR of gyrA (n = 52; S83F, S83Y, S83I, D87G, D87Y, and D87N) and parE (n = 1; M438I). Salmonella strains with mutations within QRDR of gyrA are generally more resistant to nalidixic acid (MIC 16 > 256 μg/mL). Mutations were uncommon within the QRDR of gyrB, parC, and parE genes. In the HRM assay, mutants can be distinguished from the wild-type strains based on the transition of melt curves, which is more prominent when the profiles are displayed in difference plot. In conclusion, HRM analysis allows for rapid screening for mutations at the QRDRs of gyrase and topoisomerase IV genes in Salmonella. This assay markedly reduced the sequencing effort involved in mutational studies of quinolone-resistance genes. PMID:25371903

High resolution melting analysis for rapid mutation screening in gyrase and Topoisomerase IV genes in quinolone-resistant Salmonella enterica.

PubMed

Ngoi, Soo Tein; Thong, Kwai Lin

2014-01-01

The increased Salmonella resistance to quinolones and fluoroquinolones is a public health concern in the Southeast Asian region. The objective of this study is to develop a high resolution melt curve (HRM) assay to rapidly screen for mutations in quinolone-resistant determining region (QRDR) of gyrase and topoisomerase IV genes. DNA sequencing was performed on 62 Salmonella strains to identify mutations in the QRDR of gyrA, gyrB, parC, and parE genes. Mutations were detected in QRDR of gyrA (n = 52; S83F, S83Y, S83I, D87G, D87Y, and D87N) and parE (n = 1; M438I). Salmonella strains with mutations within QRDR of gyrA are generally more resistant to nalidixic acid (MIC 16 > 256 μg/mL). Mutations were uncommon within the QRDR of gyrB, parC, and parE genes. In the HRM assay, mutants can be distinguished from the wild-type strains based on the transition of melt curves, which is more prominent when the profiles are displayed in difference plot. In conclusion, HRM analysis allows for rapid screening for mutations at the QRDRs of gyrase and topoisomerase IV genes in Salmonella. This assay markedly reduced the sequencing effort involved in mutational studies of quinolone-resistance genes.

Evaluation and comparison of rapid methods for the detection of Salmonella in naturally contaminated pine nuts using different pre enrichment media.

PubMed

Wang, Hua; Gill, Vikas S; Cheng, Chorng-Ming; Gonzalez-Escalona, Narjol; Irvin, Kari A; Zheng, Jie; Bell, Rebecca L; Jacobson, Andrew P; Hammack, Thomas S

2015-04-01

Foodborne outbreaks, involving pine nuts and peanut butter, illustrate the need to rapidly detect Salmonella in low moisture foods. However, the current Bacteriological Analytical Manual (BAM) culture method for Salmonella, using lactose broth (LB) as a pre enrichment medium, has not reliably supported real-time quantitative PCR (qPCR) assays for certain foods. We evaluated two qPCR assays in LB and four other pre enrichment media: buffered peptone water (BPW), modified BPW (mBPW), Universal Pre enrichment broth (UPB), and BAX(®) MP media to detect Salmonella in naturally-contaminated pine nuts (2011 outbreak). A four-way comparison among culture method, Pathatrix(®) Auto, VIDAS(®) Easy SLM, and qPCR was conducted. Automated DNA extraction techniques were compared with manual extraction methods (boiling or InstaGene™). There were no significant differences (P > 0.05) among the five pre enrichment media for pine nuts using the culture method. While both qPCR assays produced significantly (P ≤ 0.05) higher false negatives in 24 h pre enriched LB than in the other four media, they were as sensitive as the culture method in BPW, mBPW, UPB, and BAX media. The VIDAS Easy and qPCR were equivalent; Pathatrix was the least effective method. The Automatic PrepSEQ™ DNA extraction, using 1000 μL of pre enrichment, was as effective as manual extraction methods. Published by Elsevier Ltd.

Evaluation of the 3M™ Petrifilm™ Salmonella express system for the detection of Salmonella species in selected foods: collaborative study.

PubMed

Bird, Patrick; Flannery, Jonathan; Crowley, Erin; Agin, James; Goins, David; Jechorek, Robert

2014-01-01

The 3M™ Petriflm™ Salmonella Express (SALX) System is a simple, ready-to-use chromogenic culture medium system for the rapid qualitative detection and biochemical confirmation of Salmonella spp. in food and food process environmental samples. The 3M Petrifilm SALX System was compared using an unpaired study design in a multilaboratory collaborative study to the U.S. Department of Agriculture/Food Safety and Inspection Service (USDA/FSIS) Microbiology Laboratory Guidebook (MLG) 4.07 (2013) Isolation and Identification of Salmonella from Meat, Poultry, Pasteurized Egg and Catfish Products and Carcass and Environmental Sponges for raw ground beef and the U.S. Food and Drug Administration Bacteriological Analytical Manual (FDA/BAM) Chapter 5, Salmonella (2011) reference method for dry dog food following the current AOAC validation guidelines. For this study, a total of 17 laboratories located throughout the continental United States evaluated 1872 test portions. For the 3M Petrifilm SALX System, raw ground beef was analyzed using 25 g test portions, and dry dog food was analyzed using 375 g test portions. For the reference methods, 25 g test portions of each inatrix were analyzed. The two matrices were artificially contaminated with Salmonella at three inoculation levels: an uninoculated control level (0 CFU/test portion), a low inoculum level (0.2-2 CFU/test portion), and a high inoculum level (2-5 CFU/test portion). Each inoculation level was statistically analyzed using the probability of detection statistical model. For the raw ground beef and dry dog food test portions, no significant differences at the 95% confidence interval were observed in the number of positive samples detected by the 3M Petrifilm SALX System versus either the USDA/FSIS-MLG or FDA/BAM methods.

Simultaneous and Rapid Detection of Salmonella typhi, Bacillus anthracis, and Yersinia pestis by Using Multiplex Polymerase Chain Reaction (PCR)

PubMed Central

Safari Foroshani, Nargess; Karami, Ali; Pourali, Fatemeh

2013-01-01

Background Salmonella typhi, Bacillus anthracis, and Yersinia pestis are some serious human pathogens, which their early diagnosis is of great importance. Salmonella typhi, Bacillus anthracis, and Yersinia pestis cause typhoid fever, anthrax, and plague respectively. These bacteria can be used to make biologic weapons. Objectives In this study, we designed a new and rapid diagnostic method based on Uniplex and Multiplex PCR method. Materials and Methods Uniplex and multiplex Polymerase Chain Reaction (PCR) were conducted on virulent genes of hp and invA of Salmonella typhimurium, Pa and chr of Bacillus anthracis, and pla of Yersinia pestis. A genome from other bacteria was used to study the specificity of the primer and the PCR test. Results Standard strains used in this study showed that primers were specific. As for sensitivity, it was shown that this method can diagnose 1-10 copies of the genome, or 1-10 Colony Forming Units (CFU) for each of the bacteria. All pieces except anthrax were sequenced in PCR to validate the product. DNA fragment resulted from Bacillus anthracis was confirmed by restriction enzyme digestions. Conclusion The designed methods are accurate, rapid, and inexpensive to find and differentiate these bacteria from similar bacteria. They can be applied for rapid diagnosis of these agents in different specimens, and bioterrorism cases. PMID:24719692

PCR method based on the ogdH gene for the detection of Salmonella spp. from chicken meat samples.

PubMed

Jin, Un-Ho; Cho, Sung-Hak; Kim, Min-Gon; Ha, Sang-Do; Kim, Keun-Sung; Lee, Kyu-Ho; Kim, Kwang-Yup; Chung, Duck Hwa; Lee, Young-Choon; Kim, Cheorl-Ho

2004-09-01

In a previous paper, the ogdH gene that encodes 2-oxoglutarate dehydrogenase was isolated from Salmonella typhimurium. The catalytic N-terminal region in the enzyme was found to be very specific for the Salmonella species. Therefore, the aim of the present study was to detect S. typhimurium in food sources using primers designed for OGDH-1 and OGDH-2 which were based on the salmonella-specific region of the ogdH gene. A simple polymerase chain reaction (PCR) detection method was developed to detect low numbers of S. typhimurium in a chicken meat microbial consortium. Using the ogdH-specific primers under stringent amplification conditions and for gene probe analysis, fewer than 100 colony-forming units (CFUs) were detectable when pure cultures were employed. When the PCR assay was run on S. typhimurium-contaminated meat contents, only the positive meat samples containing as few as 200 CFUs reacted to the assay. The method employed for sample processing is simple and it was determined to provide a sensitive means of detecting trace amounts of S. typhimurium-specific sequences in the presence of mixed meat microbial populations. When compared with six representative intestinal gram-negative bacterial strains in foods, including Vibrio parahaemolyticus, V. vulnificus, Enterobacter cloacae, E. coli O157:H7, Pseudomonas aeruginosa, and Proteus sp., S. typhimurium had a unique and distinct PCR product (796 bp). In conclusion, the two OGDH primers were found to be rapid and sensitive detectors of Salmonella spp for the PCR method. Copyright 2004 The Microbiological Society of Korea

Applications of immunomagnetic capture and time-resolved fluorescence to detect outbreak Escherichia coli O157 and Salmonella in alfalfa sprouts

NASA Astrophysics Data System (ADS)

Tu, Shu-I.; Gordon, Marsha; Fett, William F.; Gehring, Andrew G.; Irwin, Peter L.

2004-03-01

Commercially available alfalfa seeds were inoculated with low levels (~ 4 CFU/g) of pathogenic bacteria. The inoculated seeds were then allowed to sprout in sterile tap water at 22°C. After 48 hours, the irrigation water and sprouts were separately transferred to bovine heart infusion (BHI) media. The microbes in the BHI samples were allowed to grow for 4 hours at 37°C and 160 rpm. Specific immunomagnetic beads (IMB) were then applied to capture the E.coli O157 and/or Salmonella in the growth media. Separation and concentration of IMB-captured pathogens were achieved using magnetic separators. The captured E. coli O157:H7 and Salmonella spp were further tagged with europium (Eu) labeled anti-E. coli O157 antibodies and samarium (Sm) labeled anti-Salmonella antibodies, respectively. After washing, the lanthanide labels were extracted out from the complexes by specific chelators to form strongly fluorescent chelates. The specific time-resolved fluorescence (TRF) associated with Eu or Sm was measured to estimate the extent of capture of the E. coli O157 and Salmonella, respectively. The results indicated that the approach could detect E. coli O157 and Salmonella enterica from the seeds inoculated with ~ 4 CFU/g of the pathogens. Non-targeted bacteria, e.g., Aeromonas and Citrobacter exhibited essentially no cross reactivity. Since the pathogen detection from the sprouts was achieved within 6 hours, the developed methodology could be use as a rapid, sensitive and specific screening process for E. coli O157 and Salmonella enterica in this popular salad food.

Evaluation of rapid SYS system as screen for Yersinia enterocolitica in the United States.

PubMed Central

Mele, L; Nadler, H; Gomez, S

1987-01-01

Clinical isolates (n = 150) from stool specimens were selected for evaluation of the Rapid SYS system (Analytab Products, Plainview, N.Y.) as a screening test for Shigella spp., Yersinia enterocolitica, and Salmonella spp. The Gram-Negative Identification Card (Vitek Systems, Inc., Hazelwood, Mo.) was used for identification. Although acceptable performance of the Rapid SYS system was described, the interpretative criteria provided by the vendor for previous studies led to inappropriate screening for Y. enterocolitica, particularly biotype 1. When corrected screening criteria were used for the present study, the sensitivity for the detection of 76 enteric pathogens was 98.7%. Of the 76 pathogens, 1 of 21 Shigella spp. was not detected. However, specificity was only 16.6% when 72 selected nonpathogens frequently encountered in stools were eliminated. Although the Rapid SYS system can identify Shigella spp., Y. enterocolitica, and Salmonella spp., only phenylalanine deaminase-producing and cytochrome oxidase-producing organisms can be eliminated from additional testing. Therefore, the Rapid SYS system cannot be used as a three-pathogen screen in the United States or in other geographic locales where Y. enterocolitica biotype 1 may be encountered. PMID:3323232

Validation of a Salmonella loop-mediated isothermal amplification assay in animal food.

PubMed

Domesle, Kelly J; Yang, Qianru; Hammack, Thomas S; Ge, Beilei

2018-01-02

Loop-mediated isothermal amplification (LAMP) has emerged as a promising alternative to PCR for pathogen detection in food testing and clinical diagnostics. This study aimed to validate a Salmonella LAMP method run on both turbidimetry (LAMP I) and fluorescence (LAMP II) platforms in representative animal food commodities. The U.S. Food and Drug Administration (FDA)'s culture-based Bacteriological Analytical Manual (BAM) method was used as the reference method and a real-time quantitative PCR (qPCR) assay was also performed. The method comparison study followed the FDA's microbiological methods validation guidelines, which align well with those from the AOAC International and ISO. Both LAMP assays were 100% specific among 300 strains (247 Salmonella of 185 serovars and 53 non-Salmonella) tested. The detection limits ranged from 1.3 to 28 cells for six Salmonella strains of various serovars. Six commodities consisting of four animal feed items (cattle feed, chicken feed, horse feed, and swine feed) and two pet food items (dry cat food and dry dog food) all yielded satisfactory results. Compared to the BAM method, the relative levels of detection (RLODs) for LAMP I ranged from 0.317 to 1 with a combined value of 0.610, while those for LAMP II ranged from 0.394 to 1.152 with a combined value of 0.783, which all fell within the acceptability limit (2.5) for an unpaired study. This also suggests that LAMP was more sensitive than the BAM method at detecting low-level Salmonella contamination in animal food and results were available 3days sooner. The performance of LAMP on both platforms was comparable to that of qPCR but notably faster, particularly LAMP II. Given the importance of Salmonella in animal food safety, the LAMP assays validated in this study holds great promise as a rapid, reliable, and robust method for routine screening of Salmonella in these commodities. Published by Elsevier B.V.

Evaluation of Molecular Methods for Identification of Salmonella Serovars

PubMed Central

Gurnik, Simone; Ahmad, Aaminah; Blimkie, Travis; Murphy, Stephanie A.; Kropinski, Andrew M.; Nash, John H. E.

2016-01-01

Classification by serotyping is the essential first step in the characterization of Salmonella isolates and is important for surveillance, source tracking, and outbreak detection. To improve detection and reduce the burden of salmonellosis, several rapid and high-throughput molecular Salmonella serotyping methods have been developed. The aim of this study was to compare three commercial kits, Salm SeroGen (Salm Sero-Genotyping AS-1 kit), Check&Trace (Check-Points), and xMAP (xMAP Salmonella serotyping assay), to the Salmonella genoserotyping array (SGSA) developed by our laboratory. They were assessed using a panel of 321 isolates that represent commonly reported serovars from human and nonhuman sources globally. The four methods correctly identified 73.8% to 94.7% of the isolates tested. The methods correctly identified 85% and 98% of the clinically important Salmonella serovars Enteritidis and Typhimurium, respectively. The methods correctly identified 75% to 100% of the nontyphoidal, broad host range Salmonella serovars, including Heidelberg, Hadar, Infantis, Kentucky, Montevideo, Newport, and Virchow. The sensitivity and specificity of Salmonella serovars Typhimurium and Enteritidis ranged from 85% to 100% and 99% to 100%, respectively. It is anticipated that whole-genome sequencing will replace serotyping in public health laboratories in the future. However, at present, it is approximately three times more expensive than molecular methods. Until consistent standards and methodologies are deployed for whole-genome sequencing, data analysis and interlaboratory comparability remain a challenge. The use of molecular serotyping will provide a valuable high-throughput alternative to traditional serotyping. This comprehensive analysis provides a detailed comparison of commercial kits available for the molecular serotyping of Salmonella. PMID:27194688

DNA aptamer-based colorimetric detection platform for Salmonella Enteritidis.

PubMed

Bayraç, Ceren; Eyidoğan, Füsun; Avni Öktem, Hüseyin

2017-12-15

Food safety is a major issue to protect public health and a key challenge is to find detection methods for identification of hazards in food. Food borne infections affects millions of people each year and among pathogens, Salmonella Enteritidis is most widely found bacteria causing food borne diseases. Therefore, simple, rapid, and specific detection methods are needed for food safety. In this study, we demonstrated the selection of DNA aptamers with high affinity and specificity against S. Enteritidis via Cell Systematic Evolution of Ligands by Exponential Enrichment (Cell-SELEX) and development of sandwich type aptamer-based colorimetric platforms for its detection. Two highly specific aptamers, crn-1 and crn-2, were developed through 12 rounds of selection with K d of 0.971µM and 0.309µM, respectively. Both aptamers were used to construct sandwich type capillary detection platforms. With the detection limit of 10 3 CFU/mL, crn-1 and crn-2 based platforms detected target bacteria specifically based on color change. This platform is also suitable for detection of S. Enteritidis in complex food matrix. Thus, this is the first to demonstrate use of Salmonella aptamers for development of the colorimetric aptamer-based detection platform in its identification and detection with naked eye in point-of-care. Copyright © 2017 Elsevier B.V. All rights reserved.

Differentiation of live and dead salmonella cells using fourier transform infrared (FTIR) spectroscopy and principle component analysis (PCA) technique

USDA-ARS?s Scientific Manuscript database

Various technologies have been developed for pathogen detection using optical, electrochemical, biochemical and physical properties. Conventional microbiological methods need time from days to week to get the result. Though this method is very sensitive and accurate, a rapid detection of pathogens i...

An Improved Multiplex Real-Time SYBR Green PCR Assay for Analysis of 24 Target Genes from 16 Bacterial Species in Fecal DNA Samples from Patients with Foodborne Illnesses.

PubMed

Kawase, Jun; Etoh, Yoshiki; Ikeda, Tetsuya; Yamaguchi, Keiji; Watahiki, Masanori; Shima, Tomoko; Kameyama, Mitsuhiro; Horikawa, Kazumi; Fukushima, Hiroshi; Goto, Ryoichi; Shirabe, Komei

2016-05-20

Here, we developed a new version of our original screening system (Rapid Foodborne Bacterial Screening 24; RFBS24), which can simultaneously detect 24 genes of foodborne pathogens in fecal DNA samples. This new version (RFBS24 ver. 5) detected all known stx2 subtypes, enterotoxigenic Escherichia coli (STh genotype), and Vibrio parahaemolyticus (trh2), which were not detected by the original RFBS24 assay. The detection limits of RFBS24 ver. 5 were approximately 5.6 × 10(-2)-5.6 × 10(-5) (ng DNA)/reaction, significantly lower (10- to 100-fold) than those of the original RFBS24 for the 22 target genes analyzed here. We also tested the new assay on fecal DNA samples from patients infected with Salmonella, Campylobacter, or enterohemorrhagic E. coli. The number of bacterial target genes detected by RFBS24 ver. 5 was greater than that detected by RFBS24. RFBS24 ver. 5 combined with an Ultra Clean Fecal DNA Isolation Kit showed adequate performance (sensitivity and specificity 89% and 100%, respectively, for Salmonella spp. and 100% and 83%, respectively, for Campylobacter jejuni) in terms of rapid detection of a causative pathogen during foodborne-illness outbreaks. Thus, RFBS24 ver. 5 is more useful than the previous assay system for detection of foodborne pathogens and offers quick simultaneous analysis of many targets and thus facilitates rapid dissemination of information to public health officials.

New Application of Hyperspectral Imaging for Bacterial Cell Classification

USDA-ARS?s Scientific Manuscript database

Hyperspectral microscopy has shown potential as a method for rapid detection of foodborne pathogenic bacteria with spectral characteristics from bacterial cells. Hyperspectral microscope images (HMIs) are collected from broiler chicken isolates of Salmonella serotypes Enteritidis, Typhimurium, Infa...

Rapid and simultaneous detection of Salmonella spp., Escherichia coli O157, and Listeria monocytogenes by magnetic capture hybridization and multiplex real-time PCR.

PubMed

Carloni, Elisa; Rotundo, Luca; Brandi, Giorgio; Amagliani, Giulia

2018-05-25

The application of rapid, specific, and sensitive methods for pathogen detection and quantification is very advantageous in diagnosis of human pathogens in several applications, including food analysis. The aim of this study was the evaluation of a method for the multiplexed detection and quantification of three significant foodborne pathogenic species (Escherichia coli O157, Salmonella spp., and Listeria monocytogenes). The assay combines specific DNA extraction by multiplex magnetic capture hybridization (mMCH) with multiplex real-time PCR. The amplification assay showed linearity in the range 10 6 -10 genomic units (GU)/PCR for each co-amplified species. The sensitivity corresponded to 1 GU/PCR for E. coli O157 and L. monocytogenes, and 10 GU/PCR for Salmonella spp. The immobilization process and the hybrid capture of the MCH showed good efficiency and reproducibility for all targets, allowing the combination in equal amounts of the different nanoparticle types in mMCH. MCH and mMCH efficiencies were similar. The detection limit of the method was 10 CFU in samples with individual pathogens and 10 2  CFU in samples with combination of the three pathogens in unequal amounts (amount's differences of 2 or 3 log). In conclusion, this multiplex molecular platform can be applied to determine the presence of target species in food samples after culture enrichment. In this way, this method could be a time-saving and sensitive tool to be used in routine diagnosis.

Rapid analysis of foodborne pathogens by surface-enhanced Raman spectroscopy

NASA Astrophysics Data System (ADS)

Sengupta, Atanu; Shende, Chetan; Huang, Hermes; Farquharson, Stuart; Inscore, Frank

2012-05-01

Foodborne diseases resulting from Campylobacter, Escherichia, Listeria, Salmonella, Shigella and Vibrio species affect as many as 76 million persons in the United States each year, resulting in 325,000 hospitalizations and 5,000 deaths. The challenge to preventing distribution and consumption of contaminated foods lies in the fact that just a few bacterial cells can rapidly multiply to millions, reaching infectious doses within a few days. Unfortunately, current methods used to detect these few cells rely on lengthy growth enrichment steps that take a similar amount of time (1 to 4 days). Consequently, there is a critical need for an analyzer that can rapidly extract and detect foodborne pathogens in 1-2 hours (not days), at 100 colony forming units per gram of food, and with a specificity that differentiates from indigenous microflora, so that false alarms are eliminated. In an effort to meet this need, we have been developing a sample system that extracts such pathogens from food, selectively binds these pathogens, and produces surface-enhanced Raman spectra (SERS). Here we present preliminary SERS measurements of Listeria and Salmonella.

Single-pipetting microfluidic assay device for rapid detection of Salmonella from poultry package.

PubMed

Fronczek, Christopher F; You, David J; Yoon, Jeong-Yeol

2013-02-15

A direct, sensitive, near-real-time, handheld optical immunoassay device was developed to detect Salmonella typhimurium in the naturally occurring liquid from fresh poultry packages (hereafter "chicken matrix"), with just single pipetting of sample (i.e., no filtration, culturing and/or isolation, thus reducing the assay time and the error associated with them). Carboxylated, polystyrene microparticles were covalently conjugated with anti-Salmonella, and the immunoagglutination due to the presence of Salmonella was detected by reading the Mie scatter signals from the microfluidic channels using a handheld device. The presence of chicken matrix did not affect the light scatter signal, since the optical parameters (particle size d, wavelength of incident light λ and scatter angle θ) were optimized to minimize the effect of sample matrix (animal tissues and blood proteins, etc.). The sample was loaded into a microfluidic chip that was split into two channels, one pre-loaded with vacuum-dried, antibody-conjugated particles and the other with vacuum-dried, bovine serum albumin-conjugated particles. This eliminated the need for a separate negative control, effectively minimizing chip-to-chip and sample-to-sample variations. Particles and the sample were diffused in-channel through chemical agitation by Tween 80, also vacuum-dried within the microchannels. Sequential mixing of the sample to the reagents under a strict laminar flow condition synergistically improved the reproducibility and linearity of the assay. In addition, dried particles were shown to successfully detect lower Salmonella concentrations for up to 8 weeks. The handheld device contains simplified circuitry eliminating unnecessary adjustment stages, providing a stable signal, thus maximizing sensitivity. Total assay time was 10 min, and the detection limit 10 CFU mL(-1) was observed in all matrices, demonstrating the suitability of this device for field assays. Copyright © 2012 Elsevier B.V. All rights reserved.

Report of data on children with non-typhi Salmonella gastroenteritis in a three-year period.

PubMed

Küçük, Öznur; Biçer, Suat; Ugraş, Meltem; Çöl, Defne; Giray, Tuba; Çiler Erdag, Gülay; Gürol, Yeşim; Yilmaz, Gülden; Yalvaç, Zerrin; Vitrinel, Ayça; Kaspar, Çigdem

2016-09-01

The purpose of this study was to evaluate the clinical and laboratory data of children with acute gastroenteritis caused by non-typhoid Salmonella spp. infections. Clinical (demographic data, symptoms and findings) and laboratory data (stool microscopy, rapid antigen tests, culture, multiplex polymerase chain reaction and blood test results) of children with acute gastroenteritis caused by non-typhoid Salmonella spp. between January 2010 and October 2012 were evaluated. Differences between the groups for categorical variables were estimated with a chi-square or Fisher exact test; for continuous variables with two independent samples a t test was used. P values < 0.05 were considered statistically significant. Sixty-seven children, 39 (58.2%) males and 28 (41.8%) females aged between 1 - 16 years (mean ± SD: 4.64 ± 2.91), were diagnosed with acute bacterial gastroenteritis caused by non-typhoid Salmonella spp. The main serotypes are Salmonella enteritidis (85%) and Salmonella typhimurium (7.5%). The presenting symptoms were diarrhoea (95.5%), fever (61.1%), vomiting (34.3%), abdominal pain (32.8%), loss of appetite (7.4%) and malaise (7.4%). Fever and dehydration (moderate and/or severe) were detected in 11 (16.4%) patients. The mean leukocyte count was 10.930/μL [95% confidence interval (CI), SD: ± 5.710/μL], neutrophil count was 7.880/μL (95% CI, SD: ± 4.960/μL), CRP was 64.16 mg/L (95% CI, SD: ± 76.24 mg/L), and erythrocyte sedimentation rate was 34.72 mm/hour (95% CI, SD: ± 13.64 mm/h). Stool microscopy was positive for leukocytes in 18 patients (26.8%). The definitive diagnosis was made with positive stool culture (n = 65) and/or PCR test (n = 4). Viral antigen positivity was detected in 10 patients (14.9%), evaluated as viral co-infection and false positive results. Antibiotic therapy and hospitalization were required in 26 (38.8%) and 23 (34.3%) patients, respectively. Salmonella carriage was detected in one patient (1.5%). Bloody diarrhoea, leukocytes in stool with an increased erythrocyte sedimentation rate and a CRP level without overt leukocytosis may indicate Salmonella infection. Viral antigens may cause false positive results in fast antigen tests in cases where clinical and laboratory findings indicate bacterial aetiology. Stool culture is a reference method in diagnosis whereas some agents may be detected via molecular techniques (polymerase chain reaction) in spite of negative culture. Multiplex polymerase chain reaction may be used to detect Salmonella spp. and may reveal false positivity for viruses as well as the detection of other bacteria.

Development of a one-step immunochromatographic strip test using gold nanoparticles for the rapid detection of Salmonella typhi in human serum.

PubMed

Preechakasedkit, Pattarachaya; Pinwattana, Kulwadee; Dungchai, Wijitar; Siangproh, Weena; Chaicumpa, Wanpen; Tongtawe, Pongsri; Chailapakul, Orawon

2012-01-15

An immunochromatographic strip test using gold nanoparticles was developed for the rapid detection of Salmonella typhi (S. typhi) in human serum. The strip test based on the principle of sandwich immunoassay by the specific binding of antigens from S. typhi O901 and antibody of S. typhi O901 on a nitrocellulose membrane. Antibody-gold nanoparticle conjugate was used as the label and was coated onto a glass fiber membrane, which was used as a conjugate pad. To create a test and control zone, antibody of S. typhi O901 and an anti-IgG were dotted on the nitrocellulose membrane, respectively. Positive samples were displayed as red dots at the test and control zones of the nitrocellulose membrane, while negative samples resulted in a red dot only in the control zone. The limit of detection (LOD) was found to be 1.14×10(5) cfu mL(-1), which could be visually detected by the naked eye within 15 min. This strip test provided a lower detection limit and analysis time than a dot blot immunoassay (8.88×10(6) cfu mL(-1) for LOD and 110 min for reaction time). In addition, our immunochromatographic strip test was employed to detect S. typhi in human serum effectively, with high accuracy. This strip test offers great promise for a rapid, simple and low-cost analysis of S. typhi. Copyright © 2011 Elsevier B.V. All rights reserved.

Simultaneous, specific and real-time detection of biothreat and frequently encountered food-borne pathogens

PubMed Central

Woubit, Abdela Salah; Yehualaeshet, Teshome; Habtemariam, Tsegaye; Samuel, Temesgen

2012-01-01

The bacterial genera Escherichia, Salmonella, Shigella, Vibrio, Yersinia and Francisella include important food safety and biothreat agents causing food-related and other human illnesses worldwide. We aimed to develop rapid methods with the capability to simultaneously and differentially detect all six pathogens in one run. Our initial experiments to use previously reported sets of primers revealed non-specificity of some of the sequences when tested against a broader array of pathogens, or proved not optimal for simultaneous detection parameters. By extensive mining of the whole genome and protein databases of diverse closely and distantly related bacterial species and strains, we have identified unique genome regions, which we utilized to develop a detection platform. Twelve of the specific genomic targets we have identified to design the primers in F. tularensis ssp. tularensis, F. tularensis ssp. novicida, S. dysentriae, S. typhimurium, V. cholera, Y. pestis, and Y. pseudotuberculosis contained either hypothetical or putative proteins, the functions of which have not been clearly defined. Corresponding primer sets were designed from the target regions for use in real-time PCR assays to detect specific biothreat pathogens at species or strain levels. The primer sets were first tested by in-silico PCR against whole genome sequences of different species, sub-species, or strains and then by in vitro PCR against genomic DNA preparations from 23 strains representing six biothreat agents (E.coli O157:H7 strain EDL 933, Shigella dysentriae, Salmonella typhi, Francisella tularensis ssp. tularensis, Vibrio cholera, and Yersinia pestis) and six foodborne pathogens (Salmonella typhimurium, Salmonella saintpaul, Shigella sonnei, Francisella novicida, Vibrio parahemolytica and Yersinia pseudotuberculosis). Each pathogen was specifically identifiable at the genus and species levels. Sensitivity assays performed using purified DNA showed the lowest detection limit of 640 fg DNA/µl for F. tularensis. A preliminary test done to detect Shigella organisms in a milk matrix showed that 6–60 colony forming units of the bacterium per milliliter of milk could be detected in about an hour. Therefore, we have developed a platform to simultaneously detect foodborne pathogen and biothreat agents specifically and in real-time. Such a platform could enable rapid detection or confirmation of contamination by these agents. PMID:22488053

Evaluation of commercial kit based on loop-mediated isothermal amplification for rapid detection of low levels of uninjured and injured Salmonella on duck meat, bean sprouts, and fishballs in Singapore.

PubMed

Lim, Hazel Sin Yue; Zheng, Qianwang; Miks-Krajnik, Marta; Turner, Matthew; Yuk, Hyun-Gyun

2015-06-01

The objective of this study was to evaluate performance of the commercial kit based on loop-mediated isothermal amplification (LAMP) in comparison with the International Organization for Standardization method for detecting uninjured and sublethally injured Salmonella cells artificially inoculated at levels of 10(0) and 10(1) CFU/25 g on raw duck wing, raw mung bean sprouts, and processed fishballs. Injured cells were prepared by a heat treatment for duck wings and fishball samples and a chlorine treatment for bean sprout samples. Additionally, a validation study was performed on naturally contaminated food samples sold in Singapore. A total of 110 samples of each commodity were analyzed in this study. Regardless of inoculum levels, the detection by the commercial LAMP kit showed 100% sensitivity and specificity for both inoculated and uninoculated samples compared with the International Organization for Standardization method, with the exception of bean sprout samples. Only 20% of bean sprout samples inoculated with 10(0) CFU/25 g injured Salmonella cells were positive by using the commercial LAMP-based kit. However, all negative samples became positive following a secondary enrichment in Rappaport-Vassiliadis medium with soy broth or after concentration by centrifugation. These results suggest that secondary enrichment or centrifugation should be considered as an additional step to increase the sensitivity of the commercial LAMP-based kit with low numbers of injured target cells in samples with high background microflora (such as mung bean sprouts). The validation study also showed that the commercial LAMP-based kit provided 91% sensitivity and 95% specificity for naturally contaminated samples. Thus, this study demonstrates that the commercial LAMP-based kit might be a cost-effective method, as this system could provide rapid, accurate detection of both uninjured and injured Salmonella cells on raw duck wings, raw mung bean sprouts, and processed fishballs in less than 26 h.

Novel genomic tools for specific and real-time detection of biothreat and frequently encountered foodborne pathogens.

PubMed

Woubit, Abdela; Yehualaeshet, Teshome; Habtemariam, Tsegaye; Samuel, Temesgen

2012-04-01

The bacterial genera Escherichia, Salmonella, Shigella, Vibrio, Yersinia, and Francisella include important food safety and biothreat agents. By extensive mining of the whole genome and protein databases of diverse, closely and distantly related bacterial species and strains, we have identified novel genome regions, which we utilized to develop a rapid detection platform for these pathogens. The specific genomic targets we have identified to design the primers in Francisella tularensis subsp. tularensis, F. tularensis subsp. novicida, Shigella dysenteriae, Salmonella enterica serovar Typhimurium, Vibrio cholerae, Yersinia pestis, and Yersinia pseudotuberculosis contained either known genes or putative proteins. Primer sets were designed from the target regions for use in real-time PCR assays to detect specific biothreat pathogens at species or strain levels. The primer sets were first tested by in silico PCR against whole-genome sequences of different species, subspecies, or strains and then by in vitro PCR against genomic DNA preparations from 23 strains representing six biothreat agents (Escherichia coli O157:H7 strain EDL 933, Shigella dysenteriae, S. enterica serovar Typhi, F. tularensis subsp. tularensis, V. cholerae, and Y. pestis) and six foodborne pathogens (Salmonella Typhimurium, Salmonella Saintpaul, Shigella sonnei, F. tularensis subsp. novicida, Vibrio parahaemolyticus, and Y. pseudotuberculosis). Each pathogen was specifically identifiable at the genus and species levels. Sensitivity assays performed with purified DNA showed the lowest detection limit of 128 fg of DNA/μl for F. tularensis subsp. tularensis. A preliminary test to detect Shigella organisms in a milk matrix also enabled the detection of 6 to 60 CFU/ml. These new tools could ultimately be used to develop platforms to simultaneously detect these pathogens.

Surface enhaced raman scattering (SERS) with biopolymer encapsulated silver nanosubstrates for rapid detection of foodborne pathogens

USDA-ARS?s Scientific Manuscript database

A biopolymer encapsulated with silver nanoparticles was prepared using polyvinyl alcohol (PVA) solution, silver nitrate, and trisodium citrate. Biopolymer based nanosubstrates were deposited on a mica sheet for SERS. Fresh cultures of Salmonella Typhimurium, Escherichia coli, Staphylococcus aureus a...

APTES Functionalized Iron Oxide-Silver Magnetic Hetero-Nanocomposites for Selective Capture and Rapid Removal of Salmonella enteritidis from Aqueous Solution

NASA Astrophysics Data System (ADS)

Trang, Vu Thi; Dinh, Ngo Xuan; Lan, Hoang; Tam, Le Thi; Huy, Tran Quang; Tuan, Pham Anh; Phan, Vu Ngoc; Le, Anh-Tuan

2018-02-01

Magnetic nanomaterials, as a promising platform for the fast and sensitive detection of bacterial pathogens, have attracted increasing interest from researchers in recent years. In this work, by utilizing a two-step synthetic technique consisting of co-precipitation and subsequent hydrothermal reaction, followed by functionalization steps with (3-aminopropyl)triethoxysilane (APTES) and the antibody against Salmonella enteritidis, antibody-conjugated Fe3O4-Ag@APTES hetero-nanocomposites were successfully prepared. Due to the specific antibody, the developed Fe3O4-Ag@APTES@SE-Ab conjugates are capable of selectively capturing S. enteritidis at a low concentration of about 101 CFU/mL. Moreover, the prepared magnetic conjugates also revealed that the S. enteritidis could be rapidly removed from water solution in 20 min by using an external magnetic field with a removal efficiency obtained of ˜ 91.36%. These results indicated that the Fe3O4-Ag@APTES@SE-Ab conjugates are promising for the rapid selective capture and removal of bacterial pathogens from aqueous environments, and can be used for improving the detection quality of pathogens in water samples using immunosensor-based diagnostic tests.

Multiplex quantification of Escherichia coli, Salmonella typhi and Vibrio cholera with three DNA targets in single reaction assay.

PubMed

Jangampalli Adi, Pradeepkiran; Naidu, Jagadish R; Matcha, Bhaskar

2017-09-01

Escherichia coli (E. coli), Salmonella typhi and Vibrio cholera harmful pathogens, which causes various diseases in humans. Rapid diagnosis of bacterial infection is an important for patient management and appropriate therapy during the early phase of the bacterial infected diseases. Among the existing techniques for identifying pathogens were less sensitive and time-consuming processes. In the present study total, 48 clinical 31 blood and 17 urine samples of patients suspected with the infections were collected from SVRR Hospital and used to detect the pathogens. Multiplex polymerase chain reaction (PCR) assay was set to design for the identification of Escherichia coli, Salmonella typhi and Vibrio cholera from the different clinical samples. Rapid diagnosis of Escherichia coli (E. coli), Salmonella and Vibrio cholera pathogens can be done with simultaneously in a single multiplex PCR assay by using specific primers with adjusted PCR conditions. Through this approach, the results represented with out of 31 blood samples 1-15 shows the positive with E. coli and remaining 14 only 11 were correlated with multiplex results of Vibrio cholera, remaining the urine samples all are positive with 17 samples correlate with the Salmonella typhi. Through the high specificity benefits of excellent sensitivity, with high resolution and reproducibility. This method of results proved and illustrates the best potential system for diagnosing the infectious disease with modern trendy. Copyright © 2017 Elsevier Ltd. All rights reserved.

[Microbiology--laboratory examinations for bacterias].

PubMed

Hen, Renjun; Imafuku, Yuji; Yoshida, Hiroshi

2002-11-01

As it has been required to identify pathogenic microbes in shorter times, simple and rapid methods have been developed and used. Here, we summarized the present situation of rapid diagnostic testing in clinical microbiology in Japan, and also presented our results on PBP2' detection. The rapid test kits available in Japan for E. coli, Helicobacter pylori, Salmonella, Streptococcus and Staphylococcus aureus were described. Rapid examination methods are based mainly on immunologic reactions, which included slide agglutination using latex particle, immunochromatography and ELISA. Times required for the identification are 10 to 15 minutes. Moreover, rapid test kits employing PCR are also marketed. Further, we evaluated MRSA-LA "Seiken" which is a rapid detection kit for PBP2' produced by MRSA. The test was shown to be highly sensitive and specific. For the rapid identification of pathogenic microbes, simple and rapid test kits described here will be used more in clinical diagnosis.

Gold nanoparticle-based enzyme-linked antibody-aptamer sandwich assay for detection of Salmonella Typhimurium.

PubMed

Wu, Wenhe; Li, Jun; Pan, Dun; Li, Jiang; Song, Shiping; Rong, Mingge; Li, Zixi; Gao, Jimin; Lu, Jianxin

2014-10-08

Enzyme-linked immunosorbent assay (ELISA) provides a convenient means for the detection of Salmonella enterica serovar Typhimurium (STM), which is important for rapid diagnosis of foodborne pathogens. However, conventional ELISA is limited by antibody-antigen immunoreactions and suffers from poor sensitivity and tedious sample pretreatment. Therefore, development of novel ELISA remains challenging. Herein, we designed a comprehensive strategy for rapid, sensitive, and quantitative detection of STM with high specificity by gold nanoparticle-based enzyme-linked antibody-aptamer sandwich (nano-ELAAS) method. STM was captured and preconcentrated from samples with aptamer-modified magnetic particles, followed by binding with detector antibodies. Then nanoprobes carrying a large amount of reporter antibodies and horseradish peroxidase molecules were used for colorimetric signal amplification. Under the optimized reaction conditions, the nano-ELAAS assay had a quantitative detection range from 1 × 10(3) to 1 × 10(8) CFU mL(-1), a limit of detection of 1 × 10(3) CFU mL(-1), and a selectivity of >10-fold for STM in samples containing other bacteria at higher concentration with an assay time less than 3 h. In addition, the developed nanoprobes were improved in terms of detection range and/or sensitivity when compared with two commercial enzyme-labeled antibody signal reporters. Finally, the nano-ELAAS method was demonstrated to work well in milk samples, a common source of STM contamination.

9 CFR 113.30 - Detection of Salmonella contamination.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Detection of Salmonella contamination... REQUIREMENTS Standard Procedures § 113.30 Detection of Salmonella contamination. The test for detection of Salmonella contamination provided in this section shall be conducted when such a test is prescribed in an...

9 CFR 113.30 - Detection of Salmonella contamination.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Detection of Salmonella contamination... REQUIREMENTS Standard Procedures § 113.30 Detection of Salmonella contamination. The test for detection of Salmonella contamination provided in this section shall be conducted when such a test is prescribed in an...

Comparative performance of three sampling techniques to detect airborne Salmonella species in poultry farms.

PubMed

Adell, Elisa; Moset, Verónica; Zhao, Yang; Jiménez-Belenguer, Ana; Cerisuelo, Alba; Cambra-López, María

2014-01-01

Sampling techniques to detect airborne Salmonella species (spp.) in two pilot scale broiler houses were compared. Broilers were inoculated at seven days of age with a marked strain of Salmonella enteritidis. The rearing cycle lasted 42 days during the summer. Airborne Salmonella spp. were sampled weekly using impaction, gravitational settling, and impingement techniques. Additionally, Salmonella spp. were sampled on feeders, drinkers, walls, and in the litter. Environmental conditions (temperature, relative humidity, and airborne particulate matter (PM) concentration) were monitored during the rearing cycle. The presence of Salmonella spp. was determined by culture-dependent and molecular methods. No cultivable Salmonella spp. were recovered from the poultry houses' surfaces, the litter, or the air before inoculation. After inoculation, cultivable Salmonella spp. were recovered from the surfaces and in the litter. Airborne cultivable Salmonella spp. Were detected using impaction and gravitational settling one or two weeks after the detection of Salmonella spp. in the litter. No cultivable Salmonella spp. were recovered using impingement based on culture-dependent techniques. At low airborne concentrations, the use of impingement for the quantification or detection of cultivable airborne Salmonella spp. is not recommended. In these cases, a combination of culture-dependent and culture-independent methods is recommended. These data are valuable to improve current measures to control the transmission of pathogens in livestock environments and for optimising the sampling and detection of airborne Salmonella spp. in practical conditions.

Transient sensitivity to nisin in cold-shocked Gram negatives.

PubMed

Boziaris, I S; Adams, M R

2000-09-01

Rapid chilling in the presence of nisin caused a dose-dependent reduction in the populations of several Gram-negative bacteria, despite the fact that appreciable structural injury to the outer membrane was not detected. Pseudomonas aeruginosa was most affected, followed by Pseudomonas fragi, Salmonella enteritidis PT4, PT7 and Escherichia coli, respectively. Addition of nisin after the chilling treatment had no effect. The results are ascribed to a transient susceptibility caused by phase changes in the lipids associated with the outer membrane, which are rapidly reversed when the cells return to higher temperatures. Combinations of chilling shock, nisin and EDTA gave much lower reductions of Salmonella and Pseudomonas on chicken skin in comparison with broths. This is attributed to a buffering of the temperature shock experienced by adherent bacteria and binding of the nisin by food particles.

MALDI-TOF mass spectrometry provides high accuracy in identification of Salmonella at species level but is limited to type or subtype Salmonella serovars.

PubMed

Kang, Lin; Li, Nan; Li, Ping; Zhou, Yang; Gao, Shan; Gao, Hongwei; Xin, Wenwen; Wang, Jinglin

2017-04-01

Salmonella can cause global foodborne illnesses in humans and many animals. The current diagnostic gold standard used for detecting Salmonella infection is microbiological culture followed by serological confirmation tests. However, these methods are complicated and time-consuming. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis offers some advantages in rapid identification, for example, simple and fast sample preparation, fast and automated measurement, and robust and reliable identification up to genus and species levels, possibly even to the strain level. In this study, we established a reference database for species identification using whole-cell MALDI-TOF MS; the database consisted of 12 obtained main spectra of the Salmonella culture collection strains belonged to seven serotypes. Eighty-two clinical isolates of Salmonella were identified using established database, and partial 16S rDNA gene sequencing and serological method were used as comparison. We found that MALDI-TOF mass spectrometry provided high accuracy in identification of Salmonella at species level but was limited to type or subtype Salmonella serovars. We also tried to find serovar-specific biomarkers and failed. Our study demonstrated that (a) MALDI-TOF MS was suitable for identification of Salmonella at species level with high accuracy and (b) that MALDI-TOF MS method presented in this study was not useful for serovar assignment of Salmonella currently, because of its low matching with serological method and (c) MALDI-TOF MS method presented in this study was not suitable to subtype S. typhimurium because of its low discriminatory ability.

High-throughput biosensors for multiplexed foodborne pathogen detection

USDA-ARS?s Scientific Manuscript database

Incidental contamination of foods by harmful bacteria (such as E. coli and Salmonella) and the toxins that they produce is a serious threat to public health and the economy in the United States. The presence of such bacteri and toxins in foods must be rapidly determined at various stages of food pr...

Rapid and Specific Detection of Salmonella spp. in Animal Feed Samples by PCR after Culture Enrichment

PubMed Central

Löfström, Charlotta; Knutsson, Rickard; Axelsson, Charlotta Engdahl; Rådström, Peter

2004-01-01

A PCR procedure has been developed for routine analysis of viable Salmonella spp. in feed samples. The objective was to develop a simple PCR-compatible enrichment procedure to enable DNA amplification without any sample pretreatment such as DNA extraction or cell lysis. PCR inhibition by 14 different feed samples and natural background flora was circumvented by the use of the DNA polymerase Tth. This DNA polymerase was found to exhibit a high level of resistance to PCR inhibitors present in these feed samples compared to DyNAzyme II, FastStart Taq, Platinum Taq, Pwo, rTth, Taq, and Tfl. The specificity of the Tth assay was confirmed by testing 101 Salmonella and 43 non-Salmonella strains isolated from feed and food samples. A sample preparation method based on culture enrichment in buffered peptone water and DNA amplification with Tth DNA polymerase was developed. The probability of detecting small numbers of salmonellae in feed, in the presence of natural background flora, was accurately determined and found to follow a logistic regression model. From this model, the probability of detecting 1 CFU per 25 g of feed in artificially contaminated soy samples was calculated and found to be 0.81. The PCR protocol was evaluated on 155 naturally contaminated feed samples and compared to an established culture-based method, NMKL-71. Eight percent of the samples were positive by PCR, compared with 3% with the conventional method. The reasons for the differences in sensitivity are discussed. Use of this method in the routine analysis of animal feed samples would improve safety in the food chain. PMID:14711627

Simultaneous Detection of Escherichia coli O157:H7, Salmonella enteritidis, and Listeria monocytogenes at a Very Low Level Using Simultaneous Enrichment Broth and Multichannel SPR Biosensor.

PubMed

Zhang, Xiaoguang; Tsuji, Sachiko; Kitaoka, Hayato; Kobayashi, Hiroshi; Tamai, Mitsuru; Honjoh, Ken-Ichi; Miyamoto, Takahisa

2017-10-01

Detection of foodborne pathogens at very low levels is still a challenge. A custom-built multichannel surface plasmon resonance (SPR) biosensor and simultaneous enrichment broth (SEB) were used to develop a simultaneous detection method for 3 important foodborne pathogens, Escherichia coli O157:H7 (O157:H7), Salmonella enteritidis, and Listeria monocytogenes, at a very low level. These 3 foodborne pathogens at a very low level (14, 6, and 28 CFU/25 g (mL) for O157:H7, S. enteritidis, and L. monocytogenes, respectively) were inoculated in SEB and incubated at 37 ˚C for 24 h. Sample prepared from the simultaneous enrichment culture was analyzed using the multichannel SPR biosensor and sensor chip immobilized with polyclonal antibodies specific to each of the target pathogens. O157:H7, S. enteritidis, and L. monocytogenes in chicken were detected simultaneously at an inoculum dose of 14, 6, and 28 CFU/25 g, respectively. Our method using a custom-built multichannel SPR biosensor and enrichment in SEB is expected as a rapid and simultaneous detection method for low levels of O157:H7, S. enteritidis, and L. monocytogenes in food. Our method is expected as a rapid and simultaneous detection method for pathogens at very low levels. It has great potential for safety control of food and microbiological detection applications. © 2017 Institute of Food Technologists®.

Salmonella detection in poultry samples. Comparison of two commercial real-time PCR systems with culture methods for the detection of Salmonella spp. in environmental and fecal samples of poultry.

PubMed

Sommer, D; Enderlein, D; Antakli, A; Schönenbrücher, H; Slaghuis, J; Redmann, T; Lierz, M

2012-01-01

The efficiency of two commercial PCR methods based on real-time technology, the foodproof® Salmonella detection system and the BAX® PCR Assay Salmonella system was compared to standardized culture methods (EN ISO 6579:2002 - Annex D) for the detection of Salmonella spp. in poultry samples. Four sample matrices (feed, dust, boot swabs, feces) obtained directly from poultry flocks, as well as artificially spiked samples of the same matrices, were used. All samples were tested for Salmonella spp. using culture methods first as the gold standard. In addition samples spiked with Salmonella Enteridis were tested to evaluate the sensitivity of both PCR methods. Furthermore all methods were evaluated in an annual ring-trial of the National Salmonella Reference Laboratory of Germany. Salmonella detection in the matrices feed, dust and boot swabs were comparable in both PCR systems whereas the results from feces differed markedly. The quality, especially the freshness, of the fecal samples had an influence on the sensitivity of the real-time PCR and the results of the culture methods. In fresh fecal samples an initial spiking level of 100cfu/25g Salmonella Enteritidis was detected. Two-days-dried fecal samples allowed the detection of 14cfu/25g. Both real- time PCR protocols appear to be suitable for the detection of Salmonella spp. in all four matrices. The foodproof® system detected eight samples more to be positive compared to the BAX® system, but had a potential false positive result in one case. In 7-days-dried samples none of the methods was able to detect Salmonella likely through letal cell damage. In general the advantage of PCR analyses over the culture method is the reduction of working time from 4-5 days to only 2 days. However, especially for the analysis of fecal samples official validation should be conducted according to the requirement of EN ISO6579:2002 - Annex D.

A sandwich-type optical immunosensor based on the alkaline phosphatase enzyme for Salmonella thypimurium detection

NASA Astrophysics Data System (ADS)

Widyastuti, E.; Puspitasari Schonherr, M. F.; Masruroh, A.; Anggraeni, R. A.; Nisak, Y. K.; Mursidah, S.

2018-03-01

Salmonella is pathogenic bacteria that caused foodborne diseases which being called Salmonellosis. Prevalence of Salmonellosis that being caused by Salmonella thypimurium in Indonesia is quite high. However, detection of Salmonella bacteria in food still limited, complicated, and required a lot time. Sensitive optical assay for Salmonella thypimurium paper based detection has been developed by integrating sandwich assay between antibody-antigen complex and alkaline phosphatase enzyme that produce visible bluish-purple colour with presence of NBT-BCIP substrate. The results showed that Limit of Quantitation of detection is 105 CFU mL-1 with detection time 15 minutes. Linearity test between Colour intensity that produced from Salmonella concentration presence on samples showed that detection has good linearity. Selectivity test exhibited excellent sensitivity with good discrimination against Escherichia coli.

The economic burden of a Salmonella Thompson outbreak caused by smoked salmon in the Netherlands, 2012-2013.

PubMed

Suijkerbuijk, Anita W M; Bouwknegt, Martijn; Mangen, Marie-Josee J; de Wit, G Ardine; van Pelt, Wilfrid; Bijkerk, Paul; Friesema, Ingrid H M

2017-04-01

In 2012, the Netherlands experienced the most extensive food-related outbreak of Salmonella ever recorded. It was caused by smoked salmon contaminated with Salmonella Thompson during processing. In total, 1149 cases of salmonellosis were laboratory confirmed and reported to RIVM. Twenty percent of cases was hospitalised and four cases were reported to be fatal. The purpose of this study was to estimate total costs of the Salmonella Thompson outbreak. Data from a case-control study were used to estimate the cost-of-illness of reported cases (i.e. healthcare costs, patient costs and production losses). Outbreak control costs were estimated based on interviews with staff from health authorities. Using the Dutch foodborne disease burden and cost-of-illness model, we estimated the number of underestimated cases and the associated cost-of-illness. The estimated number of cases, including reported and underestimated cases was 21 123. Adjusted for underestimation, the total cost-of-illness would be €6.8 million (95% CI €2.5-€16.7 million) with productivity losses being the main cost driver. Adding outbreak control costs, the total outbreak costs are estimated at €7.5 million. In the Netherlands, measures are taken to reduce salmonella concentrations in food, but detection of contamination during food processing remains difficult. As shown, Salmonella outbreaks have the potential for a relatively high disease and economic burden for society. Early warning and close cooperation between the industry, health authorities and laboratories is essential for rapid detection, control of outbreaks, and to reduce disease and economic burden. © The Author 2016. Published by Oxford University Press on behalf of the European Public Health Association. All rights reserved.

Assessment of Meat and Poultry Product Recalls Due to Salmonella Contamination: Product Recovery and Illness Prevention.

PubMed

Seys, Scott A; Sampedro, Fernando; Hedberg, Craig W

2017-08-01

Data from the recalls of meat and poultry products from 2000 through 2012 due to Salmonella contamination were used to assess the factors associated with the recovery of the recalled product and to develop quantitative models to estimate the number of illnesses prevented by recalls. The percentage of product recovered following a recall action was not dependent on establishment size, recall expansions, complexity of the distribution chain, type of distribution, amount of time between the production and recall dates, or number of pounds of product recalled. However, illness-related recalls were associated with larger amounts of recalled product, smaller percentages of recalled product recovered, a greater number of days between the production date and recall date, and nationwide distribution than were recalls that were not illness related. In addition, the detection of recall-associated illnesses appeared to be enhanced in states with strong foodborne illness investigation systems. The number of Salmonella illnesses prevented by recalls was based on the number of illnesses occurring relative to the number of pounds consumed, which was then extrapolated to the number of pounds of recalled product recovered. A simulation using a program evaluation and review technique probability distribution with illness-related recalls from 2003 through 2012 estimated that there were 19,000 prevented Salmonella illnesses, after adjusting for underdiagnosis. Recalls not associated with illnesses from 2000 through 2012 prevented an estimated additional 8,300 Salmonella illnesses, after adjusting for underdiagnosis. Although further improvements to ensure accurate and complete reporting should be undertaken, our study demonstrates that recalls are an important tool for preventing additional Salmonella illnesses. Moreover, additional training resources dedicated to public health agencies for enhancing foodborne illness detection, investigations, and rapid response and reporting would further prevent illnesses.

Comprehensive analysis of Salmonella sequence polymorphisms and development of a LDR-UA assay for the detection and characterization of selected serotypes.

PubMed

Lauri, Andrea; Castiglioni, Bianca; Mariani, Paola

2011-07-01

Salmonella is a major cause of food-borne disease, and Salmonella enterica subspecies I includes the most clinically relevant serotypes. Salmonella serotype determination is important for the disease etiology assessment and contamination source tracking. This task will be facilitated by the disclosure of Salmonella serotype sequence polymorphisms, here annotated in seven genes (sefA, safA, safC, bigA, invA, fimA, and phsB) from 139 S. enterica strains, of which 109 belonging to 44 serotypes of subsp. I. One hundred nineteen polymorphic sites were scored and associated to single serotypes or to serotype groups belonging to S. enterica subsp. I. A diagnostic tool was constructed based on the Ligation Detection Reaction-Universal Array (LDR-UA) for the detection of polymorphic sites uniquely associated to serotypes of primary interest (Salmonella Hadar, Salmonella Infantis, Salmonella Enteritidis, Salmonella Typhimurium, Salmonella Gallinarum, Salmonella Virchow, and Salmonella Paratyphi B). The implementation of promiscuous probes allowed the diagnosis of ten further serotypes that could be associated to a unique hybridization pattern. Finally, the sensitivity and applicability of the tool was tested on target DNA dilutions and with controlled meat contamination, allowing the detection of one Salmonella CFU in 25 g of meat.

Detection of Salmonella invA gene in shrimp enrichment culture by polymerase chain reaction.

PubMed

Upadhyay, Bishnu Prasad; Utrarachkij, Fuangfa; Thongshoob, Jarinee; Mahakunkijcharoen, Yuvadee; Wongchinda, Niracha; Suthienkul, Orasa; Khusmith, Srisin

2010-03-01

Contamination of seafood with salmonellae is a major public health concern. Detection of Salmonella by standard culture methods is time consuming. In this study, an enrichment culture step prior to polymerase chain reaction (PCR) was applied to detect 284 bp fragment of Salmonella invA in comparison with the conventional culture method in 100 shrimp samples collected from four different shrimp farms and fresh food markets around Bangkok. Samples were pre-enriched in non-selective lactose broth (LB) and selective tetrathionate broth (TTB). PCR detection limit was 10 pg and 10(4) cfu/ml of viable salmonellae with 100% specificity. PCR assay detected 19 different Salmonella serovars belonging to 8 serogroups (B, C1, C2-C3, D1, E1, E4 and K) commonly found in clinical and environmental samples in Thailand. The detection rate of PCR following TTB enrichment (24%) was higher than conventional culture method (19%). PCR following TTB, but not in LB enrichment allowed salmonella detection with 84% sensitivity, 90% specificity and 89% accuracy. Shrimp samples collected from fresh food markets had higher levels of contaminated salmonellae than those from shrimp farms. The results indicated that incorporation of an enrichment step prior to PCR has the potential to be applied for detection of naturally contaminated salmonellae in food, environment and clinical samples.

Evaluation of the performance of the IQ-Check kits and the USDA Microbiology Laboratory Guidebook methods for detection of Shiga toxin-producing Escherichia coli (STEC) and STEC and Salmonella simultaneously in ground beef.

PubMed

Baranzoni, G M; Fratamico, P M; Boccia, F; Bagi, L K; Kim, G-H; Anastasio, A; Pepe, T

2017-03-01

To evaluate the performance of the IQ-Check kits and the USDA Microbiology Laboratory Guidebook (MLG) methods for detection of the top seven Shiga toxin-producing Escherichia coli (STEC) (O157:H7, O26, O45, O103, O111, O121 and O145) in ground beef and both STEC and Salmonella in co-inoculated samples. Ground beef samples inoculated with ~10 CFU of STEC or both STEC and Salmonella Typhimurium were stored at 4°C for 72 h, followed by screening with the IQ-Check and BAX System kit (MLG) methods that employ different enrichment media. STEC and S. Typhimurium were detected after 12 and 18 h and their presence was confirmed by colony isolation. Both methods were able to detect STEC in ground beef after 12 h of enrichment in samples inoculated with low levels of the pathogen. STEC and S. Typhimurium can be detected and isolated in co-inoculated ground beef samples. The IQ-Check methods are comparable to the MLG methods for detection of STEC and simultaneous detection of STEC and S. Typhimurium in seeded ground beef after a short enrichment time, thus the IQ-Check method can be useful for the food industry for rapid detection of these pathogens. Published 2016. This article has been contributed to by US Government employees and their work is in the public domain in the USA.

Adaptation of red blood cell lysis represents a fundamental breakthrough that improves the sensitivity of Salmonella detection in blood

PubMed Central

Boyd, MA; Tennant, SM; Melendez, JH; Toema, D; Galen, JE; Geddes, CD; Levine, MM

2015-01-01

Aims Isolation of Salmonella Typhi from blood culture is the standard diagnostic for confirming typhoid fever but it is unavailable in many developing countries. We previously described a Microwave Accelerated Metal Enhanced Fluorescence (MAMEF)-based assay to detect Salmonella in medium. Attempts to detect Salmonella in blood were unsuccessful, presumably due to the interference of erythrocytes. The objective of this study was to evaluate various blood treatment methods that could be used prior to PCR, real-time PCR or MAMEF to increase sensitivity of detection of Salmonella. Methods and Results We tested ammonium chloride and erythrocyte lysis buffer, water, Lymphocyte Separation Medium, BD Vacutainer® CPT™ Tubes and dextran. Erythrocyte lysis buffer was the best isolation method as it is fast, inexpensive and works with either fresh or stored blood. The sensitivity of PCR- and real-time PCR detection of Salmonella in spiked blood was improved when whole blood was first lysed using erythrocyte lysis buffer prior to DNA extraction. Removal of erythrocytes and clotting factors also enabled reproducible lysis of Salmonella and fragmentation of DNA, which are necessary for MAMEF sensing. Conclusions Use of the erythrocyte lysis procedure prior to DNA extraction has enabled improved sensitivity of Salmonella detection by PCR and real-time PCR and has allowed lysis and fragmentation of Salmonella using microwave radiation (for future detection by MAMEF). Significance and Impact of the Study Adaptation of the blood lysis method represents a fundamental breakthrough that improves the sensitivity of DNA-based detection of Salmonella in blood. PMID:25630831

Detection of Salmonella bacterium in drinking water using microring resonator.

PubMed

Bahadoran, Mahdi; Noorden, Ahmad Fakhrurrazi Ahmad; Mohajer, Faeze Sadat; Abd Mubin, Mohamad Helmi; Chaudhary, Kashif; Jalil, Muhammad Arif; Ali, Jalil; Yupapin, Preecha

2016-01-01

A new microring resonator system is proposed for the detection of the Salmonella bacterium in drinking water, which is made up of SiO2-TiO2 waveguide embedded inside thin film layer of the flagellin. The change in refractive index due to the binding of the Salmonella bacterium with flagellin layer causes a shift in the output signal wavelength and the variation in through and drop port's intensities, which leads to the detection of Salmonella bacterium in drinking water. The sensitivity of proposed sensor for detecting of Salmonella bacterium in water solution is 149 nm/RIU and the limit of detection is 7 × 10(-4)RIU.

Electrochemical genosensing of Salmonella, Listeria and Escherichia coli on silica magnetic particles.

PubMed

Liébana, Susana; Brandão, Delfina; Cortés, Pilar; Campoy, Susana; Alegret, Salvador; Pividori, María Isabel

2016-01-21

A magneto-genosensing approach for the detection of the three most common pathogenic bacteria in food safety, such as Salmonella, Listeria and Escherichia coli is presented. The methodology is based on the detection of the tagged amplified DNA obtained by single-tagging PCR with a set of specific primers for each pathogen, followed by electrochemical magneto-genosensing on silica magnetic particles. A set of primers were selected for the amplification of the invA (278 bp), prfA (217 bp) and eaeA (151 bp) being one of the primers for each set tagged with fluorescein, biotin and digoxigenin coding for Salmonella enterica, Listeria monocytogenes and E. coli, respectively. The single-tagged amplicons were then immobilized on silica MPs based on the nucleic acid-binding properties of silica particles in the presence of the chaotropic agent as guanidinium thiocyanate. The assessment of the silica MPs as a platform for electrochemical magneto-genosensing is described, including the main parameters to selectively attach longer dsDNA fragments instead of shorter ssDNA primers based on their negative charge density of the sugar-phosphate backbone. This approach resulted to be a promising detection tool with sensing features of rapidity and sensitivity very suitable to be implemented on DNA biosensors and microfluidic platforms. Copyright © 2015 Elsevier B.V. All rights reserved.

Recombinant Phage Probes for Salmonella Typhimurium Detection

DTIC Science & Technology

2005-03-23

food safety analysis that are slower, labor-intensive, and cost-inefficient. Confirmation of presence in food products can take as long as 48 hours by conventional culture. Current rapid detection initiatives include biosensors that routinely incorporate antibodies as the biorecognition unit. Although sensitive and specific, antibodies are costly and may degrade under unfavorable environmental conditions. We believe that a stable, inexpensive substitute for antibodies is filamentous phage manipulated through phage display technique then affinity selected for specificity to

Detection of Salmonella by indicator agar media and PCR as affected by alfalfa seed homogenates and native bacteria.

PubMed

Liao, C-H; Shollenberger, L M

2003-01-01

To investigate and prevent the undesirable effect of native bacteria and alfalfa seed homogenates on detection of Salmonella in alfalfa seeds by indicator agar media and polymerase chain reaction (PCR). The relative sensitivity of five indicator agar media, including modified semisolid RV (MSRV), xylose-lysine-Tergitol 4 (XLT4), Hektoen enteric agar (HEA), brilliant green agar (BGA) and bismuth sulphite agar (BSA), for detection of Salmonella in the presence of a large number of native bacteria from alfalfa seeds was examined. The detection limit as measured by the ratio between the numbers of native bacteria and Salmonella was estimated to be 10(6) to 1 for MSRV and 10(3) to 1 for XLT4, HEA, BGA or BSA. Presence of alfalfa seed homogenates markedly reduced the sensitivity of Salmonella detection by PCR. The minimal number of Salmonella detectable by PCR was determined to be 1-10 and 100-1000 CFU in the absence and presence of seed homogenate, respectively. Application of anti-Salmonella immunomagnetic beads permitted detection of 2-5 CFU of heat-injured cells in 25 g of seeds within 24 h by PCR. The MSRV medium is more sensitive than other indicator agars for detecting a small number of motile Salmonella in samples containing a large number of native bacteria. Application of immunomagnetic beads eliminates the PCR-inhibitory activity of seed homogenates and improves the detection of Salmonella in inoculated seeds. The results generated from this study will aid the seed distributors, sprout growers and public health officials to identify and recall the Salmonella-contaminated seed lots to be used for sprout production.

The design of a microfluidic biochip for the rapid, multiplexed detection of foodborne pathogens by surface plasmon resonance imaging

NASA Astrophysics Data System (ADS)

Zordan, Michael D.; Grafton, Meggie M. G.; Park, Kinam; Leary, James F.

2010-02-01

The rapid detection of foodborne pathogens is increasingly important due to the rising occurrence of contaminated food supplies. We have previously demonstrated the design of a hybrid optical device that has the capability to perform realtime surface plasmon resonance (SPR) and epi-fluorescence imaging. We now present the design of a microfluidic biochip consisting of a two-dimensional array of functionalized gold spots. The spots on the array have been functionalized with capture peptides that specifically bind E. coli O157:H7 or Salmonella enterica. This array is enclosed by a PDMS microfluidic flow cell. A magnetically pre-concentrated sample is injected into the biochip, and whole pathogens will bind to the capture array. The previously constructed optical device is being used to detect the presence and identity of captured pathogens using SPR imaging. This detection occurs in a label-free manner, and does not require the culture of bacterial samples. Molecular imaging can also be performed using the epi-fluorescence capabilities of the device to determine pathogen state, or to validate the identity of the captured pathogens using fluorescently labeled antibodies. We demonstrate the real-time screening of a sample for the presence of E. coli O157:H7 and Salmonella enterica. Additionally the mechanical properties of the microfluidic flow cell will be assessed. The effect of these properties on pathogen capture will be examined.

DuPont Qualicon BAX System polymerase chain reaction assay. Performance Tested Method 100201.

PubMed

Tice, George; Andaloro, Bridget; Fallon, Dawn; Wallace, F Morgan

2009-01-01

A recent outbreak of Salmonella in peanut butter has highlighted the need for validation of rapid detection methods. A multilaboratory study for detecting Salmonella in peanut butter was conducted as part of the AOAC Research Institute Emergency Response Validation program for methods that detect outbreak threats to food safety. Three sites tested spiked samples from the same master mix according to the U.S. Food and Drug Administration's Bacteriological Analytical Manual (FDA-BAM) method and the BAX System method. Salmonella Typhimurium (ATCC 14028) was grown in brain heart infusion for 24 h at 37 degrees C, then diluted to appropriate levels for sample inoculation. Master samples of peanut butter were spiked at high and low target levels, mixed, and allowed to equilibrate at room temperature for 2 weeks. Spike levels were low [1.08 most probable number (MPN)/25 g]; high (11.5 MPN/25 g) and unspiked to serve as negative controls. Each master sample was divided into 25 g portions and coded to blind the samples. Twenty portions of each spiked master sample and five portions of the unspiked sample were tested at each site. At each testing site, samples were blended in 25 g portions with 225 mL prewarmed lactose broth until thoroughly homogenized, then allowed to remain at room temperature for 55-65 min. Samples were adjusted to a pH of 6.8 +/- 0.2, if necessary, and incubated for 22-26 h at 35 degrees C. Across the three reporting laboratories, the BAX System detected Salmonella in 10/60 low-spike samples and 58/60 high-spike samples. The reference FDA-BAM method yielded positive results for 11/60 low-spike and 58/60 high-spike samples. Neither method demonstrated positive results for any of the 15 unspiked samples.

The evaluation and application of multilocus variable number tandem repeat analysis (MLVA) for the molecular epidemiological study of Salmonella enterica subsp. enterica serovar Enteritidis infection.

PubMed

Liu, Yao; Shi, Xiaolu; Li, Yinghui; Chen, Qiongcheng; Jiang, Min; Li, Wanli; Qiu, Yaqun; Lin, Yiman; Jiang, Yixiang; Kan, Biao; Sun, Qun; Hu, Qinghua

2016-01-29

Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis) is one of the most prevalent Salmonella serotypes that cause gastroenteritis worldwide and the most prevalent serotype causing Salmonella infections in China. A rapid molecular typing method with high throughput and good epidemiological discrimination is urgently needed for detecting the outbreaks and finding the source for effective control of S. Enteritidis infections. In this study, 194 strains which included 47 from six outbreaks that were well-characterized epidemiologically were analyzed with pulse field gel electrophoresis (PFGE) and multilocus variable number tandem repeat analysis (MLVA). Seven VNTR loci published by the US Center for Disease Control and Prevention (CDC) were used to evaluate and develop MLVA scheme for S. Enteritidis molecular subtyping by comparing with PFGE, and then MLVA was applied to the suspected outbreaks detection. All S. Enteritidis isolates were analyzed with MLVA to establish a MLVA database in Shenzhen, Guangdong province, China to facilitate the detection of S. Enteritidis infection clusters. There were 33 MLVA types and 29 PFGE patterns among 147 sporadic isolates. These two measures had Simpson indices of 0.7701 and 0.8043, respectively, which did not differ significantly. Epidemiological concordance was evaluated by typing 47 isolates from six epidemiologically well-characterized outbreaks and it did not differ for PFGE and MLVA. We applied the well established MLVA method to detect two S. Enteritidis foodborne outbreaks and find their sources successfully in 2014. A MLVA database of 491 S. Enteritidis strains isolated from 2004 to 2014 was established for the surveillance of clusters in the future. MLVA typing of S. Enteritidis would be an effective tool for early warning and epidemiological surveillance of S. Enteritidis infections.

An Electrochemical DNA Biosensor for the Detection of Salmonella Using Polymeric Films and Electrochemical Labels

NASA Astrophysics Data System (ADS)

Diaz Serrano, Madeline

Waterborne and foodborne diseases are one of the principal public health problems worldwide. Microorganisms are the major agents of foodborne illness: pathogens such as Salmonella, Campylobacter jejuni and Escherichia coli, and parasites such as cryptosporidium. The most popular methods to detect Salmonella are based on culture and colony counting methods, ELISA, Gel electrophoresis and the polymerase chain reaction. Conventional detection methods are laborious and time-consuming, allowing for portions of the food to be distributed, marketed, sold and eaten before the analysis is done and the problem even detected. By these reasons, the rapid, easy and portable detection of foodborne organisms will facilitate the disease treatment. Our particular interest is to develop a nucleic acid biosensor (NAB) for the detection of pathogenic microorganisms in food and water samples. In this research, we report on the development of a NAB prototype using a polymer modified electrode surface together with sequences of different lengths for the OmpC gene from Salmonella as probes and Ferrocene-labeled target (Fc-ssDNA), Ferrocene-labeled tri(ethylene glycol) (Fc-PEG) and Ruthenium-Ferrocene (Ru-Fe) bimetallic complex as an electrochemical labels. We have optimized several PS films and anchored nucleic acid sequences with different lengths at gold and carbon surfaces. Non contact mode AFM and XPS were used to monitor each step of the NAB preparation, from polymer modification to oligos hybridization (conventional design). The hybridization reaction was followed electrochemically using a Fc-ssDNA and Fc-PEG in solution taking advantage of the morphological changes generated upon hybridization. We observed a small current at the potential for the Fe oxidation without signal amplification at +296 mV vs. Ag/AgCl for the Fc-ssDNA strategy and a small current at +524 mV for the Fc-PEG strategy. The immobilization, hybridization and signal amplification of Biotin- OmpC Salmonella genes generated by E.Z. Vega were obtained on NHS-PS-NHS 10.3 KD and detected by SWV and CC using Ru-Fe bimetallic complex as a redox label and GOx/glucose in PBS buffer. Calibration curves of biotin-OmpC probe hybridization were performed by CC, a catalytic charge was observed due to the presence of Ru-Fe bimetallic complex, GOx-A and glucose. The lowest target sequence detectable concentration was 0.41 microM.

Development of a Loop Mediated Isothermal Amplification (LAMP) - Surface Enhanced Raman spectroscopy (SERS) Assay for the Detection of Salmonella Enterica Serotype Enteritidis.

PubMed

Draz, Mohamed Shehata; Lu, Xiaonan

2016-01-01

As a major foodborne pathogen, Salmonella enterica serotype Enteritidis is increasingly rising as a global health concern. Here, we developed an integrated assay that combines loop mediated isothermal amplification (LAMP) and surface enhanced Raman spectroscopy (SERS) for DNA detection of S. Enteritidis using specifically designed Raman active Au-nanoprobes. The target DNA was amplified by LAMP and then labeled with Au-nanoprobes comprised of gold nanoparticle-modified with specific cy5/DNA probes to allow the detection by SERS. The sensitivity of the developed LAMP-SERS detection assay (66 CFU/mL) was ~100-fold higher than the conventional polymerase chain reaction (PCR) method. Significantly, this technique allowed highly specific detection of the target DNA of S. Enteritidis and could differentiate it from the DNA of closely related bacterial species or non-specific contamination, making it more accurate and reliable than the standard LAMP technique. The applicability of detection of S. Enteritidis in milk samples using LAMP-SERS assay was validated as well. In sum, the developed LAMP-SERS assay is highly specific and sensitive, and has the potential to be applied for rapid detection of different foodborne pathogens and other microbial contaminants.

Survival and Filamentation of Salmonella enterica Serovar Enteritidis PT4 and Salmonella enterica Serovar Typhimurium DT104 at Low Water Activity

PubMed Central

Mattick, K. L.; Jørgensen, F.; Legan, J. D.; Cole, M. B.; Porter, J.; Lappin-Scott, H. M.; Humphrey, T. J.

2000-01-01

In this study we investigated the long-term survival of and morphological changes in Salmonella strains at low water activity (aw). Salmonella enterica serovar Enteritidis PT4 and Salmonella enterica serovar Typhimurium DT104 survived at low aw for long periods, but minimum humectant concentrations of 8% NaCl (aw, 0.95), 96% sucrose (aw, 0.94), and 32% glycerol (aw, 0.92) were bactericidal under most conditions. Salmonella rpoS mutants were usually more sensitive to bactericidal levels of NaCl, sucrose, and glycerol. At a lethal aw, incubation at 37°C resulted in more rapid loss of viability than incubation at 21°C. At aw values of 0.93 to 0.98, strains of S. enterica serovar Enteritidis and S. enterica serovar Typhimurium formed filaments, some of which were at least 200 μm long. Filamentation was independent of rpoS expression. When the preparations were returned to high-aw conditions, the filaments formed septa, and division was complete within approximately 2 to 3 h. The variable survival of Salmonella strains at low aw highlights the importance of strain choice when researchers produce modelling data to simulate worst-case scenarios or conduct risk assessments based on laboratory data. The continued increase in Salmonella biomass at low aw (without a concomitant increase in microbial count) would not have been detected by traditional microbiological enumeration tests if the tests had been performed immediately after low-aw storage. If Salmonella strains form filaments in food products that have low aw values (0.92 to 0.98), there are significant implications for public health and for designing methods for microbiological monitoring. PMID:10742199

Physical Covering for Control of Escherichia coli O157:H7 and Salmonella spp. in Static and Windrow Composting Processes

PubMed Central

Yossa, Irene; Macarisin, Dumitru; Millner, Patricia

2015-01-01

This study investigated the effect of a 30-cm covering of finished compost (FC) on survival of Escherichia coli O157:H7 and Salmonella spp. in active static and windrow composting systems. Feedstocks inoculated with E. coli O157:H7 (7.41 log CFU/g) and Salmonella (6.46 log CFU/g) were placed in biosentry tubes (7.5-cm diameter, 30-cm height) at three locations: (i and ii) two opposing sides at the interface between the FC cover layer (where present) and the feedstock material (each positioned approximately 10 cm below the pile's surface) and (iii) an internal location (top) (approximately 30 cm below the surface). On specific sampling days, surviving populations of inoculated E. coli O157:H7 and Salmonella, generic E. coli, and coliforms in compost samples were determined. Salmonella spp. were reduced significantly within 24 h in windrow piles and were below the detection limit after 3 and 7 days at internal locations of windrow and static piles containing FC covering, respectively. Likewise, E. coli O157:H7 was undetectable after 1 day in windrow piles covered with finished compost. Use of FC as a covering layer significantly increased the number of days that temperatures in the windrows remained ≥55°C at all locations and in static piles at internal locations. These time-temperature exposures resulted in rapid reduction of inoculated pathogens, and the rate of bacterial reduction was rapid in windrow piles. The sample location significantly influenced the survival of these pathogens at internal locations compared to that at interface locations of piles. Finished compost covering of compost piles aids in the reduction of pathogens during the composting process. PMID:25576620

Comparison of Real-Time PCR, Reverse Transcriptase Real-Time PCR, Loop-Mediated Isothermal Amplification, and the FDA Conventional Microbiological Method for the Detection of Salmonella spp. in Produce ▿ †

PubMed Central

Zhang, Guodong; Brown, Eric W.; González-Escalona, Narjol

2011-01-01

Contamination of foods, especially produce, with Salmonella spp. is a major concern for public health. Several methods are available for the detection of Salmonella in produce, but their relative efficiency for detecting Salmonella in commonly consumed vegetables, often associated with outbreaks of food poisoning, needs to be confirmed. In this study, the effectiveness of three molecular methods for detection of Salmonella in six produce matrices was evaluated and compared to the FDA microbiological detection method. Samples of cilantro (coriander leaves), lettuce, parsley, spinach, tomato, and jalapeno pepper were inoculated with Salmonella serovars at two different levels (105 and <101 CFU/25 g of produce). The inoculated produce was assayed by the FDA Salmonella culture method (Bacteriological Analytical Manual) and by three molecular methods: quantitative real-time PCR (qPCR), quantitative reverse transcriptase real-time PCR (RT-qPCR), and loop-mediated isothermal amplification (LAMP). Comparable results were obtained by these four methods, which all detected as little as 2 CFU of Salmonella cells/25 g of produce. All control samples (not inoculated) were negative by the four methods. RT-qPCR detects only live Salmonella cells, obviating the danger of false-positive results from nonviable cells. False negatives (inhibition of either qPCR or RT-qPCR) were avoided by the use of either a DNA or an RNA amplification internal control (IAC). Compared to the conventional culture method, the qPCR, RT-qPCR, and LAMP assays allowed faster and equally accurate detection of Salmonella spp. in six high-risk produce commodities. PMID:21803916

Comparison of CHROMagar Salmonella Medium and Xylose-Lysine-Desoxycholate and Salmonella-Shigella Agars for Isolation of Salmonella Strains from Stool Samples

PubMed Central

Maddocks, Susan; Olma, Tom; Chen, Sharon

2002-01-01

The growth and appearance of 115 stock Salmonella isolates on a new formulation of CHROMagar Salmonella (CAS) medium were compared to those on xylose-lysine-desoxycholate agar (XLD), Salmonella-Shigella agar (SS), and Hektoen enteric agar (HEA) media. CAS medium was then compared prospectively to XLD and SS for the detection and presumptive identification of Salmonella strains in 500 consecutive clinical stool samples. All stock Salmonella isolates produced typical mauve colonies on CAS medium. Nine Salmonella strains were isolated from clinical specimens. The sensitivities for the detection of salmonellae after primary plating on CAS medium and the combination of XLD and SS after enrichment were 100%. The specificity for the detection of salmonellae after primary plating on CAS medium (83%) was significantly (P < 0.0001) higher than that after primary plating on the combination of SS and XLD media (55%) (a 28% difference in rates; 95% confidence interval, 23.0 to 34%). Twenty-nine non-Salmonella organisms produced mauve colonies on CAS medium, including 17 Candida spp. (59%) and 8 Pseudomonas spp. (28%). These were easily excluded as salmonellae by colony morphology, microscopic examination of a wet preparation, or oxidase testing. One biochemically inert Escherichia coli isolate required further identification to differentiate it from Salmonella spp. The use of plating on CAS medium demonstrated high levels of sensitivity and specificity and reduced the time to final identification of Salmonella spp., resulting in substantial cost savings. It can be recommended for use for the primary isolation of Salmonella spp. from stool specimens. Other media (e.g., XLD) are required to detect Shigella spp. concurrently. PMID:12149365

Development and use of polymerase chain reaction for the specific detection of Salmonella Typhimurium in stool and food samples.

PubMed

Lin, J S; Tsen, H Y

1999-10-01

Salmonella Typhimurium is one of the most important Salmonella serovars that may cause foodborne disease and human salmonellosis infection. Detection of this organism in the clinical samples of persons with gastroenteritis and the food samples associated with such persons may allow us to trace the cause of disease. Because malic acid dehydrogenase, an enzyme of the citric acid cycle, is common to organisms, the gene (mdh) coding for this enzyme was selected for the design of Salmonella Typhimurium-specific polymerase chain reaction (PCR) primers. By comparison of the mdh gene sequences of Salmonella Typhimurium and other Salmonella serotypes and of some isolates of other genera, two oligonucleotides were designed and used as PCR primers for the specific detection of Salmonella Typhimurium. The molecular weight of the PCR product was 261 bp as expected. Salmonella serovars other than Salmonella Typhimurium and isolates of other genera in the Enterobacteriaceae that is closely related to Salmonella did not generate any false-positive results. When this primer pair was used for the detection of Salmonella Typhimurium cells artificially inoculated into human stool specimens and food samples, such as milk and raw chicken meat, levels as low as 10(0) CFU per 0.1 g of stool specimen or per ml of milk or food homogenate could be detected if an 8- to 12-h preculture step using combined lactose-tetrathionate broth was performed prior to the PCR.

Detection and classification of salmonella serotypes using spectral signatures collected by fourier transform infrared (FT-IR) spectroscopy

USDA-ARS?s Scientific Manuscript database

Spectral signatures of Salmonella serotypes namely Salmonella Typhimurium, Salmonella Enteritidis, Salmonella Infantis, Salmonella Heidelberg and Salmonella Kentucky were collected using Fourier transform infrared spectroscopy (FT-IR). About 5-10 µL of Salmonella suspensions with concentrations of 1...

Limitations of a localized surface plasmon resonance sensor on Salmonella detection

USDA-ARS?s Scientific Manuscript database

We have designed a localized surface plasmon resonance (LSPR) biosensor to perform the whole cell detection of Salmonella using gold nanoparticls fabricated by oblique angle deposition technique. The LSPR sensor showed a plasmon peak shift due to the Salmonella antigen and anti-Salmonella antibody r...

A simple nucleic acid hybridization/latex agglutination assay for the rapid detection of polymerase chain reaction amplicons.

PubMed

Vollenhofer-Schrumpf, Sabine; Buresch, Ronald; Schinkinger, Manfred

2007-03-01

We have developed a new method for the detection of nucleic acid hybridization, based on a simple latex agglutination test that can be evaluated by the unaided eye. Nucleic acid, e.g., a polymerase chain reaction (PCR) product, is denatured and incubated with polystyrene beads carrying covalently bound complementary oligonucleotide sequences. Hybridization of the nucleic acids leads to aggregation of the latex particles, thereby verifying the presence of target sequence. The test is performed at room temperature, and results are available within 10 min. As a proof of principle, the hybridization/latex agglutination assay was applied to the detection of purified PCR fragments either specific for Salmonella spp. or a synthetic sequence, and to the detection of Salmonella enterica in artificially contaminated chicken samples. A few nanograms of purified PCR fragments were detectable. In artificially contaminated chicken samples, 3 colony-forming units (cfu)/25 g were detected in one of three replicates, and 30 cfu/25 g were detected in both of two replicates when samples for PCR were taken directly from primary enrichment, demonstrating the practical applicability of this test system. Even multiplex detection might be achievable. This novel kind of assay could be useful for a range of applications where hybridization of nucleic acids, e.g., PCR fragments, is to be detected.

Formation of mutagens in cooked foods. I. Beef.

PubMed

Spingarn, N E; Weisburger, J H

1979-09-01

Mutagens detectable by Salmonella typhimurium TA98, after activation by liver S-9 fraction, are formed when meat is cooked by frying, broiling and boiling. High levels of mutagenic activity are formed rapidly when frying, or more slowly during broiling. Formation of mutagens in boiled beef stock requires several days under reflux, but shows a strong concentration dependence. Time curves suggest that a period exists during which mutagens are not readily formed; however, after this period mutagen production is rapid. Hamburgers from commercial franchises were frequently mutagenically active.

Rapid real-time PCR methods to distinguish Salmonella Enteritidis wildtype field isolates from vaccine strains Salmovac SE/Gallivac SE and AviPro SALMONELLA VAC E.

PubMed

Maurischat, Sven; Szabo, Istvan; Baumann, Beatrice; Malorny, Burkhard

2015-05-01

Salmonella enterica serovar Enteritidis is a major non-typhoid Salmonella serovar causing human salmonellosis mainly associated with the consumption of poultry and products thereof. To reduce infections in poultry, S. Enteritidis live vaccine strains AviPro SALMONELLA VAC E and Salmovac SE/Gallivac SE have been licensed and used in several countries worldwide. To definitively diagnose a S. Enteritidis contamination in vaccinated herds a reliable and fast method for the differentiation between vaccine and wildtype field isolates is required. In this study, we developed and validated real-time PCR (qPCR) assays to distinguish those variants genetically. Suitable target sequences were identified by whole genome sequencing (WGS) using the Illumina MiSeq system. SNP regions in kdpA and nhaA proved to be most useful for differentiation of AviPro SALMONELLA VAC E and Salmovac SE/Gallivac SE, respectively, from wildtype strains. For each vaccine strain one TaqMan-qPCR assay and one alternative approach using High Resolution Melting (HRM) analysis was designed. All 30 Salmovac SE and 7 AviPro SALMONELLA VAC E vaccine strain reisolates tested were correctly identified by both approaches (100% inclusivity). Furthermore, all 137 (TaqMan) and 97 (HRM) Salmonella non-vaccine and related Enterobacteriaceae strains tested were excluded (100% exclusivity). The analytical detection limits were determined to be approx. 10(2) genome copies/reaction for the TaqMan and 10(4) genome copies/reaction for the HRM approach. The real-time PCR assays proved to be a reliable and fast alternative to the cultural vaccine strain identification tests helping decision makers in control measurements to take action within a shorter period of time. Copyright © 2015 Elsevier B.V. All rights reserved.

Assessment of Salmonella survival in dry-cured Italian salami.

PubMed

Bonardi, S; Bruini, I; Bolzoni, L; Cozzolino, P; Pierantoni, M; Brindani, F; Bellotti, P; Renzi, M; Pongolini, S

2017-12-04

The inactivation of Salmonella during curing of Italian traditional pork salami was investigated. A total of 150 batches of ground raw meat (GRM) used for salami manufacturing by four producers were tested for Salmonella by real-time PCR followed by ISO 6579 cultural confirmation and MPN enumeration. Salami produced with Salmonella positive GRMs were re-tested at the end of their curing period. Aw, pH and NaCl content were also measured. Detection of Salmonella was performed testing both 25 and 50g of the samples. By Real-Time PCR 37% of the GRMs resulted positive, but cultural detection of Salmonella was obtained in 14% of the samples only. Salmonella enumeration ranged from 31 MPN/g to <1.3 MPN/g. The difference between testing 50g and 25g of the samples was statistically significant (p value≤0.01). In particular, ISO-50g detected Salmonella in 100% of all positive samples, vs. 62% of ISO-25g. Salami made of the contaminated GRMs were 29% Salmonella-positive, as most batches of salami produced with Salmonella-positive GRMs resulted negative after regular curing (20-48days). Overall, 13% of salami produced with Salmonella-contaminated GRMs were positive. They belonged to six batches, which turned out negative after prolonged curing ranging between 49 and 86days. Salmonella enumeration in salami ranged from 8.7 MPN/g to <1.3 MPN/g. Unlike GRMs, no significant difference was observed between the ISO-50g and the ISO-25g in detecting Salmonella in cured salami (p value: >0.05). The most common Salmonella serovars in GRMs were Derby (52%), Typhimurium monophasic variant 4, (Barbuti et al., 1993), 12:i:- (19%) and Stanley (10%). Salmonella Derby (56%), London, Branderup, Panama (13%, respectively) and Goldcoast (6%) were most frequent in cured salami. The study showed negative correlation between real-time CT values and cultural confirmation of Salmonella, as well as the importance of sample size for Salmonella detection. Among considered factors with possible effect on the occurrence of Salmonella in salami, statistical analysis revealed a role for aw in salami and for Salmonella load in GRMs, while pH and NaCl content did not significantly affect the probability of finding Salmonella in dry-cured salami in the context of this study. In particular the lower aw values due to longer curing were associated with lower Salmonella presence in traditional dry-cured salami. Copyright © 2017 Elsevier B.V. All rights reserved.

Amplification of ST50 gene using dry-reagent-based polymerase chain reaction for the detection of Salmonella typhi.

PubMed

Aziah, Ismail; Ravichandran, Manickam; Ismail, Asma

2007-12-01

Conventional polymerase chain reaction (PCR) testing requires many pipetting steps and has to be transported and stored in cold chain. To overcome these limitations, we designed a ready-to-use PCR test for Salmonella typhi using PCR reagents, primers against the ST50 gene of S. typhi, a built-in internal amplification control (IAC), and gel loading dye mixed and freeze-dried in a single tube. The 2-step dry-reagent-based assay was used to amplify a 1238-bp target gene and an 810-bp IAC gene from 73 BACTEC blood culture broths (33 true positives for S. typhi and 40 true negatives for non-S. typhi). The sensitivity, specificity, positive predictive value, and negative predictive value of the PCR assay were 87.9%, 100%, 100%, and 90.9%, respectively. We suggest that this rapid 2-step PCR test could be used for the rapid diagnosis of typhoid fever.

False positive malaria rapid diagnostic test in returning traveler with typhoid fever.

PubMed

Meatherall, Bonnie; Preston, Keith; Pillai, Dylan R

2014-07-09

Rapid diagnostic tests play a pivotal role in the early diagnosis of malaria where microscopy or polymerase chain reaction are not immediately available. We report the case of a 39 year old traveler to Canada who presented with fever, headache, and abdominal pain after visiting friends and relatives in India. While in India, the individual was not ill and had no signs or symptoms of malaria. Laboratory testing upon his return to Canada identified a false positive malaria rapid diagnostic (BinaxNOW® malaria) result for P. falciparum with coincident Salmonella Typhi bacteraemia without rheumatoid or autoimmune factors. Rapid diagnostic test false positivity for malaria coincided with the presence or absence of Salmonella Typhi in the blood. Clinicians should be aware that Salmonella Typhi infection may result in a false positive malaria rapid diagnostic test. The mechanism of this cross-reactivity is not clear.

Customizable PCR-microplate array for differential identification of multiple pathogens

PubMed Central

Woubit, Abdela; Yehualaeshet, Teshome; Roberts, Sherrelle; Graham, Martha; Kim, Moonil; Samuel, Temesgen

2014-01-01

Customizable PCR-microplate arrays were developed for the rapid identification of Francisella tularensis subsp. tularensis, Salmonella Typhi, Shigella dysenteriae, Yersinia pestis, Vibrio cholerae Escherichia coli O157:H7, Salmonella Typhimurium, Salmonella Saintpaul, Francisella tularensis subsp. novicida, Vibrio parahaemolyticus, and Yersinia pseudotuberculosis. Previously, we identified highly specific primers targeting each of the pathogens above. Here, we report the development of customizable PCR-microplate arrays for simultaneous identification of the pathogens using the primers. A mixed aliquot of genomic DNA from 38 different strains was used to validate three PCR-microplate array formats. Identical PCR conditions were used to run all the samples on the three formats. Results show specific amplifications on all the three custom plates. In a preliminary test to evaluate the sensitivity of these assays in laboratory-inoculated samples, detection limits as low as 9 cfu/g/ml S. Typhimurium were obtained from beef hot dog, and 78 cfu/ml from milk. Such microplate arrays could serve as valuable tools for initial identification or secondary confirmation of these pathogens. PMID:24215700

Investigation of Outbreaks of Salmonella enterica Serovar Typhimurium and Its Monophasic Variants Using Whole-Genome Sequencing, Denmark

PubMed Central

Gymoese, Pernille; Sørensen, Gitte; Litrup, Eva; Olsen, John Elmerdal; Nielsen, Eva Møller

2017-01-01

Whole-genome sequencing is rapidly replacing current molecular typing methods for surveillance purposes. Our study evaluates core-genome single-nucleotide polymorphism analysis for outbreak detection and linking of sources of Salmonella enterica serovar Typhimurium and its monophasic variants during a 7-month surveillance period in Denmark. We reanalyzed and defined 8 previously characterized outbreaks from the phylogenetic relatedness of the isolates, epidemiologic data, and food traceback investigations. All outbreaks were identified, and we were able to exclude unrelated and include additional related human cases. We were furthermore able to link possible food and veterinary sources to the outbreaks. Isolates clustered according to sequence types (STs) 19, 34, and 36. Our study shows that core-genome single-nucleotide polymorphism analysis is suitable for surveillance and outbreak investigation for Salmonella Typhimurium (ST19 and ST36), but whole genome–wide analysis may be required for the tight genetic clone of monophasic variants (ST34). PMID:28930002

[The fundamental role of stage control technology on the detectability for Salmonella networking laboratory].

PubMed

Zhou, Yong-ming; Chen, Xiu-hua; Xu, Wen; Jin, Hui-ming; Li, Chao-qun; Liang, Wei-li; Wang, Duo-chun; Yan, Mei-ying; Lou, Jing; Kan, Biao; Ran, Lu; Cui, Zhi-gang; Wang, Shu-kun; Xu, Xue-bin

2013-11-01

To evaluated the fundamental role of stage control technology (SCT) on the detectability for Salmonella networking laboratories. Appropriate Salmonella detection methods after key point control being evaluated, were establishment and optimized. Our training and evaluation networking laboratories participated in the World Health Organization-Global Salmonella Surveillance Project (WHO-GSS) and China-U.S. Collaborative Program on Emerging and Re-emerging infectious diseases Project (GFN) in Shanghai. Staff members from the Yunnan Yuxi city Center for Disease Control and Prevention were trained on Salmonella isolation from diarrhea specimens. Data on annual Salmonella positive rates was collected from the provincial-level monitoring sites to be part of the GSS and GFN projects from 2006 to 2012. The methodology was designed based on the conventional detection procedure of Salmonella which involved the processes as enrichment, isolation, species identification and sero-typing. These methods were simultaneously used to satisfy the sensitivity requirements on non-typhoid Salmonella detection for networking laboratories. Public Health Laboratories in Shanghai had developed from 5 in 2006 to 9 in 2011, and Clinical laboratories from 8 to 22. Number of clinical isolates, including typhoid and non-typhoid Salmonella increased from 196 in 2006 to 1442 in 2011. The positive rate of Salmonella isolated from the clinical diarrhea cases was 2.4% in Yuxi county, in 2012. At present, three other provincial monitoring sites were using the SBG technique as selectivity enrichment broth for Salmonella isolation, with Shanghai having the most stable positive baseline. The method of SCT was proved the premise of the network laboratory construction. Based on this, the improvement of precise phenotypic identification and molecular typing capabilities could reach the level equivalent to the national networking laboratory.

Evaluation of PCR detection of Salmonella in alfalfa sprouts and spent irrigation water collected during sprouting of naturally contaminated seed.

PubMed

Maks, Nicole; Fu, Tong-Jen

2013-02-01

This study evaluated the efficacy of a PCR-based system (DuPont Qualicon BAX) for detection of Salmonella in sprouts and spent irrigation water collected during sprouting of seeds naturally contaminated with Salmonella. Alfalfa seeds were grown in Mason jars at 20 and 30°C for 3 days. Levels of Salmonella present in the water and sprouts were determined by most-probable-number (MPN) analysis. Background microflora levels were also determined. Samples of spent irrigation water and sprouts were enriched overnight individually in tetrathionate broth and in buffered peptone water with novobiocin at 42°C and then run in the BAX system. Samples were also enriched according to the U.S. Food and Drug Administration's Bacteriological Analytical Manual (FDA BAM) method for Salmonella as a comparison. Salmonella levels were lower at 20°C compared with 30°C for some trials, and background microflora levels ranged from 10(7) to 10(8) CFU/g or ml at 20°C and 10(8) to 10(9) CFU/g or ml at 30°C. In trials with a Salmonella level >1.1 MPN/g or ml, both the BAX and FDA BAM methods were able to detect Salmonella in all samples. In trials with lower levels (0.21 MPN/g or ml or lower) of Salmonella, BAX was able to detect more positive samples than FDA BAM. For one trial with <0.003 MPN/g or ml of Salmonella, the presence of the pathogen was not indicated by either the BAX or the FDA BAM method. The results suggest that PCR detected low levels of Salmonella in sprouts or spent irrigation water collected from sprouting of naturally contaminated seeds.

Survival and growth of Salmonella in salsa and related ingredients.

PubMed

Ma, Li; Zhang, Guodong; Gerner-Smidt, Peter; Tauxe, Robert V; Doyle, Michael P

2010-03-01

A large outbreak of Salmonella Saintpaul associated with raw jalapeño peppers, serrano peppers, and possibly tomatoes was reported in the United States in 2008. During the outbreak, two clusters of illness investigated among restaurant patrons were significantly associated with eating salsa. Experiments were performed to determine the survival and growth characteristics of Salmonella in salsa and related major ingredients, i.e., tomatoes, jalapeño peppers, and cilantro. Intact and chopped vegetables and different formulations of salsas were inoculated with a five-strain mixture of Salmonella and then stored at 4, 12, and 21 degrees C for up to 7 days. Salmonella populations were monitored during storage. Salmonella did not grow, but survived on intact tomatoes and jalapeño peppers, whereas significant growth at 12 and 21 degrees C was observed on intact cilantro. In general, growth of Salmonella occurred in all chopped vegetables when stored at 12 and 21 degrees C, with chopped jalapeño peppers being the most supportive of Salmonella growth. Regardless of differences in salsa formulation, no growth of Salmonella (initial inoculation ca. 3 log CFU/g) was observed in salsa held at 4 degrees C; however, rapid or gradual decreases in Salmonella populations were only observed in formulations that contained both fresh garlic and lime juice. Salmonella grew at 12 and 21 degrees C in salsas, except for those formulations that contained both fresh garlic and lime juice, in which salmonellae were rapidly or gradually inactivated, depending on salsa formulation. These results highlight the importance of preharvest pathogen contamination control of fresh produce and proper formulation and storage of salsa.

Comparison of Cultivation and PCR-Hybridization for Detection of Salmonella in Porcine Fecal and Water Samples†

PubMed Central

Feder, Ingrid; Nietfeld, Jerome C.; Galland, John; Yeary, Teresa; Sargeant, Jan M.; Oberst, Richard; Tamplin, Mark L.; Luchansky, John B.

2001-01-01

A total of 150 fecal and water samples from four swine farms were tested for the presence of Salmonella enterica using different enrichment techniques as follows: (i) 92 fecal samples from nursery and farrowing barns at three swine farms were preenriched overnight in tryptic soy broth (TSB) at 37°C followed by overnight enrichment in Rappaport-Vassiliadis 10 broth (RV10) at 42°C; (ii) 24 water samples from the third farm were preenriched overnight in 3MC broth at 37°C followed by overnight enrichment in RV10 at 42°C; and (iii) 34 fecal samples from a fourth farm, a finishing farm, were enriched overnight in RV10 at 42°C with no additional enrichment. Following each of the enrichment techniques, samples were subcultured onto modified semisolid Rappaport-Vassiliadis (MSRV) agar prior to transfer to Hektoen Enteric agar plates for the recovery of viable Salmonella bacteria. Presumptive Salmonella isolates were biochemically and serologically confirmed. For the PCR detection of Salmonella, a 1-ml portion was removed from each sample after the first overnight enrichment and the DNA was extracted using a Sepharose CL-6B spin column. Amplicons (457 bp) derived from primers to the invA and invE genes were confirmed as Salmonella specific on ethidium bromide-stained agarose gels by Southern hybridization with a 20-mer oligonucleotide probe specific for the Salmonella invA gene. Neither the standard microbiological method nor the molecular method detected all of the 65 samples that tested positive by both methods or either method alone. Salmonella bacteria were detected by both cultivation and PCR-hybridization in 68% (17 of 25) of the positive samples that were preenriched in TSB, in 73% (11 of 15) of the positive samples preenriched in 3MC broth, and in 24% (6 of 25) of the positive samples enriched in RV10. Agreement between Salmonella detection using cultivation with preenrichment and detection by PCR was 76% using the kappa statistic. However, agreement between Salmonella detection using cultivation without preenrichment and detection by PCR was about 6%; the PCR assay detected 80% (20 of 25) of the 25 positive samples, while Salmonella bacteria were recovered from only 44% (11 of 25) by cultivation. Our results indicate that the PCR-hybridization approach is equivalent to or better than cultivation for detecting Salmonella in swine feces or water samples from swine farms when using the medium combinations evaluated in this study. PMID:11427557

Comparative examination and validation of ELISA test systems for Salmonella typhimurium diagnosis of slaughtering pigs.

PubMed

Szabó, I; Scherer, K; Roesler, U; Appel, B; Nöckler, K; Hensel, A

2008-05-10

The most frequently isolated Salmonella serotype from pork in Germany is S. typhimurium, especially phagetype DT 104. The monitoring programs on Salmonella in swine are based on enzyme-linked immunoadsorbent assay (ELISA) detecting antibodies in serum or meat juice. These serological results are used to classify swine herds in three categories to assess the hygienic status of farm regarding Salmonella infection in pigs. The object of this study was the comparative evaluation of four indirect Salmonella ELISA tests approved in Germany to detect Salmonella typhimurium infection of swine. Three tests (A-C) are based on LPS-antigen and directed against specific IgG-antibodies. The fourth test (D) bases on a whole-cell-lysate antigen and discriminates between Salmonella specific IgA-, IgM- and IgG-antibodies. In a longitudinal study sixteen 6 weeks old weaning pigs were orally infected with S. typhimurium DT 104. During an observation period of 138d clinical and bacteriological parameters were monitored and serum samples obtained at regular intervals as well as meat juice samples taken at slaughter were examined by the respective ELISA systems. Study results reveal that all tested ELISA systems are able to detect S. typhimurium infection in pigs in both sample matrices, blood serum and meat juice whereas test D showed the highest sensitivity to detect Salmonella antibodies in pigs. The sensitivity to detect Salmonella antibodies varied between tests A and C according to the used cut-off (test specific cut-off vs. recommended surveillance cut-off) resulting in a change of seroprevalence and hence may influence the Salmonella status of the farm.

Comparison of four sampling methods for the detection of Salmonella in broiler litter.

PubMed

Buhr, R J; Richardson, L J; Cason, J A; Cox, N A; Fairchild, B D

2007-01-01

Experiments were conducted to compare litter sampling methods for the detection of Salmonella. In experiment 1, chicks were challenged orally with a suspension of naladixic acid-resistant Salmonella and wing banded, and additional nonchallenged chicks were placed into each of 2 challenge pens. Nonchallenged chicks were placed into each nonchallenge pen located adjacent to the challenge pens. At 7, 8, 10, and 11 wk of age the litter was sampled using 4 methods: fecal droppings, litter grab, drag swab, and sock. For the challenge pens, Salmonella-positive samples were detected in 3 of 16 fecal samples, 6 of 16 litter grab samples, 7 of 16 drag swabs samples, and 7 of 16 sock samples. Samples from the nonchallenge pens were Salmonella positive in 2 of 16 litter grab samples, 9 of 16 drag swab samples, and 9 of 16 sock samples. In experiment 2, chicks were challenged with Salmonella, and the litter in the challenge and adjacent nonchallenge pens were sampled at 4, 6, and 8 wk of age with broilers remaining in all pens. For the challenge pens, Salmonella was detected in 10 of 36 fecal samples, 20 of 36 litter grab samples, 14 of 36 drag swab samples, and 26 of 36 sock samples. Samples from the adjacent nonchallenge pens were positive for Salmonella in 6 of 36 fecal droppings samples, 4 of 36 litter grab samples, 7 of 36 drag swab samples, and 19 of 36 sock samples. Sock samples had the highest rates of Salmonella detection. In experiment 3, the litter from a Salmonella-challenged flock was sampled at 7, 8, and 9 wk by socks and drag swabs. In addition, comparisons with drag swabs that were stepped on during sampling were made. Both socks (24 of 36, 67%) and drag swabs that were stepped on (25 of 36, 69%) showed significantly more Salmonella-positive samples than the traditional drag swab method (16 of 36, 44%). Drag swabs that were stepped on had comparable Salmonella detection level to that for socks. Litter sampling methods that incorporate stepping on the sample material while in contact with the litter appear to detect Salmonella in greater incidence than traditional sampling methods of dragging swabs over the litter surface.

[Use of new immunoglobulin isotype-specific ELISA-systems to detect Salmonella infections in pigs].

PubMed

Ehlers, Joachim; Alt, Michael; Trepnau, Daniela; Lehmann, Jörg

2006-01-01

In Germany, the program for controlling salmonella infections in pigs is based on tests detecting salmonella-lipopolysaccharide (LPS) induced antibodies in meat-juice or blood. These conventional tests which are based on the technology of enzyme-linked immunosorbent assay (ELISA) detect exclusively or mainly immunoglobulin(lg)G antibodies. Meanwhile, novel ELISA systems (WCE-ELISA, 3-Isotype-Screening-ELISA) have been developed, which additionally detect the antibody classes IgM and IgA.This fact enables the registration of fresh salmonella infections (starting with day 5 p.i.) and thus, the distinction between early and older infections. The results show that animals with early salmonella infections appear significantly more often in herds with a high than with a low prevalence. With the newly developed tests this group of animals can be detected much more efficiently and precisely than with the tests used so far. Due to their clearly improved sensitivity the application of the WCE-ELISA and the 3-Isotype-Screening-ELISA in terms of the QS-Salmonella-Monitoring program can therefore significantly improve the selection of farms with potential salmonella excretors. Additionally, the WCE-ELISA can be applied very suitable for the examination of individual animals.

Horizontal Dissemination of Antimicrobial Resistance Determinants in Multiple Salmonella Serotypes following Isolation from the Commercial Swine Operation Environment after Manure Application.

PubMed

Pornsukarom, Suchawan; Thakur, Siddhartha

2017-10-15

The aim of this study was to characterize the plasmids carrying antimicrobial resistance (AMR) determinants in multiple Salmonella serotypes recovered from the commercial swine farm environment after manure application on land. Manure and soil samples were collected on day 0 before and after manure application on six farms in North Carolina, and sequential soil samples were recollected on days 7, 14, and 21 from the same plots. All environmental samples were processed for Salmonella , and their plasmid contents were further characterized. A total of 14 isolates including Salmonella enterica serotypes Johannesburg ( n = 2), Ohio ( n = 2), Rissen ( n = 1), Typhimurium var5- ( n = 5), Worthington ( n = 3), and 4,12:i:- ( n = 1), representing different farms, were selected for plasmid analysis. Antimicrobial susceptibility testing was done by broth microdilution against a panel of 14 antimicrobials on the 14 confirmed transconjugants after conjugation assays. The plasmids were isolated by modified alkaline lysis, and PCRs were performed on purified plasmid DNA to identify the AMR determinants and the plasmid replicon types. The plasmids were sequenced for further analysis and to compare profiles and create phylogenetic trees. A class 1 integron with an ANT(2″)-Ia- aadA2 cassette was detected in the 50-kb IncN plasmids identified in S Worthington isolates. We identified 100-kb and 90-kb IncI1 plasmids in S Johannesburg and S Rissen isolates carrying the bla CMY-2 and tet (A) genes, respectively. An identical 95-kb IncF plasmid was widely disseminated among the different serotypes and across different farms. Our study provides evidence on the importance of horizontal dissemination of resistance determinants through plasmids of multiple Salmonella serotypes distributed across commercial swine farms after manure application. IMPORTANCE The horizontal gene transfer of antimicrobial resistance (AMR) determinants located on plasmids is considered to be the main reason for the rapid proliferation and spread of drug resistance. The deposition of manure generated in swine production systems into the environment is identified as a potential source of AMR dissemination. In this study, AMR gene-carrying plasmids were detected in multiple Salmonella serotypes across different commercial swine farms in North Carolina. The plasmid profiles were characterized based on Salmonella serotype donors and incompatibility (Inc) groups. We found that different Inc plasmids showed evidence of AMR gene transfer in multiple Salmonella serotypes. We detected an identical 95-kb plasmid that was widely distributed across swine farms in North Carolina. These conjugable resistance plasmids were able to persist on land after swine manure application. Our study provides strong evidence of AMR determinant dissemination present in plasmids of multiple Salmonella serotypes in the environment after manure application. Copyright © 2017 American Society for Microbiology.

Detection of Viable Cryptosporidium parvum in Soil by Reverse Transcription–Real-Time PCR Targeting hsp70 mRNA ▿

PubMed Central

Liang, Zhanbei; Keeley, Ann

2011-01-01

Extraction of high-quality mRNA from Cryptosporidium parvum is a key step in PCR detection of viable oocysts in environmental samples. Current methods for monitoring oocysts are limited to water samples; therefore, the goal of this study was to develop a rapid and sensitive procedure for Cryptosporidium detection in soil samples. The efficiencies of five RNA extraction methods were compared (mRNA extraction with the Dynabeads mRNA Direct kit after chemical and physical sample treatments, and total RNA extraction methods using the FastRNA Pro Soil-Direct, PowerSoil Total RNA, E.Z.N.A. soil RNA, and Norgen soil RNA purification kits) for the direct detection of Cryptosporidium with oocyst-spiked sandy, loamy, and clay soils by using TaqMan reverse transcription-PCR. The study also evaluated the presence of inhibitors by synthesis and incorporation of an internal positive control (IPC) RNA into reverse transcription amplifications, used different facilitators (bovine serum albumin, yeast RNA, salmon DNA, skim milk powder, casein, polyvinylpyrrolidone, sodium hexametaphosphate, and Salmonella enterica serovar Typhi) to mitigate RNA binding on soil components, and applied various treatments (β-mercaptoethanol and bead beating) to inactivate RNase and ensure the complete lysis of oocysts. The results of spiking studies showed that Salmonella cells most efficiently relieved binding of RNA. With the inclusion of Salmonella during extraction, the most efficient mRNA method was Dynabeads, with a detection limit of 6 × 102 oocysts g−1 of sandy soil. The most efficient total RNA method was PowerSoil, with detection limits of 1.5 × 102, 1.5 × 103, and 1.5 × 104 C. parvum oocysts g−1 soil for sandy, loamy, and clay samples, respectively. PMID:21803904

Efficient and Specific Detection of Salmonella in Food Samples Using a stn-Based Loop-Mediated Isothermal Amplification Method

PubMed Central

2015-01-01

The Salmonella enterotoxin (stn) gene exhibits high homology among S. enterica serovars and S. bongori. A set of 6 specific primers targeting the stn gene were designed for detection of Salmonella spp. using the loop-mediated isothermal amplification (LAMP) method. The primers amplified target sequences in all 102 strains of 87 serovars of Salmonella tested and no products were detected in 57 non-Salmonella strains. The detection limit in pure cultures was 5 fg DNA/reaction when amplified at 65°C for 25 min. The LAMP assay could detect Salmonella in artificially contaminated food samples as low as 220 cells/g of food without a preenrichment step. However, the sensitivity was increased 100-fold (~2 cells/g) following 5 hr preenrichment at 35°C. The LAMP technique, with a preenrichment step for 5 and 16 hr, was shown to give 100% specificity with food samples compared to the reference culture method in which 67 out of 90 food samples gave positive results. Different food matrixes did not interfere with LAMP detection which employed a simple boiling method for DNA template preparation. The results indicate that the LAMP method, targeting the stn gene, has great potential for detection of Salmonella in food samples with both high specificity and high sensitivity. PMID:26543859

Detection of Salmonella species in chicken carcasses using genus specific primer belong to invA gene in Sohag city, Egypt.

PubMed

Abdel-Aziz, Nahed Mahmoud

2016-10-01

This study aimed to detect Salmonella species found as contaminants in chicken carcass (thigh, breast, wings, liver, and gizzard). A total of 75 chicken samples including thigh, breast, wings, liver, and gizzard (15 of each) were collected from different markets in Sohag city for detection of Salmonella species by culture methods, biochemical tests, serology, and polymerase chain reaction. The overall incidence of Salmonella contamination of 75 examined samples was found to be 6.6% with the higher percentage of Salmonella being isolated from liver samples (13.3%) followed by thigh, wings, gizzard (6.6%) while breast show negative result. Results in this study indicate that contamination of chicken carcass with Salmonella needs strict hygienic measures to prevent their transmission to human.

A Portable Impedance Immunosensing System for Rapid Detection of Salmonella Typhimurium

PubMed Central

Wen, Tao; Wang, Ronghui; Sotero, America; Li, Yanbin

2017-01-01

Salmonella Typhimurium is one of the most dangerous foodborne pathogens and poses a significant threat to human health. The objective of this study was to develop a portable impedance immunosensing system for rapid and sensitive detection of S. Typhimurium in poultry. The developed portable impedance immunosensing system consisted of a gold interdigitated array microelectrode (IDAM), a signal acquisitive interface and a laptop computer with LabVIEW software. The IDAM was first functionalized with 16-Mercaptohexadecanoic acid, and streptavidin was immobilized onto the electrode surface through covalent bonding. Then, biotin-labelled S. Typhimurium-antibody was immobilized onto the IDAM surface. Samples were dropped on the surface of the IDAM and the S. Typhimurium cells in the samples were captured by the antibody on the IDAM. This resulted in impedance changes that were measured and displayed with the LabVIEW software. An equivalent circuit of the immunosensor demonstrated that the largest change in impedance was due to the electron-transfer resistance. The equivalent circuit showed an increase of 35% for the electron-transfer resistance value compared to the negative control. The calibration result indicated that the portable impedance immunosensing system could be used to measure the standard impedance elements, and it had a maximum error of measurement of approximately 13%. For pure culture detection, the system had a linear relationship between the impedance change and the logarithmic value of S. Typhimurium cells ranging from 76 to 7.6 × 106 CFU (colony-forming unit) (50 μL)−1. The immunosensor also had a correlation coefficient of 0.98, and a high specificity for detection of S. Typhimurium cells with a limit of detection (LOD) of 102 CFU (50 μL)−1. The detection time from the moment a sample was introduced to the display of the results was 1 h. To conclude, the portable impedance immunosensing system for detection of S. Typhimurium achieved an LOD that is comparable with commercial electrochemical impedance instruments. The developed impedance immunosensor has advantages in portability, low cost, rapid detection and label-free features showing a great potential for in-field detection of foodborne pathogens. PMID:28846643

A Portable Impedance Immunosensing System for Rapid Detection of Salmonella Typhimurium.

PubMed

Wen, Tao; Wang, Ronghui; Sotero, America; Li, Yanbin

2017-08-28

Salmonella Typhimurium is one of the most dangerous foodborne pathogens and poses a significant threat to human health. The objective of this study was to develop a portable impedance immunosensing system for rapid and sensitive detection of S . Typhimurium in poultry. The developed portable impedance immunosensing system consisted of a gold interdigitated array microelectrode (IDAM), a signal acquisitive interface and a laptop computer with LabVIEW software. The IDAM was first functionalized with 16-Mercaptohexadecanoic acid, and streptavidin was immobilized onto the electrode surface through covalent bonding. Then, biotin-labelled S . Typhimurium -antibody was immobilized onto the IDAM surface. Samples were dropped on the surface of the IDAM and the S . Typhimurium cells in the samples were captured by the antibody on the IDAM. This resulted in impedance changes that were measured and displayed with the LabVIEW software. An equivalent circuit of the immunosensor demonstrated that the largest change in impedance was due to the electron-transfer resistance. The equivalent circuit showed an increase of 35% for the electron-transfer resistance value compared to the negative control. The calibration result indicated that the portable impedance immunosensing system could be used to measure the standard impedance elements, and it had a maximum error of measurement of approximately 13%. For pure culture detection, the system had a linear relationship between the impedance change and the logarithmic value of S . Typhimurium cells ranging from 76 to 7.6 × 10⁶ CFU (colony-forming unit) (50 μL) -1 . The immunosensor also had a correlation coefficient of 0.98, and a high specificity for detection of S . Typhimurium cells with a limit of detection (LOD) of 10² CFU (50 μL) -1 . The detection time from the moment a sample was introduced to the display of the results was 1 h. To conclude, the portable impedance immunosensing system for detection of S . Typhimurium achieved an LOD that is comparable with commercial electrochemical impedance instruments. The developed impedance immunosensor has advantages in portability, low cost, rapid detection and label-free features showing a great potential for in-field detection of foodborne pathogens.

Detection of Salmonella sp in chicken cuts using immunomagnetic separation

PubMed Central

de Cássia dos Santos da Conceição, Rita; Moreira, Ângela Nunes; Ramos, Roberta Juliano; Goularte, Fabiana Lemos; Carvalhal, José Beiro; Aleixo, José Antonio Guimarães

2008-01-01

The immunomagnetic separation (IMS) is a technique that has been used to increase sensitivity and specificity and to decrease the time required for detection of Salmonella in foods through different methodologies. In this work we report on the development of a method for detection of Salmonella in chicken cuts using in house antibody-sensitized microspheres associated to conventional plating in selective agar (IMS-plating). First, protein A-coated microspheres were sensitized with polyclonal antibodies against lipopolysacharide and flagella from salmonellae and used to standardize a procedure for capturing Salmonella Enteritidis from pure cultures and detection in selective agar. Subsequently, samples of chicken meat experimentally contaminated with S. Enteritidis were analyzed immediately after contamination and after 24h of refrigeration using three enrichment protocols. The detection limit of the IMS-plating procedure after standardization with pure culture was about 2x10 CFU/mL. The protocol using non-selective enrichment for 6-8h, selective enrichment for 16-18h and a post-enrichment for 4h gave the best results of S. Enteritidis detection by IMS-plating in experimentally contaminated meat. IMS-plating using this protocol was compared to the standard culture method for salmonellae detection in naturally contaminated chicken cuts and yielded 100% sensitivity and 94% specificity. The method developed using in house prepared magnetic microespheres for IMS and plating in selective agar was able to diminish by at least one day the time required for detection of Salmonella in chicken products by the conventional culture method. PMID:24031199

Salmonella spp. contamination in commercial layer hen farms using different types of samples and detection methods.

PubMed

Soria, M C; Soria, M A; Bueno, D J; Godano, E I; Gómez, S C; ViaButron, I A; Padin, V M; Rogé, A D

2017-08-01

The performance of detection methods (culture methods and polymerase chain reaction assay) and plating media used in the same type of samples were determined as well as the specificity of PCR primers to detected Salmonella spp. contamination in layer hen farms. Also, the association of farm characteristics with Salmonella presence was evaluated. Environmental samples (feces, feed, drinking water, air, boot-swabs) and eggs were taken from 40 layer hen houses. Salmonella spp. was most detected in boot-swabs taken around the houses (30% and 35% by isolation and PCR, respectively) follow by fecal samples (15.2% and 13.6% by isolation and PCR, respectively). Eggs, drinking water, and air samples were negative for Salmonella detection. Salmonella Schwarzengrund and S. Enteritidis were the most isolated serotypes. For plating media, relative specificity was 1, and the relative sensitivity was greater for EF-18 agar than XLDT agar in feed and fecal samples. However, relative sensitivity was greater in XLDT agar than EF-18 agar for boot-swab samples. Agreement was between fair to good depending on the sample, and it was good between isolation and PCR (feces and boot-swabs), without agreement for feed samples. Salmonella spp. PCR was positive for all strains, while S. Typhimurium PCR was negative. Salmonella Enteritidis PCR used was not specific. Based in the multiple logistic regression analyses, categorization by counties was significant for Salmonella spp. presence (P-value = 0.010). This study shows the importance of considering different types of samples, plating media and detection methods during a Salmonella spp. monitoring study. In addition, it is important to incorporate the sampling of floors around the layer hen houses to learn if biosecurity measures should be strengthened to minimize the entry and spread of Salmonella in the houses. Also, the performance of some PCR methods and S. Enteritidis PCR should be improved, and biosecurity measures in hen farms must be reinforced in the region of more concentrated layer hen houses to reduce the probability of Salmonella spp. presence. © 2017 Poultry Science Association Inc.

Culture- and molecular-based detection of swine-adapted Salmonella shed by avian scavengers.

PubMed

Blanco, Guillermo; Díaz de Tuesta, Juan A

2018-09-01

Salmonella can play an important role as a disease agent in wildlife, which can then act as carriers and reservoirs of sanitary importance at the livestock-human interface. Transmission from livestock to avian scavengers can occur when these species consume contaminated carcasses and meat remains in supplementary feeding stations and rubbish dumps. We compared the performance of PCR-based detection with conventional culture-based methods to detect Salmonella in the faeces of red kites (Milvus milvus) and griffon vultures (Gyps fulvus) in central Spain. The occurrence of culturable Salmonella was intermediate in red kites (1.9%, n=52) and high in griffon vultures (26.3%, n=99). These proportions were clearly higher with PCR-based detection (13.5% and 40.4%, respectively). Confirmation cultures failed to grow Salmonella in all faecal samples positive by the molecular assay but negative by the initial conventional culture in both scavenger species, indicating the occurrence of false (non-culturable) positives by PCR-based detection. This suggests that the molecular assay is highly sensitive to detecting viable Salmonella in cultures, but also partial genomes and dead or unviable bacteria from past infections or contamination. Thus, the actual occurrence of Salmonella in a particular sampling time period can be underestimated when using only culture detection. The serovars found in the scavenger faeces were among the most frequently isolated in pigs from Spain and other EU countries, especially those generally recognized as swine-adapted monophasic variants of S. Typhimurium. Because the studied species obtain much of their food from pig carcasses, this livestock may be the primary source of Salmonella via direct ingestion of infected carcasses and indirectly via contamination due to the unsanitary conditions found in supplementary feeding stations established for scavenger conservation. Combining culture- and molecular-based detection is encouraged to understand the epidemiology and impact of Salmonella in wildlife populations. Copyright © 2018 Elsevier B.V. All rights reserved.

The global establishment of a highly-fluoroquinolone resistant Salmonella enterica serotype Kentucky ST198 strain.

PubMed

Le Hello, Simon; Bekhit, Amany; Granier, Sophie A; Barua, Himel; Beutlich, Janine; Zając, Magdalena; Münch, Sebastian; Sintchenko, Vitali; Bouchrif, Brahim; Fashae, Kayode; Pinsard, Jean-Louis; Sontag, Lucile; Fabre, Laetitia; Garnier, Martine; Guibert, Véronique; Howard, Peter; Hendriksen, Rene S; Christensen, Jens P; Biswas, Paritosh K; Cloeckaert, Axel; Rabsch, Wolfgang; Wasyl, Dariusz; Doublet, Benoit; Weill, François-Xavier

2013-01-01

While the spread of Salmonella enterica serotype Kentucky resistant to ciprofloxacin across Africa and the Middle-East has been described recently, the presence of this strain in humans, food, various animal species (livestock, pets, and wildlife) and in environment is suspected in other countries of different continents. Here, we report results of an in-depth molecular epidemiological study on a global human and non-human collection of S. Kentucky (n = 70). We performed XbaI-pulsed field gel electrophoresis and multilocus sequence typing, assessed mutations in the quinolone resistance-determining regions, detected β-lactam resistance mechanisms, and screened the presence of the Salmonella genomic island 1 (SGI1). In this study, we highlight the rapid and extensive worldwide dissemination of the ciprofloxacin-resistant S. Kentucky ST198-X1-SGI1 strain since the mid-2000s in an increasingly large number of contaminated sources, including the environment. This strain has accumulated an increasing number of chromosomal and plasmid resistance determinants and has been identified in the Indian subcontinent, Southeast Asia and Europe since 2010. The second substitution at position 87 in GyrA (replacing the amino acid Asp) appeared helpful for epidemiological studies to track the origin of contamination. This global study provides evidence leading to the conclusion that high-level resistance to ciprofloxacin in S. Kentucky is a simple microbiological trait that facilitates the identification of the epidemic clone of interest, ST198-X1-SGI1. Taking this into account is essential in order to detect and monitor it easily and to take rapid measures in livestock to ensure control of this infection.

The global establishment of a highly-fluoroquinolone resistant Salmonella enterica serotype Kentucky ST198 strain

PubMed Central

Le Hello, Simon; Bekhit, Amany; Granier, Sophie A.; Barua, Himel; Beutlich, Janine; Zając, Magdalena; Münch, Sebastian; Sintchenko, Vitali; Bouchrif, Brahim; Fashae, Kayode; Pinsard, Jean-Louis; Sontag, Lucile; Fabre, Laetitia; Garnier, Martine; Guibert, Véronique; Howard, Peter; Hendriksen, Rene S.; Christensen, Jens P.; Biswas, Paritosh K.; Cloeckaert, Axel; Rabsch, Wolfgang; Wasyl, Dariusz; Doublet, Benoit; Weill, François-Xavier

2013-01-01

While the spread of Salmonella enterica serotype Kentucky resistant to ciprofloxacin across Africa and the Middle-East has been described recently, the presence of this strain in humans, food, various animal species (livestock, pets, and wildlife) and in environment is suspected in other countries of different continents. Here, we report results of an in-depth molecular epidemiological study on a global human and non-human collection of S. Kentucky (n = 70). We performed XbaI-pulsed field gel electrophoresis and multilocus sequence typing, assessed mutations in the quinolone resistance-determining regions, detected β-lactam resistance mechanisms, and screened the presence of the Salmonella genomic island 1 (SGI1). In this study, we highlight the rapid and extensive worldwide dissemination of the ciprofloxacin-resistant S. Kentucky ST198-X1-SGI1 strain since the mid-2000s in an increasingly large number of contaminated sources, including the environment. This strain has accumulated an increasing number of chromosomal and plasmid resistance determinants and has been identified in the Indian subcontinent, Southeast Asia and Europe since 2010. The second substitution at position 87 in GyrA (replacing the amino acid Asp) appeared helpful for epidemiological studies to track the origin of contamination. This global study provides evidence leading to the conclusion that high-level resistance to ciprofloxacin in S. Kentucky is a simple microbiological trait that facilitates the identification of the epidemic clone of interest, ST198-X1-SGI1. Taking this into account is essential in order to detect and monitor it easily and to take rapid measures in livestock to ensure control of this infection. PMID:24385975

Selectively Reduced Intracellular Proliferation of Salmonella enterica Serovar Typhimurium within APCs Limits Antigen Presentation and Development of a Rapid CD8 T Cell Response1

PubMed Central

Albaghdadi, Homam; Robinson, Nirmal; Finlay, Brett; Krishnan, Lakshmi; Sad, Subash

2014-01-01

Ag presentation to CD8+ T cells commences immediately after infection, which facilitates their rapid expansion and control of pathogen. This paradigm is not followed during infection with virulent Salmonella enterica serovar Typhimurium (ST), an intracellular bacterium that causes mortality in susceptible C57BL/6J mice within 7 days and a chronic infection in resistant mice (129 × 1SvJ). Infection of mice with OVA-expressing ST results in the development of a CD8+ T cell response that is detectable only after the second week of infection despite the early detectable bacterial burden. The mechanism behind the delayed CD8+ T cell activation was evaluated, and it was found that dendritic cells/macrophages or mice infected with ST-OVA failed to present Ag to OVA-specific CD8+ T cells. Lack of early Ag presentation was not rescued when mice or dendritic cells/macrophages were infected with an attenuated aroA mutant of ST or with mutants having defective Salmonella pathogenicity island I/II genes. Although extracellular ST proliferated extensively, the replication of ST was highly muted once inside macrophages. This muted intracellular proliferation of ST resulted in the generation of poor levels of intracellular Ag and peptide-MHC complex on the surface of dendritic cells. Additional experiments revealed that ST did not actively inhibit Ag presentation, rather it inhibited the uptake of another intracellular pathogen, Listeria monocytogenes, thereby causing inhibition of Ag presentation against L. monocytogenes. Taken together, this study reveals a dichotomy in the proliferation of ST and indicates that selectively reduced intra-cellular proliferation of virulent pathogens may be an important mechanism of immune evasion. PMID:19692639

Factors Associated with Salmonella Prevalence in U.S. Swine Grower-Finisher Operations, 2012.

PubMed

Bjork, Kathe E; Fields, Victoria; Garber, Lindsey P; Kopral, Christine A

2018-05-15

Nontyphoidal Salmonella is an important foodborne pathogen with diverse serotypes occurring in animal and human populations. The prevalence of the organism on swine farms has been associated with numerous risk factors, and although there are strong veterinary public health controls for preventing Salmonella from entering food, there remains interest in eradicating or controlling the organism in the preharvest environment. In this study, using data collected via the U.S. Department of Agriculture (USDA) National Animal Health Monitoring System Swine 2012 study, we describe nontyphoidal Salmonella and specific serotype prevalence on U.S. grower-finisher swine operations and investigate associations between Salmonella detection and numerous factors via multiple correspondence analysis (MCA) and regression analysis. MCA plots, complementary to univariate analyses, display relationships between covariates and Salmonella detection at the farm level. In the univariate analysis, Salmonella detection varied with feed characteristics and farm management practices, reports of diseases on farms and vaccinations administered, and administration of certain antimicrobials. Results from the univariate analysis reinforce the importance of biosecurity in managing diseases and pathogens such as Salmonella on farms. All multivariable regression models for the likelihood of Salmonella detection were strongly affected by multicollinearity among variables, and only one variable, pelleted feed preparation, remained in the final model. The study was limited by its cross-sectional nature, timelines of data collection, and reliance on operator-reported data via a convenience sample.

Whole-bacterium SELEX of DNA aptamers for rapid detection of E.coli O157:H7 using a QCM sensor.

PubMed

Yu, Xiaofan; Chen, Fang; Wang, Ronghui; Li, Yanbin

2018-01-20

The rapid detection of foodborne pathogens is critical to ensure food safety. The objective of this study is to select aptamers specifically bound to Escherichia coli O157:H7 using the whole-bacterium SELEX (Systematic Evolution of Ligands by Exponential Enrichment) and apply the selected aptamer to a QCM (quartz crystal microbalance) sensor for rapid and sensitive detection of target bacteria. A total of 19 rounds of selection against live E. coli O157:H7 and 6 rounds of counter selection against a mixture of Staphylococcus aureus, Listeria monocytogenes, and Salmonella Typhimurium, were performed. The aptamer pool from the last round was cloned and sequenced. One sequence S1 that appeared 16 times was characterized and a dissociation constant (K d ) of 10.30nM was obtained. Subsequently, a QCM aptasensor was developed for the rapid detection of E. coli O157:H7. The limit of detection (LOD) and the detection time of the aptasensor was determined to be 1.46×10 3 CFU/ml and 50min, respectively. This study demonstrated that the ssDNA aptamer selected by the whole-bacterium SELEX possessed higher sensitivity than previous work and the potential use of the constructed QCM aptasensor in rapid screening of foodborne pathogens. Copyright © 2017 Elsevier B.V. All rights reserved.

A novel strategy to obtain quantitative data for modelling: combined enrichment and real-time PCR for enumeration of salmonellae from pig carcasses.

PubMed

Krämer, Nadine; Löfström, Charlotta; Vigre, Håkan; Hoorfar, Jeffrey; Bunge, Cornelia; Malorny, Burkhard

2011-03-01

Salmonella is a major zoonotic pathogen which causes outbreaks and sporadic cases of gastroenteritis in humans worldwide. The primary sources for Salmonella are food-producing animals such as pigs and poultry. For risk assessment and hazard analysis and critical control point (HACCP) concepts, it is essential to produce large amounts of quantitative data, which is currently not achievable with the standard cultural based methods for enumeration of Salmonella. This study presents the development of a novel strategy to enumerate low numbers of Salmonella in cork borer samples taken from pig carcasses as a first concept and proof of principle for a new sensitive and rapid quantification method based on combined enrichment and real-time PCR. The novelty of the approach is in the short pre-enrichment step, where for most bacteria, growth is in the log phase. The method consists of an 8h pre-enrichment of the cork borer sample diluted 1:10 in non-selective buffered peptone water, followed by DNA extraction, and Salmonella detection and quantification by real-time PCR. The limit of quantification was 1.4 colony forming units (CFU)/20 cm(2) (approximately 10 g) of artificially contaminated sample with 95% confidence interval of ± 0.7 log CFU/sample. The precision was similar to the standard reference most probable number (MPN) method. A screening of 200 potentially naturally contaminated cork borer samples obtained over seven weeks in a slaughterhouse resulted in 25 Salmonella-positive samples. The analysis of salmonellae within these samples showed that the PCR method had a higher sensitivity for samples with a low contamination level (<6.7 CFU/sample), where 15 of the samples negative with the MPN method was detected with the PCR method and 5 were found to be negative by both methods. For the samples with a higher contamination level (6.7-310 CFU/sample) a good agreement between the results obtained with the PCR and MPN methods was obtained. The quantitative real-time PCR method can easily be applied to other food and environmental matrices by adaptation of the pre-enrichment time and media. Copyright © 2010 Elsevier B.V. All rights reserved.

AOAC Official MethodSM Matrix Extension Validation Study of Assurance GDSTM for the Detection of Salmonella in Selected Spices.

PubMed

Feldsine, Philip; Kaur, Mandeep; Shah, Khyati; Immerman, Amy; Jucker, Markus; Lienau, Andrew

2015-01-01

Assurance GDSTM for Salmonella Tq has been validated according to the AOAC INTERNATIONAL Methods Committee Guidelines for Validation of Microbiological Methods for Food and Environmental Surfaces for the detection of selected foods and environmental surfaces (Official Method of AnalysisSM 2009.03, Performance Tested MethodSM No. 050602). The method also completed AFNOR validation (following the ISO 16140 standard) compared to the reference method EN ISO 6579. For AFNOR, GDS was given a scope covering all human food, animal feed stuff, and environmental surfaces (Certificate No. TRA02/12-01/09). Results showed that Assurance GDS for Salmonella (GDS) has high sensitivity and is equivalent to the reference culture methods for the detection of motile and non-motile Salmonella. As part of the aforementioned validations, inclusivity and exclusivity studies, stability, and ruggedness studies were also conducted. Assurance GDS has 100% inclusivity and exclusivity among the 100 Salmonella serovars and 35 non-Salmonella organisms analyzed. To add to the scope of the Assurance GDS for Salmonella method, a matrix extension study was conducted, following the AOAC guidelines, to validate the application of the method for selected spices, specifically curry powder, cumin powder, and chili powder, for the detection of Salmonella.

Detection of Salmonella spp. from chevon, mutton and its environment in retail meat shops in Anand city (Gujarat), India

PubMed Central

Makwana, P. P.; Nayak, J. B.; Brahmbhatt, M. N.; Chaudhary, J. H.

2015-01-01

Aim: The aim of this study was (i) To attempt isolation and identification of Salmonella species from samples. (ii) Serotyping of Salmonella isolates. (iii) Detection of virulence factor associated genes by polymerase chain reaction (PCR). Materials and Methods: A total of 284 samples comprised of chevon and mutton (112 samples each) as well as 60 samples (20 each of retail meat shops environment samples viz. Butchers’ hands, knives and log swabs) were collected from the retail meat shops in and around Anand City under aseptic precautions. Rappaport-vassiliadis soy bean meal broth and tetrathionate broth was used for the enrichment of all the samples and inoculation was done on brilliant green agar and xylose lysine deoxycholate agar. This was followed by the confirmation of isolates using biochemical tests. For the serotyping, isolates were sent to the National Salmonella and Escherichia Centre, Central Research Institute, Kasauli, Himachal Pradesh. Detection of virulence genes was performed by PCR technique using previously reported primer. Result: Of 284 meats and retail meat shops environment samples, 13 (4.58%) samples were found positive for Salmonella. It was interesting to know that incidence of Salmonella was more in mutton (6.25%) than chevon (3.57%). In case of meat shop environmental samples 1 (5.00%) sample observed positive for Salmonella separately among the butchers’ hands and knives swabs (Each of 20 samples) examined. Out of 13, eleven isolates detected as Salmonella Typhimurium, whereas only two isolates were detected as Salmonella Enteritidis. All Salmonella isolates possess invA and stn genes, whereas nine isolates had a presence of spvR gene while only five of the isolates revealed the presence of spvC gene as shown by in vitro detection of virulence genes by PCR. Conclusion: Therefore, might be suggested that the good hygiene practices and effective control measures should be taken to encourage clean meat production with prolonged shelf-life. PMID:27047102

Survey of Salmonella contamination of non-United Kingdom-produced raw shell eggs on retail sale in the northwest of England and London, 2005 to 2006.

PubMed

Little, C L; Walsh, S; Hucklesby, L; Surman-Lee, S; Pathak, K; Gatty, Y; Greenwood, M; De Pinna, E; Threlfall, E J; Maund, A; Chan, C H

2007-10-01

This survey was prompted by a change in the epidemiology of Salmonella Enteritidis infections in England and Wales and elsewhere in Europe and, to our knowledge, is the first survey to provide information on Salmonella contamination of non-United Kingdom eggs on retail sale. Based on 10,464 non-United Kingdom eggs (1744 pooled samples of six eggs) purchased between March 2005 and July 2006, the total weighted prevalence estimate for all Salmonella detected in non-United Kingdom eggs was 3.3%. Of the eggs sampled, most were produced in Spain (66.3%), France (20.0%), or The Netherlands (7.4%). Salmonella was detected from 4.4 and 0.3% of eggs produced in Spain and France, respectively, with weighted prevalence estimates. Eight different Salmonella serotypes were recovered from non-United Kingdom eggs, of which Salmonella Enteritidis predominated, with an estimated prevalence of 2.6%. Salmonella Enteritidis was obtained only from Spanish eggs. Nine different phage types of Salmonella Enteritidis were identified, with phage type 1 found to be the predominant phage type. Most of the Salmonella Enteritidis isolates obtained from Spanish eggs in the survey were resistant to nalidixic acid with concomitant decreased susceptibility to ciprofloxacin (0.125 to 1.0 mg/liter) or ampicillin (8.0 mg/liter). Salmonella Enteritidis phage type 1 until now had not been detected in eggs examined as part of previous United Kingdom egg surveys but has been detected in eggs of Spanish origin examined during recent national outbreaks of Salmonella Enteritidis non-phage type 4 infections in England and Wales. Eggs are a commonly consumed food that may occasionally be contaminated with Salmonella. The rates of contamination may be linked to the origin of the eggs. Consumers and caterers need to be aware of this continuing hazard, adopt appropriate control measures, and follow advice provided by national food agencies in order to reduce the risk of infection.

Development of an immunochromatographic test with anti-LipL32-coupled gold nanoparticles for Leptospira detection.

PubMed

Chirathaworn, Chintana; Janwitthayanan, Weena; Sereemaspun, Amornpun; Lertpocasombat, Kanchalee; Rungpanich, Utane; Ekpo, Pattama; Suwancharoen, Duangjai

2014-04-01

Detection of antibody specific to Leptospira by various immunological techniques has been used for leptospirosis diagnosis. However, the sensitivity of antibody detection during the first few days after infection is low. Molecular techniques are suggested to provide earlier diagnosis than antibody detection, but a rapid and easy to perform assay for Leptospira antigen detection would provide an additional useful tool for disease diagnosis. In this study, we coupled gold nanoparticles with antibody to LipL32, a protein commonly found in pathogenic Leptospira. This coupled gold reagent was used in the immunochromatographic strip for Leptospira detection. We demonstrated that the sensitivity of Leptospira detection by this strip was 10(3) ml(-1). There was no positive result detected when strips were tested with non-pathogenic Leptospira, Staphylococcus aureus, Streptococcus group B, Acinetobacter baumannii, Escherichia coli, Salmonella typhi, Klebsiella pneumoniae, Enterococcus faecalis or Enterococcus faecium. These data suggest that gold nanoparticles coupled with antibody to LipL32 could be used for Leptospira detection by a rapid test based on an immunochromatographic technique.

A Single-Step Enrichment Medium for Nonchromogenic Isolation of Healthy and Cold-Injured Salmonella spp. from Fresh Vegetables.

PubMed

Kim, Hong-Seok; Choi, Dasom; Kang, Il-Byeong; Kim, Dong-Hyeon; Yim, Jin-Hyeok; Kim, Young-Ji; Chon, Jung-Whan; Oh, Deog-Hwan; Seo, Kun-Ho

2017-02-01

Culture-based detection of nontyphoidal Salmonella spp. in foods requires at least four working days; therefore, new detection methods that shorten the test time are needed. In this study, we developed a novel single-step Salmonella enrichment broth, SSE-1, and compared its detection capability with that of commercial single-step ONE broth-Salmonella (OBS) medium and a conventional two-step enrichment method using buffered peptone water and Rappaport-Vassiliadis soy broth (BPW-RVS). Minimally processed lettuce samples were artificially inoculated with low levels of healthy and cold-injured Salmonella Enteritidis (10 0 or 10 1 colony-forming unit/25 g), incubated in OBS, BPW-RVS, and SSE-1 broths, and streaked on xylose lysine deoxycholate (XLD) agar. Salmonella recoverability was significantly higher in BPW-RVS (79.2%) and SSE-1 (83.3%) compared to OBS (39.3%) (p < 0.05). Our data suggest that the SSE-1 single-step enrichment broth could completely replace two-step enrichment with reduced enrichment time from 48 to 24 h, performing better than commercial single-step enrichment medium in the conventional nonchromogenic Salmonella detection, thus saving time, labor, and cost.

Prevalence of Extended-Spectrum β-Lactamases CTX-M-8 and CTX-M-2-Producing Salmonella Serotypes from Clinical and Nonhuman Isolates in Brazil.

PubMed

Fernandes, Sueli Aparecida; Camargo, Carlos Henrique; Francisco, Gabriela Rodrigues; Bueno, Maria Fernanda Campagnari; Garcia, Doroti Oliveira; Doi, Yohei; Casas, Monique Ribeiro Tiba

2017-07-01

We characterized extended-spectrum β-lactamases (ESBL) enzymes among Salmonella strains isolated in Brazil from 2009 to 2014. Salmonella recovered from both clinical and nonhuman (food, poultry, and environment) sources were subjected to antimicrobial susceptibility testing. β-lactamases genes were detected by polymerase chain reaction/sequencing; plasmid profiles and transferability were assessed by S1-pulsed field gel electrophoresis (PFGE). Genetic diversity was evaluated by XbaI-PFGE. Out of 630 Salmonella strains screened, 46 displayed ESBL phenotype, distributed across 11 different serotypes. bla CTX-M-8 and bla CTX-M-2 genes were detected at frequencies of 47% and 41%, respectively. bla SHV-5 and bla SHV-2 were also detected but in lower frequencies (4%, 2%). bla TEM-1 gene was detected in 22% of the strains. Most of the ESBL genes were transferable by conjugation, and the respective bla ESBL gene was detected in the recipient strain, indicating the location of ESBL determinants on transferable plasmids. XbaI-PFGE revealed genomic diversity of Salmonella Typhimurium bearing bla CTX-M-2 , bla CTX-M-8 , bla TEM-1 , and bla SHV-2 genes. Salmonella Muenchen (harboring bla CTX-M-2 ) and Salmonella Corvallis (bla CTX-M-8 and bla SHV-5 ) showed clonal relatedness within respective serotypes. Our findings underscore the occurrence of diverse ESBL genes in several Salmonella serotypes, reinforcing the need for continuous surveillance of resistance genes circulating in human and nonhuman sources.

Enumeration of Salmonella and Campylobacter spp. in environmental farm samples and processing plant carcass rinses from commercial broiler chicken flocks.

PubMed

Berghaus, Roy D; Thayer, Stephan G; Law, Bibiana F; Mild, Rita M; Hofacre, Charles L; Singer, Randall S

2013-07-01

A prospective cohort study was performed to evaluate the prevalences and loads of Salmonella and Campylobacter spp. in farm and processing plant samples collected from 55 commercial broiler chicken flocks. Environmental samples were collected from broiler houses within 48 h before slaughter, and carcass rinses were performed on birds from the same flocks at 4 different stages of processing. Salmonella was detected in farm samples of 50 (90.9%) flocks and in processing samples of 52 (94.5%) flocks. Campylobacter was detected in farm samples of 35 (63.6%) flocks and in processing samples of 48 (87.3%) flocks. There was a significant positive relationship between environmental farm samples and processing plant carcass rinses with respect to both Salmonella and Campylobacter prevalences and loads. Campylobacter loads were significantly higher than Salmonella loads, and the correlations between samples collected from the same flocks were higher for Campylobacter than they were for Salmonella. Boot socks were the most sensitive sample type for detection of Salmonella on the farm, whereas litter samples had the strongest association with Salmonella loads in pre- and postchill carcass rinses. Boot socks, drag swabs, and fecal samples all had similar sensitivities for detecting Campylobacter on the farm, and all were more strongly associated with Campylobacter loads in carcass rinses than were litter samples. Farm samples explained a greater proportion of the variability in carcass rinse prevalences and loads for Campylobacter than they did for Salmonella. Salmonella and Campylobacter prevalences and loads both decreased significantly as birds progressed through the processing plant.

Enumeration of Salmonella and Campylobacter spp. in Environmental Farm Samples and Processing Plant Carcass Rinses from Commercial Broiler Chicken Flocks

PubMed Central

Thayer, Stephan G.; Law, Bibiana F.; Mild, Rita M.; Hofacre, Charles L.; Singer, Randall S.

2013-01-01

A prospective cohort study was performed to evaluate the prevalences and loads of Salmonella and Campylobacter spp. in farm and processing plant samples collected from 55 commercial broiler chicken flocks. Environmental samples were collected from broiler houses within 48 h before slaughter, and carcass rinses were performed on birds from the same flocks at 4 different stages of processing. Salmonella was detected in farm samples of 50 (90.9%) flocks and in processing samples of 52 (94.5%) flocks. Campylobacter was detected in farm samples of 35 (63.6%) flocks and in processing samples of 48 (87.3%) flocks. There was a significant positive relationship between environmental farm samples and processing plant carcass rinses with respect to both Salmonella and Campylobacter prevalences and loads. Campylobacter loads were significantly higher than Salmonella loads, and the correlations between samples collected from the same flocks were higher for Campylobacter than they were for Salmonella. Boot socks were the most sensitive sample type for detection of Salmonella on the farm, whereas litter samples had the strongest association with Salmonella loads in pre- and postchill carcass rinses. Boot socks, drag swabs, and fecal samples all had similar sensitivities for detecting Campylobacter on the farm, and all were more strongly associated with Campylobacter loads in carcass rinses than were litter samples. Farm samples explained a greater proportion of the variability in carcass rinse prevalences and loads for Campylobacter than they did for Salmonella. Salmonella and Campylobacter prevalences and loads both decreased significantly as birds progressed through the processing plant. PMID:23624481

Evaluation of a Multiplex PCR Assay for the Identification of Salmonella Serovars Enteritidis and Typhimurium Using Retail and Abattoir Samples.

PubMed

Ogunremi, Dele; Nadin-Davis, Susan; Dupras, Andrée Ann; Márquez, Imelda Gálvan; Omidi, Katayoun; Pope, Louise; Devenish, John; Burke, Teresa; Allain, Ray; Leclair, Daniel

2017-02-01

A multiplex PCR was developed to identify the two most common serovars of Salmonella causing foodborne illness in Canada, namely, serovars Enteritidis and Typhimurium. The PCR was designed to amplify DNA fragments from four Salmonella genes, namely, invA gene (211-bp fragment), iroB gene (309-bp fragment), Typhimurium STM 4497 (523-bp fragment), and Enteritidis SE147228 (612-bp fragment). In addition, a 1,026-bp ribosomal DNA (rDNA) fragment universally present in bacterial species was included in the assay as an internal control fragment. The detection rate of the PCR was 100% among Salmonella Enteritidis (n = 92) and Salmonella Typhimurium (n = 33) isolates. All tested Salmonella isolates (n = 194) were successfully identified based on the amplification of at least one Salmonella -specific DNA fragment. None of the four Salmonella DNA amplicons were detected in any of the non- Salmonella isolates (n = 126), indicating an exclusivity rate of 100%. When applied to crude extracts of 2,001 field isolates of Salmonella obtained during the course of a national microbiological baseline study in broiler chickens and chicken products sampled from abattoir and retail outlets, 163 isolates, or 8.1%, tested positive for Salmonella Enteritidis and another 80 isolates, or 4.0%, tested as Salmonella Typhimurium. All isolates identified by serological testing as Salmonella Enteritidis in the microbiological study were also identified by using the multiplex PCR. The new test can be used to identify or confirm pure isolates of the two serovars and is also amenable for integration into existing culture procedures for accurate detection of Salmonella colonies.

A systematic review of the clinical, public health and cost-effectiveness of rapid diagnostic tests for the detection and identification of bacterial intestinal pathogens in faeces and food.

PubMed

Abubakar, I; Irvine, L; Aldus, C F; Wyatt, G M; Fordham, R; Schelenz, S; Shepstone, L; Howe, A; Peck, M; Hunter, P R

2007-09-01

To determine the diagnostic accuracy of tests for the rapid diagnosis of bacterial food poisoning in clinical and public health practice and to estimate the cost-effectiveness of these assays in a hypothetical population in order to inform policy on the use of these tests. Studies evaluating diagnostic accuracy of rapid tests were retrieved using electronic databases and handsearching reference lists and key journals. Hospital laboratories and test manufacturers were contacted for cost data, and clinicians involved in the care of patients with food poisoning were invited to discuss the conclusions of this review using the nominal group technique. A systematic review of the current medical literature on assays used for the rapid diagnosis of bacterial food poisoning was carried out. Specific organisms under review were Salmonella, Campylobacter, Escherichia coli O157, Staphylococcus aureus, Clostridium perfringens and Bacillus cereus. Data extraction was undertaken using standardised data extraction forms. Where a sufficient number of studies evaluating comparable tests were identified, meta-analysis was performed. A decision analytic model was developed, using effectiveness data from the review and cost data from hospitals and manufacturers, which contributed to an assessment of the cost-effectiveness of rapid tests in a hypothetical UK population. Finally, diagnostic accuracy and cost-effectiveness results were presented to a focus group of GPs, microbiologists and consultants in communicable disease control, to assess professional opinion on the use of rapid tests in the diagnosis of food poisoning. Good test performance levels were observed with rapid test methods, especially for polymerase chain reaction (PCR) assays. The estimated levels of diagnostic accuracy using the area under the curve of the summary receiver operating characteristic curve was very high. Indeed, although traditional culture is the natural reference test to use for comparative statistical analysis, on many occasions the rapid test outperforms culture, detecting additional 'truly' positive cases of food-borne illness. The significance of these additional positives requires further investigation. Economic modelling suggests that adoption of rapid tests in combination with routine culture is unlikely to be cost-effective, however, as the cost of rapid technologies decreases; total replacement with rapid technologies may be feasible. Despite the relatively poor quality of reporting of studies evaluating rapid detection methods, the reviewed evidence shows that PCR for Campylobacter, Salmonella and E. coli O157 is potentially very successful in identifying pathogens, possibly detecting more than the number currently reported using culture. Less is known about the benefits of testing for B. cereus, C. perfringens and S. aureus. Further investigation is needed on how clinical outcomes may be altered if test results are available more quickly and at a greater precision than in the current practice of bacterial culture.

Development of a rapid method to quantify Salmonella Typhimurium using a combination of MPN with qPCR and a shortened time incubation.

PubMed

Kim, Sun Ae; Park, Si Hong; Lee, Sang In; Ricke, Steven C

2017-08-01

A novel method was developed for the specific quantification of S. Typhimurium using a most-probable-number (MPN) combined with qPCR and a shortened incubation time (MPN-qPCR-SIT). For S. Typhimurium enumeration, dilutions of samples were transferred into three wells on a microtiter plate and the plate was incubated for 4 h. The S. Typhimurium presence in the wells was identified using a qPCR and populations were determined based on an MPN calculation. The R 2 between the MPN-qPCR-SIT and conventional MPN exhibited a high level of correlation (0.9335-0.9752), suggesting that the MPN-qPCR-SIT offers a reliable alternative method for S. Typhimurium quantification. Although plating and qPCR were limited in their ability to detect low levels of S. Typhimurium (e.g. 0.18 log MPN/ml), these levels could be successfully detected with the MPN-qPCR-SIT. Chicken breast samples inoculated with S. Typhimurium were incubated at 0, 4, and 24 h and incubated samples were subjected to microbiome analysis. Levels of Salmonella and Enterobacteriaceae increased significantly with incubation time. The obvious benefits of the MPN-qPCR-SIT are: 1) a further confirmation step is not required, 2) the detection limit is as low as conventional MPN, but 3) is more rapid, requiring approximately 7 h to simultaneously complete quantification. Copyright © 2017 Elsevier Ltd. All rights reserved.

The consequences of a sudden demographic change on the seroprevalence pattern, virulence genes, identification and characterisation of integron-mediated antibiotic resistance in the Salmonella enterica isolated from clinically diarrhoeic humans in Egypt.

PubMed

Osman, K M; Hassan, W M M; Mohamed, R A H

2014-08-01

The present study was undertaken to identify and characterise integrons and integrated resistance gene cassettes among eight multidrug-resistant (MDR) Salmonella serovars isolated from humans in Egypt. Virulotyping by polymerase chain reaction (PCR) was used for the detection of the presence of virulence genes. Integron PCR was used to detect the presence of class 1 in the MDR strains. The associated individual resistance gene cassettes were identified using specific PCRs. The isolated serovars were Salmonella Grampian (C1; 2/5), Larose (C1; 1/5), Hato (B; 1/5) and Texas (B; 1/5). Among the Salmonella serovars, five Salmonella isolates showed the highest resistance to amoxicillin, ampicillin, chloramphenicol, lincomycin, gentamicin, nalidixic acid, streptomycin and trimethoprim (100%), followed by neomycin, norfloxacin and tetracycline (80%), while the lowest resistance was recorded to colistin sulphate and ciprofloxacin in percentages of 20 and 40%, respectively. The invA, avrA, ssaQ, mgtC, siiD and sopB genes were detected in all isolates (100%), while the spvC and gipA genes were totally (100%) absent from all isolates. The remaining three virulence genes were diversely distributed as follows: the bcfC gene was detected in all isolates except Salmonella Hato (80%); the sodC1 gene was detected only in Salmonella Grampian and Salmonella Texas (60%); and the sopE1 gene was detected only in Salmonella Grampian, Hato and Texas (60%). Class 1 integrons were detected in 90% of the MDR isolates, comprising serovars Muenster, Florian, Noya, Grampian, Larose, Hato and Texas. Of the class 1 integron-positive isolates, 45% harboured Salmonella genomic island 1 (SGI1) either right junction or right and left junction having an A-C-S-T phenotype. Of the class 1 integron-positive isolates, 44% harboured integron gene cassette aadA2, while 11% harboured the floR gene present in multidrug resistance flanked by two integrons of SGI1. The results of the present study indicate that class 1 integrons carrying gene cassettes conferring resistance mainly to aminoglycosides are widespread among the MDR Salmonella serovars isolated from humans in Egypt, indicating the important role of these genetic elements in the dissemination of multidrug resistance.

[Infection risks associated with importation of fresh food in Iceland].

PubMed

Kristinsson, Karl G; Georgsson, Franklín

2015-06-01

Access to safe food is a privilege for people living in Iceland. Rapid increase in antimicrobial resistance, related to factory farming and antimicrobial use in agriculture, is a major threat to public health. Increasing food trade between countries and continents facilitates global spread of pathogens and resistance. Icelandic agriculture has benefitted from its isolation and small size. After interventions to reduce the prevalence of Campylobacter and Salmonella at poultry farms, the incidence of human campylobacteriolsis is 17-43/100.000, of which about half is domestically acquired and Salmonella infections 10-15/100.000 mainly acquired abroad. Since Enterohaemorrhagic E. coli (EHEC) has not been detected in domestic cattle, the low incidence of infections is not surprising (0-0.6/100.000/year). A recent outbreak due to a multiresistant EHEC strain was traced to imported lettuce. Antimicrobial use in Icelandic agriculture is among the lowest in Europe and domestic infections caused by Salmonella and Campylobacter are rarely caused by resistant strains. Carbapenemase producing Enterobacteriaceae have not been found in Iceland. Low use of antimicrobials in Icelandic agriculture and actions to limit the spread of Campylobacter and Salmonella have been successful. The public should be informed of the importance of the origin of food and that Icelandic food products are among the safest.

Highly sensitive surface-scanning detector for the direct bacterial detection using magnetoelastic (ME) biosensors

NASA Astrophysics Data System (ADS)

Liu, Yuzhe; Horikawa, Shin; Chen, I.-Hsuan; Du, Songtao; Wikle, Howard C.; Suh, Sang-Jin; Chin, Bryan A.

2017-05-01

This paper demonstrates a highly sensitive surface-scanning detector used for magnetoelastic (ME) biosensors for the detection of Salmonella on the surface of a polyethylene (PE) food preparation surface. The design and fabrication methods of the new planar spiral coil are introduced. Different concentrations of Salmonella were measured on the surface of a PE board. The efficacy of Salmonella capture and detection is discussed.

Occurrence of foodborne bacteria in Alberta feedlots.

PubMed

Van Donkersgoed, Joyce; Bohaychuk, Valerie; Besser, Thomas; Song, Xin-Ming; Wagner, Bruce; Hancock, Dale; Renter, David; Dargatz, David

2009-02-01

The occurrence of generic Escherichia coli, E. coli O157, Salmonella, and Campylobacter in cattle manure, beef carcasses, catch basin water, and soils receiving manure application was determined in 21 Alberta feedlots. In cattle manure, generic E. coli (98%, 2069/2100) and Campylobacter (76%, 1590/2100) were frequently detected; E. coli O157 (7%, 143/2100) and Salmonella (1%, 20/2100) were less frequently detected. Samples from beef carcasses in the cooler following Hazard Analysis Critical Control Point interventions yielded only 1 isolate each of generic E. coli and Campylobacter (1/1653) and no Salmonella (0/1653). Catch basin water specimens were positive for generic E. coli in both the spring (62%, 13/21) and the fall (52%, 11/21). Other bacteria were detected only in the spring water specimens, including E. coli O157 (29%, 6/21), Salmonella (5%, 1/21), and Campylobacter (52%, 11/21). Generic E. coli was frequently isolated from soil specimens (30%, 27/88), but E. coli O157 was not found in soil samples obtained in the spring and was only occasionally detected in the fall samples (9%, 3/32). Salmonella were occasionally found in the soil specimens collected in the spring (3%, 2/56), but not in the fall season (0/32). Campylobacter jejuni was frequent in cattle manure (66%, 1070/1623), but rare in carcass and environmental samples. E. coli O157 and Salmonella were rarely detected in cattle or the environment. Generic E. coli and Salmonella were rarely detected on carcasses.

Comparing validation of four ELISA-systems for detection of Salmonella derby- and Salmonella infantis-infected pigs.

PubMed

Roesler, Uwe; Szabo, Istvan; Matthies, Claudia; Albrecht, Kerstin; Leffler, Martin; Scherer, Kathrin; Nöckler, Karsten; Lehmann, Jörg; Methner, Ulrich; Hensel, Andreas; Truyen, Uwe

2011-01-01

The objective of this study was the comparative evaluation of four indirect Salmonella ELISA tests at study time approved in Germany to detect Salmonella infection in pigs.Three tests are based on a LPS-antigen mix and directed against specific IgG antibodies. The fourth test is based on a purified S. Typhimurium whole-cell lysate antigen and discriminates between Salmonella-specific IgM-, IgA-, and IgG- antibodies. In a longitudinal study, two groups of six weeks old hybrid piglets were orally infected with a porcine S. Infantis or S. Derby strain. Clinical and bacteriological parameters were monitored weekly during an observation period of 130 days after infection and serum samples were investigated in parallel with the respective ELISAs. Apparently, the LPS-based ELISA systems used in this study failed to recognize S. Infantis-infected pigs although those animals shed the pathogen in high amounts throughout the study until day 81 post infection (p. i.). In contrast, the isotype-specific Salmonella Typhimurium whole-cell-lysate based ELISA was capable of detecting Salmonella-infected pigs from day ten p. i. at all tested serotypes and revealed the highest sensitivity in detection of S. Infantis-infected pigs. Furthermore, it became apparent that the often used surveillance cut-off value of 40 OD% is not appropriate for intra-vitam detection of S. Infantis- and S. Derby-infected pigs. In contrast, the cut-off values of the ELISAs given by the suppliers result in considerable higher detection rates.

Isolation and characterization of Salmonella enterica in day-old ducklings in Egypt

PubMed Central

Osman, Kamelia M; Marouf, Sherif H; Zolnikov, Tara R; AlAtfeehy, Nayerah

2014-01-01

Importing day-old ducklings (DOD) unknowingly infected with non-typhoid Salmonella (NTS) may be associated with disease risk. Domestic and international trade may enhance this risk. Salmonella enterica serovars, their virulence genes combinations and antibiotic resistance, garner attention for their potentiality to contribute to the adverse health effects on populations throughout the world. The aim of this study was to estimate the risk of imported versus domestic DOD as potential carriers of NTS. The results confirm the prevalence of salmonellosis in imported ducklings was 18.5% (25/135), whereas only 12% (9/75) of cases were determined in the domestic ducklings. Fourteen serovars (Salmonella enteritidis, Salmonella kisii, Salmonella typhimurium, Salmonella gaillac, Salmonella uno, Salmonella eingedi, Salmonella shubra, Salmonella bardo, Salmonella inganda, Salmonella kentucky, Salmonella stanley, Salmonella virchow, Salmonella haifa, and Salmonella anatum) were isolated from the imported ducklings, whereas only S. enteritidis, S. typhimurium, S. virchow, and S. shubra were isolated from the domestic ducklings. The isolated Salmonella serovars were 100% susceptible to only colistin sulphate and 100% resistant to lincomycin. The 14 Salmonella serovars were screened for 11 virulence genes (invA, avrA, ssaQ, mgtC, siiD, sopB, gipA, sodC1, sopE1, spvC, and bcfC) by PCR. The invA, sopB, and bcfC genes were detected in 100% of the Salmonella serovars; alternatively, the gipA gene was absent in all of the isolated Salmonella serovars. The 11 virulent genes were not detected in either of S. stanley or S. haifa serovars. The results confirm an association between antibiotic resistance and virulence of Salmonella in the DOD. This study confirms the need for a country adherence to strict public health and food safety regimes. PMID:24548159

Evaluation of corn oil as an additive in the pre-enrichment step to increase recovery of Salmonella enterica from oregano.

PubMed

Jean-Gilles Beaubrun, Junia; Flamer, Marie-Laure; Addy, Nicole; Ewing, Laura; Gopinath, Gopal; Jarvis, Karen; Grim, Chris; Hanes, Darcy E

2016-08-01

Phenolic compounds associated with essential oils of spices and herbs possess a variety of antioxidant and antimicrobial properties that interfere with Salmonella detection from fresh and dried products. Finding a compound to neutralize the effect of these antimicrobial compounds, while allowing Salmonella growth during pre-enrichment, is a crucial step in both traditional pathogen isolation and molecular detection from these foods. This study evaluated the effectiveness of corn oil as a component of the pre-enrichment broth to counteract antimicrobial compounds properties and increase the recovery of Salmonella from spices. Oregano samples artificially contaminated with Salmonella enterica were pre-enriched in modified Buffered Peptone Water (mBPW) supplemented with and without 2% (vol/vol) corn oil respectively. Samples were incubated overnight at 37 °C. The results showed that recovery of Salmonella from oregano samples was increased by ≥50% when pre-enriched with corn oil. Serovars were confirmed using a PCR serotyping method. In addition, shot-gun metagenomics analyses demonstrated bacterial diversity and the effect of corn oil on the relative prevalence of Salmonella in the oregano samples. Modifying pre-enrichment broths with corn oil improved the detection and isolation of Salmonella from oregano, and may provide an alternative method for pathogen detection in dried food matrices such as spices. Published by Elsevier Ltd.

The effect of pre-enrichment media on the recovery and detection of Salmonella in feed

USDA-ARS?s Scientific Manuscript database

Current methodology for detecting Salmonella in feeds and feed ingredients are adapted from food safety methods. These methods do not take into account the stressed state of Salmonella in feed, presence of competing microorganisms nor the sample matrix. The objective was to evaluate four pre-enrichm...

Salmonella epidemiology: A whirlwind of change.

PubMed

Besser, John M

2018-05-01

The field of infectious disease epidemiology for Salmonella and other enteric pathogens is undergoing some of the most profound changes since the time of Kauffman and White. Rapid advances in "big data" technologies such as genomics and metagenomics are making it possible to monitor and control salmonellosis in new and exciting ways. Epidemiological methods are becoming increasingly robust through the routine use of standardized hypothesis-generating questionnaires, iterative open-ended interviewing, informational trace-backs and new modeling techniques for describing the attribution of disease to food sources. In addition, Salmonella epidemiology is facing important challenges and new opportunities due to the rapid adoption of culture independent diagnostic test panels by clinical laboratories. Where is this unprecedented wave of change taking us? This chapter will examine emerging trends in Salmonella epidemiology, and take a peek into the not-so-distant future. Published by Elsevier Ltd.

Distribution and Antimicrobial Susceptibility of Foodborne Salmonella Serovars in Eight Provinces in China from 2007 to 2012 (Except 2009).

PubMed

Wang, Yin; Cao, Chenyang; Alali, Walid Q; Cui, Shenghui; Li, Fengqin; Zhu, Jianghui; Wang, Xin; Meng, Jianghong; Yang, Baowei

2017-07-01

One thousand four hundred ninety-one Salmonella isolates recovered from retail foods including chicken, beef, fish, pork, dumplings, and cold dishes in China in 2007, 2008, 2010, 2011, and 2012 were analyzed for distribution of serotype and antimicrobial susceptibility. A total of 129 Salmonella serotypes were detected among 1491 isolates. Salmonella Enteritidis (21.5%), Typhimurium (11.0%), Indiana (10.8%), Thompson (5.4%), Derby (5.1%), Agona (3.8%), and Shubra (3.0%) were the seven most important serotypes in 1491 isolates. For antibiotic susceptibility, except 16 (1.1%) isolates were susceptible to all tested antibiotics, 131 (8.8%) resisted 1-2 and 1344 (90.1%) resisted three or more antibiotics. One thousand forty-six (70.2%) of 1491 Salmonella isolates were identified as multidrug-resistant (MDR) isolates, which could resist three or more categories of antibiotics. Resistance to sulfisoxazole (78.1%) was most common among the tested Salmonella, followed by tetracycline (70.6%), trimethoprim/sulfamethoxazole (68.0%), and nalidixic acid (63.4%). Resistances to amikacin (20.0%), levofloxacin (18.7%), gatifloxacin (17.9%), ceftriaxone (17.7%), and cefoxitin (13.2%) were less frequently detected. Resistance to fluoroquinolones was most common among Salmonella Shubra and Indiana isolates, while resistance to cephalosporins was frequently detected among Salmonella Thompson isolates. The results highlighted the diversity of Salmonella serotypes and the high prevalence of Salmonella MDR isolates in China. Compared with Salmonella Enteritidis and Typhimurium isolates, the higher fluoroquinolones and cephalosporins resistance rates of some individual serotypes (Salmonella Shubra, Indiana, and Thompson) also provided more information for further study related to fluoroquinolones or cephalosporin-resistant Salmonella.

Aptamer-based impedimetric sensor for bacterial typing.

PubMed

Labib, Mahmoud; Zamay, Anna S; Kolovskaya, Olga S; Reshetneva, Irina T; Zamay, Galina S; Kibbee, Richard J; Sattar, Syed A; Zamay, Tatiana N; Berezovski, Maxim V

2012-10-02

The development of an aptamer-based impedimetric sensor for typing of bacteria (AIST-B) is presented. Highly specific DNA aptamers to Salmonella enteritidis were selected via Cell-SELEX technique. Twelve rounds of selection were performed; each comprises a positive selection step against S. enteritidis and a negative selection step against a mixture of related pathogens, including Salmonella typhimurium, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Citrobacter freundii, to ensure the species-specificity of the selected aptamers. After sequencing of the pool showing the highest binding affinity to S. enteritidis, a DNA sequence of high affinity to the bacteria was integrated into an impedimetric sensor via self-assembly onto a gold nanoparticles-modified screen-printed carbon electrode (GNPs-SPCE). Remarkably, this aptasensor is highly selective and can successfully detect S. enteritidis down to 600 CFU mL(-1) (equivalent to 18 CFU in 30 μL assay volume) in 10 min and distinguish it from other Salmonella species, including S. typhimurium and S. choleraesuis. This report is envisaged to open a new venue for the aptamer-based typing of a variety of microorganisms using a rapid, economic, and label-free electrochemical platform.

Prevalence and counts of Salmonella spp. in minimally processed vegetables in São Paulo, Brazil.

PubMed

Sant'Ana, Anderson S; Landgraf, Mariza; Destro, Maria Teresa; Franco, Bernadette D G M

2011-09-01

Minimally processed vegetables (MPV) may be important vehicles of Salmonella spp. and cause disease. This study aimed at detecting and enumerating Salmonella spp. in MPV marketed in the city of São Paulo, Brazil. A total of 512 samples of MPV packages collected in retail stores were tested for Salmonella spp. and total coliforms and Escherichia coli as indication of the hygienic status. Salmonella spp. was detected in four samples, two using the detection method and two using the counting method, where the results were 8.8 × 10(2) CFU/g and 2.4 × 10(2) CFU/g. The serovars were Salmonella Typhimurium (three samples) and Salmonella enterica subsp. enterica O:47:z4,z23:- (one sample). Fourteen samples (2.7%) presented counts of E. coli above the maximum limit established by the Brazilian regulation for MPV (10(2) CFU/g). Therefore, tightened surveillance and effective intervention strategies are necessary in order to address consumers and governments concerns on safety of MPV. Copyright © 2011 Elsevier Ltd. All rights reserved.

Detection of Salmonellae in the Environment

PubMed Central

Thomason, Berenice M.; Biddle, James W.; Cherry, William B.

1975-01-01

The incidence of salmonellae in contrasting environments was compared in this study. Samples collected from or near surface waters in a lush hardwood forest yielded four salmonellae serotypes from six culturally positive samples. A total of 76 samples collected from the top of a granite outcropping over a 3-month period yielded 10 positive samples. Only two salmonellae serotypes were isolated, and one of these was isolated only once. The nature of the sample material had no significant effect on the detection of salmonellae from the two sampling sites. However, the presence or absence of visible moisture in the sample significantly affected the recovery of salmonellae. The results showed that even a harsh environment such as that found on top of Stone Mountain may serve as an ecological niche for the survival and transmission of salmonellae. PMID:1106319

DNA hybridization assay for detection of Salmonella in foods: collaborative study.

PubMed

Flowers, R S; Klatt, M J; Mozola, M A; Curiale, M S; Gabis, D A; Silliker, J H

1987-01-01

A collaborative study was performed in 11 laboratories to validate a DNA hybridization (DNAH) procedure for detection of Salmonella in foods. The DNAH procedure was compared to the standard culture method for detection of Salmonella in 6 foods: ground pepper, soy flour, dry whole egg, milk chocolate, nonfat dry milk, and raw deboned turkey. With the exception of turkey which was naturally contaminated, uninoculated and inoculated samples of each food group were analyzed. Results for the DNAH method were significantly better than for the standard culture method at the 5% probability level for the detection of Salmonella in turkey. There was no significant difference between the methods for the other 5 foods. The method has been adopted official first action.

Prevalence, resistance patterns, and risk factors for antimicrobial resistance in bacteria from retail chicken meat in Colombia.

PubMed

Donado-Godoy, Pilar; Byrne, Barbara A; León, Maribel; Castellanos, Ricardo; Vanegas, Consuelo; Coral, Adriana; Arevalo, Alejandra; Clavijo, Viviana; Vargas, Mercedes; Romero Zuñiga, Juan J; Tafur, McAllister; Pérez-Gutierrez, Enrique; Smith, Woutrina A

2015-04-01

As a step toward implementing the Colombian Integrated Program for Antimicrobial Resistance Surveillance (COIPARS), this study aimed to establish the baseline antimicrobial resistance patterns of Salmonella serovars, Escherichia coli, and Enterococcus spp. isolates in retail poultry meat from independent stores and from a main chain distributor center. MICs of the isolates were determined for antimicrobials used both in humans and animals, using an automated system. Salmonella serovars were isolated from 26% of the meat samples and E. coli from 83%, whereas Enterococcus faecalis and Enterococcus faecium were detected in 81 and 13% of the meat samples, respectively. A principal finding of concern in this study was that almost 98% of isolates tested were multidrug resistant. Ceftiofur, enrofloxacin, nalidixic acid, and tetracycline were the antimicrobials that showed the highest frequency of resistance among Salmonella and E. coli isolates. For enterococci, 61.5% of E. faecium isolates were found to be resistant to quinupristin-dalfopristin; this is significant because it is used to treat nosocomial infections when vancomycin resistance is present. Vancomycin resistance was detected in 4% of the E. faecalis isolates. The results of our study highlight the need for rapid implementation of an integrated program for surveillance of antimicrobial resistance by the Colombian authorities in order to monitor trends, raise awareness, and help promote practices to safeguard later generation antimicrobial agents.

Development of gold nanoparticle-aptamer-based LSPR sensing chips for the rapid detection of Salmonella typhimurium in pork meat.

PubMed

Oh, Seo Yeong; Heo, Nam Su; Shukla, Shruti; Cho, Hye-Jin; Vilian, A T Ezhil; Kim, Jinwoon; Lee, Sang Yup; Han, Young-Kyu; Yoo, Seung Min; Huh, Yun Suk

2017-08-31

A non-labeled, portable plasmonic biosensor-based device was developed to enable the ultra-sensitive and selective detection of Salmonella typhimurium in pork meat samples. Specifically, a plasmonic sensor, using the self-assembly of gold nanoparticles (AuNPs) to achieve a regulated diameter of 20 nm for the AuNP monolayers, was used to conduct high-density deposition on a transparent substrate, which produced longitudinal wavelength extinction shifts via a localized surface plasmon resonance (LSPR) signal. The developed aptamers conjugated to the LSPR sensing chips revealed an ultra-sensitive upper limit of detection (LOD) of approximately 10 4 cfu/mL for S. typhimurium in pure culture under the optimal assay conditions, with a total analysis time of 30-35 min. When the LSPR sensing chips were applied on artificially contaminated pork meat samples, S. typhimurium in the spiked pork meat samples was also detected at an LOD of 1.0 × 10 4 cfu/mL. The developed method could detect S. typhimurium in spiked pork meat samples without a pre-enrichment step. Additionally, the LSPR sensing chips developed against S. typhimurium were not susceptible to any effect of the food matrix or background contaminant microflora. These findings confirmed that the developed gold nanoparticle-aptamer-based LSPR sensing chips could facilitate sensitive detection of S. typhimurium in food samples.

A multiplex real-time PCR assay, based on invA and pagC genes, for the detection and quantification of Salmonella enterica from cattle lymph nodes.

PubMed

Bai, Jianfa; Trinetta, Valentina; Shi, Xiaorong; Noll, Lance W; Magossi, Gabriela; Zheng, Wanglong; Porter, Elizabeth P; Cernicchiaro, Natalia; Renter, David G; Nagaraja, Tiruvoor G

2018-05-01

Cattle lymph nodes can harbor Salmonella and potentially contaminate beef products. We have developed and validated a new real-time PCR (qPCR) assay for the detection and quantification of Salmonella enterica in cattle lymph nodes. The assay targets both the invA and pagC genes, the most conserved molecular targets in Salmonella enterica. An 18S rRNA gene assay that amplifies from cattle and other animal species was also included as an internal control. Available DNA sequences for invA, pagC and 18S rRNA genes were used for primer and probe selections. Three Salmonella serotypes, S. Typhimurium, S. Anatum, and S. Montevideo, were used to assess the assay's analytical sensitivity. Correlation coefficients of standard curves generated for each target and for all three serotypes were >99% and qPCR amplification efficiencies were between 93% and 110%. Assay sensitivity was also determined using standard curve data generated from Salmonella-negative cattle lymph nodes spiked with 10-fold dilutions of the three Salmonella serotypes. Assay specificity was determined using Salmonella culture method, and qPCR testing on 36 Salmonella strains representing 33 serotypes, 38 Salmonella strains of unknown serotypes, 252 E. coli strains representing 40 serogroups, and 31 other bacterial strains representing 18 different species. A collection of 647 cattle lymph node samples from steers procured from the Midwest region of the US were tested by the qPCR, and compared to culture-method of detection. Salmonella prevalence by qPCR for pre-enriched and enriched lymph nodes was 19.8% (128/647) and 94.9% (614/647), respectively. A majority of qPCR positive pre-enriched samples (105/128) were at concentrations between 10 4 and 10 5  CFU/mL. Culture method detected Salmonella in 7.7% (50/647) and 80.7% (522/647) of pre- and post-enriched samples, respectively; 96.0% (48/50) of pre-enriched and 99.4% (519/522) of post-enriched culture-positive samples were also positive by qPCR. More samples tested positive by qPCR than by culture method, indicating that the real-time PCR assay was more sensitive. Our data indicate that this triplex qPCR can be used to accurately detect and quantify Salmonella enterica strains from cattle lymph node samples. The assay may serve as a useful tool to monitor the prevalence of Salmonella in beef production systems. Copyright © 2018 Elsevier B.V. All rights reserved.

Evaluation of the Impact of Varied Carvacrol Concentrations on Salmonella Recovery in Oregano and How Corn Oil Can Minimize the Effect of Carvacrol during Preenrichment.

PubMed

Beaubrun, Junia Jean-Gilles; Addy, Nicole; Keltner, Zachary; Farris, Samantha; Ewing, Laura; Gopinath, Gopal; Hanes, Darcy E

2018-06-01

Phenolic compounds, like carvacrol, in oregano interfere with the detection of foodborne pathogens such as Salmonella enterica. Carvacrol concentration varies based on plant cultivars and growth region. Six oregano cultivars were used to compare the impact of carvacrol concentration on Salmonella and to evaluate the effectiveness of corn oil to help increase Salmonella survival for detection. The results of Agilent 1200 series high-performance liquid chromatography analysis showed that carvacrol concentration in the six oregano cultivars ranged from 64 to 11,200 ppm. Oregano samples were artificially contaminated with S. enterica and were preenriched in Trypticase soy broth with or without 2% (v/v) corn oil. After 18 to 24 h at 37°C, aliquots were transferred to selective enrichment broths. Salmonella was recovered onto xylose lysine Tergitol 4 agar. Six Salmonella serovars were compared, and recovery varied based on carvacrol concentration and serovar. Samples with higher concentrations of carvacrol showed Salmonella recovery only when they were preenriched with corn oil. Based on metagenomic analysis, the microflora associated with the oregano also varied per cultivar. The results show that, as carvacrol levels increased, Salmonella survival decreased. However, the addition of corn oil to the preenrichment broth can minimize the antimicrobial effects of the phenolic compounds, thus allowing for increased detection of Salmonella from oregano cultivars.

Prevention of Salmonella cross-contamination in an oilmeal manufacturing plant.

PubMed

Morita, T; Kitazawa, H; Iida, T; Kamata, S

2006-08-01

The mechanisms of Salmonella contamination in an oilmeal plant were investigated and the basic data were collected in order to achieve control of Salmonella in oilmeal. Salmonella was detected in all contamination vectors and environmental factors investigated, namely: operators, processing floor, dust in the air and rodents. In particular, high concentrations of Salmonella were detected on the processing floor of the manufacturing area, which has high oil content. Steam was the most effective disinfection method used for the processing floor, as the effects of heat sterilization and disinfection may work in tandem. In addition, restricting the movement of operators of the production chain remarkably reduced Salmonella contamination, even in areas of otherwise high contamination. Within the oilmeal plant, high Salmonella contamination rates for the processing floor represent the greatest risk of contamination of oilmeal via operators, dust in the air and rodents. Therefore, control of the processing floor is the most important means for reducing the oilmeal contamination rate. Specific Salmonella control methods for oilmeal plants have been established.

Modification of the BAX System PCR assay for detecting Salmonella in beef, produce, and soy protein isolate. Performance Tested Method 100201.

PubMed

Peng, Linda X; Wallace, Morgan; Andaloro, Bridget; Fallon, Dawn; Fleck, Lois; Delduco, Dan; Tice, George

2011-01-01

The BAX System PCR assay for Salmonella detection in foods was previously validated as AOAC Research Institute (RI) Performance Tested Method (PTM) 100201. New studies were conducted on beef and produce using the same media and protocol currently approved for the BAX System PCR assay for E. coli O157:H7 multiplex (MP). Additionally, soy protein isolate was tested for matrix extension using the U.S. Food and Drug Administration-Bacteriological Analytical Manual (FDA-BAM) enrichment protocols. The studies compared the BAX System method to the U.S. Department of Agriculture culture method for detecting Salmonella in beef and the FDA-BAM culture method for detecting Salmonella in produce and soy protein isolate. Method comparison studies on low-level inoculates showed that the BAX System assay for Salmonella performed as well as or better than the reference method for detecting Salmonella in beef and produce in 8-24 h enrichment when the BAX System E. coli O157:H7 MP media was used, and soy protein isolate in 20 h enrichment with lactose broth followed by 3 h regrowth in brain heart infusion broth. An inclusivity panel of 104 Salmonella strains with diverse serotypes was tested by the BAX System using the proprietary BAX System media and returned all positive results. Ruggedness factors involved in the enrichment phase were also evaluated by testing outside the specified parameters, and none of the factors examined affected the performance of the assay.

Rapid electrochemical quantification of Salmonella Pullorum and Salmonella Gallinarum based on glucose oxidase and antibody-modified silica nanoparticles.

PubMed

Luo, Yiheng; Dou, Wenchao; Zhao, Guangying

2017-07-01

In this article, a facile and sensitive electrochemical method for quantification of Salmonella Pullorum and Salmonella Gallinarum (S. Pullorum and S. Gallinarum) was established by monitoring glucose consumption with a personal glucose meter (PGM). Antibody-functionalized magnetic nanoparticles (IgG-MNPs) were used to capture and enrich S. Pullorum and S. Gallinarum, and IgG-MNPs-S. Pullorum and IgG-MNPs-S. Gallinarum complexes were magnetically separated from a sample using a permanent magnet. The trace tag was prepared by loading polyclonal antibodies and high-content glucose oxidase on amino-functionalized silica nanoparticles (IgG-SiNPs-GOx). With a sandwich-type immunoassay format, IgG-SiNPs-GOx were added into the above mixture solution and conjugated to the complexes, forming sandwich composites IgG-MNPs/S. Pullorum and S. Gallinarum/IgG-SiNPs-GOx. The above sandwich composites were dispersed in glucose solution. Before and after the hydrolysis of glucose, the concentration of glucose was measured using PGM. Under optimal conditions, a linear relationship between the decrease of glucose concentration and the logarithm of S. Pullorum and S. Gallinarum concentration was obtained in the concentration range from 1.27 × 10 2 to 1.27 × 10 5  CFU mL -1 , with a detection limit of 7.2 × 10 1  CFU mL -1 (S/N = 3). This study provides a portable, low-cost, and quantitative analytical method for bacteria detection; thus, it has a great potential in the prevention of disease caused by S. Pullorum and S. Gallinarum in poultry. Graphical abstract A schematic illustration of the fabrication process of IgG-SiNPs-GOD nanomaterials (A) and IgG-MNPs (B) and experimental procedure of detection of S. Pullorum and S. Gallinarum using GOD-functionalized silica nanospheres as trace tags based on PGM (C).

Development of protective immunity to Salmonella, a mucosal pathogen with a systemic agenda

PubMed Central

Griffin, Amanda J.; McSorley, Stephen J.

2014-01-01

Salmonella infections can cause a range of intestinal and systemic disease in human and animal hosts. While some Salmonella serovars initiate a localized intestinal inflammatory response, others use the intestine as a portal of entry to initiate a systemic infection. Considerable progress has been made in understanding bacterial invasion and dissemination strategies and the nature of the Salmonella-specific immune response to oral infection. Innate and adaptive immunity are rapidly initiated after oral infection but these effector responses can also be hindered by bacterial evasion strategies. Furthermore, although Salmonella resides within intramacrophage phagosomes, recent studies highlight a surprising collaboration of CD4 Th1, Th17, and B cell responses in mediating resistance to Salmonella infection. PMID:21307847

Highly specific and cost-efficient detection of Salmonella Paratyphi A combining aptamers with single-walled carbon nanotubes.

PubMed

Yang, Ming; Peng, Zhihui; Ning, Yi; Chen, Yongzhe; Zhou, Qin; Deng, Le

2013-05-22

In this paper, a panel of single-stranded DNA aptamers with high affinity and specificity against Salmonella Paratyphi A was selected from an enriched oligonucleotide pool by a whole-cell-Systematic Evolution of Ligands by Exponential Enrichment (SELEX) procedure, during which four other Salmonella serovars were used as counter-selection targets. It was determined through a fluorescence assay that the selected aptamers had high binding ability and specificity to this pathogen. The dissociation constant of these aptamers were up to nanomolar range, and aptamer Apt22 with the lowest Kd (47 ± 3 nM) was used in cell imaging experiments. To detect this bacteria with high specificity and cost-efficiently, a novel useful detection method was also constructed based on the noncovalent self-assembly of single-walled carbon nanotubes (SWNTs) and DNAzyme-labeled aptamer detection probes. The amounts of target bacteria could be quantified by exploiting chemoluminescence intensity changes at 420 nm and the detection limit of the method was 103 cfu/mL. This study demonstrated the applicability of Salmonella specific aptamers and their potential for use in the detection of Salmonella in food, clinical and environmental samples.

[Antibody response after immunization with a Salmonella Typhimurium live vaccine in dependence on the way of application].

PubMed

Trepnau, Daniela; Ulrich, Evelyn; Uhlig, Reinhard; Lindner, Thomas; Selbitz, Hans-Joachim; Rösler, Uwe; Gabert, Jörg; Bergfeld, Uwe; Fehlhaber, Karsten; Brabetz, Werner; Lehmann, Jörg

2008-01-01

In Germany now, the recognition of Salmonella infections in pig herds is based on three different commercial tests detecting antibodies against Salmonella-derived lipopolysaccharide (LPS). However, a serious disadvantage of these tests, used so far, is the restricted detection of antibodies belonging predominantly to the immunoglobulin class g (IgG). Therefore, a new test was developed to detect three Ig classes (IgM, IgG and IgA). Different constellations between the three Ig classes allow the evaluation of the current infection status of each pig. Under field conditions, this was proved in three different vaccination trials using a commercial Salmonella Typhimurium live vaccine.

First detection of oqxAB in Salmonella spp. isolated from food.

PubMed

Wong, Marcus Ho Yin; Chen, Sheng

2013-01-01

Food-borne salmonellosis is an important public health problem worldwide and the second leading cause of food-borne illnesses in Hong Kong. In this study, the prevalence and antimicrobial resistance of Salmonella in meat products in Hong Kong were determined. Interestingly, a plasmid-mediated quinolone resistance (PMQR) gene combination, oqxAB, which mediates resistance to nalidixic acid, chloramphenicol, and olaquindox, was for the first time detectable on the chromosomes of two Salmonella enterica serovar Derby isolates. Further surveillance of oqxAB in Salmonella will be needed.

Cold plasma rapid decontamination of food contact surfaces contaminated with Salmonella biofilms.

PubMed

Niemira, Brendan A; Boyd, Glenn; Sites, Joseph

2014-05-01

Cross-contamination of foods from persistent pathogen reservoirs is a known risk factor in processing environments. Industry requires a rapid, waterless, zero-contact, chemical-free method for removing pathogens from food contact surfaces. Cold plasma was tested for its ability to inactivate Salmonella biofilms. A 3-strain Salmonella culture was grown to form adherent biofilms for 24, 48, or 72 h on a test surface (glass slides). These were placed on a conveyor belt and passed at various line speeds to provide exposure times of 5, 10, or 15 s. The test plate was either 5 or 7.5 cm under a plasma jet emitter operating at 1 atm using filtered air as the feed gas. The frequency of high-voltage electricity was varied from 23 to 48 kHz. At the closer spacing (5 cm), cold plasma reduced Salmonella biofilms by up to 1.57 log CFU/mL (5 s), 1.82 log CFU/mL (10 s), and 2.13 log CFU/mL (15 s). Increasing the distance to 7.5 cm generally reduced the efficacy of the 15 s treatment, but had variable effects on the 5 and 10 s treatments. Variation of the high-voltage electricity had a greater effect on 10 and 15 s treatments, particularly at the 7.5 cm spacing. For each combination of time, distance, and frequency, Salmonella biofilms of 24, 48, and 72 h growth responded consistently with each other. The results show that short treatments with cold plasma yielded up to a 2.13 log reduction of a durable form of Salmonella contamination on a model food contact surface. This technology shows promise as a possible tool for rapid disinfection of materials associated with food processing. Pathogens such as Salmonella can form chemical-resistant biofilms, making them difficult to remove from food contact surfaces. A 15 s treatment with cold plasma reduced mature Salmonella biofilms by up to 2.13 log CFU/mL (99.3%). This contact-free, waterless method uses no chemical sanitizers. Cold plasma may therefore have a practical application for conveyor belts, equipment, and other food contact surfaces where a rapid, dry antimicrobial process is required. © 2014 Institute of Food Technologists®

[Study on simultaneous contamination of Salmonella and Campylobacter in retail chicken carcasses in Beijing].

PubMed

Hu, Yujie; Wang, Yeru; Li, Fengqin

2015-01-01

To elucidate the simultaneous contamination of Salmonella and Campylobacter in retail chicken carcasses in Beijing and to carry out the serological typing of all Salmonella isolates as well as the identification of Campylobacter at the species level. A total of 33 chicken carcasses were collected from Beijing supermarkets and farm's trade markets from May to July. All samples were enumerated for Salmonella and Campylobacter. All Salmonella isolates obtained were further serotyped and Campylobacter were identified at the species level. Totally, 19 samples (19/33, 57.6%) and 5 samples (5/33, 15.2%) were positive for Salmonella with the mean level of 119.4 MPN/100g and Campylobacter with the mean level of 58.6 CFU/g, respectively. In terms of Salmonella, 166 isolates with 14 serotypes were obtained. Salmonella Enteritidis was the most common serovar detected followed by S. Indiana. Serovar diversity was very high in all Salmonella isolates and various Salmonella serovars were detected in the same chicken carcass. A total of 11 serovar distribution spectrums were found and S. Enteritidis in combination with S. Indiana was the predominant. The retail chicken carcasses in Beijing collected from May to July were heavily contaminated by Salmonella with high serovar diversity.

Biocontrol and Rapid Detection of Food-Borne Pathogens Using Bacteriophages and Endolysins

PubMed Central

Bai, Jaewoo; Kim, You-Tae; Ryu, Sangryeol; Lee, Ju-Hoon

2016-01-01

Bacteriophages have been suggested as natural food preservatives as well as rapid detection materials for food-borne pathogens in various foods. Since Listeria monocytogenes-targeting phage cocktail (ListShield) was approved for applications in foods, numerous phages have been screened and experimentally characterized for phage applications in foods. A single phage and phage cocktail treatments to various foods contaminated with food-borne pathogens including E. coli O157:H7, Salmonella enterica, Campylobacter jejuni, Listeria monocytogenes, Staphylococcus aureus, Cronobacter sakazakii, and Vibrio spp. revealed that they have great potential to control various food-borne pathogens and may be alternative for conventional food preservatives. In addition, phage-derived endolysins with high host specificity and host lysis activities may be preferred to food applications rather than phages. For rapid detection of food-borne pathogens, cell-wall binding domains (CBDs) from endolysins have been suggested due to their high host-specific binding. Fluorescence-tagged CBDs have been successfully evaluated and suggested to be alternative materials of expensive antibodies for various detection applications. Most recently, reporter phage systems have been developed and tested to confirm their usability and accuracy for specific detection. These systems revealed some advantages like rapid detection of only viable pathogenic cells without interference by food components in a very short reaction time, suggesting that these systems may be suitable for monitoring of pathogens in foods. Consequently, phage is the next-generation biocontrol agent as well as rapid detection tool to confirm and even identify the food-borne pathogens present in various foods. PMID:27092128

Impact of Season, Demographic and Environmental Factors on Salmonella Occurrence in Raccoons (Procyon lotor) from Swine Farms and Conservation Areas in Southern Ontario

PubMed Central

Pearl, David L.; Janecko, Nicol; Boerlin, Patrick; Reid-Smith, Richard J.; Parmley, Jane; Jardine, Claire M.

2016-01-01

Salmonella has been detected in the feces of many wildlife species, including raccoons (Procyon lotor), but little is known about the epidemiology of Salmonella in wildlife living in different habitat types. Our objective was to investigate demographic, temporal, and climatic factors associated with the carriage of Salmonella in raccoons and their environment on swine farms and conservation areas. Using a repeated cross-sectional study design, we collected fecal samples from raccoons and environmental samples (soil, manure pits, dumpsters) on 5 swine farms and 5 conservation areas in Ontario, Canada once every five weeks from May to November, 2011–2013. Salmonella was detected in 26% (279/1093; 95% CI 22.9–28.2) of raccoon fecal samples, 6% (88/1609; 95% CI 4.5–6.8) of soil samples, 30% (21/69; 95% CI 20.0–42.7) of manure pit samples, and 23% (7/31; 95% CI 9.6–41.0) of dumpster samples. Of samples testing positive for Salmonella, antimicrobial resistance was detected in 5% (14/279; 95% CI 2.8–8.3) of raccoon fecal, 8% (7/89; 95% CI 3.2–15.5) of soil, 10% (2/21; 95% CI 1.2–30.4) of manure pit, and 0/7 dumpster samples. Using multi-level multivariable logistic regression analyses, we found location type (swine farm or conservation area) was not a significant explanatory variable for Salmonella occurrence in raccoon feces or soil (p > 0.05). However, detection of Salmonella in raccoon feces was associated with rainfall, season, and sex with various interaction effects among these variables. We detected a variety of Salmonella serovars that infect humans and livestock in the feces of raccoons indicating that raccoons living near humans, regardless of location type, may play a role in the epidemiology of salmonellosis in livestock and humans in southwestern Ontario. PMID:27611198

Impact of Season, Demographic and Environmental Factors on Salmonella Occurrence in Raccoons (Procyon lotor) from Swine Farms and Conservation Areas in Southern Ontario.

PubMed

Bondo, Kristin J; Pearl, David L; Janecko, Nicol; Boerlin, Patrick; Reid-Smith, Richard J; Parmley, Jane; Jardine, Claire M

2016-01-01

Salmonella has been detected in the feces of many wildlife species, including raccoons (Procyon lotor), but little is known about the epidemiology of Salmonella in wildlife living in different habitat types. Our objective was to investigate demographic, temporal, and climatic factors associated with the carriage of Salmonella in raccoons and their environment on swine farms and conservation areas. Using a repeated cross-sectional study design, we collected fecal samples from raccoons and environmental samples (soil, manure pits, dumpsters) on 5 swine farms and 5 conservation areas in Ontario, Canada once every five weeks from May to November, 2011-2013. Salmonella was detected in 26% (279/1093; 95% CI 22.9-28.2) of raccoon fecal samples, 6% (88/1609; 95% CI 4.5-6.8) of soil samples, 30% (21/69; 95% CI 20.0-42.7) of manure pit samples, and 23% (7/31; 95% CI 9.6-41.0) of dumpster samples. Of samples testing positive for Salmonella, antimicrobial resistance was detected in 5% (14/279; 95% CI 2.8-8.3) of raccoon fecal, 8% (7/89; 95% CI 3.2-15.5) of soil, 10% (2/21; 95% CI 1.2-30.4) of manure pit, and 0/7 dumpster samples. Using multi-level multivariable logistic regression analyses, we found location type (swine farm or conservation area) was not a significant explanatory variable for Salmonella occurrence in raccoon feces or soil (p > 0.05). However, detection of Salmonella in raccoon feces was associated with rainfall, season, and sex with various interaction effects among these variables. We detected a variety of Salmonella serovars that infect humans and livestock in the feces of raccoons indicating that raccoons living near humans, regardless of location type, may play a role in the epidemiology of salmonellosis in livestock and humans in southwestern Ontario.

Label-free detection of salmonella typhimurium with ssDNA aptamers

USDA-ARS?s Scientific Manuscript database

Foodborne pathogen Salmonella enterica is one of the major causes of gastrointestinal infections in human and animals. Conventional detection methods are time consuming and not effective enough under emergency circumstances to control outbreaks immediately. Therefore, biosensors that can detect Salm...

Reduced salmonella fecal shedding in swine administered porcine granulocyte-colony stimulating factor (G-CSF)

USDA-ARS?s Scientific Manuscript database

Salmonella colonization of food animals is a concern for animal health, food safety and public health. Key objectives of pre-harvest food safety programs are to detect asymptomatic Salmonella carriage in food animals, reduce colonization, and prevent transmission of Salmonella to other animals and ...

Application of bacterial reverse mutation assay for detection of non-genotoxic carcinogens.

PubMed

Kanode, Rewan; Chandra, Saurabh; Sharma, Sharad

2017-06-01

Non-genotoxic carcinogens may play a significant role in development of cancer. Currently short-term assays for mutagenicity classify genotoxic carcinogens and lack the abilities to detect epigenetic carcinogens. The need to develop an endpoint always remains to recognize potentially carcinogenic agents employing rapid and practical bioassays. For this, the present study utilized TA98 and TA1537 tester strains of Salmonella typhimurium to evaluate four non-genotoxic carcinogenic agents (Coumarin, β-Myrcene, Bis(2-ethylhexyl) phthalate and trans-anethole). These chemicals were tested individually and in combination with promutagens 2-aminoanthracene (2AA) and benzo(a)pyrene (BP) in presence of metabolic activation system (S9) by plate incorporation method. Exposure to all four test chemicals revealed marked increase of revertant colonies in promutagen combined groups as compared to promutagens alone. However significantly greater fold responses were observed with 2AA combination groups (Coumarin +2AA, β-Myrcene +2AA, Bis(2-ethylhexyl) phthalate +2AA and trans-anethole +2AA) with TA98 strain as compared with TA1537, which seems to have enhanced the mutagenic response of 2AA in metabolically activated conditions. It is concluded that out of both tester strains TA98 strain of Salmonella typhimurium has the potential to detect non-genotoxic carcinogens when combined with potent promutgens either by inhibiting or modulating activities of liver microsomal enzymes biochemically which may indirectly contribute to neoplastic alterations. Further this simple, short-term alternative assay may provide rapid information during extrapolative toxicology for differentiating genotoxic and non-genotoxic carcinogens.

Quantification and molecular characterization of Salmonella isolated from food samples involved in salmonellosis outbreaks in Rio Grande do Sul, Brazil

PubMed Central

Mürmann, Lisandra; dos Santos, Maria Cecília; Longaray, Solange Mendes; Both, Jane Mari Corrêa; Cardoso, Marisa

2008-01-01

Data concerning the prevalence and populations of Salmonella in foods implicated in outbreaks may be important to the development of quantitative microbial risk assessments of individual food products. In this sense, the objective of the present study was to assess the amount of Salmonella sp. in different foods implicated in foodborne outbreaks in Rio Grande do Sul occurred in 2005 and to characterize the isolated strains using phenotypic and genotypic methods. Nineteen food samples involved in ten foodborne outbreaks occurred in 2005, and positive on Salmonella isolation at the Central Laboratory of the Health Department of the State of Rio Grande do Sul, were included in this study. Food samples were submitted to estimation of Salmonella using the Most Probable Number (MPN) technique. Moreover, one confirmed Salmonella colony of each food sample was serotyped, characterized by its XbaI-macrorestriction profile, and submitted to antimicrobial resistance testing. Foods containing eggs, mayonnaise or chicken were contaminated with Salmonella in eight outbreaks. Higher counts (>107 MPN.g-1) of Salmonella were detected mostly in foods containing mayonnaise. The isolation of Salmonella from multiple food items in five outbreaks probably resulted from the cross-contamination, and the high Salmonella counts detected in almost all analyzed samples probably resulted from storing in inadequate temperature. All strains were identified as S. Enteritidis, and presented a unique macrorestriction profile, demonstrating the predominance of one clonal group in foods involved in the salmonellosis outbreaks. A low frequency of antimicrobial resistant S. Enteritidis strains was observed and nalidixic acid was the only resistance marker detected. PMID:24031261

3M™ Molecular detection system versus MALDI-TOF mass spectrometry and molecular techniques for the identification of Escherichia coli 0157:H7, Salmonella spp. &Listeria spp.

PubMed

Loff, Marché; Mare, Louise; de Kwaadsteniet, Michele; Khan, Wesaal

2014-06-01

The aim of this study was to compare standard selective plating, conventional PCR (16S rRNA and species specific primers), MALDI-TOF MS and the 3M™ Molecular Detection System for the routine detection of the pathogens Listeria, Salmonella and Escherichia coli 0157:H7 in wastewater and river water samples. MALDI-TOF MS was able to positively identify 20/21 (95%) of the E. coli isolates obtained at genus and species level, while 16S rRNA sequencing only correctly identified 6/21 (28%) as E. coli strains. None of the presumptive positive Listeria spp. and Salmonella spp. isolates obtained by culturing on selective media were positively identified by MALDI-TOF and 16S rRNA analysis. The species-specific E. coli 0157:H7 PCR described in this present study, was not able to detect any E. coli 0157:H7 strains in the wastewater and river water samples analysed. However, E. coli strains, Listeria spp., L. monocytogenes and Salmonella spp. were detected using species specific PCR. Escherichia coli 0157:H7, Listeria spp. and Salmonella spp. were also sporadically detected throughout the sampling period in the wastewater and river water samples analysed by the 3M™ Molecular Detection System. MALDI-TOF MS, which is a simple, accurate and cost-effective detection method, efficiently identified the culturable organisms, while in the current study both species specific PCR (Listeria spp. and Salmonella spp.) and 3M™ Molecular Detection System could be utilised for the direct routine analysis of pathogens in water sources. Copyright © 2014 Elsevier B.V. All rights reserved.

Comparison of a novel strategy for the detection and isolation of Salmonella in shell eggs with the Food and Drug Administration Bacteriological Analytical Manual method.

PubMed

Zhang, Guodong; Thau, Eve; Brown, Eric W; Hammack, Thomas S

2013-12-01

The current FDA Bacteriological Analytical Manual (BAM) method for the detection of Salmonella in eggs requires 2 wk to complete. The objective of this project was to improve the BAM method for the detection and isolation of Salmonella in whole shell eggs. A novel protocol, using 1,000 g of liquid eggs for direct preenrichment with 2 L of tryptic soy broth (TSB) followed by enrichment using Rappaport-Vassiliadis and Tetrathionate broths, was compared with the standard BAM method, which requires 96 h room temperature incubation of whole shell egg samples followed by preenrichment in TSB supplemented with FeSO4. Four Salmonella ser. Enteritidis (4 phage types) and one Salmonella ser. Heidelberg isolates were used in the study. Bulk inoculated pooled liquid eggs, weighing 52 or 56 kg (approximately 1,100 eggs) were used in each trial. Twenty 1,000-g test portions were withdrawn from the pooled eggs for both the alternative and the reference methods. Test portions were inoculated with Salmonella at 1 to 5 cfu/1,000 g eggs. Two replicates were performed for each isolate. In the 8 trials conducted with Salmonella ser. Enteritidis, the alternative method was significantly (P < 0.05) more productive than the reference method in 3 trials, and significantly (P < 0.05) less productive than the reference method in 1 trial. There were no significant (P < 0.05) differences between the 2 methods for the other 4 trials. For Salmonella ser. Heidelberg, combined data from 2 trials showed the alternative method was significantly (P < 0.05) more efficient than the BAM method. We have concluded that the alternative method, described herein, has the potential to replace the current BAM culture method for detection and isolation of Salmonella from shell eggs based on the following factors: 1) the alternative method is 4 d shorter than the reference method; 2) it uses regular TSB instead of the more complicated TSB supplemented with FeSO4; and 3) it was equivalent or superior to the reference method in 9 out of 10 trials for the detection of Salmonella in shell eggs.

Rapid Emergence and Clonal Dissemination of CTX-M-15-Producing Salmonella enterica Serotype Virchow, South Korea.

PubMed

Kim, Jin Seok; Yun, Young-Sun; Kim, Soo Jin; Jeon, Se-Eun; Lee, Deog-Yong; Chung, Gyung Tae; Yoo, Cheon-Kwon; Kim, Junyoung

2016-01-01

The prevalence of cefotaxime-resistant Salmonella enterica serotype Virchow has dramatically increased in South Korea since the first isolation in 2011. Of 68 isolates collected over 10 years, 28 cefotaxime-resistant isolates harbored the bla(CTX-M-15) extended-spectrum β-lactamase gene and were closely related genetically, demonstrating the clonal dissemination of CTX-M-15-producing Salmonella Virchow in South Korea.

Detection of biological contaminants on foods and food surfaces using laser-induced breakdown spectroscopy (LIBS).

PubMed

Multari, Rosalie A; Cremers, David A; Dupre, Jo Anne M; Gustafson, John E

2013-09-11

The rapid detection of biological contaminants, such as Escherichia coli O157:H7 and Salmonella enterica , on foods and food-processing surfaces is important to ensure food safety and streamline the food-monitoring process. Laser-induced breakdown spectroscopy (LIBS) is an ideal candidate technology for this application because sample preparation is minimal and results are available rapidly (seconds to minutes). Here, multivariate regression analysis of LIBS data is used to differentiate the live bacterial pathogens E. coli O157:H7 and S. enterica on various foods (eggshell, milk, bologna, ground beef, chicken, and lettuce) and surfaces (metal drain strainer and cutting board). The type (E. coli or S. enterica) of bacteria could be differentiated in all cases studied along with the metabolic state (viable or heat killed). This study provides data showing the potential of LIBS for the rapid identification of biological contaminants using spectra collected directly from foods and surfaces.

Detection of cell surface hydrophobicity, biofilm and fimbirae genes in salmonella isolated from tunisian clinical and poultry meat.

PubMed

Ben Abdallah, Fethi; Lagha, Rihab; Said, Khaled; Kallel, Héla; Gharbi, Jawhar

2014-04-01

The aim of this study was to evaluate the ability of 15 serotypes of Salmonella to form biofilm on polystyrene, polyvinyl chloride (PVC) and glass surfaces. . Initially slime production was assessed on CRA agar and hydrophobicity of 20 Salmonella strains isolated from poultry and human and two Salmonella enterica serovar Typhimurium references strains was achieved by microbial adhesion to n-hexadecane. In addition, biofilm formation on polystyrene, PVC and glass surfaces was also investigated by using MTT and XTT colorimetric assay. Further, distribution of Salmonella enterotoxin (stn), Salmonella Enteritidis fimbrial (sef) and plasmid encoded fimbrial (pef) genes among tested strains was achieved by PCR. Salmonella strains developed red and white colonies on CRA and they are considered as hydrophilic with affinity values to n-hexadecane ranged between 0.29% and 29.55%. Quantitative biofilm assays showed that bacteria are able to form biofilm on polystyrene with different degrees and 54.54% of strains produce a strong biofilm on glass. In addition, all the strains form only a moderate (54.54%) and weak (40.91%) biofilm on PVC. PCR detection showed that only S. Enteritidis harbour Sef gene, whereas Pef and stn genes were detected in S. Kentucky, S. Amsterdam, S. Hadar, S. Enteritidis and S. Typhimurium. Salmonella serotypes are able to form biofilm on hydrophobic and hydrophilic industrial surfaces. Biofilm formation of Salmonella on these surfaces has an increased potential to compromise food safety and potentiate public health risk.

The role of defeathering in the contamination of turkey skin by Salmonella species and Listeria monocytogenes.

PubMed

Clouser, C S; Doores, S; Mast, M G; Knabel, S J

1995-04-01

This study was undertaken to determine whether the incidence of either Salmonella spp. or Listeria monocytogenes on turkeys at three commercial processors could be related to the type of defeathering system: 1) conventional, 58 C common bath scald; 2) kosher, 7 C common bath scald; or 3) steam-spray, 62 C nonimmersion scald. Flocks were sampled before defeathering, after defeathering, and after chill at each facility. The incidence of Salmonella-positive turkeys significantly increased subsequent to conventional defeathering (10 positive out of 14) as compared with before defeathering (3/14). The number of Salmonella-positive carcasses following kosher (0/14) and steam-spray (2/14) defeathering were similar to the number of Salmonella-positive carcasses found prior to defeathering (1/14 and 3/14, respectively). The incidence of Salmonella-positive carcasses following chill was slightly lower, but not significantly different than the number of Salmonella-positive carcasses found immediately following defeathering at all processors (8/14, 0/14, 1/14 for conventional, kosher, and steam-spray processors, respectively). Although L. monocytogenes was detected on turkeys sampled before chilling (2/10, kosher) and after chilling (8/14, kosher; 1/14, conventional), no L. monocytogenes was detected on turkeys at any of the processors prior to the evisceration process. Flocks with high aerobic plate counts prior to processing were more likely to contain Salmonella-positive birds throughout processing. Aerobic plate counts of all flocks were similar after chill whether or not Salmonella spp. and L. monocytogenes were detected.

Carbon nanotube-based aptasensor for sensitive electrochemical detection of whole-cell Salmonella.

PubMed

Hasan, Md Rakibul; Pulingam, Thiruchelvi; Appaturi, Jimmy Nelson; Zifruddin, Anis Nadyra; Teh, Swe Jyan; Lim, Teck Wei; Ibrahim, Fatimah; Leo, Bey Fen; Thong, Kwai Lin

2018-08-01

In this study, an amino-modified aptasensor using multi-walled carbon nanotubes (MWCNTs)-deposited ITO electrode was prepared and evaluated for the detection of pathogenic Salmonella bacteria. An amino-modified aptamer (ssDNA) which binds selectively to whole-cell Salmonella was immobilised on the COOH-rich MWCNTs to produce the ssDNA/MWCNT/ITO electrode. The morphology of the MWCNT before and after interaction with the aptamers were observed using scanning electron microscopy (SEM). Cyclic voltammetry and electrochemical impedance spectroscopy techniques were used to investigate the electrochemical properties and conductivity of the aptasensor. The results showed that the impedance measured at the ssDNA/MWCNT/ITO electrode surface increased after exposure to Salmonella cells, which indicated successful binding of Salmonella on the aptamer-functionalised surface. The developed ssDNA/MWCNT/ITO aptasensor was stable and maintained linearity when the scan rate was increased from 10 mV s -1 to 90 mV s -1 . The detection limit of the ssDNA/MWCNT/ITO aptasensor, determined from the sensitivity analysis, was found to be 5.5 × 10 1  cfu mL -1 and 6.7 × 10 1  cfu mL -1 for S. Enteritidis and S. Typhimurium, respectively. The specificity test demonstrated that Salmonella bound specifically to the ssDNA/MWCNT/ITO aptasensor surface, when compared with non-Salmonella spp. The prepared aptasensor was successfully applied for the detection of Salmonella in food samples. Copyright © 2018 Elsevier Inc. All rights reserved.

Verification and large scale clinical evaluation of a national standard protocol for Salmonella spp./Shigella spp. screening using real-time PCR combined with guided culture.

PubMed

Tang, Xi-Jun; Yang, Ze; Chen, Xin-Bin; Tian, Wen-Fang; Tu, Cheng-Ning; Wang, Hai-Bo

2018-02-01

Salmonella spp./Shigella spp. are often associated with food poisoning and fecal-oral transmission of acute gastroenteritis that requires strict monitoring, especially among people who would handle food and water. In 2014, the National Health and Family Planning Commission of the P. R. China issued a national standard protocol (recommendatory) for the screening of Salmonella spp./Shigella spp.. However, its performance has not been fully studied. Whether it was suitable for use in our laboratory was still unknown. In the current study, the new protocol was first verified by various experiments and then its clinical performance was evaluated in about 20,000 stool samples over a three-year period. Verification results showed that the new protocol was highly specific and reproducible. Sensitivity (as defined as the lower limit of detection) of the new protocol at the PCR step was 10 3 CFU/mL and 10 1 CFU/mL for Salmonella spp. and Shigella spp., while that at the guided culture step was 10 4 CFU/mL and 10 3 CFU/mL, respectively. The large scale clinical evaluation indicated that the new protocol could increase the positivity rate by two fold and decrease the workload/median turnaround time significantly. In conclusion, the protocol was verified and evaluated and was proven to be a valuable platform for the rapid, specific, sensitive and high-throughput screening of Salmonella spp./Shigella spp. Copyright © 2017 Elsevier B.V. All rights reserved.

Determination of viable Salmonellae from potable and source water through PMA assisted qPCR.

PubMed

Singh, Gulshan; Vajpayee, Poornima; Bhatti, Saurabh; Ronnie, Nirmala; Shah, Nimish; McClure, Peter; Shanker, Rishi

2013-07-01

Resource constrained countries identified as endemic zones for pathogenicity of Salmonella bear an economic burden due to recurring expenditure on medical treatment. qPCR used for Salmonella detection could not discriminate between viable and nonviable cells. Propidium monoazide (PMA) that selectively penetrates nonviable cells to cross-link their DNA, was coupled with ttr gene specific qPCR for quantifying viable salmonellae in source/potable waters collected from a north Indian city. Source water (raw water for urban potable water supply) and urban potable water exhibited viable salmonellae in the range of 2.1×10(4)-2.6×10(6) and 2-7160CFU/100mL, respectively. Potable water at water works exhibited DNA from dead cells but no viable cells were detected. PMA assisted qPCR could specifically detect low numbers of live salmonellae in Source and potable waters. This strategy can be used in surveillance of urban potable water distribution networks to map contamination points for better microbial risk management. Copyright © 2013 Elsevier Inc. All rights reserved.

Prophage Integrase Typing Is a Useful Indicator of Genomic Diversity in Salmonella enterica

PubMed Central

Colavecchio, Anna; D’Souza, Yasmin; Tompkins, Elizabeth; Jeukens, Julie; Freschi, Luca; Emond-Rheault, Jean-Guillaume; Kukavica-Ibrulj, Irena; Boyle, Brian; Bekal, Sadjia; Tamber, Sandeep; Levesque, Roger C.; Goodridge, Lawrence D.

2017-01-01

Salmonella enterica is a bacterial species that is a major cause of illness in humans and food-producing animals. S. enterica exhibits considerable inter-serovar diversity, as evidenced by the large number of host adapted serovars that have been identified. The development of methods to assess genome diversity in S. enterica will help to further define the limits of diversity in this foodborne pathogen. Thus, we evaluated a PCR assay, which targets prophage integrase genes, as a rapid method to investigate S. enterica genome diversity. To evaluate the PCR prophage integrase assay, 49 isolates of S. enterica were selected, including 19 clinical isolates from clonal serovars (Enteritidis and Heidelberg) that commonly cause human illness, and 30 isolates from food-associated Salmonella serovars that rarely cause human illness. The number of integrase genes identified by the PCR assay was compared to the number of integrase genes within intact prophages identified by whole genome sequencing and phage finding program PHASTER. The PCR assay identified a total of 147 prophage integrase genes within the 49 S. enterica genomes (79 integrase genes in the food-associated Salmonella isolates, 50 integrase genes in S. Enteritidis, and 18 integrase genes in S. Heidelberg). In comparison, whole genome sequencing and PHASTER identified a total of 75 prophage integrase genes within 102 intact prophages in the 49 S. enterica genomes (44 integrase genes in the food-associated Salmonella isolates, 21 integrase genes in S. Enteritidis, and 9 integrase genes in S. Heidelberg). Collectively, both the PCR assay and PHASTER identified the presence of a large diversity of prophage integrase genes in the food-associated isolates compared to the clinical isolates, thus indicating a high degree of diversity in the food-associated isolates, and confirming the clonal nature of S. Enteritidis and S. Heidelberg. Moreover, PHASTER revealed a diversity of 29 different types of prophages and 23 different integrase genes within the food-associated isolates, but only identified four different phages and integrase genes within clonal isolates of S. Enteritidis and S. Heidelberg. These results demonstrate the potential usefulness of PCR based detection of prophage integrase genes as a rapid indicator of genome diversity in S. enterica. PMID:28740489

Prophage Integrase Typing Is a Useful Indicator of Genomic Diversity in Salmonella enterica.

PubMed

Colavecchio, Anna; D'Souza, Yasmin; Tompkins, Elizabeth; Jeukens, Julie; Freschi, Luca; Emond-Rheault, Jean-Guillaume; Kukavica-Ibrulj, Irena; Boyle, Brian; Bekal, Sadjia; Tamber, Sandeep; Levesque, Roger C; Goodridge, Lawrence D

2017-01-01

Salmonella enterica is a bacterial species that is a major cause of illness in humans and food-producing animals. S. enterica exhibits considerable inter-serovar diversity, as evidenced by the large number of host adapted serovars that have been identified. The development of methods to assess genome diversity in S. enterica will help to further define the limits of diversity in this foodborne pathogen. Thus, we evaluated a PCR assay, which targets prophage integrase genes, as a rapid method to investigate S. enterica genome diversity. To evaluate the PCR prophage integrase assay, 49 isolates of S. enterica were selected, including 19 clinical isolates from clonal serovars (Enteritidis and Heidelberg) that commonly cause human illness, and 30 isolates from food-associated Salmonella serovars that rarely cause human illness. The number of integrase genes identified by the PCR assay was compared to the number of integrase genes within intact prophages identified by whole genome sequencing and phage finding program PHASTER. The PCR assay identified a total of 147 prophage integrase genes within the 49 S. enterica genomes (79 integrase genes in the food-associated Salmonella isolates, 50 integrase genes in S . Enteritidis, and 18 integrase genes in S . Heidelberg). In comparison, whole genome sequencing and PHASTER identified a total of 75 prophage integrase genes within 102 intact prophages in the 49 S. enterica genomes (44 integrase genes in the food-associated Salmonella isolates, 21 integrase genes in S . Enteritidis, and 9 integrase genes in S . Heidelberg). Collectively, both the PCR assay and PHASTER identified the presence of a large diversity of prophage integrase genes in the food-associated isolates compared to the clinical isolates, thus indicating a high degree of diversity in the food-associated isolates, and confirming the clonal nature of S . Enteritidis and S . Heidelberg. Moreover, PHASTER revealed a diversity of 29 different types of prophages and 23 different integrase genes within the food-associated isolates, but only identified four different phages and integrase genes within clonal isolates of S. Enteritidis and S. Heidelberg. These results demonstrate the potential usefulness of PCR based detection of prophage integrase genes as a rapid indicator of genome diversity in S. enterica .

Injury and death of various Salmonella serotypes due to acidic conditions

USDA-ARS?s Scientific Manuscript database

Acid injury of Salmonella could prevent detection of Salmonella in feed and feed-type samples. A previous study showed that after incubation in commonly used pre-enrichment media, mixed feeds and feed ingredients reached a pH (4.0 to 5.0) capable of injuring or killing Salmonella. Approximately 10...

Comparison of individual, pooled, and composite fecal sampling methods for detection of Salmonella on U.S. dairy operations

USDA-ARS?s Scientific Manuscript database

The objectives of this study were to estimate the prevalence of Salmonella for individual, pooled, and composite fecal samples and to compare culture results from each sample type for determining herd Salmonella infection status and identifying Salmonella serotype(s). The USDA’s National Animal Hea...

Prevalence and antimicrobial resistance of Salmonella isolated from broiler farms, chicken carcasses, and street-vended restaurants in Casamance, Senegal.

PubMed

Dione, Michel M; Ieven, Margareta; Garin, Benoît; Marcotty, Tanguy; Geerts, Stanny

2009-11-01

This study was undertaken to determine the prevalence and distribution of Salmonella on 57 randomly selected broiler farms at the end of the rearing period and in chicken products in urban and periurban areas in Casamance, Senegal, and to evaluate the antimicrobial resistance profiles of the Salmonella serovars. Salmonella was detected in chicken feces, on carcass skin, and in muscle on 35.1, 38.6, and 29.8% of farms, respectively. Salmonella was found in chicken meat servings from 14.3% of the 42 street restaurants and in 40.4% of the 285 chicken carcasses examined. The prevalence on skin and in muscle was significantly associated with the detection of Salmonella in feces (P

Induction of prophage lambda by chlorinated organics: Detection of some single-species/single-site carcinogens

DOE Office of Scientific and Technical Information (OSTI.GOV)

DeMarini, D.M.; Brooks, H.G.

1992-01-01

Twenty-eight chlorinated organic compounds were evaluated for their ability to induce DNA damage using the Microscreen prophage-induction assay in Escherichia coli. Comparison of the performance characteristics of the prophage-induction and Salmonella assays to rodent carcinogenicity assays showed that the prophage-induction assay had a somewhat higher specificity than did the Salmonella assay (70% vs. 50%); sensitivity, concordance, and positive and negative predictivity were similar for the two microbial assays. The Microscreen prophage-induction assay failed to detect eight carcinogens, perhaps due to toxicity or other unknown factors; five of these eight carcinogens were detected by the Salmonella assay. However, the prophage-induction assaymore » did detect six carcinogens that were not detected by the Salmonella assay, and five of these were single-species, single-site carcinogens, mostly mouse liver carcinogens. Some of these carcinogens, such as the chloroethanes, produce free radicals, which may be the basis for their carcinogenicity and ability to induce prophage. The prophage-induction (or other SOS) assay may be useful in identifying some genotoxic chlorinated carcinogens that induce DNA damage that do not revert the standard Salmonella tester strains.« less

Label-free as-grown double wall carbon nanotubes bundles for Salmonella typhimurium immunoassay.

PubMed

Punbusayakul, Niramol; Talapatra, Saikat; Ajayan, Pulickel M; Surareungchai, Werasak

2013-01-01

A label-free immunosensor from as-grown double wall carbon nanotubes (DW) bundles was developed for detecting Salmonella typhimurium. The immunosensor was fabricated by using the as-grown DW bundles as an electrode material with an anti-Salmonella impregnated on the surface. The immunosensor was electrochemically characterized by cyclic voltammetry. The working potential (100, 200, 300 and 400 mV vs. Ag/AgCl) and the anti-Salmonella concentration (10, 25, 50, 75, and 100 μg/mL) at the electrode were subsequently optimized. Then, chronoamperometry was used with the optimum potential of 100 mV vs. Ag/AgCl) and the optimum impregnated anti-Salmonella of 10 μg/mL to detect S. typhimurium cells (0-10(9) CFU/mL). The DW immunosensor exhibited a detection range of 10(2) to 10(7) CFU/mL for the bacteria with a limit of detection of 8.9 CFU/mL according to the IUPAC recommendation. The electrode also showed specificity to S. typhimurium but no current response to Escherichia coli. These findings suggest that the use of a label-free DW immunosensor is promising for detecting S. typhimurium.

Sources of Salmonella on broiler carcasses during transportation and processing: modes of contamination and methods of control.

PubMed

Corry, Janet E L; Allen, V M; Hudson, W R; Breslin, M F; Davies, R H

2002-01-01

The prevalence and types of salmonella in broiler chickens during transportation and during slaughter and dressing were studied. This was part of a comprehensive investigation of salmonellas in two UK poultry companies, which aimed to find the origins and mechanisms of salmonella contamination. Salmonellas were isolated using cultural methods. Serovars of Salmonella detected during rearing were usually also found in a small proportion of birds on the day of slaughter and on the carcasses at various points during processing. There was little evidence of salmonellas spreading to large numbers of carcasses during processing. Many serovars found in the feedmills or hatcheries were also detected in the birds during rearing and/or slaughter. Transport crates were contaminated with salmonellas after washing and disinfection. Prevalence of salmonellas fell in the two companies during this survey. A small number of serovars predominated in the processing plants of each company. These serovars originated from the feed mills. Reasons for transport crate contamination were: (1) inadequate cleaning, resulting in residual faecal soiling; (2) disinfectant concentration and temperature of disinfectant too low; (3) contaminated recycled flume water used to soak the crates. Efforts to control salmonella infection in broilers need to concentrate on crate cleaning and disinfection and hygiene in the feed mills.

Molecular-Based Identification and Detection of Salmonella in Food Production Systems: Current Perspectives.

PubMed

Ricke, Steven C; Kim, Sun Ae; Shi, Zhaohao; Park, Si Hong

2018-04-19

Salmonella remains a prominent cause of foodborne illnesses and can originate from a wide range of food products. Given the continued presence of pathogenic Salmonella in food production systems, there is a consistent need to improve identification and detection methods that can identify this pathogen at all stages in food systems. Methods for subtyping have evolved over the years, and the introduction of whole genome sequencing and advancements in PCR technologies has greatly improved the resolution for differentiating strains within a particular serovar. This, in turn, has led to the continued improvement in Salmonella detection technologies for utilization in food production systems. In this review, the focus will be on recent advancements in these technologies, as well as potential issues associated with the application of these tools in food production. In addition, the recent and emerging research developments on Salmonella detection and identification methodologies and their potential application in food production systems will be discussed. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

A novel multiplex PCR for the simultaneous detection of Salmonella enterica and Shigella species.

PubMed

Radhika, M; Saugata, Majumder; Murali, H S; Batra, H V

2014-01-01

Salmonella enterica and Shigella species are commonly associated with food and water borne infections leading to gastrointestinal diseases. The present work was undertaken to develop a sensitive and reliable PCR based detection system for simultaneous detection of Salmonella enterica and Shigella at species level. For this the conserved regions of specific genes namely ipaH1, ipaH, wbgZ, wzy and invA were targeted for detection of Shigella genus, S. flexneri, S. sonnei, S. boydii and Salmonella enterica respectively along with an internal amplification control (IAC). The results showed that twenty Salmonella and eleven Shigella spp., were accurately identified by the assay without showing non-specificity against closely related other Enterobacteriaceae organisms and also against other pathogens. Further evaluation of multiplex PCR was undertaken on 50 natural samples of chicken, eggs and poultry litter and results compared with conventional culture isolation and identification procedure. The multiplex PCR identified the presence of Salmonella and Shigella strains with a short pre-enrichment step of 5 h in peptone water and the same samples were processed by conventional procedures for comparison. Therefore, this reported multiplex PCR can serve as an alternative to the tedious time-consuming procedure of culture and identification in food safety laboratories.

A Rapid and Simple Real-Time PCR Assay for Detecting Foodborne Pathogenic Bacteria in Human Feces.

PubMed

Hanabara, Yutaro; Ueda, Yutaka

2016-11-22

A rapid, simple method for detecting foodborne pathogenic bacteria in human feces is greatly needed. Here, we examined the efficacy of a method that employs a combination of a commercial PCR master mix, which is insensitive to PCR inhibitors, and a DNA extraction method which used sodium dodecyl benzene sulfonate (SDBS), and Tween 20 to counteract the inhibitory effects of SDBS on the PCR assay. This method could detect the target genes (stx1 and stx2 of enterohemorrhagic Escherichia coli, invA of Salmonella Enteritidis, tdh of Vibrio parahaemolyticus, gyrA of Campylobacter jejuni, ceuE of Campylobacter coli, SEA of Staphylococcus aureus, ces of Bacillus cereus, and cpe of Clostridium perfringens) in a fecal suspension containing 1.0 × 10 1 to 1.0 × 10 3 CFU/ml. Furthermore, the assay was neither inhibited nor influenced by individual differences among the fecal samples of 10 subjects or fecal concentration (40-160 mg/ml in the fecal suspension). When we attempted to detect the genes of pathogenic bacteria in 4 actual clinical cases, we found that this method was more sensitive than standard culture method. These results showed that this assay is a rapid, simple detection method for foodborne pathogenic bacteria in human feces.

Assessment of the microbiological safety of edible dried seeds from retail premises in the United Kingdom with a focus on Salmonella spp.

PubMed

Willis, Caroline; Little, Christine L; Sagoo, Satnam; de Pinna, Elizabeth; Threlfall, John

2009-12-01

Sesame seed products have recently been associated with a number of Salmonella outbreaks in the UK and elsewhere. Aside from sesame seeds, there is little published information on the prevalence of Salmonella spp. in edible seeds. A study of 3735 samples of retail edible dried seeds in the UK was therefore carried out between October 2007 and March 2008 to assess their microbiological safety in relation to Salmonella contamination and levels of Escherichia coli, an indicator of faecal contamination. Overall, Salmonella was detected in 23 samples (0.6%), of which over half (57%) were sesame seeds. Other seeds contaminated with Salmonella were linseed (1 sample), sunflower (1 sample), alfalfa (1 sample), melon (4 samples) and mixed seeds (3 samples). E. coli was detected in 9% of samples, with 1.5% containing unsatisfactory levels (> or = 10(2)/g). These included melon, pumpkin, sesame, hemp, poppy, linseed, sunflower and mixed seeds. The UK retailers affected by the detection of Salmonella in their products recalled the contaminated batches, and Food Standards Agency food alerts were issued to advise against the consumption of affected seed products. This study highlights the importance of good hygiene practices and effective decontamination procedures during the production of these products.

Evaluation of three commercial enzyme-linked immunosorbent assays for the detection of antibodies against Salmonella spp. in meat juice from finishing pigs in Spain.

PubMed

Vico, J P; Engel, B; Buist, W G; Mainar-Jaime, R C

2010-11-01

The control of animal salmonellosis is considered as a major objective in Europe and indirect ELISAs will be important tools for the implementation of control programs for this infection in pigs. We analyse the results yielded by three commercial ELISAs (Herdcheck Swine Salmonella, SALMOTYPE Pig Screen, and PrioCHECK Salmonella) on meat juice samples from a population of slaughter pigs of Aragon, NW Spain, to assess their efficacy using traditional and latent-class approaches. Overall, the Herdcheck Swine Salmonella detected more Salmonella-infected pigs than the other two tests, but its relative sensitivity was low (65.9%). A similar result was observed when only serotypes detectable by this test were considered (69.1%). When a Bayesian approach was used the Herdcheck Swine Salmonella showed also the highest overall accuracy (sensitivity = 88% and specificity = 74%). Our results suggest that a relatively small proportion of the observed prevalence in herds would be explained by using these ELISAs. Also, this study points out that when different ELISA tests are used within the same herd, results may differ substantially. Thus, caution is advised if it is decided to use these assays for herd health classification in Spanish Salmonella control programs. © 2010 Blackwell Verlag GmbH.

Aptamer-based viability impedimetric sensor for bacteria.

PubMed

Labib, Mahmoud; Zamay, Anna S; Kolovskaya, Olga S; Reshetneva, Irina T; Zamay, Galina S; Kibbee, Richard J; Sattar, Syed A; Zamay, Tatiana N; Berezovski, Maxim V

2012-11-06

The development of an aptamer-based viability impedimetric sensor for bacteria (AptaVISens-B) is presented. Highly specific DNA aptamers to live Salmonella typhimurium were selected via the cell-systematic evolution of ligands by exponential enrichment (SELEX) technique. Twelve rounds of selection were performed; each comprises a positive selection step against viable S. typhimurium and a negative selection step against heat killed S. typhimurium and a mixture of related pathogens, including Salmonella enteritidis, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Citrobacter freundii to ensure the species specificity of the selected aptamers. The DNA sequence showing the highest binding affinity to the bacteria was further integrated into an impedimetric sensor via self-assembly onto a gold nanoparticle-modified screen-printed carbon electrode (GNP-SPCE). Remarkably, this aptasensor is highly selective and can successfully detect S. typhimurium down to 600 CFU mL(-1) (equivalent to 18 live cells in 30 μL of assay volume) and distinguish it from other Salmonella species, including S. enteritidis and S. choleraesuis. This report is envisaged to open a new venue for the aptamer-based viability sensing of a variety of microorganisms, particularly viable but nonculturable (VBNC) bacteria, using a rapid, economic, and label-free electrochemical platform.

Salmonella biofilm formation on Aspergillus niger involves cellulose - chitin interactions

USDA-ARS?s Scientific Manuscript database

Salmonella cycles between host and nonhost environments, where it can become an active member of complex microbial communities. The role of fungi in the environmental adaptation of enteric pathogens remains relatively unexplored. We have discovered that S. enterica Typhimurium rapidly attaches to an...

Diazonium-based impedimetric aptasensor for the rapid label-free detection of Salmonella typhimurium in food sample.

PubMed

Bagheryan, Zahra; Raoof, Jahan-Bakhsh; Golabi, Mohsen; Turner, Anthony P F; Beni, Valerio

2016-06-15

Fast and accurate detection of microorganisms is of key importance in clinical analysis and in food and water quality monitoring. Salmonella typhimurium is responsible for about a third of all cases of foodborne diseases and consequently, its fast detection is of great importance for ensuring the safety of foodstuffs. We report the development of a label-free impedimetric aptamer-based biosensor for S. typhimurium detection. The aptamer biosensor was fabricated by grafting a diazonium-supporting layer onto screen-printed carbon electrodes (SPEs), via electrochemical or chemical approaches, followed by chemical immobilisation of aminated-aptamer. FTIR-ATR, contact angle and electrochemical measurements were used to monitor the fabrication process. Results showed that electrochemical immobilisation of the diazonium-grafting layer allowed the formation of a denser aptamer layer, which resulted in higher sensitivity. The developed aptamer-biosensor responded linearly, on a logarithm scale, over the concentration range 1 × 10(1) to 1 × 10(8)CFU mL(-1), with a limit of quantification (LOQ) of 1 × 10(1) CFU mL(-1) and a limit of detection (LOD) of 6 CFU mL(-1). Selectivity studies showed that the aptamer biosensor could discriminate S. typhimurium from 6 other model bacteria strains. Finally, recovery studies demonstrated its suitability for the detection of S. typhimurium in spiked (1 × 10(2), 1 × 10(4) and 1 × 10(6) CFU mL(-1)) apple juice samples. Copyright © 2016 Elsevier B.V. All rights reserved.

Detection of Cell Surface Hydrophobicity, Biofilm and Fimbirae Genes in Salmonella Isolated from Tunisian Clinical and Poultry Meat

PubMed Central

BEN ABDALLAH, Fethi; LAGHA, Rihab; SAID, Khaled; KALLEL, Héla; GHARBI, Jawhar

2014-01-01

Abstract Background The aim of this study was to evaluate the ability of 15 serotypes of Salmonella to form biofilm on polystyrene, polyvinyl chloride (PVC) and glass surfaces. . Methods Initially slime production was assessed on CRA agar and hydrophobicity of 20 Salmonella strains isolated from poultry and human and two Salmonella enterica serovar Typhimurium references strains was achieved by microbial adhesion to n-hexadecane. In addition, biofilm formation on polystyrene, PVC and glass surfaces was also investigated by using MTT and XTT colorimetric assay. Further, distribution of Salmonella enterotoxin (stn), Salmonella Enteritidis fimbrial (sef) and plasmid encoded fimbrial (pef) genes among tested strains was achieved by PCR. Results Salmonella strains developed red and white colonies on CRA and they are considered as hydrophilic with affinity values to n-hexadecane ranged between 0.29% and 29.55%. Quantitative biofilm assays showed that bacteria are able to form biofilm on polystyrene with different degrees and 54.54% of strains produce a strong biofilm on glass. In addition, all the strains form only a moderate (54.54%) and weak (40.91%) biofilm on PVC. PCR detection showed that only S. Enteritidis harbour Sef gene, whereas Pef and stn genes were detected in S. Kentucky, S. Amsterdam, S. Hadar, S. Enteritidis and S. Typhimurium. Conclusion Salmonella serotypes are able to form biofilm on hydrophobic and hydrophilic industrial surfaces. Biofilm formation of Salmonella on these surfaces has an increased potential to compromise food safety and potentiate public health risk. PMID:26005652

The SPR detection of Salmonella enteritidis in food using aptamers as recongnition elements

NASA Astrophysics Data System (ADS)

Di, W. T.; Du, X. W.; Pan, M. F.; Wang, J. P.

2017-09-01

In this experiment, a fast, accurate, non-destructive, unmarked and simple-operation detection method for Salmonella enteritidis in food was established by the BI-3000 plasma resonance biosensor (SPR). This article establishes a method of using nucleic acid aptamer as immune recognition element in SPR which can be employed to detect Salmonella enteritidis in food for the first time. The experimental conditions were screened and the experimental scheme was validated and applied. The best flow rate was 5μL/min, the best concentration of the aptamers was 180mM, and the best regenerating solution was the 20mM NaOH. This method had almost no cross-reactivity. Besides, we established a standard curve of Salmonella enteritidis and SPR signal, with the detection limit of 2 cfu/mL. Finally, we tested the samples of chicken, pork, shrimp and fish purchased from supermarkets. The method has the advantages of short time, low detection limit and easy operation, which can be used for a large number of food samples.

Prevalence, seasonal occurrence and antimicrobial resistance of Salmonella in poultry retail products in Greece.

PubMed

Zdragas, A; Mazaraki, K; Vafeas, G; Giantzi, V; Papadopoulos, T; Ekateriniadou, L

2012-10-01

To detect the prevalence, the seasonal occurrence and distribution of Salmonella serotypes in poultry products and to determine the resistance profile of Salmonella isolates. A total of 96 skin-on chicken carcasses and 30 liver samples were analysed between May 2007 and May 2009 from twenty-two different commercial farm brands found in retail market countrywide. Salmonella was isolated from 38 (39·5%) of 96 chicken carcasses and from 10 (33·3%) of 30 liver samples. Higher isolation rate (60·4%) was observed in carcasses detected during summer (May to October), and lower isolation rate (18·7%) was observed in carcasses detected during winter (November to April); in liver samples, the positive rates were 53·4 and 13·2%, respectively. Twelve serotypes were detected with the serotypes Hadar, Enteritidis and Blockley being the most prevalent at 29·2, 22·9 and 12·5%, respectively. Nine of 11 Salm. Enteritidis isolates occurred during summer. Of 48 isolates, 38 (79%) were resistant to one or more of the antimicrobial agents used. The highest resistance rates were found to the following antimicrobials: streptomycin (64·5%), tetracycline (56·2%), nalidixic acid (39·5%), ampicillin and rifampicin (33·3%). The relatively high Salmonella spp. contamination rates of raw chicken meat and liver have been detected. Salm. Enteritidis isolates peaked in summer, increasing the risk to human health. Antibiotic resistance of Salmonella still remains a threat as resistance plasmids may be extensively shared between animal and humans. The study enabled us to improve the data on the seasonal occurrence of Salmonella and to determine the antimicrobial pattern profile and trends in Salmonella strains isolated from poultry retail products in Greece. © 2012 The Authors. Letters in Applied Microbiology © 2012 The Society for Applied Microbiology.

Interpretations of Antibody Responses to Salmonella enterica Serotype Enteritidis gm Flagellin in Poultry Flocks Are Enhanced by a Kinetics-Based Enzyme-Linked Immunosorbent Assay

PubMed Central

McDonough, Patrick L.; Jacobson, Richard H.; Timoney, John F.; Mutalib, Ahmed; Kradel, David C.; Chang, Yung-fu; Shin, Sang J.; Lein, Donald H.; Trock, Susan; Wheeler, Kaye

1998-01-01

Many regulatory and diagnostic programs for the detection of Salmonella enterica serotype Enteritidis infection in commercial poultry flocks have relied on rapid Pullorum agglutination tests to screen birds because of the shared antigens of S. enterica Enteritidis and S. enterica Pullorum and Gallinarum; however, the use of the enzyme-linked immunosorbent assay (ELISA) format affords better analytical sensitivity than crude agglutination tests. In this study, we adapted our earlier conventional indirect ELISA, using gm flagellin as the antigen, to a kinetics-based, computer-controlled ELISA (KELA). The KELA was used to screen for flagellin antibody from three commercial flocks: (i) a large flock involved in a U.S. Department of Agriculture trace back from a human S. enterica Enteritidis foodborne outbreak (n = 3,209), (ii) a flock infected with the endemic S. enterica Enteritidis serotype but which also had multiple other salmonella serotypes (n = 65), and (iii) an S. enterica Pullorum-infected flock (n = 12). The first flock (S. enterica Enteritidis prevalence of 2.45% based on culture) provided a field test of the KELA and allowed the calculation of diagnostic sensitivity (D-Sn) and diagnostic specificity (D-Sp). With a cutoff of 10 (used for screening flocks [i.e., high sensitivity]), the KELA has a D-Sn of 95.2% and a D-Sp of 18.5%; with a cutoff of 140 (used in confirmatory flock testing [i.e., high specificity]), the KELA has a D-Sn of 28.0% and a D-Sp of 99.1%. We found that with a cutoff of 60 (D-Sn = 63.1%; D-Sp = 91.6%), we could eliminate reactions in the KELA caused by other non-S. enterica Enteritidis salmonellae. The KELA was also compared to two commercial rapid Pullorum tests, the Solvay (D-Sn = 94.9%; D-Sp = 55.5%) and the Vineland (D-Sn = 62.0%; D-Sp = 75.3%). PMID:9665965

Interpretations of antibody responses to Salmonella enterica serotype enteritidis gm flagellin in poultry flocks are enhanced by a kinetics-based enzyme-linked immunosorbent assay.

PubMed

McDonough, P L; Jacobson, R H; Timoney, J F; Mutalib, A; Kradel, D C; Chang, Y F; Shin, S J; Lein, D H; Trock, S; Wheeler, K

1998-07-01

Many regulatory and diagnostic programs for the detection of Salmonella enterica serotype Enteritidis infection in commercial poultry flocks have relied on rapid Pullorum agglutination tests to screen birds because of the shared antigens of S. enterica Enteritidis and S. enterica Pullorum and Gallinarum; however, the use of the enzyme-linked immunosorbent assay (ELISA) format affords better analytical sensitivity than crude agglutination tests. In this study, we adapted our earlier conventional indirect ELISA, using gm flagellin as the antigen, to a kinetics-based, computer-controlled ELISA (KELA). The KELA was used to screen for flagellin antibody from three commercial flocks: (i) a large flock involved in a U.S. Department of Agriculture trace back from a human S. enterica Enteritidis foodborne outbreak (n = 3,209), (ii) a flock infected with the endemic S. enterica Enteritidis serotype but which also had multiple other Salmonella serotypes (n = 65), and (iii) an S. enterica Pullorum-infected flock (n = 12). The first flock (S. enterica Enteritidis prevalence of 2.45% based on culture) provided a field test of the KELA and allowed the calculation of diagnostic sensitivity (D-Sn) and diagnostic specificity (D-Sp). With a cutoff of 10 (used for screening flocks [i.e., high sensitivity]), the KELA has a D-Sn of 95.2% and a D-Sp of 18.5%; with a cutoff of 140 (used in confirmatory flock testing [i.e., high specificity]), the KELA has a D-Sn of 28.0% and a D-Sp of 99.1%. We found that with a cutoff of 60 (D-Sn = 63.1%; D-Sp = 91.6%), we could eliminate reactions in the KELA caused by other non-S. enterica Enteritidis salmonellae. The KELA was also compared to two commercial rapid Pullorum tests, the Solvay (D-Sn = 94.9%; D-Sp = 55.5%) and the Vineland (D-Sn = 62.0%; D-Sp = 75.3%).

Label-free impedimetric biosensor for Salmonella Typhimurium detection based on poly [pyrrole-co-3-carboxyl-pyrrole] copolymer supported aptamer.

PubMed

Sheikhzadeh, E; Chamsaz, M; Turner, A P F; Jager, E W H; Beni, V

2016-06-15

The Gram-negative bacterium, Salmonella Typhimurium (S. Typhimurium) is a food borne pathogen responsible for numerous hospitalisations and deaths all over the world. Conventional detection methods for pathogens are time consuming and labour-intensive. Hence, there is considerable interest in faster and simpler detection methods. Polypyrrole-based polymers, due to their intrinsic chemical and electrical properties, have been demonstrated to be valuable candidates for the fabrication of chemo/biosensors and functional surfaces. Similarly aptamers have been shown to be good alternatives to antibodies in the development of affinity biosensors. In this study, we report on the combination of poly [pyrrole-co-3-carboxyl-pyrrole] copolymer and aptamer for the development of a label-less electrochemical biosensor suitable for the detection of S. Typhimurium. Impedimetric measurements were facilitated by the effect of the aptamer/target interaction on the intrinsic conjugation of the poly [pyrrole-co-3-carboxyl-pyrrole] copolymer and subsequently on its electrical properties. The aptasensor detected S. Typhimurium in the concentration range 10(2)-10(8) CFU mL(-1) with high selectivity over other model pathogens and with a limit of quantification (LOQ) of 100 CFU mL(-1) and a limit of detection (LOD) of 3 CFU mL(-1). The suitability of the aptasensor for real sample detection was demonstrated via recovery studies performed in spiked apple juice samples. We envisage this to be a viable approach for the inexpensive and rapid detection of pathogens in food, and possibly in other environmental samples. Copyright © 2016 Elsevier B.V. All rights reserved.

Salmonella enterica isolates from pasture-raised poultry exhibit antimicrobial resistance and class I integrons.

PubMed

Melendez, S N; Hanning, I; Han, J; Nayak, R; Clement, A R; Wooming, A; Hererra, P; Jones, F T; Foley, S L; Ricke, S C

2010-12-01

While considerable foodborne pathogen research has been conducted on conventionally produced broilers and turkeys, few studies have focused on free-range (organic) or pastured poultry. The current surveillance study was designed to isolate, identify and genetically characterize Salmonella from pastured poultry farm environment and from retail samples. In this study, 59 isolates were collected from two pastured poultry farms (n = 164; pens, feed, water and insect traps) and retail carcasses (n = 36) from a local natural foods store and a local processing plant. All isolates were serotyped and analysed phenotypically (antimicrobial resistance profiles) and genotypically (DNA fingerprints, plasmid profiles and integron analysis). Salmonella enterica was detected using standard microbiological methods. Salmonella Kentucky was the most prevalent serotype detected from the sampled sources (53%), followed by Salmonella Enteritidis (24%), Bareilly (10%), Mbandaka (7%), Montevideo (5%) or Newport (2%). All isolates were resistant to sulfisoxazole and novobiocin, and the majority (40/59) possessed class I integrons shown by PCR detection. Each Salmonella serotype elicited a distinct pulsed-field gel electrophoresis fingerprint profile, and unique differences were observed among the serotypes.  The findings of this study show that Salmonella serotypes isolated from pasture-raised poultry exhibit antimicrobial resistance and class I integrons.  This study demonstrates that despite the cessation of antibiotic usage in poultry production, antibiotic resistant Salmonella may still be recovered from the environment and poultry products. © 2010 The Authors. Journal of Applied Microbiology © 2010 The Society for Applied Microbiology.

Growth dynamics of Salmonella enterica strains on alfalfa sprouts and in waste seed irrigation water.

PubMed

Howard, Michael B; Hutcheson, Steven W

2003-01-01

Alfalfa sprouts and other seed sprouts have been implicated in numerous outbreaks of salmonellosis. The source of these epidemics appears to have been low-level contamination of seeds by Salmonella bacteria that developed into clinically significant populations during the seed germination process. To test the possibility that Salmonella enterica strains carry host range determinants that allow them to grow on alfalfa, strains isolated from alfalfa or other sources were surveyed for their ability to grow on germinating alfalfa seeds. An S. enterica serovar Cubana strain originally isolated from contaminated alfalfa sprouts multiplied most rapidly during the initial 24 h of the seed germination process. Germinating alfalfa seeds supported the multiplication of S. enterica cells prior to the emergence of the root radicle at 72 h. Thereafter, much lower rates of multiplication were apparent. The ability of S. enterica to grow on germinating alfalfa seeds was independent of the serovar, isolation source, or virulence of the strain. Isolates obtained from alfalfa attained population levels similar to those observed for strains isolated from contaminated meat products or stools. Each of the strains could be detected in the waste irrigation water, with populations being strongly correlated with those detected on the germinating alfalfa seeds. The S. enterica strains were capable of utilizing the waste irrigation water as a sole carbon and nitrogen source. S. enterica strains thus appear to grow saprophytically on soluble organics released from seeds during early phases of germination. The ability to detect S. enterica in the waste irrigation water early in the germination process indicates that this method may be used as a simple way to monitor the contamination of sprouts during commercial operations.

Label-free screening of foodborne Salmonella using surface plasmon resonance imaging

USDA-ARS?s Scientific Manuscript database

Since 15 pathogens cause approximately 95% of the foodborne infections, it is desirable to develop rapid and simultaneous screening methods for these major pathogens. In this study, we developed an immunoassay for Salmonella based on surface plasmon resonance imaging (SPRi). The sensor surface modif...

Method for the detection of Salmonella enterica serovar Enteritidis

DOEpatents

Agron, Peter G.; Andersen, Gary L.; Walker, Richard L.

2008-10-28

Described herein is the identification of a novel Salmonella enterica serovar Enteritidis locus that serves as a marker for DNA-based identification of this bacterium. In addition, three primer pairs derived from this locus that may be used in a nucleotide detection method to detect the presence of the bacterium are also disclosed herein.

Sensitive Monitoring of Enterobacterial Contamination of Food Using Self-Propelled Janus Microsensors.

PubMed

Pacheco, M; Jurado-Sánchez, B; Escarpa, A

2018-02-20

Food poisoning caused by bacteria is a major cause of disease and death worldwide. Herein we describe the use of Janus micromotors as mobile sensors for the detection of toxins released by enterobacteria as indicators of food contamination. The micromotors are prepared by a Pickering emulsion approach and rely on the simultaneous encapsulation of platinum nanoparticles for enhanced bubble-propulsion and receptor-functionalized quantum dots (QDs) for selective binding with the 3-deoxy-d-manno-oct-2-ulosonic acid target in the endotoxin molecule. Lipopolysaccharides (LPS) from Salmonella enterica were used as target endotoxins, which upon interaction with the QDs induce a rapid quenching of the native fluorescence of the micromotors in a concentration-dependent manner. The micromotor assay can readily detect concentrations as low as 0.07 ng mL -1 of endotoxin, which is far below the level considered toxic to humans (275 μg mL -1 ). Micromotors have been successfully applied for the detection of Salmonella toxin in food samples in 15 min compared with several hours required by the existing Gold Standard method. Such ultrafast and reliable approach holds considerable promise for food contamination screening while awaiting the results of bacterial cultures in a myriad of food safety and security defense applications.

Clinical and economic evaluation of BBL CHROMagar Salmonella (CHROMSal) versus subculture after selenite broth enrichment to CHROMSal and Hektoen enteric agars to detect enteric Salmonella in a large regional microbiology laboratory.

PubMed

Church, Deirdre L; Emshey, Diana; Lloyd, Tracie; Pitout, Johann

2010-09-01

Stool culture for enteric pathogens is one of the most labor-intensive clinical microbiology procedures. Direct plating of stool to BBL CHROMagar Salmonella (CHROMSal) (BD Diagnostics, Sparks, MD) versus subculture after selenite broth enrichment (Sel) to CHROMSal (Sel-CHROMSal) and Hektoen enteric agar (Sel-Hek) (PML Microbiologicals, Eugene, OR) to detect Salmonella were compared. The number of colony picks and biochemical/serotyping tests per plate was recorded. A cost comparison was done. Fifty-one of 2999 (1.7%) stools yielded Salmonella sp., and 80% of isolates grew on CHROMSal by 24 h. CHROMSal demonstrated much less false-positive growth compared to Sel-Hek (P < 0.0001), which reduced biochemical and serotyping tests by 85% and 20%, respectively. Sel-CHROMSal and CHROMSal versus Sel-Hek improved enteric Salmonella detection when compared to a true positive "gold standard" (i.e., recovery by any culture method) with a sensitivity, specificity, positive predictive value, and negative predictive value of 100% and 94.12%, 100% and 99.97%, 100% and 97.96%, and 100% and 99.90%, respectively. CHROMSal use would result in substantial cost and labor savings.

A label-free electrochemical impedance immunosensor based on AuNPs/PAMAM-MWCNT-Chi nanocomposite modified glassy carbon electrode for detection of Salmonella typhimurium in milk.

PubMed

Dong, Jing; Zhao, Han; Xu, Minrong; Ma, Qiang; Ai, Shiyun

2013-12-01

A sensitive and stable label-free electrochemical impedance immunosensor for the detection of Salmonella typhimurium was developed by immobilising anti-Salmonella antibodies onto the gold nanoparticles and poly(amidoamine)-multiwalled carbon nanotubes-chitosan nanocomposite film modified glassy carbon electrode (AuNPs/PAMAM-MWCNT-Chi/GCE). Electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) were used to verify the stepwise assembly of the immunosensor. Co-addition of MWCNT, PAMAM and AuNPs greatly enhanced the sensitivity of the immunosensor. The immobilisation of antibodies and the binding of Salmonella cells to the modified electrode increased the electron-transfer resistance (Ret), which was directly measured with EIS using [Fe(CN)6](3-/4-) as a redox probe. A linear relationship of Ret and Salmonella concentration was obtained in the Salmonella concentration range of 1.0×10(3) to 1.0×10(7) CFU mL(-1) with a detection limit of 5.0×10(2) CFU mL(-1). Additionally, the proposed method was successfully applied to determine S. typhimurium content in milk samples with satisfactory results. Copyright © 2013 Elsevier Ltd. All rights reserved.

Assessment of Consumer Exposure to Salmonella spp., Campylobacter spp., and Shiga Toxin-Producing Escherichia coli in Meat Products at Retail in the City of Sao Paulo, Brazil.

PubMed

Ristori, Christiane Asturiano; Rowlands, Ruth Estela Gravato; Martins, Cecília Geraldes; Barbosa, Maria Luisa; Dos Santos, Luis Fernando; Jakabi, Miyoko; de Melo Franco, Bernadette Dora Gombossy

2017-08-01

Meat products may be vehicles of bacterial pathogens to humans, and Salmonella spp., Campylobacter spp., and Shiga toxin-producing Escherichia coli (STEC) are the most relevant. The aim of this study was to generate data on prevalence of these three pathogens in 552 samples of meat products (hot dogs, pork sausages, raw ground beef, and raw chicken legs) sold at retail in the city of Sao Paulo, Brazil. Salmonella spp. was detected in 5.8% (32/552) of samples, comprising pork sausages 62.5% (20/32) and chicken legs 37.5% (12/32). The counts of Salmonella spp. were low, ranging from < 0.3 to 9.3 × 10 most probable number per gram and the most frequent serovars were Salmonella Typhimurium (28.1%), Salmonella I 4,[5],12:i:- (15.6%), Salmonella Enteritidis (12.5%), Salmonella Derby, and Salmonella Brandenburg (9.4%). Campylobacter spp. was detected in 33 samples (6.0%), comprising chicken legs (82%) and ground beef (18%). All samples were negative for STEC. These results suggest that meat products when subjected to inadequate cooking and/or cross-contamination with other products ready for consumption can lead to occurrence of outbreaks, highlighting the risks associated with them.

Improving Salmonella determination in Sinaloa rivers with ultrafiltration and most probable number methods.

PubMed

Jimenez, Maribel; Chaidez, Cristobal

2012-07-01

Monitoring of waterborne pathogens is improved by using concentration methods prior to detection; however, direct microbial enumeration is desired to study microbial ecology and human health risks. The aim of this work was to determine Salmonella presence in river water with an ultrafiltration system coupled with the ISO 6579:1993 isolation standard method (UFS-ISO). Most probable number (MPN) method was used directly in water samples to estimate Salmonella populations. Additionally, the effect between Salmonella determination and water turbidity was evaluated. Ten liters or three tenfold dilutions (1, 0.1, and 0.01 mL) of water were processed for Salmonella detection and estimation by the UFS-ISO and MPN methods, respectively. A total of 84 water samples were tested, and Salmonella was confirmed in 64/84 (76%) and 38/84 (44%) when UFS-ISO and MPN were used, respectively. Salmonella populations were less than 5 × 10(3) MPN/L in 73/84 of samples evaluated (87%), and only three (3.5%) showed contamination with numbers greater than 4.5 × 10(4) MPN/L. Water turbidity did not affect Salmonella determination regardless of the performed method. These findings suggest that Salmonella abundance in Sinaloa rivers is not a health risk for human infections in spite of its persistence. Thus, choosing the appropriate strategy to study Salmonella in river water samples is necessary to clarify its behavior and transport in the environment.

Detection of Salmonella spp. in Retail Raw Food Samples from Vietnam and Characterization of Their Antibiotic Resistance▿

PubMed Central

Van, Thi Thu Hao; Moutafis, George; Istivan, Taghrid; Tran, Linh Thuoc; Coloe, Peter J.

2007-01-01

A study was conducted to examine the levels of Salmonella spp. contamination in raw food samples, including chicken, beef, pork, and shellfish, from Vietnam and to determine their antibiotic resistance characteristics. A total of 180 samples were collected and examined for the presence of Salmonella spp., yielding 91 Salmonella isolates. Sixty-one percent of meat and 18% of shellfish samples were contaminated with Salmonella spp. Susceptibility of all isolates to a variety of antimicrobial agents was tested, and resistance to tetracycline, ampicillin/amoxicillin, nalidixic acid, sulfafurazole, and streptomycin was found in 40.7%, 22.0%, 18.7%, 16.5%, and 14.3% of the isolates, respectively. Resistance to enrofloxacin, trimethoprim, chloramphenicol, kanamycin, and gentamicin was also detected (8.8 to 2.2%). About half (50.5%) of the isolates were resistant to at least one antibiotic, and multiresistant Salmonella isolates, resistant to at least three different classes of antibiotics, were isolated from all food types. One isolate from chicken (serovar Albany) contained a variant of the Salmonella genomic island 1 antibiotic resistance gene cluster. The results show that antibiotic resistance in Salmonella spp. in raw food samples from Vietnam is significant. PMID:17766455

A rapid method for the detection of foodborne pathogens by extraction of a trace amount of DNA from raw milk based on amino-modified silica-coated magnetic nanoparticles and polymerase chain reaction.

PubMed

Bai, Yalong; Song, Minghui; Cui, Yan; Shi, Chunlei; Wang, Dapeng; Paoli, George C; Shi, Xianming

2013-07-17

A method based on amino-modified silica-coated magnetic nanoparticles (ASMNPs) and polymerase chain reaction (PCR) was developed to rapidly and sensitively detect foodborne pathogens in raw milk. After optimizing parameters such as pH, temperature, and time, a trace amount of genomic DNA of pathogens could be extracted directly from complex matrices such as raw milk using ASMNPs. The magnetically separated complexes of genomic DNA and ASMNPs were directly subjected to single PCR (S-PCR) or multiplex PCR (M-PCR) to detect single or multiple pathogens from raw milk samples. Salmonella Enteritidis (Gram-negative) and Listeria monocytogenes (Gram-positive) were used as model organisms to artificially contaminate raw milk samples. After magnetic separation and S-PCR, the detection sensitivities were 8 CFU mL(-1) and 13 CFU mL(-1) respectively for these two types of pathogens. Furthermore, this method was successfully used to detect multiple pathogens (S. Enteritidis and L. monocytogenes) from artificially contaminated raw milk using M-PCR at sensitivities of 15 CFU mL(-1) and 25 CFU mL(-1), respectively. This method has great potential to rapidly and sensitively detect pathogens in raw milk or other complex food matrices. Copyright © 2013 Elsevier B.V. All rights reserved.

A PDMS/paper/glass hybrid microfluidic biochip integrated with aptamer-functionalized graphene oxide nano-biosensors for one-step multiplexed pathogen detection.

PubMed

Zuo, Peng; Li, XiuJun; Dominguez, Delfina C; Ye, Bang-Ce

2013-10-07

Infectious pathogens often cause serious public health concerns throughout the world. There is an increasing demand for simple, rapid and sensitive approaches for multiplexed pathogen detection. In this paper we have developed a polydimethylsiloxane (PDMS)/paper/glass hybrid microfluidic system integrated with aptamer-functionalized graphene oxide (GO) nano-biosensors for simple, one-step, multiplexed pathogen detection. The paper substrate used in this hybrid microfluidic system facilitated the integration of aptamer biosensors on the microfluidic biochip, and avoided complicated surface treatment and aptamer probe immobilization in a PDMS or glass-only microfluidic system. Lactobacillus acidophilus was used as a bacterium model to develop the microfluidic platform with a detection limit of 11.0 cfu mL(-1). We have also successfully extended this method to the simultaneous detection of two infectious pathogens - Staphylococcus aureus and Salmonella enterica. This method is simple and fast. The one-step 'turn on' pathogen assay in a ready-to-use microfluidic device only takes ~10 min to complete on the biochip. Furthermore, this microfluidic device has great potential in rapid detection of a wide variety of different other bacterial and viral pathogens.

A PDMS/paper/glass hybrid microfluidic biochip integrated with aptamer-functionalized graphene oxide nano-biosensors for one-step multiplexed pathogen detection

PubMed Central

Zuo, Peng; Dominguez, Delfina C.; Ye, Bang-Ce

2014-01-01

Infectious pathogens often cause serious public health concerns throughout the world. There is an increasing demand for simple, rapid and sensitive approaches for multiplexed pathogen detection. In this paper we have developed a polydimethylsiloxane (PDMS)/paper/glass hybrid microfluidic system integrated with aptamer-functionalized graphene oxide (GO) nano-biosensors for simple, one-step, multiplexed pathogen detection. The paper substrate used in this hybrid microfluidic system facilitated the integration of aptamer biosensors on the microfluidic biochip, and avoided complicated surface treatment and aptamer probe immobilization in a PDMS or glass-only microfluidic system. Lactobacillus acidophilus was used as a bacterium model to develop the microfluidic platform with a detection limit of 11.0 cfu mL−1. We have also successfully extended this method to the simultaneous detection of two infectious pathogens - Staphylococcus aureus and Salmonella enterica. This method is simple and fast. The one-step ‘turn on’ pathogen assay in a ready-to-use microfluidic device only takes ~10 min to complete on the biochip. Furthermore, this microfluidic device has great potential in rapid detection of a wide variety of different other bacterial and viral pathogens. PMID:23929394

The new ISO 6579-1: A real horizontal standard for detection of Salmonella, at last!

PubMed

Mooijman, Kirsten A

2018-05-01

Up to 2016, three international standard methods existed for the detection of Salmonella spp. in food, animal feed and samples from the primary production stage: ISO 6785:2001 for milk and milk products, ISO 6579:2002 for (other) food and animal feed and Annex D of ISO 6579:2007 for samples from the primary production stage. In 2009, an ISO/CEN working group started with the revision of ISO 6579:2002 with two main aims: combining the three aforementioned standards in one document and improving the information in ISO 6579:2002. Additionally it was decided to split ISO 6579 into three parts, where part 1 describes the detection, part 2 the enumeration by mini-MPN (published in 2012) and part 3 the serotyping of Salmonella (published in 2014). This paper describes the experiments and choices made for improving the part on detection of Salmonella (ISO 6579-1). The final voting stage on (draft) ISO 6579-1 was finished by the end of December 2016, with a positive outcome. Finally, a real horizontal standard became available for detection of Salmonella in food, animal feed, environmental samples in the area of food production and food handling and in samples from the primary production stage in 2017. Copyright © 2017 Elsevier Ltd. All rights reserved.

Survival of Salmonella during baking of peanut butter cookies.

PubMed

Lathrop, Amanda A; Taylor, Tiffany; Schnepf, James

2014-04-01

Peanuts and peanut-based products have been the source of recent Salmonella outbreaks worldwide. Because peanut butter is commonly used as an ingredient in baked goods, such as cookies, the potential risk of Salmonella remaining in these products after baking needs to be assessed. This research examines the potential hazard of Salmonella in peanut butter cookies when it is introduced via the peanut-derived ingredient. The survival of Salmonella during the baking of peanut butter cookies was determined. Commercial, creamy-style peanut butter was artificially inoculated with a five-strain Salmonella cocktail at a target concentration of 10(8) CFU/g. The inoculated peanut butter was then used to prepare peanut butter cookie dough following a standard recipe. Cookies were baked at 350 °F (177 °C) and were sampled after 10, 11, 12, 13, 14, and 15 min. Temperature profiles of the oven and cookies were monitored during baking. The water activity and pH of the inoculated and uninoculated peanut butter, raw dough, and baked cookies were measured. Immediately after baking, cookies were cooled, and the survival of Salmonella was determined by direct plating or enrichment. After baking cookies for 10 min, the minimum reduction of Salmonella observed was 4.8 log. In cookies baked for 13 and 14 min, Salmonella was only detectable by enrichment reflecting a Salmonella reduction in the range of 5.2 to 6.2 log. Cookies baked for 15 min had no detectable Salmonella. Results of this study showed that proper baking will reduce Salmonella in peanut butter cookies by 5 log or more.

Phage therapy reduces lairage-induced increases in Salmonella colonization in market weight pigs

USDA-ARS?s Scientific Manuscript database

Contamination of meat and meat products with foodborne pathogens usually results from the carcass coming in contact with the feces of an infected animal during processing. In the case of Salmonella, several recent studies have reported that pigs become rapidly infected with the organism during tran...

Development of a rapid serotyping method for Salmonella enterica using serotype-specific single-nucleotide polymorphisms

USDA-ARS?s Scientific Manuscript database

Salmonella enterica subsp. enterica serotype Enteriditis (S. Enteriditis) is the leading cause of salmonellosis worldwide, including the USA. Many S. enterica serotypes known to cause foodborne disease are associated with broiler meat contamination. While some serotypes are specific to birds (S. e...

Novel DNA binding and regulatory activities for s54 (RpoN) in Salmonella Typhimurium 14028s

USDA-ARS?s Scientific Manuscript database

Salmonella enterica subspecies enterica serovar Typhimurium, the causative agent of gastrointestinal disease and septicemia in humans, has been extensively studied, providing a comprehensive model system in which to study how pathogens respond to the rapidly changing conditions during persistence in...

Rapid identification of Salmonella serotypes through hyperspectral microscopy with different lighting sources

USDA-ARS?s Scientific Manuscript database

Hyperspectral microscope imaging (HMI) has the potential to classify foodborne pathogenic bacteria at cell level by combining microscope images with a spectrophotometer. In this study, the spectra generated from HMIs of five live Salmonella serovars from two light sources, metal halide (MH) and tun...

Pen-on-paper strategy for point-of-care testing: Rapid prototyping of fully written microfluidic biosensor.

PubMed

Li, Zedong; Li, Fei; Xing, Yue; Liu, Zhi; You, Minli; Li, Yingchun; Wen, Ting; Qu, Zhiguo; Ling Li, Xiao; Xu, Feng

2017-12-15

Paper-based microfluidic biosensors have recently attracted increasing attentions in point-of-care testing (POCT) territories benefiting from their affordable, accessible and eco-friendly features, where technologies for fabricating such biosensors are preferred to be equipment free, easy-to-operate and capable of rapid prototyping. In this work, we developed a pen-on-paper (PoP) strategy based on two custom-made pens, i.e., a wax pen and a conductive-ink pen, to fully write paper-based microfluidic biosensors through directly writing both microfluidic channels and electrodes. Particularly, the proposed wax pen is competent to realize one-step fabrication of wax channels on paper, as the melted wax penetrates into paper during writing process without any post-treatments. The practical applications of the fabricated paper-based microfluidic biosensors are demonstrated by both colorimetric detection of Salmonella typhimurium DNA with detection limit of 1nM and electrochemical measurement of glucose with detection limit of 1mM. The developed PoP strategy for making microfluidic biosensors on paper characterized by true simplicity, prominent portability and excellent capability for rapid prototyping shows promising prospect in POCT applications. Copyright © 2017 Elsevier B.V. All rights reserved.

Early Recovery of Salmonella from Food Using a 6-Hour Non-selective Pre-enrichment and Reformulation of Tetrathionate Broth.

PubMed

Daquigan, Ninalynn; Grim, Christopher J; White, James R; Hanes, Darcy E; Jarvis, Karen G

2016-01-01

Culture based methods are commonly employed to detect pathogens in food and environmental samples. These methods are time consuming and complex, requiring multiple non-selective and selective enrichment broths, and usually take at least 1 week to recover and identify pathogens. Improving pathogen detection in foods is a primary goal for regulatory agencies and industry. Salmonella detection in food relies on a series of culture steps in broth formulations optimized to resuscitate Salmonella and reduce the abundance of competitive bacteria. Examples of non-selective pre-enrichment broths used to isolate Salmonella from food include Lactose, Universal Pre-enrichment, BPW, and Trypticase Soy broths. Tetrathionate (TT) and Rappaport-Vassiliadis (RV) broths are employed after a 24-h non-selective enrichment to select for Salmonella and hamper the growth of competitive bacteria. In this study, we tested a new formulation of TT broth that lacks brilliant green dye and has lower levels of TT . We employed this TT broth formulation in conjunction with a 6-h non-selective pre-enrichment period and determined that Salmonella recovery was possible one day earlier than standard food culture methods. We tested the shortened culture method in different non-selective enrichment broths, enumerated Salmonella in the non-selective enrichments, and used 16S rRNA gene sequencing to determine the proportional abundances of Salmonella in the TT and RV selective enrichments. Together these data revealed that a 6-h non-selective pre-enrichment reduces the levels of competitive bacteria inoculated into the selective TT and RV broths, enabling the recovery of Salmonella 1 day earlier than standard culture enrichment methods.

The Use of a Combined Bioinformatics Approach to Locate Antibiotic Resistance Genes on Plasmids From Whole Genome Sequences of Salmonella enterica Serovars From Humans in Ghana.

PubMed

Kudirkiene, Egle; Andoh, Linda A; Ahmed, Shahana; Herrero-Fresno, Ana; Dalsgaard, Anders; Obiri-Danso, Kwasi; Olsen, John E

2018-01-01

In the current study, we identified plasmids carrying antimicrobial resistance genes in draft whole genome sequences of 16 selected Salmonella enterica isolates representing six different serovars from humans in Ghana. The plasmids and the location of resistance genes in the genomes were predicted using a combination of PlasmidFinder, ResFinder, plasmidSPAdes and BLAST genomic analysis tools. Subsequently, S1-PFGE was employed for analysis of plasmid profiles. Whole genome sequencing confirmed the presence of antimicrobial resistance genes in Salmonella isolates showing multidrug resistance phenotypically. ESBL, either bla TEM52-B or bla CTX-M15 were present in two cephalosporin resistant isolates of S . Virchow and S . Poona, respectively. The systematic genome analysis revealed the presence of different plasmids in different serovars, with or without insertion of antimicrobial resistance genes. In S . Enteritidis, resistance genes were carried predominantly on plasmids of IncN type, in S . Typhimurium on plasmids of IncFII(S)/IncFIB(S)/IncQ1 type. In S . Virchow and in S . Poona, resistance genes were detected on plasmids of IncX1 and TrfA/IncHI2/IncHI2A type, respectively. The latter two plasmids were described for the first time in these serovars. The combination of genomic analytical tools allowed nearly full mapping of the resistance plasmids in all Salmonella strains analyzed. The results suggest that the improved analytical approach used in the current study may be used to identify plasmids that are specifically associated with resistance phenotypes in whole genome sequences. Such knowledge would allow the development of rapid multidrug resistance tracking tools in Salmonella populations using WGS.

Growth of Staphylococcus and Salmonella on Frankfurters With and Without Sodium Nitrite

PubMed Central

Bayne, Henry G.; Michener, H. David

1975-01-01

Conventional and nitrite-free frankfurters in loosely wrapped packages were compared as to their ability to support growth of Salmonella, Staphylococcus, and their naturally occurring spoilage flora at 7 C (simulating refrigerated storage) and 20 C (simulating possible temperature abuse). At 7 C Salmonella did not grow in either type of frankfurter; Staphylococcus and the natural spoilage flora sometimes grew more rapidly in the absence of nitrite, but the difference was not significant. At 20 C growth of Salmonella, Staphylococcus, and of the spoilage flora was, at most, only slightly faster on nitrite-free frankfurters. Salmonella was not suppressed in broth culture experiments at the pH and nitrite content found in frankfurters. Although either type of frankfurter can become hazardous due to growth of Salmonella or Staphylococcus, no unusual or additional hazard resulted from the omission of nitrite from frankfurters. PMID:952

Temporal analyses of Salmonellae in a headwater spring ecosystem reveals the effects of precipitation and runoff events.

PubMed

Gaertner, James P; Garres, Tiffany; Becker, Jesse C; Jimenez, Maria L; Forstner, Michael R J; Hahn, Dittmar

2009-03-01

Sediments and water from the spring and slough arm of Spring Lake, the pristine headwaters of the San Marcos River, Texas, were analyzed for Salmonellae by culture and molecular techniques before and after three major precipitation events, each with intermediate dry periods. Polymerase chain reaction (PCR)-assisted analyses of enrichment cultures detected Salmonellae in samples after all three precipitation events, but failed to detect them immediately prior to the rainfall events. Detection among individual locations differed with respect to the precipitation event analyzed, and strains isolated were highly variable with respect to serovars. These results demonstrate that rainwater associated effects, most likely surface runoff, provide an avenue for short-term pollution of aquatic systems with Salmonellae that do not, however, appear to establish for the long-term in water nor sediments.

Au/Si Hetero-Nanorod-based Biosensor for Salmonella Detection

USDA-ARS?s Scientific Manuscript database

Technical Abstract Among several potentials of nanotechnology applications for food industry, development of nanoscale sensors for food safety and biosecurity measurement are emerging. A novel biosensor for Salmonella detection was developed using Au/Si nanorods. The Si nanorods were fabricated by...

Fate and survival of Salmonella Typhimurium and Escherichia coli O157:H7 in repacked soil lysimeters after application of cattle slurry and human urine.

PubMed

Nyberg, Karin A; Ottoson, Jakob R; Vinnerås, Björn; Albihn, Ann

2014-09-01

Use of cattle slurry as a fertiliser is common practice around the world. Human urine use is not as common, but owing to its fertiliser value this might change in the future. It is essential to minimise the transfer of enteric pathogens through fertilisation, with respect to both animal and public health. Therefore the objective of this research was to study the survival and transport of Salmonella Typhimurium and Escherichia coli O157:H7 in two agricultural soils when applied to soil along with either cattle slurry or human urine over a period of 180 days. Both Salmonella and E. coli O157:H7 were more rapidly reduced when applied together with human urine than when applied with cattle slurry. However, both pathogens persisted in low amounts at 20 and 50 cm depth in both soils throughout the whole study period. No Salmonella or E. coli O157:H7 was detected in the leachate over the 180 day study. The risk of disease transmission is higher when cattle slurry is used as fertiliser compared with human urine. However, the risk of groundwater infiltration would be low as long as water velocity through the soil is moderate. Increased knowledge of pathogen persistence in soil after fertiliser application is a valuable tool for improving risk evaluations and formulating guidelines for the use of cattle and/or human wastes in cropping soils. © 2014 Society of Chemical Industry.

Fecal prevalence, serotype distribution and antimicrobial resistance of Salmonellae in dairy cattle in central Ethiopia.

PubMed

Eguale, Tadesse; Engidawork, Ephrem; Gebreyes, Wondwossen A; Asrat, Daniel; Alemayehu, Haile; Medhin, Girmay; Johnson, Roger P; Gunn, John S

2016-02-16

Salmonellae are major worldwide zoonotic pathogens infecting a wide range of vertebrate species including humans. Consumption of contaminated dairy products and contact with dairy cattle represent a common source of non-typhoidal Salmonella infection in humans. Despite a large number of small-scale dairy farms in Addis Ababa and its surrounding districts, little is known about the status of Salmonella in these farms. Salmonella was recovered from the feces of at least one animal in 7.6% (10/132) of the dairy farms. Out of 1203 fecal samples examined, 30 were positive for Salmonella resulting in a weighted animal level prevalence of 2.3%. Detection of diarrhea in an animal and in a farm was significantly associated with animal level (p = 0.012) and herd level (p < 0.001) prevalence of Salmonella. Animal level prevalence of Salmonella was significantly associated with age (p = 0.023) and study location; it was highest among those under 6 months of age and in farms from Adaa district and Addis Ababa (p < 0.001). Nine different serotypes were identified using standard serological agglutination tests. The most frequently recovered serotypes were Salmonella Typhimurium (23.3%), S. Saintpaul (20%), S. Kentucky (16.7%) and S. Virchow (16.7%). All isolates were resistant or intermediately resistant to at least one of the 18 drugs tested. Twenty-six (86.7%), 19 (63.3 %), 18 (60%), 16 (53.3%) of the isolates were resistant to streptomycin, nitrofurantoin, sulfisoxazole and tetracycline , respectively. Resistance to 2 drugs was detected in 27 (90%) of the isolates. Resistance to 3 or more drugs was detected in 21 (70%) of the isolates, while resistance to 7 or more drugs was detected in 11 (36.7%) of the isolates. The rate of occurrence of multi-drug resistance (MDR) in Salmonella strains isolated from dairy farms in Addis Ababa was significantly higher than those isolated from farms outside of Addis Ababa (p = 0.009). MDR was more common in S. Kentucky, S. Virchow and S. Saintpaul. Isolation of Salmonella serotypes commonly known for causing human salmonellosis that are associated with an MDR phenotype in dairy farms in close proximity with human population is a major public health concern. These findings imply the need for a strict pathogen reduction strategy.

Effects of climate change on Salmonella infections.

PubMed

Akil, Luma; Ahmad, H Anwar; Reddy, Remata S

2014-12-01

Climate change and global warming have been reported to increase spread of foodborne pathogens. To understand these effects on Salmonella infections, modeling approaches such as regression analysis and neural network (NN) were used. Monthly data for Salmonella outbreaks in Mississippi (MS), Tennessee (TN), and Alabama (AL) were analyzed from 2002 to 2011 using analysis of variance and time series analysis. Meteorological data were collected and the correlation with salmonellosis was examined using regression analysis and NN. A seasonal trend in Salmonella infections was observed (p<0.001). Strong positive correlation was found between high temperature and Salmonella infections in MS and for the combined states (MS, TN, AL) models (R(2)=0.554; R(2)=0.415, respectively). NN models showed a strong effect of rise in temperature on the Salmonella outbreaks. In this study, an increase of 1°F was shown to result in four cases increase of Salmonella in MS. However, no correlation between monthly average precipitation rate and Salmonella infections was observed. There is consistent evidence that gastrointestinal infection with bacterial pathogens is positively correlated with ambient temperature, as warmer temperatures enable more rapid replication. Warming trends in the United States and specifically in the southern states may increase rates of Salmonella infections.

First detection and characterization of Salmonella spp. in poultry and swine raised in backyard production systems in central Chile.

PubMed

Alegria-Moran, R; Rivera, D; Toledo, V; Moreno-Switt, A I; Hamilton-West, C

2017-11-01

Little is known about Salmonella serovars circulating in backyard poultry and swine populations worldwide. Backyard production systems (BPS) that raise swine and/or poultry are distributed across Chile, but are more heavily concentrated in central Chile, where industrialized systems are in close contact with BPS. This study aims to detect and identify circulating Salmonella serovars in poultry and swine raised in BPS. Bacteriological Salmonella isolation was carried out for 1744 samples collected from 329 BPS in central Chile. Faecal samples were taken from swine, poultry, geese, ducks, turkeys and peacocks, as well as environmental faecal samples. Confirmation of Salmonella spp. was performed using invA-polymerase chain reaction (PCR). Identification of serovars was carried out using a molecular serotyping approach, where serogroups were confirmed by a multiplex PCR of Salmonella serogroup genes for five Salmonella O antigens (i.e., D, B, C1, C2-C3, and E1), along with two PCR amplifications, followed by sequencing of fliC and fljB genes. A total of 25 samples (1·4% of total samples) from 15 BPS (4·6 % of total sampled BPS) were found positive for Salmonella. Positive samples were found in poultry (chickens and ducks), swine and environmental sources. Molecular prediction of serovars on Salmonella isolated showed 52·0% of S. Typhimurium, 16·0% of S. Infantis, 16·0% S. Enteritidis, 8·0% S. Hadar, 4·0% S. Tennessee and 4·0% S. Kentucky. Poor biosecurity measures were found on sampled BPS, where a high percentage of mixed confinement systems (72·8%); and almost half of the sampled BPS with improper management of infected mortalities (e.g. selling the carcasses of infected animals for consumption). Number of birds other than chickens (P = 0·014; OR = 1·04; IC (95%) = 1·01-1·07), mixed productive objective (P = 0·030; OR = 5·35; IC (95%) = 1·24-27·59) and mixed animal replacement origin (P = 0017; OR = 5·19; IC (95%) = 1·35-20·47) were detected as risk factors for BPS positivity to Salmonella spp. This is the first evidence of serovars of Salmonella spp. circulating in BPS from central Chile. Detected serovars have been linked to human and animal clinical outbreaks worldwide and in Chile, highlighting the importance of BPS on the control and dissemination of Salmonella serovars potentially hazardous to public health.

Prevalence, antibiotic resistance, and extended-spectrum and AmpC β-lactamase productivity of Salmonella isolates from raw meat and seafood samples in Ho Chi Minh City, Vietnam.

PubMed

Nguyen, Dao Thi Anh; Kanki, Masashi; Nguyen, Phuc Do; Le, Hien Thi; Ngo, Phong Thanh; Tran, Doan Nguyen Minh; Le, Ninh Hoang; Dang, Chinh Van; Kawai, Takao; Kawahara, Ryuji; Yonogi, Shinya; Hirai, Yuji; Jinnai, Michio; Yamasaki, Shinji; Kumeda, Yuko; Yamamoto, Yoshimasa

2016-11-07

Salmonellosis is a type of foodborne disease caused by Salmonella enterica and is a frequent cause of childhood diarrhea in Vietnam. Of particular concern is the dissemination of multidrug-resistant Salmonella, as extended-spectrum β-lactamase (ESBL)-positive isolates were recently detected in children in Vietnam. In the present study, the prevalence and antibiotic resistance of Salmonella isolates obtained from 409 raw meat and seafood samples collected between October 2012 and March 2015 from slaughterhouses, wholesale fish market, and retail markets in Ho Chi Minh City, Vietnam were examined. A high rate of Salmonella contamination was detected in the pork (69.7%), poultry (65.3%), beef (58.3%), shrimp (49.1%), and farmed freshwater fish samples (36.6%). A total of 53 Salmonella serovars were found, of which S. Rissen, S. Weltevreden, S. London, S. Anatum, S. Typhimurium, and S. Corvallis were the most prevalent. In addition, 4 monophasic S. Typhimurium strains were identified using a PCR method for the detection of a specific IS200 fragment within the fliB-fliA intergenic region. The Salmonella isolates had a high prevalence (62.2%) of resistance to antimicrobial agents, particularly tetracycline (53.3%), ampicillin (43.8%), chloramphenicol (37.5%), and trimethoprim/sulfamethoxazole (31.3%). Isolates with resistance to three or more classes of antimicrobials were found (41.1%). Especially, isolates such as S. monophasic Typhimurium, S. Schwarzengrund, S. Indiana, S. Newport, S. Saintpaul and S. Bovismorbificans exhibited resistance to 6 classes of antimicrobials (3.3%). All 7 S. Indiana strains were resistant to between 4 and 6 classes of antimicrobials, including ciprofloxacin, which is commonly used for the treatment of human Salmonella infections. Two fish isolates were confirmed to be CTX-M-55 ESBL-producing Salmonella serovars Bovismorbificans and Newport, and five CMY-2 AmpC-producing Salmonella isolates of serovars Braenderup (4) and Typhimurium (1) were detected in poultry samples. The findings from this study, which is the first report of ESBL- and AmpC-producing Salmonella isolates from food in Vietnam, indicate that multidrug-resistant Salmonella are widely disseminated not only in meats, but also in seafood, within the food distribution system of Vietnam. The presence of these multidrug-resistant strains is a public health concern and suggests that the use of antimicrobial agents in both humans and animals in Vietnam should be tightly controlled.

Effect of ionizing radiation on the quantitative detection of Salmonella using real-time PCR

NASA Astrophysics Data System (ADS)

Lim, Sangyong; Jung, Jinwoo; Kim, Minjeong; Ryu, Sangryeol; Kim, Dongho

2008-09-01

Food irradiation is an economically viable technology for inactivating foodborne pathogens, but irradiation can mask pathogens in unhygienically prepared food. The aim of this study was to investigate the effect of irradiation treatment on the detection of Salmonella using real-time PCR. Three commercially available kits were tested, of which the InstaGene Matrix procedure was most effective in preparing template DNA from Salmonella exposed to radiation in broth culture. The minimum level of detection by real-time PCR combined with InstaGene Matrix was 3 log units of Salmonella per milliliter. However, when pure cultures of Salmonella were irradiated at 3 and 5 kGy, the cycle threshold ( CT) increased 1-1.5-fold compared to irradiation at 0 and 1 kGy. This indicated that irradiation treatment may result in an underestimation of bacterial counts due to radiation-induced DNA lesions. We also compared CT values in inoculated chicken homogenates before and after irradiation, which in this model caused a 1.3-3.3-fold underestimation of bacterial counts with respect to irradiation dose.

Detection of Brucellosis in Sika Deer ( Cervus nippon ) through Loop-mediated Isothermal Amplification (LAMP).

PubMed

Liu, Qianhong; Wei, Jie; Sun, Qingsong; Wang, Ben; Wang, Yuting; Hu, Ying; Wu, Wenrong

2017-07-01

Brucellosis (Brucella bovis) in sika deer ( Cervus nippon ) can cause enormous losses to stag breeding, especially in areas in which stag breeding has become an important industry. It also poses a threat to humans because it is a zoonotic disease. Use of the loop-mediated isothermal amplification (LAMP) assay has been poorly described in the diagnosis of brucellosis in deer. We developed a LAMP assay targeting the omp25 gene sequence to detect brucellosis in sika deer. The reaction can be completed in 60 min at 63 C and, with a detection limit of 17 pg, it was more sensitive than conventional PCR, with its detection limit of 1.7 ng. No cross-reactivity was observed with four bacteria: Escherichia coli , Salmonella enterica subsp. enterica, Clostridium pasteurianum , and Pseudomonas aeruginosa . We used 263 samples of blood to evaluate the reaction. The percentage of agreement between LAMP and PCR reached 91%; relative specificity reached 87%, and relative sensitivity reached 100%. The results indicate LAMP can be a simple and rapid diagnostic tool for detecting brucellosis in sika deer, particularly in the field, where it is essential to control brucellosis in deer with a rapid and accurate diagnosis for removal of positive animals.

Isolation, amplification and characterization of foodborne pathogen disease bacteria gene for rapid kit test development

NASA Astrophysics Data System (ADS)

Nurjayadi, M.; Santoso, I.; Kartika, I. R.; Kurniadewi, F.; Saamia, V.; Sofihan, W.; Nurkhasanah, D.

2017-07-01

There is a lot of public concern over food safety. Food-safety cases recently, including many food poisoning cases in both the developed and developing countries, considered to be the national security threats which involved police investigation. Quick and accurate detection methods are needed to handle the food poisoning cases with a big number of sufferers at the same time. Therefore, the research is aimed to develop a specific, sensitive, and rapid result molecular detection tool for foodborne pathogen bacteria. We, thus, propose genomic level approach with Polymerase Chain Reaction. The research has successfully produced a specific primer to perform amplification to fim-C S. typhi, E. coli, and pef Salmonella typhimurium genes. The electrophoresis result shows that amplification products are 95 base pairs, 121 base pairs, and 139 base pairs; and all three genes are in accordance with the size of the in silico to third genes bacteria. In conclusion, the research has been successfully designed a specific detection tool to three foodborne pathogen bacteria genes. Further stages test and the uses of Real-time PCR in the detection are still in the trial process for better detection method.

Rapid molecular pathotyping of major salmonella enterica serotypes based on single-nucleotide polymorphisms (SNPs) in the adenylate cyclase (cyaA) gene

USDA-ARS?s Scientific Manuscript database

Introduction: Salmonella enterica subsp. enterica serotype Enteriditis (S. Enteriditis) is the leading cause of salmonellosis worldwide, including the USA. Many S. enterica serotypes known to cause foodborne disease are associated with broiler meat contamination. While some serotypes are specific...

Comparative study of shell swab and shell crush methods for the recovery of Salmonella from shell eggs

USDA-ARS?s Scientific Manuscript database

Egg associated Salmonella Enteritidis outbreaks have been a major cause of foodborne illness in Japan as well as in the United States and several European countries. Researchers have been attempting to develop a rapid and highly sensitive method for the recovery of microorganisms from shell eggs. ...

Salmonella in the pork production chain and its impact on human health in the European Union.

PubMed

Bonardi, S

2017-06-01

Salmonella spp. comprise the second most common food-borne pathogens in the European Union (EU). The role of pigs as carriers of Salmonella has been intensively studied both on farm and at slaughter. Salmonella infection in pigs may cause fever, diarrhoea, prostration and mortality. However, most infected pigs remain healthy carriers, and those infected at the end of the fattening period could pose a threat to human health. Contamination of pig carcasses can occur on the slaughter line, and it is linked to cross-contamination from other carcasses and the presence of Salmonella in the environment. Therefore, Salmonella serovars present on pig carcasses can be different from those detected in the same bathes on the farm. In recent years, S. Typhimurium, S. Derby and S. serotype 4,[5],12:i:- (a monophasic variant of S. Typhimurium) have been the most common serovars to be detected in pigs in EU countries, but S. Rissen, S. Infantis, S. Enteritidis and S. Brandenburg have also been reported. In humans, several cases of salmonellosis have been linked to the consumption of raw or undercooked pork and pork products. Among the main serovars of porcine origin detected in confirmed human cases, S. Typhimurium, the monophasic variant S. 4,[5],12:i:- and S. Derby are certainly the most important.

Culture versus PCR for Salmonella Species Identification in Some Dairy Products and Dairy Handlers with Special Concern to Its Zoonotic Importance.

PubMed

Gwida, Mayada M; Al-Ashmawy, Maha A M

2014-01-01

A total of 200 samples of milk and dairy products as well as 120 samples of dairy handlers were randomly collected from different dairy farms and supermarkets in Dakahlia Governorate, Egypt. The conventional cultural and serotyping methods for detection of Salmonella in dairy products were applied and the results were compared with those obtained by molecular screening assay using (ttr sequence). The obtained results revealed that 21% of milk and dairy products (42/200) were positive for Salmonella species using enrichment culture-based PCR method, while 12% of different dairy samples (24/200) were found to be positive for Salmonella species by using the conventional culture methods. Two stool specimens out of 40 apparently healthy dairy handlers were positive by the PCR method. Serotyping of Salmonella isolates revealed that 58.3% (14/24) from different dairy products were contaminated with Salmonella Typhimurium. We conclude that the enrichment culture-based PCR assay has high sensitivity and specificity for detection of Salmonella species in dairy products and handlers. High incidence of Salmonella Typhimurium in the examined dairy samples highlights the important role played by milk and dairy products as a vehicle in disease prevalence. Great effort should be applied for reducing foodborne risk for consumers.

9 CFR 113.30 - Detection of Salmonella contamination.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 9 Animals and Animal Products 1 2013-01-01 2013-01-01 false Detection of Salmonella contamination. 113.30 Section 113.30 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE... collected from the bulk suspension before bacteriostatic or bactericidal agents have been added. When tissue...

9 CFR 113.30 - Detection of Salmonella contamination.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false Detection of Salmonella contamination. 113.30 Section 113.30 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE... collected from the bulk suspension before bacteriostatic or bactericidal agents have been added. When tissue...

9 CFR 113.30 - Detection of Salmonella contamination.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false Detection of Salmonella contamination. 113.30 Section 113.30 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE... collected from the bulk suspension before bacteriostatic or bactericidal agents have been added. When tissue...

Evaluation of a polymerase chain reaction-based system for detection of Salmonella enteritidis, Escherichia coli O157:H7, Listeria spp., and Listeria monocytogenes on fresh fruits and vegetables.

PubMed

Shearer, A E; Strapp, C M; Joerger, R D

2001-06-01

A polymerase chain reaction (PCR)-based detection system, BAX, was evaluated for its sensitivity in detecting Salmonella Enteritidis, Escherichia coli O157:H7, Listeria sp., and Listeria monocytogenes on fresh produce. Fifteen different types of produce (alfalfa sprouts, green peppers, parsley, white cabbage, radishes, onions, carrots, mushrooms, leaf lettuce, tomatoes, strawberries, cantaloupe, mango, apples, and oranges) were inoculated, in separate studies, with Salmonella Enteritidis, E. coli O157:H7, and L. monocytogenes down to the predicted level of 1 CFU per 25-g sample. Detection by BAX was compared to recovery of the inoculated bacteria by culture methods according to the Food and Drug Administration's (FDA) Bacteriological Analytical Manual (BAM). BAX was essentially as sensitive as the culture-based method in detecting Salmonella Enteritidis and L. monocytogenes and more sensitive than the culture-based method for the detection of E. coli O157:H7 on green pepper, carrot, radish, and sprout samples. Detection of the pathogenic bacteria in samples spiked with a predicted number of less than 10 CFU was possible for most produce samples, but both methods failed to detect L. monocytogenes on carrot samples and one of two mushroom and onion samples spiked with less than 100 CFU. Both BAX and the culture method were also unable to consistently recover low numbers of E. coli O157:H7 from alfalfa sprouts. The PCR method allowed detection of Salmonella Enteritidis, E. coli O157:H7, and L. monocytogenes at least 2 days earlier than the conventional culture methods.

Influence of commercial laying hen housing systems on the incidence and identification of Salmonella and Campylobacter1

PubMed Central

Jones, D. R.; Guard, J.; Gast, R. K.; Buhr, R. J.; Fedorka-Cray, P. J.; Abdo, Z.; Plumblee, J. R.; Bourassa, D. V.; Cox, N. A.; Rigsby, L. L.; Robison, C. I.; Regmi, P.; Karcher, D. M.

2016-01-01

The housing of laying hens is important for social, industrial, and regulatory aspects. Many studies have compared hen housing systems on the research farm, but few have fully examined commercial housing systems and management strategies. The current study compared hens housed in commercial cage-free aviary, conventional cage, and enriched colony cage systems. Environmental and eggshell pool samples were collected from selected cages/segments of the housing systems throughout the production cycle and monitored for Salmonella and Campylobacter prevalence. At 77 wk of age, 120 hens per housing system were examined for Salmonella and Campylobacter colonization in the: adrenal glands, spleen, ceca, follicles, and upper reproductive tract. All isolates detected from environmental swabs, eggshell pools, and tissues were identified for serotype. Two predominant Salmonella were detected in all samples: S. Braenderup and S. Kentucky. Campylobacter coli and C. jejuni were the only Campylobacter detected in the flocks. Across all housing systems, approximately 7% of hens were colonized with Salmonella, whereas > 90% were colonized with Campylobacter. Salmonella Braenderup was the isolate most frequently detected in environmental swabs (P < 0.0001) and housing system impacted Salmonella spp. shedding (P < 0.0001). Campylobacter jejuni was the isolate most frequently found in environmental swabs (P < 0.01), while housing system impacted the prevalence of C. coli and jejuni in ceca (P < 0.0001). The results of this study provide a greater understanding of the impact of hen housing systems on hen health and product safety. Additionally, producers and academia can utilize the findings to make informed decisions on hen housing and management strategies to enhance hen health and food safety. PMID:26976901

Detection of Salmonella sp., Vibrio sp. and total plate count bacteria on blood cockle (Anadara granosa)

NASA Astrophysics Data System (ADS)

Ekawati, ER; Yusmiati, S. N. H.

2018-01-01

Blood cockle (Anadara granosa) has high level of zinc and protein, which is beneficial for therapeutic function for malnourished particularly stunting case in children. Zinc in animal foods is more absorbable than that from vegetable food. Blood cockle (Anadara granosa) is rich in nutrient and an excellent environment for the growth of microorganisms. This research aimed to identify the contamination of Salmonella sp., Vibrio sp. and total plate count bacteria on blood cockle (Anadara granosa). This was observation research with laboratory analysis. Salmonella sp. and Vibrio sp. were detected from blood cockle. Total plate count was determine of the total amount of the bacteria. Results detected from 20 samples of blood cockle showed that all samples were negative of Salmonella sp. and 1 sample positive Vibrio sp. The result of total plate count bacteria was < 5 x 105 colony/g sample.

The effect of enrichment broth and temperature on the recovery of Salmonella

USDA-ARS?s Scientific Manuscript database

Statement of the Problem: No single enrichment broth or temperature is used consistently throughout the research, regulatory or industry laboratories for the detection of Salmonella. This lack of a single methodology leads to confusion and possible bias both for and against Salmonella serotypes. The...

Testing Feeds for Salmonella.

USDA-ARS?s Scientific Manuscript database

Human salmonellosis outbreaks have been linked to contamination of animal feeds. Thus it is crucial to employ sensitive Salmonella detection methods for animal feeds. Based on a review of the literature, Salmonella sustains acid injury at about pH 4.0 to5.0. Low pH can also alter the metabolism of S...

Analysis of Antimicrobial Resistance Genes Detected in MDR Salmonella enterica Serovar Typhimurium animal isolates from the National Antimicrobial Resistance Monitoring System

USDA-ARS?s Scientific Manuscript database

Background: The presence of Multi-Drug Resistant (MDR) Salmonella in food animals is concerning. To understand how antimicrobial resistance (AR) develops, the genetic elements responsible for MDR phenotypes in Salmonella animal isolates were investigated. National Antimicrobial Resistance Monitoring...

Serotype Distribution, Antimicrobial Resistance, and Class 1 Integrons Profiles of Salmonella from Animals in Slaughterhouses in Shandong Province, China

PubMed Central

Zhao, Xiaonan; Ye, Chaoqun; Chang, Weishan; Sun, Shuhong

2017-01-01

The current study aimed to analyze the prevalence and characterization of Salmonella enterica isolated from animals in slaughterhouses before slaughter. A total of 143 non-duplicate Salmonella were recovered from 1,000 fresh fecal swabs collected from four major pig slaughterhouses (49/600, 8.2%) and four major chicken slaughterhouses (94/400, 23.5%) between March and July 2016. Among Salmonella isolates from pigs, the predominant serovars were Salmonella Rissen (28/49, 57.1%) and Typhimurium (14/49, 28.6%), and high antimicrobial resistance rates were observed for tetracycline (44/49, 89.8%) and ampicillin (16/49, 32.7%). Class 1 integrons were detected in 10.2% (5/49) of these isolates and all contained gene cassettes aadA2 (0.65 kb). Two β-lactamase genes were detected among these isolates, and most of these isolates carried blaTEM-1 (46/49), followed by blaOXA-1(4/49). Seven STs (MLST/ST, multilocus sequence typing) were detected in these isolates, and the predominant type was ST469 (19.6%). Among Salmonella isolates from chickens, the predominant serovars were Salmonella Indiana (67/94, 71.3%) and Enteritidis (23/94, 24.5%), and high antimicrobial resistance rates were observed for nalidixic acid (89/94, 94.7%), ampicillin (88/94, 93.6%) and tetracycline (81/94, 86.2%). Class 1 integrons were detected in 23 isolates (23/94, 24.5%), which contained empty integrons (0.15 kb, n = 6) or gene cassettes drfA17-aadA5 (1.7 kb, n = 6), aadA2 (1.2 kb, n = 5), drfA16-blaPSE-1-aadA2-ereA2 (1.6 kb, n = 5) or drfA1-aadA1 (1.4 kb, n = 1). Three β-lactamase genes were detected, and all 94 isolates carried blaTEM-1, followed by blaCTX-M-55 (n = 19) and blaSPE−1 (n = 3). Five STs were found in these isolates, and the predominant type was ST17 (71.3%). Our findings indicated that Salmonella was widespread in animals at slaughter and may be transmitted from animal to fork. PMID:28680418

Identification of a Plasmid-Mediated Quinolone Resistance Gene in Salmonella Isolates from Texas Dairy Farm Environmental Samples.

PubMed

Cummings, K J; Rodriguez-Rivera, L D; Norman, K N; Ohta, N; Scott, H M

2017-06-01

A recent increase in plasmid-mediated quinolone resistance (PMQR) has been detected among Salmonella isolated from humans in the United States, and it is necessary to determine the sources of human infection. We had previously isolated Salmonella from dairy farm environmental samples collected in Texas, and isolates were tested for anti-microbial susceptibility. Two isolates, serotyped as Salmonella Muenster, showed the discordant pattern of nalidixic acid susceptibility and intermediate susceptibility to ciprofloxacin. For this project, whole-genome sequencing of both isolates was performed to detect genes associated with quinolone resistance. The plasmid-mediated qnrB19 gene and IncR plasmid type were identified in both isolates. To our knowledge, this is the first report of PMQR in Salmonella isolated from food animals or agricultural environments in the United States. © 2016 Blackwell Verlag GmbH.

The Salmonella In Silico Typing Resource (SISTR): An Open Web-Accessible Tool for Rapidly Typing and Subtyping Draft Salmonella Genome Assemblies.

PubMed

Yoshida, Catherine E; Kruczkiewicz, Peter; Laing, Chad R; Lingohr, Erika J; Gannon, Victor P J; Nash, John H E; Taboada, Eduardo N

2016-01-01

For nearly 100 years serotyping has been the gold standard for the identification of Salmonella serovars. Despite the increasing adoption of DNA-based subtyping approaches, serotype information remains a cornerstone in food safety and public health activities aimed at reducing the burden of salmonellosis. At the same time, recent advances in whole-genome sequencing (WGS) promise to revolutionize our ability to perform advanced pathogen characterization in support of improved source attribution and outbreak analysis. We present the Salmonella In Silico Typing Resource (SISTR), a bioinformatics platform for rapidly performing simultaneous in silico analyses for several leading subtyping methods on draft Salmonella genome assemblies. In addition to performing serovar prediction by genoserotyping, this resource integrates sequence-based typing analyses for: Multi-Locus Sequence Typing (MLST), ribosomal MLST (rMLST), and core genome MLST (cgMLST). We show how phylogenetic context from cgMLST analysis can supplement the genoserotyping analysis and increase the accuracy of in silico serovar prediction to over 94.6% on a dataset comprised of 4,188 finished genomes and WGS draft assemblies. In addition to allowing analysis of user-uploaded whole-genome assemblies, the SISTR platform incorporates a database comprising over 4,000 publicly available genomes, allowing users to place their isolates in a broader phylogenetic and epidemiological context. The resource incorporates several metadata driven visualizations to examine the phylogenetic, geospatial and temporal distribution of genome-sequenced isolates. As sequencing of Salmonella isolates at public health laboratories around the world becomes increasingly common, rapid in silico analysis of minimally processed draft genome assemblies provides a powerful approach for molecular epidemiology in support of public health investigations. Moreover, this type of integrated analysis using multiple sequence-based methods of sub-typing allows for continuity with historical serotyping data as we transition towards the increasing adoption of genomic analyses in epidemiology. The SISTR platform is freely available on the web at https://lfz.corefacility.ca/sistr-app/.

All-carbon suspended nanowire sensors as a rapid highly-sensitive label-free chemiresistive biosensing platform.

PubMed

Thiha, Aung; Ibrahim, Fatimah; Muniandy, Shalini; Dinshaw, Ignatius Julian; Teh, Swe Jyan; Thong, Kwai Lin; Leo, Bey Fen; Madou, Marc

2018-06-01

Nanowire sensors offer great potential as highly sensitive electrochemical and electronic biosensors because of their small size, high aspect ratios, and electronic properties. Nevertheless, the available methods to fabricate carbon nanowires in a controlled manner remain limited to expensive techniques. This paper presents a simple fabrication technique for sub-100 nm suspended carbon nanowire sensors by integrating electrospinning and photolithography techniques. Carbon Microelectromechanical Systems (C-MEMS) fabrication techniques allow fabrication of high aspect ratio carbon structures by patterning photoresist polymers into desired shapes and subsequent carbonization of resultant structures by pyrolysis. In our sensor platform, suspended nanowires were deposited by electrospinning while photolithography was used to fabricate support structures. We have achieved suspended carbon nanowires with sub-100 nm diameters in this study. The sensor platform was then integrated with a microfluidic chip to form a lab-on-chip device for label-free chemiresistive biosensing. We have investigated this nanoelectronics label-free biosensor's performance towards bacterial sensing by functionalization with Salmonella-specific aptamer probes. The device was tested with varying concentrations of Salmonella Typhimurium to evaluate sensitivity and various other bacteria to investigate specificity. The results showed that the sensor is highly specific and sensitive in detection of Salmonella with a detection limit of 10 CFU mL -1 . Moreover, this proposed chemiresistive assay has a reduced turnaround time of 5 min and sample volume requirement of 5 µL which are much less than reported in the literature. Copyright © 2018 Elsevier B.V. All rights reserved.

Analysis of antimicrobial resistance genes detected in multidrug-resistant Salmonella enterica serovar Typhimurium isolated from food animals.

PubMed

Glenn, LaShanda M; Lindsey, Rebecca L; Frank, Joseph F; Meinersmann, Richard J; Englen, Mark D; Fedorka-Cray, Paula J; Frye, Jonathan G

2011-09-01

Multidrug-resistant (MDR) Salmonella enterica serovar Typhimurium is the most prevalent penta-resistant serovar isolated from animals by the U.S. National Antimicrobial Resistance Monitoring System. Penta-resistant isolates are often resistant to ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, and tetracycline. To investigate MDR in Salmonella Typhimurium (including variant 5-), one isolate each from cattle, poultry, and swine with at least the ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, and tetracycline phenotype were selected for each year from 1997 to 2007 (n = 33) for microarray analysis of antimicrobial resistance, incompatibility IncA/C, and HI1 plasmid genes. Cluster analysis based on these data separated 31 of the isolates into two groups A and B (15 and 16 isolates, respectively). Isolates in group A were phage type DT104 or U302 and were mostly swine isolates (7/15). Genes detected included intI1, bla(PSE-1), floR, aadA, sulI, tet(G), and tetR, which are often found in Salmonella Genomic Island I. Isolates in group B had numerous IncA/C plasmid genes detected and were mostly cattle isolates (9/16). Genes detected included bla(CMY-2), floR, aac(3), aadA, aphA1, strA, strB, sulI, sulII, dfrA, dhf, tet(A)(B)(C)(D), and tetR, which are often found on MDR-AmpC IncA/C plasmids. The IncA/C replicon was also detected in all group B isolates. The two remaining isolates did not cluster with any others and both had many HI1 plasmid genes detected. Linkage disequilibrium analysis detected significant associations between plasmid replicon type, phage type, and animal source. These data suggest that MDR in Salmonella Typhimurium is associated with DT104/Salmonella Genomic Island I or IncA/C MDR-AmpC encoding plasmids and these genetic elements have persisted throughout the study period.

USE OF SALMONELLA MICROSUSPENSION BIOASSAY TO DETECT THE MUTGENICITY OF MUNITIONS COMPOUNDS AT LOW CONCENTRATIONS

EPA Science Inventory

Use of a Salmonella Microsuspension Bioassay to Detect the Mutagenicity of
Munitions Compounds at Low Concentrations

Abstract

Past production and handling of munitions has resulted in soil contamination at various military facilities. Depending on...

Label-free SERS detection of Salmonella Typhimurium on DNA aptamer modified AgNR substrates

USDA-ARS?s Scientific Manuscript database

A straightforward label-free method based on aptamer binding and surface enhanced Raman specstroscopy (SERS) has been developed for the detection of Salmonella Typhimurium, an important foodborne pathogen that causes gastroenteritis in both humans and animals. Surface of the SERS-active silver nanor...

DETECTION OF FRNA COLIPHAGES IN GROUNDWATER: INTERFERENCE WITH THE ASSAY BY SOMATIC SALMONELLA BACTERIOPHAGES

EPA Science Inventory

Groundwater samples from two sites in Alabama, USA were plaque assayed for F-specific RNA (FRNA) coliphages using Salmonella typhimurium WG49 as the host bacterium. While numerous plaques were detected with WG49 (a strain possessing Escherichia coli F pili), plaques were also obs...

Multi-laboratory validation study of multilocus variable-number tandem repeat analysis (MLVA) for Salmonella enterica serovar Enteritidis, 2015

PubMed Central

Peters, Tansy; Bertrand, Sophie; Björkman, Jonas T; Brandal, Lin T; Brown, Derek J; Erdõsi, Tímea; Heck, Max; Ibrahem, Salha; Johansson, Karin; Kornschober, Christian; Kotila, Saara M; Le Hello, Simon; Lienemann, Taru; Mattheus, Wesley; Nielsen, Eva Møller; Ragimbeau, Catherine; Rumore, Jillian; Sabol, Ashley; Torpdahl, Mia; Trees, Eija; Tuohy, Alma; de Pinna, Elizabeth

2017-01-01

Multilocus variable-number tandem repeat analysis (MLVA) is a rapid and reproducible typing method that is an important tool for investigation, as well as detection, of national and multinational outbreaks of a range of food-borne pathogens. Salmonella enterica serovar Enteritidis is the most common Salmonella serovar associated with human salmonellosis in the European Union/European Economic Area and North America. Fourteen laboratories from 13 countries in Europe and North America participated in a validation study for MLVA of S. Enteritidis targeting five loci. Following normalisation of fragment sizes using a set of reference strains, a blinded set of 24 strains with known allele sizes was analysed by each participant. The S. Enteritidis 5-loci MLVA protocol was shown to produce internationally comparable results as more than 90% of the participants reported less than 5% discrepant MLVA profiles. All 14 participating laboratories performed well, even those where experience with this typing method was limited. The raw fragment length data were consistent throughout, and the inter-laboratory validation helped to standardise the conversion of raw data to repeat numbers with at least two countries updating their internal procedures. However, differences in assigned MLVA profiles remain between well-established protocols and should be taken into account when exchanging data. PMID:28277220

Prevalence and antimicrobial susceptibility of Salmonella and Shigella spp. among children with gastroenteritis in an Iranian referral hospital.

PubMed

Mahmoudi, Shima; Pourakbari, Babak; Moradzadeh, Mina; Eshaghi, Hamid; Ramezani, Amitis; Haghi Ashtiani, Mohammad Taghi; Keshavarz Valian, Sepideh; Mamishi, Setareh

2017-08-01

Gastroenteritis is one of the leading cause of illnesses through the world, especially in developing countries.Salmonella and Shigella infections are considered as the main public health problems in children. The aim of this study was to detect the prevalence and antimicrobial susceptibility of Salmonella and Shigella spp. among children with gastroenteritis in an Iranian referral hospital. During April 2013 to April 2014, all medical records of children with gastroenteritis admitted to a pediatric medical center were evaluated. Positive stool cultures of children were evaluated and frequency of Salmonella and Shigella spp. and their antimicrobial susceptibility were detected. In this study, 676 patients with the mean age of 24.94 months were enrolled. Eighty-eight (42%) Salmonella spp., 85 (40%) Shigella spp., 33 (16%) E. coli and 5(2%) candida albicans were isolated from 211 positive stool cultures. Among 85 Shigella spp. isolates, S. sonnei, S. flexneri and other Shigella spp. were isolated from 39 (46%) isolates, 36(42%) and 10(12%), respectively. Among 88 isolated Salmonella spp., 36 (41%) isolates were Salmonella Serogroup D, 26 (30%) were Salmonella Serogroup B, 20 (23%) isolates were Salmonella Serogroup C and 6 (7%) were other Salmonella spp. isolates. Thirty-eight percent of Salmonella serogroup B were resistant to nalidixic acid, while higher frequency of nalidixic acid resistant was found in Salmonella serogroup C and Salmonella serogroup D. The higher frequency of ampicillin resistant was found in Shigella spp. than Salmonella spp. High frequency of cefotaxime resistant was seen in S. sonei and S. flexneri (77% and 56%, respectively), whereas more than 90% of Salmonella serogroup B, C and D were susceptible to this antibiotic. In conclusion, Shigella and Salmonella serogroups can be considered as important etiological agents of acute diarrhea in children. Since the prevalence of antibiotic resistance is increasing in recent years in Iran, further studies on the prevalence, antimicrobial susceptibility pattern and mechanisms of antibiotic resistance in these species is highly recommended. Copyright © 2017. Published by Elsevier Ltd.

Isolation of Salmonella Virchow from a fruit bat (Pteropus giganteus).

PubMed

Islam, Ausraful; Mikolon, Andrea; Mikoleit, Matthew; Ahmed, Dilruba; Khan, Salah Udddin; Sharker, M A Yushuf; Hossain, M Jahangir; Islam, Ariful; Epstein, Jonathan H; Zeidner, Nord; Luby, Stephen P

2013-12-01

Detection of zoonotic pathogens carried by bats is important both for understanding disease ecology and for developing preventive measures. Pteropus fruit bats have been identified as potential carriers of Salmonella enterica serotype Typhi. A cross-sectional study was conducted to determine the prevalence of Salmonella Typhi and other Salmonella serotypes in Pteropus giganteus fruit bats in Bangladesh. Rectal swabs were collected from 302 bats and cultured for Salmonella species. The bats were trapped in three districts (Faridpur, Rajbari, and Cox's Bazar). Salmonella Typhi was not found but one juvenile female bat from Faridpur district was positive for Salmonella Virchow. Close associations between frugivorous bats, humans, and livestock in rural Bangladesh make it likely that the bat was infected by consuming contaminated water.

Production of the Plant Hormone Auxin by Salmonella and Its Role in the Interactions with Plants and Animals.

PubMed

Cox, Clayton E; Brandl, Maria T; de Moraes, Marcos H; Gunasekera, Sarath; Teplitski, Max

2017-01-01

The ability of human enteric pathogens to colonize plants and use them as alternate hosts is now well established. Salmonella , similarly to phytobacteria, appears to be capable of producing the plant hormone auxin via an indole-3-pyruvate decarboxylase (IpdC), a key enzyme of the IPyA pathway. A deletion of the Salmonella ipdC significantly reduced auxin synthesis in laboratory culture. The Salmonella ipdC gene was expressed on root surfaces of Medicago truncatula . M. truncatula auxin-responsive GH3::GUS reporter was activated by the wild type Salmonella , and not but the ipdC mutant, implying that the bacterially produced IAA (Indole Acetic Acid) was detected by the seedlings. Seedling infections with the wild type Salmonella caused an increase in secondary root formation, which was not observed in the ipdC mutant. The wild type Salmonella cells were detected as aggregates at the sites of lateral root emergence, whereas the ipdC mutant cells were evenly distributed in the rhizosphere. However, both strains appeared to colonize seedlings well in growth pouch experiments. The ipdC mutant was also less virulent in a murine model of infection. When mice were infected by oral gavage, the ipdC mutant was as proficient as the wild type strain in colonization of the intestine, but it was defective in the ability to cross the intestinal barrier. Fewer cells of the ipdC mutant, compared with the wild type strain, were detected in Peyer's patches, spleen and in the liver. Orthologs of ipdC are found in all Salmonella genomes and are distributed among many animal pathogens and plant-associated bacteria of the Enterobacteriaceae , suggesting a broad ecological role of the IpdC-catalyzed pathway.

Detection of Salmonella spp. and Listeria monocytogenes in Suspended Organic Waste by Nucleic Acid Extraction and PCR

PubMed Central

Burtscher, Carola; Fall, Papa A.; Wilderer, Peter A.; Wuertz, Stefan

1999-01-01

A nucleic acid-based method for the detection of the bacterial pathogens Salmonella spp. and Listeria monocytogenes in biological waste was developed. The detection limits were less than 10 cells per ml of biological waste. The method does not include a phenol extraction step and can be easily performed in 1 to 2 days. PMID:10224026

Plasmid-mediated quinolone resistance in non-Typhi serotypes of Salmonella enterica.

PubMed

Gay, Kathryn; Robicsek, Ari; Strahilevitz, Jacob; Park, Chi Hye; Jacoby, George; Barrett, Timothy J; Medalla, Felicita; Chiller, Tom M; Hooper, David C

2006-08-01

Serious infections with Salmonella species are often treated with fluoroquinolones or extended-spectrum beta-lactams. Increasingly recognized in Enterobacteriaceae, plasmid-mediated quinolone resistance is encoded by qnr genes. Here, we report the presence of qnr variants in human isolates of non-Typhi serotypes of Salmonella enterica (hereafter referred to as non-Typhi Salmonella) from the United States National Antimicrobial Resistance Monitoring System for Enteric Bacteria. All non-Typhi Salmonella specimens from the United States National Antimicrobial Resistance Monitoring System for Enteric Bacteria collected from 1996 to 2003 with ciprofloxacin minimum inhibitory concentrations > or = 0.06 microg/mL (233 specimens) and a subset with minimum inhibitory concentrations < or = 0.03 microg/mL (102 specimens) were screened for all known qnr genes (A, B, and S) by polymerase chain reaction. For isolates with positive results, qnr and quinolone resistance-determining region sequences were determined. Plasmids containing qnr genes were characterized by conjugation or transformation. Conjugative plasmids harboring qnrB variants were detected in 7 Salmonella enterica serotype Berta isolates and 1 Salmonella enterica serotype Mbandaka isolate. The S. Mbandaka plasmid also had an extended-spectrum beta -lactamase. Variants of qnrS on nonconjugative plasmids were detected in isolates of Salmonella enterica serotype Anatum and Salmonella enterica serotype Bovismorbificans. Plasmid-mediated quinolone resistance appears to be widely distributed, though it is still uncommon in non-Typhi Salmonella isolates from the United States, including strains that are quinolone susceptible by the criteria of the Clinical and Laboratory Standards Institute (formerly the National Committee for Clinical Laboratory Standards). The presence of this gene in non-Typhi Salmonella that causes infection in humans suggests potential for spread through the food supply, which is a public health concern.

Distribution of Salmonella serovars in breeding, nursery, and grow-to-finish pigs, and risk factors for shedding in ten farrow-to-finish swine farms in Alberta and Saskatchewan.

PubMed

Wilkins, Wendy; Rajić, Andrijana; Waldner, Cheryl; McFall, Margaret; Chow, Eva; Muckle, Anne; Rosengren, Leigh

2010-04-01

The study objectives were to investigate Salmonella prevalence, serovar distribution, and risk factors for shedding in 10 purposively selected farrow-to-finish farms in Saskatchewan and Alberta. Pooled fecal samples from the breeding and grow-finish phases and individual fecal samples from breeding, nursery, and grow-finish pigs were cultured for Salmonella; serotyping of isolates was performed. Pig and pen characteristics were recorded for each pig and pen sampled.Overall, 407/1143 (36%) of samples were Salmonella positive; within-farm prevalence ranged from 1% to 79%. Sows, nursery, and grow-finish pigs accounted for 43%, 29%, and 28% of positive samples, respectively. More Salmonella were detected in pooled pen than individual pig samples (P < 0.001). Among 418 Salmonella isolates, there were 19 distinct serovars; the most common were S. Derby (28.5%), S. Typhimurium, var. Copenhagen (19.1%), S. Putten (11.8%), S. Infantis (6.8%), and S. Mbandaka (6.1%). Sows were more likely to shed Salmonella than nursery or grow-finisher (OR 2.9, P < 0.001) pigs. Pelleted feed (OR 8.2, P < 0.001) and nose-to-nose pig contact through pens (OR 2.2, P = 0.005) were associated with increased Salmonella prevalence. Significant differences in serovar distribution were detected among production phases. The use of pooled pen samples is recommended as a more efficient means for accurate evaluation of Salmonella status in different phases of pig production. The breeding herd might be an important source of Salmonella persistence within farrow-to-finish farms and should be targeted in control efforts. The latter might also apply to the use of pelleted feed, which remains the most consistently reported significant risk factor for Salmonella shedding in pigs.

Method for Salmonella concentration from water at pH 3.5, using micro-fiber glass filters.

PubMed Central

Block, J C; Rolland, D

1979-01-01

A method is described for the concentration of Salmonella from water. As is done with enterovirus, Salmonella bacteria were concentrated from water in two steps: by pH 3.5 adsorption on and pH 9.5 elution from 8-micron porosity micro-fiber glass filter tubes. This method worked in less than 30 min, and Salmonella typhimurium was inactivated only slightly in spite of rapid pH variations (pH 3.5 to 9.5). It was demonstrated that the retention by the filters stems from two phenomena: a low retention in the micro-fiber glass labyrinth for small filtered volumes, and a high retention by adsorption at pH 3.5 for any filtered volume (experiments done with 15- and 80-liter samples). Addition in tap water of trivalent ions like Al3+ did not increase Salmonella adsorption. In most of the trials, Salmonella recovery varied from 42 to 93%. Preliminary field investigations indicate that enterovirus and Salmonella may both be concentrated from the same water sample by this procedure. PMID:39501

Isolation, characterization, and application of bacteriophages for Salmonella spp. biocontrol in pigs.

PubMed

Albino, Luiz A A; Rostagno, Marcos H; Húngaro, Humberto M; Mendonça, Regina C S

2014-08-01

Foodborne illness due to Salmonella-contaminated pork products is an important public health problem, causing significant economic losses worldwide. The use of bacteriophages is a potential intervention tool that has attracted interest for the control of foodborne pathogens. The objective of this study was to detect the presence of Salmonella in commercial pig farms and to isolate specific autochthonous bacteriophages against Salmonella Typhimurium, to characterize them and to evaluate their lytic capacity against Salmonella Typhimurium in vivo and in vitro. Salmonella was isolated on 50% (4/8) of the farms, with serotype Typhimurium being the most prevalent, detected in 48.2% of samples (13/27). The isolated Salmonella Typhimurium bacteriophages belong to the Podoviridae family, were active against serotypes Abony, Enteritidis, Typhi, and Typhimurium, but not against serotypes Arizonae, Cholerasuis, Gallinarum, and Pullorum. In in vitro tests, bacteriophage at 10(7) PFU/mL and 10(9) PFU/mL significantly reduced (p<0.05) Salmonella Typhimurium counts in 1.6 and 2.5 log10 colony-forming units (CFU)/mL, respectively, after 24 h. Before the in vivo treatment with bacteriophages, Salmonella was identified in 93.3% (28/30) of the fecal samples from the pigs inoculated with 10(6) CFU/mL, and only in 56.6% (17/30) after the treatment consisting of oral administration of the pool of the bacteriophages after the fasting period, simulating a common preslaughter practice. These results indicate that the pool of bacteriophages administered was capable of reducing the colonization of Salmonella in pigs.

Effects of Climate Change on Salmonella Infections

PubMed Central

Akil, Luma; Reddy, Remata S.

2014-01-01

Abstract Background: Climate change and global warming have been reported to increase spread of foodborne pathogens. To understand these effects on Salmonella infections, modeling approaches such as regression analysis and neural network (NN) were used. Methods: Monthly data for Salmonella outbreaks in Mississippi (MS), Tennessee (TN), and Alabama (AL) were analyzed from 2002 to 2011 using analysis of variance and time series analysis. Meteorological data were collected and the correlation with salmonellosis was examined using regression analysis and NN. Results: A seasonal trend in Salmonella infections was observed (p<0.001). Strong positive correlation was found between high temperature and Salmonella infections in MS and for the combined states (MS, TN, AL) models (R2=0.554; R2=0.415, respectively). NN models showed a strong effect of rise in temperature on the Salmonella outbreaks. In this study, an increase of 1°F was shown to result in four cases increase of Salmonella in MS. However, no correlation between monthly average precipitation rate and Salmonella infections was observed. Conclusion: There is consistent evidence that gastrointestinal infection with bacterial pathogens is positively correlated with ambient temperature, as warmer temperatures enable more rapid replication. Warming trends in the United States and specifically in the southern states may increase rates of Salmonella infections. PMID:25496072

Detection of Salmonella serotypes by overnight incubation of entire broiler carcass

USDA-ARS?s Scientific Manuscript database

Salmonella is a human bacterial pathogen that has been associated with poultry and poultry products. There are multiple ways to sample broiler chicken carcasses for the prevalence of Salmonella. A common method in the USA is a whole carcass rinse and culture of an aliquot of the rinse. The object...

Orally incoculated Salmonella typhimurium is detected in the lymph nodes and synovial fluid of swine

USDA-ARS?s Scientific Manuscript database

Salmonella is a foodborne pathogen that has been associated with illnesses from the consumption of meat products. Traditional carcass sampling techniques fail to account for contamination via atypical carcass reservoirs such as lymph nodes and synovial fluid that may harbor Salmonella. In this two-p...

A Rapid Systematic Review and Meta-Analysis of the Efficacy of Slaughter and Processing Interventions to Control Non-Typhoidal Salmonella in Beef and Pork

PubMed Central

Young, Ian; Wilhelm, Barbara J.; Cahill, Sarah; Nakagawa, Rei; Desmarchelier, Patricia; Rajić, Andrijana

2016-01-01

Pork is one of the major food sources of human salmonellosis worldwide, while beef products have been implicated in numerous foodborne outbreaks. As a result, effective interventions to reduce Salmonella contamination during beef and pork processing are of interest to both regulators and industry. We conducted a rapid systematic review and meta-analysis of literature investigating the efficacy of slaughter and processing interventions to control Salmonella in beef and pork. Review steps included: a comprehensive search strategy; relevance screening of abstracts; relevance confirmation of articles; data extraction; risk-of-bias assessment; meta-analysis (where appropriate); and a weight-of-evidence assessment. A total of 191 relevant experimental studies were identified. Two controlled trials indicated that hot water and steam treatments are effective at reducing the prevalence of Salmonella on beef carcasses (relative risk [RR] = 0.11, 95% confidence interval [CI]: 0.02, 0.58), while four trials found that pre-chill organic acid washes are effective at reducing Salmonella on pork carcasses (RR = 0.32, 95% CI: 0.13, 0.78), with high confidence in the estimates of effect. Four quasi-experimental studies found that post-exsanguination chemical washes were effective to reduce the prevalence of Salmonella on cattle hides, with low confidence in the specific estimate of effect; moderate confidence was found for the effect estimates of scalding (RR = 0.20, 95% CI: 0.14, 0.29) and singeing (RR = 0.34, 95% CI: 0.22, 0.52) of pork carcasses. The overall evidence supported enhanced reductions of Salmonella through a multiple-hurdle approach. In conclusion, various slaughter and processing interventions can contribute to reducing Salmonella on beef and pork carcasses, depending on the context of application; an appropriate combination should be selected, validated, and verified by establishment operators within their local conditions. PMID:28104927

A Rapid Systematic Review and Meta-Analysis of the Efficacy of Slaughter and Processing Interventions to Control Non-Typhoidal Salmonella in Beef and Pork.

PubMed

Young, Ian; Wilhelm, Barbara J; Cahill, Sarah; Nakagawa, Rei; Desmarchelier, Patricia; Rajić, Andrijana

2016-12-01

Pork is one of the major food sources of human salmonellosis worldwide, while beef products have been implicated in numerous foodborne outbreaks. As a result, effective interventions to reduce Salmonella contamination during beef and pork processing are of interest to both regulators and industry. We conducted a rapid systematic review and meta-analysis of literature investigating the efficacy of slaughter and processing interventions to control Salmonella in beef and pork. Review steps included: a comprehensive search strategy; relevance screening of abstracts; relevance confirmation of articles; data extraction; risk-of-bias assessment; meta-analysis (where appropriate); and a weight-of-evidence assessment. A total of 191 relevant experimental studies were identified. Two controlled trials indicated that hot water and steam treatments are effective at reducing the prevalence of Salmonella on beef carcasses (relative risk [RR] = 0.11, 95% confidence interval [CI]: 0.02, 0.58), while four trials found that pre-chill organic acid washes are effective at reducing Salmonella on pork carcasses (RR = 0.32, 95% CI: 0.13, 0.78), with high confidence in the estimates of effect. Four quasi-experimental studies found that post-exsanguination chemical washes were effective to reduce the prevalence of Salmonella on cattle hides, with low confidence in the specific estimate of effect; moderate confidence was found for the effect estimates of scalding (RR = 0.20, 95% CI: 0.14, 0.29) and singeing (RR = 0.34, 95% CI: 0.22, 0.52) of pork carcasses. The overall evidence supported enhanced reductions of Salmonella through a multiple-hurdle approach. In conclusion, various slaughter and processing interventions can contribute to reducing Salmonella on beef and pork carcasses, depending on the context of application; an appropriate combination should be selected, validated, and verified by establishment operators within their local conditions.

Changes in the Salmonella enterica Enteritidis phenotypes in presence of acyl homoserine lactone quorum sensing signals.

PubMed

Campos-Galvão, Maria Emilene Martino; Ribon, Andrea Oliveira Barros; Araújo, Elza Fernandes; Vanetti, Maria Cristina Dantas

2016-05-01

Quorum sensing is used by bacteria to coordinate gene expression in response to population density and involves the production, detection and response to extracellular signaling molecules known as autoinducers (AIs). Salmonella does not synthesize the AI-1, acyl homoserine lactone (AHL) common to gram-negative bacteria; however, it has a receptor for AI-1, the SdiA protein. The effect of SdiA in modulating phenotypes of Salmonella has not been elucidated. In this report, we provide evidence that the AIs-1 affect Salmonella enterica serovar Enteritidis behavior by enhancing the biofilm formation and expression of virulence genes under anaerobic conditions. Biofilm formation by Salmonella was detected by the crystal violet method and by scanning electron microscopy. The presence of AHLs, particularly C12-HSL, increased biofilm formation and promoted expression of biofilm formation genes (lpfA, fimF, fliF, glgC) and virulence genes (hilA, invA, invF). Our results demonstrated that AHLs produced by other organisms played an important role in virulence phenotypes of Salmonella Enteritidis. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Method for establishing the presence of salmonella bacteria in eggs

DOEpatents

Johnston, Roger G.; Sinha, Dipen N.

1995-01-01

Measurement of the acoustical resonances in eggs is shown to provide a rapid, noninvasive technique for establishing the presence of Salmonella bacteria. The technique is also sensitive to yolk puncture, shell cracks, and may be sensitive to other yolk properties and to egg freshness. Remote characterization, potentially useful for characterizing large numbers of eggs, has been demonstrated.

Multidrug-Resistant Salmonella enterica Serovar Infantis, Israel

PubMed Central

Valinsky, Lea; Weinberger, Miriam; Guy, Sara; Jaffe, Joseph; Schorr, Yosef Ilan; Raisfeld, Abraham; Agmon, Vered; Nissan, Israel

2010-01-01

To determine whether rapid emergence of Salmonella enterica serovar Infantis in Israel resulted from an increase in different biotypes or spread of 1 clone, we characterized 87 serovar Infantis isolates on the genotypic and phenotypic levels. The emerging strain comprised 1 genetic clone with a distinct pulsed-field gel electrophoresis profile and a common antimicrobial drug resistance pattern. PMID:21029536

Multidrug-resistant Salmonella enterica serovar Infantis, Israel.

PubMed

Gal-Mor, Ohad; Valinsky, Lea; Weinberger, Miriam; Guy, Sara; Jaffe, Joseph; Schorr, Yosef Ilan; Raisfeld, Abraham; Agmon, Vered; Nissan, Israel

2010-11-01

To determine whether rapid emergence of Salmonella enterica serovar Infantis in Israel resulted from an increase in different biotypes or spread of 1 clone, we characterized 87 serovar Infantis isolates on the genotypic and phenotypic levels. The emerging strain comprised 1 genetic clone with a distinct pulsed-field gel electrophoresis profile and a common antimicrobial drug resistance pattern.

Validation of Single and Pooled Manure Drag Swabs for the Detection of Salmonella Serovar Enteritidis in Commercial Poultry Houses.

PubMed

Kinde, Hailu; Goodluck, Helen A; Pitesky, Maurice; Friend, Tom D; Campbell, James A; Hill, Ashley E

2015-12-01

Single swabs (cultured individually) are currently used in the Food and Drug Administration (FDA) official method for sampling the environment of commercial laying hens for the detection of Salmonella enterica ssp. serovar Enteritidis (Salmonella Enteritidis). The FDA has also granted provisional acceptance of the National Poultry Improvement Plan's (NPIP) Salmonella isolation and identification methodology for samples taken from table-egg layer flock environments. The NPIP method, as with the FDA method, requires single-swab culturing for the environmental sampling of laying houses for Salmonella Enteritidis. The FDA culture protocol requires a multistep culture enrichment broth, and it is more labor intensive than the NPIP culture protocol, which requires a single enrichment broth. The main objective of this study was to compare the FDA single-swab culturing protocol with that of the NPIP culturing protocol but using a four-swab pool scheme. Single and multi-laboratory testing of replicate manure drag swab sets (n  =  525 and 672, respectively) collected from a Salmonella Enteritidis-free commercial poultry flock was performed by artificially contaminating swabs with either Salmonella Enteritidis phage type 4, 8, or 13a at one of two inoculation levels: low, x¯  = 2.5 CFU (range 2.5-2.7), or medium, x¯  = 10.0 CFU (range 7.5-12). For each replicate, a single swab (inoculated), sets of two swabs (one inoculated and one uninoculated), and sets of four swabs (one inoculated and three uninoculated), testing was conducted using the FDA or NPIP culture method. For swabs inoculated with phage type 8, the NPIP method was more efficient (P < 0.05) for all swab sets at both inoculation levels than the reference method. The single swabs in the NPIP method were significantly (P < 0.05) better than four-pool swabs in detecting Salmonella Enteritidis at the lower inoculation level. In the collaborative study (n  =  13 labs) using Salmonella Enteritidis phage type 13a inoculated swabs, there was no significant difference (P > 0.05) between the FDA method (single swabs) and the pooled NPIP method (four-pool swabs). The study concludes that the pooled NPIP method is not significantly different from the FDA method for the detection of Salmonella Enteritidis in drag swabs in commercial poultry laying houses. Consequently based on the FDA's Salmonella Enteritidis rule for equivalency of different methods, the pooled NPIP method should be considered equivalent. Furthermore, the pooled NPIP method was more efficient and cost effective.

PREVALENCE OF SALMONELLA IN CAPTIVE REPTILES FROM CROATIA.

PubMed

Lukac, Maja; Pedersen, Karl; Prukner-Radovcic, Estella

2015-06-01

Salmonellosis transmitted by pet reptiles is an increasing public health issue worldwide. The aim of this study was to investigate the prevalence of Salmonella strains from captive reptiles in Croatia. From November 2009 to November 2011 a total of 292 skin, pharyngeal, cloacal, and fecal samples from 200 apparently healthy reptiles were tested for Salmonella excretions by bacteriologic culture and serotyping. These 200 individual reptiles included 31 lizards, 79 chelonians, and 90 snakes belonging to private owners or housed at the Zagreb Zoo, Croatia. Salmonella was detected in a total of 13% of the animals, among them 48.4% lizards, 8.9% snakes, and 3.8% turtles. Representatives of five of the six Salmonella enterica subspecies were identified with the following proportions in the total number of isolates: Salmonella enterica enterica 34.6%, Salmonella enterica houtenae 23.1%, Salmonella enterica arizonae 23.1%, Salmonella enterica diarizonae 15.4%, and Salmonella enterica salamae 3.8%. The 14 different serovars isolated included several rarely occurring serovars such as Salmonella Apapa, Salmonella Halle, Salmonella Kisarawe, and Salmonella Potengi. These findings confirm that the prevalence of Salmonella is considerable in captive reptiles in Croatia, indicating that these animals may harbor serovars not commonly seen in veterinary or human microbiologic practice. This should be addressed in the prevention and diagnostics of human reptile-transmitted infections.

Non-crosslinking gold nanoprobe-LAMP for simple, colorimetric, and specific detection of Salmonella typhi

NASA Astrophysics Data System (ADS)

Bozorgmehr, Ali; Yazdanparast, Razieh; Mollasalehi, Hamidreza

2016-12-01

In this study, we developed a non-crosslinking gold nanoprobe loop-mediated isothermal amplification (LAMP) method for nanodiagnosis of bacterial typhoid fever source, Salmonella typhi. Therefore, a unique region in the S. typhi genomic DNA was targeted for LAMP amplification using a specific set of four precisely designed primers. Also, for specific colorimetric visualization of the amplicons, a thiolated oligonucleotide probe, complementary to the single-stranded loop region of the amplicons between F2 and F1C segments, was designed. The probe was bound to the surface of gold nanoparticles via covalent bonds. Increasing the salt concentration in the detection reaction medium led to aggregation of nanoprobes in the blank and the negative vessels in a time-dependent form. That was followed by a change in the surface plasmon resonance (SPR) leading to blue/black color that was observable by the naked eyes after about 5 min. Meanwhile, the original pink/red color was retained in the positive sample due to the large interparticle spaces and the stability against the ionic strength elevation which persisted for about 30 min. The whole process of DNA extraction, amplification, and detection took less than 2 h with a sensitivity of 20 CFU/ml. The developed gold nanoprobe-LAMP could serve as a simple, rapid, and cost-effective method for nanodiagnosis of S. typhi in point-of-need applications.

Detection of Salmonella spp. with the BACTEC 9240 Automated Blood Culture System in 2008 - 2014 in Southern Iran (Shiraz): Biogrouping, MIC, and Antimicrobial Susceptibility Profiles of Isolates.

PubMed

Anvarinejad, Mojtaba; Pouladfar, Gholam Reza; Pourabbas, Bahman; Amin Shahidi, Maneli; Rafaatpour, Noroddin; Dehyadegari, Mohammad Ali; Abbasi, Pejman; Mardaneh, Jalal

2016-04-01

Human salmonellosis continues to be a major international problem, in terms of both morbidity and economic losses. The antibiotic resistance of Salmonella is an increasing public health emergency, since infections from resistant bacteria are more difficult and costly to treat. The aims of the present study were to investigate the isolation of Salmonella spp. with the BACTEC automated system from blood samples during 2008 - 2014 in southern Iran (Shiraz). Detection of subspecies, biogrouping, and antimicrobial susceptibility testing by the disc diffusion and agar dilution methods were performed. A total of 19 Salmonella spp. were consecutively isolated using BACTEC from blood samples of patients between 2008 and 2014 in Shiraz, Iran. The isolates were identified as Salmonella, based on biochemical tests embedded in the API-20E system. In order to characterize the biogroups and subspecies, biochemical testing was performed. Susceptibility testing (disc diffusion and agar dilution) and extended-spectrum β-lactamase (ESBL) detection were performed according to the clinical and laboratory standards institute (CLSI) guidelines. Of the total 19 Salmonella spp. isolates recovered by the BACTEC automated system, all belonged to the Salmonella enterica subsp. houtenae. Five isolates (26.5%) were resistant to azithromycin. Six (31.5%) isolates with the disc diffusion method and five (26.3%) with the agar dilution method displayed resistance to nalidixic acid (minimum inhibitory concentration [MIC] > 32 μg/mL). All nalidixic acid-resistant isolates were also ciprofloxacin-sensitive. All isolates were ESBL-negative. Twenty-one percent of isolates were found to be resistant to chloramphenicol (MIC ≥ 32 μg/mL), and 16% were resistant to ampicillin (MIC ≥ 32 μg/mL). The results indicate that multidrug-resistant (MDR) strains of Salmonella are increasing in number, and fewer antibiotics may be useful for treating S. enterica infections. Routine investigation and reporting of antibiotic MICs in patients presenting with Salmonella infections is suggested.

Development of Real Time PCR Using Novel Genomic Target for Detection of Multiple Salmonella Serovars from Milk and Chickens

USDA-ARS?s Scientific Manuscript database

Background: A highly sensitive and specific novel genomic and plasmid target-based PCR platform was developed to detect multiple Salmonella serovars (S. Heidelberg, S. Dublin, S. Hadar, S. Kentucky and S. Enteritidis). Through extensive genome mining of protein databases of these serovars and compar...

Impact of Strain Variation on the Ability of Biosensor Technology to Detect Salmonella enterica

USDA-ARS?s Scientific Manuscript database

Introduction: It is important to develop methods that can quickly and accurately detect the presence of bacteria in the food supply that cause disease. Salmonella enterica is a bacteria that is often associated with contamination of food. Strains vary in their ability to cause illness and to spread...

Microfiltration of enzyme treated egg whites for accelerated detection of viable Salmonella

USDA-ARS?s Scientific Manuscript database

We report detection of <13 CFU of Salmonella per 25 g egg white within 7 h by concentrating the bacteria using microfiltration through 0.2-lm cutoff polyethersulfone hollow fiber membranes. A combination of enzyme treatment, controlled cross-flow on both sides of the hollow fibers, and media selecti...

Development of a novel loop-mediated isothermal amplification (LAMP) assay for the detection of Salmonella ser. Enteritidis from egg products

USDA-ARS?s Scientific Manuscript database

Salmonella ser. Enteritidis is a major public health concern worldwide. Loop-mediated isothermal amplification (LAMP) is a novel simple, easy-to-operate detection technology that amplifies DNA with high speed, efficiency, and specificity under isothermal conditions. The objective of this study was t...

Evaluation of VIDAS Salmonella (SLM) easy Salmonella method for the detection of Salmonella in a variety of foods: collaborative study.

PubMed

Crowley, Erin; Bird, Patrick; Fisher, Kiel; Goetz, Katherine; Benzinger, M Joseph; Agin, James; Goins, David; Johnson, Ronald L

2011-01-01

The VIDAS Salmonella (SLM) Easy Salmonella method is a specific enzyme-linked fluorescent immunoassay performed in the automated VIDAS instrument. The VIDAS Easy Salmonella method is a simple 2-step enrichment procedure, using pre-enrichment followed by selective enrichment in a newly formulated broth, SX2 broth. This new method was compared in a multilaboratory collaborative study to the U.S. Food and Drug Administration's Bacteriological Analytical Manual, Chapter 5 method for five food matrixes (liquid egg, vanilla ice cream, spinach, raw shrimp, and peanut butter) and the U.S. Department of Agriculture's Microbiology Laboratory Guidebook 4.04 method for deli turkey. Each food type was artificially contaminated with Salmonella at three inoculation levels. A total of 15 laboratories representing government, academia, and industry, throughout the United States, participated. In this study, 1583 samples were analyzed, of which 792 were paired replicates and 791 were unpaired replicates. Of the 792 paired replicates, 285 were positive by both the VIDAS and reference methods. Of the 791 unpaired replicates, 341 were positive by the VIDAS method and 325 were positive by the cultural reference method. A Chi-square analysis of each of the six food types was performed at the three inoculation levels tested. For all foods evaluated, the VIDAS Easy SLM method demonstrated results comparable to those of the reference methods for the detection of Salmonella.

Optical immunosensors for detection of Listeria monocytogenes and Salmonella enteritidis from food

NASA Astrophysics Data System (ADS)

Bhunia, Arun K.; Geng, Tao; Lathrop, Amanda; Valadez, Angela; Morgan, Mark T.

2004-03-01

Listeria monocytogenes and Salmonella are two major foodborne pathogens of significant concern. Two optical evanescent wave immunosensors were evaluated for detection: Antibody-coupled fiber-optic biosensor and a surface plasmon resonant (SPR) immunosensor. In the fiber-optic sensor, polyclonal antibodies for the test organisms were immobilized on polystyrene fiber wave -guides using streptavidin - biotin chemistry. Cyanine 5 -labeled monoclonal antibodies C11E9 (for L. monocytogenes) and SF-11 (for Salmonella Enteritidis) were used to generate a specific fluorescent signal. Signal acquisition was performed by launching a laser-light (635 nm) from an Analyte-2000. This immunosensor was able to detect 103 - 109 cfu/ml of L. monocytogenes or 106-109 cfu/ml of Salmonella Enteritidis and the assays were conducted at near real-time with results obtained within one hour of sampling. The assays were specific and showed signal even in the presence of other microorganisms such as E. coli, Enterococcus faecalis or Salmonella Typhimurium. In the SPR system, IAsys instrument (resonant mirror sensor) was used. Monoclonal antibody-C11E9 was directly immobilized onto a carboxylate cuvette. Whole Listeria cells at various concentrations did not yield any signal while surface protein extracts did. Crude protein extracts from L. monocytogenes and L. innocua had average binding responses of around 150 arc sec (0.25 ng/mm2), which was significantly different from L. grayi, L. ivanovii, or L. welshimeri with average responses of <48 arc sec. Both fiber-optic and SPR sensors show promise in near real-time detection of foodborne L. monocytogenes and Salmonella Enteritidis.

Assessment of the microbiological safety of edible roasted nut kernels on retail sale in England, with a focus on Salmonella.

PubMed

Little, C L; Jemmott, W; Surman-Lee, S; Hucklesby, L; de Pinnal, E

2009-04-01

There is little published information on the prevalence of Salmonella in edible nut kernels. A study in early 2008 of edible roasted nut kernels on retail sale in England was undertaken to assess the microbiological safety of this product. A total of 727 nut kernel samples of different varieties were examined. Overall, Salmonella and Escherichia coli were detected from 0.2 and 0.4% of edible roasted nut kernels. Of the nut varieties examined, Salmonella Havana was detected from 1 (4.0%) sample of pistachio nuts, indicating a risk to health. The United Kingdom Food Standards Agency was immediately informed, and full investigations were undertaken. Further examination established the contamination to be associated with the pistachio kernels and not the partly opened shells. Salmonella was not detected in other varieties tested (almonds, Brazils, cashews, hazelnuts, macadamia, peanuts, pecans, pine nuts, and walnuts). E. coli was found at low levels (range of 3.6 to 4/g) in walnuts (1.4%), almonds (1.2%), and Brazils (0.5%). The presence of Salmonella is unacceptable in edible nut kernels. Prevention of microbial contamination in these products lies in the application of good agricultural, manufacturing, and storage practices together with a hazard analysis and critical control points system that encompass all stages of production, processing, and distribution.

Detection and enumeration of Salmonella enteritidis in homemade ice cream associated with an outbreak: comparison of conventional and real-time PCR methods.

PubMed

Seo, K H; Valentin-Bon, I E; Brackett, R E

2006-03-01

Salmonellosis caused by Salmonella Enteritidis (SE) is a significant cause of foodborne illnesses in the United States. Consumption of undercooked eggs and egg-containing products has been the primary risk factor for the disease. The importance of the bacterial enumeration technique has been enormously stressed because of the quantitative risk analysis of SE in shell eggs. Traditional enumeration methods mainly depend on slow and tedious most-probable-number (MPN) methods. Therefore, specific, sensitive, and rapid methods for SE quantitation are needed to collect sufficient data for risk assessment and food safety policy development. We previously developed a real-time quantitative PCR assay for the direct detection and enumeration of SE and, in this study, applied it to naturally contaminated ice cream samples with and without enrichment. The detection limit of the real-time PCR assay was determined with artificially inoculated ice cream. When applied to the direct detection and quantification of SE in ice cream, the real-time PCR assay was as sensitive as the conventional plate count method in frequency of detection. However, populations of SE derived from real-time quantitative PCR were approximately 1 log higher than provided by MPN and CFU values obtained by conventional culture methods. The detection and enumeration of SE in naturally contaminated ice cream can be completed in 3 h by this real-time PCR method, whereas the cultural enrichment method requires 5 to 7 days. A commercial immunoassay for the specific detection of SE was also included in the study. The real-time PCR assay proved to be a valuable tool that may be useful to the food industry in monitoring its processes to improve product quality and safety.

Detection of Salmonella enterica in pigs at slaughter and comparison with human isolates in Italy.

PubMed

Bonardi, Silvia; Alpigiani, Irene; Bruini, Ilaria; Barilli, Elena; Brindani, Franco; Morganti, Marina; Cavallini, Pierugo; Bolzoni, Luca; Pongolini, Stefano

2016-02-02

In 2013-2014, 201 pigs belonging to 67 batches were tested for Salmonella in their mesenteric lymph nodes (MLN) in one abattoir of Northern Italy. For each batch, faecal material was collected at lairage by swabbing the pen floor for approximately 1600 cm(2). The aim of this study was to investigate the prevalence of Salmonella in MLN of pigs at slaughter, to assess Salmonella contamination at lairage and to evaluate the effect of lairage duration on its prevalence. Serotyping, XbaI PFGE typing and antimicrobial testing of the isolates were performed. Pig and human Salmonella isolates of the same region of Italy were compared to evaluate possible correlations. Salmonella enterica was isolated from 19.9% of the MLN and 49.3% of the environmental faecal samples. Nine different serovars were identified among 75 S. enterica isolates. In MLN Salmonella Derby was the most common (52.5%), followed by S. enterica 4,[5],12:i:- (17.5%) and Salmonella Rissen (10.0%). In faecal samples S. Derby was prevalent (51.4%), followed by S. enterica 4,[5], 12:i:- (20.0%) and Salmonella Brandenburg (14.3%). Lairage holding varied between 1 and ≥ 12 h (median value: 2.5h). In pigs held for 1-3h, 14.1% were positive for Salmonella in MLN but the prevalence reached 31.8% when they were held for ≥ 12 h. The contamination of MLN was statistically different (p=0.0045) between the two groups, thus confirming the role of long-lasting lairage in Salmonella contamination of pigs. XbaI PFGE typing detected 36 PFGE types. Twenty-three PFGE types were identified among the 40 MLN isolates and 22 PFGE types among the 35 faecal isolates. A total of 11 PFGE types were shared between the MLN of pigs and the lairage environment. Among S. Derby, 6 shared PFGE types between MLN and faeces were found and among S. enterica 4,[5],12:i:- one PFGE type was common between MLN and the faecal samples. Shared profiles between human and swine isolates of S. Derby, S. enterica 4,[5],12:i:-, S. Rissen, Salmonella Manhattan, S. Brandenburg, Salmonella Livingstone, Salmonella London and Salmonella Muenchen were identified. Among S. Derby and S. enterica 4,[5],12:i:- isolates found in pigs, 6/15 profiles (40.0%) and 8/10 (80.0%) were shared with human isolates. High resistance rates to streptomycin (97.3%), sulphonamide compounds (84.0%) and tetracycline (56.0%) were observed. No resistance was detected to ertapenem and meropenem. High proportions of isolates showed intermediate sensitivity to ciprofloxacin (85.3%) and cefotaxime (66.7%). High sensitivity rates were found to chloramphenicol (96.0%) and trimethoprim/sulfamethoxazole (81.3%). Copyright © 2015 Elsevier B.V. All rights reserved.

Modeling and analysis of a microresonating biosensor for detection of Salmonella bacteria in human blood.

PubMed

Bahadoran, Mahdi; Noorden, Ahmad Fakhrurrazi Ahmad; Chaudhary, Kashif; Mohajer, Faeze Sadat; Aziz, Muhammad Safwan; Hashim, Shahrin; Ali, Jalil; Yupapin, Preecha

2014-07-18

A new photonics biosensor configuration comprising a Double-side Ring Add-drop Filter microring resonator (DR-ADF) made from SiO2-TiO2 material is proposed for the detection of Salmonella bacteria (SB) in blood. The scattering matrix method using inductive calculation is used to determine the output signal's intensities in the blood with and without presence of Salmonella. The change in refractive index due to the reaction of Salmonella bacteria with its applied antibody on the flagellin layer loaded on the sensing and detecting microresonator causes the increase in through and dropper port's intensities of the output signal which leads to the detection of SB in blood. A shift in the output signal wavelength is observed with resolution of 0.01 nm. The change in intensity and shift in wavelength is analyzed with respect to the change in the refractive index which contributes toward achieving an ultra-high sensitivity of 95,500 nm/RIU which is almost two orders higher than that of reported from single ring sensors and the limit of detection is in the order of 1 × 10(-8) RIU. In applications, such a system can be employed for a high sensitive and fast detection of bacteria.

Modeling and Analysis of a Microresonating Biosensor for Detection of Salmonella Bacteria in Human Blood

PubMed Central

Bahadoran, Mahdi; Noorden, Ahmad Fakhrurrazi Ahmad; Chaudhary, Kashif; Mohajer, Faeze Sadat; Aziz, Muhammad Safwan; Hashim, Shahrin; Ali, Jalil; Yupapin, Preecha

2014-01-01

A new photonics biosensor configuration comprising a Double-side Ring Add-drop Filter microring resonator (DR-ADF) made from SiO2-TiO2 material is proposed for the detection of Salmonella bacteria (SB) in blood. The scattering matrix method using inductive calculation is used to determine the output signal's intensities in the blood with and without presence of Salmonella. The change in refractive index due to the reaction of Salmonella bacteria with its applied antibody on the flagellin layer loaded on the sensing and detecting microresonator causes the increase in through and dropper port's intensities of the output signal which leads to the detection of SB in blood. A shift in the output signal wavelength is observed with resolution of 0.01 nm. The change in intensity and shift in wavelength is analyzed with respect to the change in the refractive index which contributes toward achieving an ultra-high sensitivity of 95,500 nm/RIU which is almost two orders higher than that of reported from single ring sensors and the limit of detection is in the order of 1 × 10−8 RIU. In applications, such a system can be employed for a high sensitive and fast detection of bacteria. PMID:25046015

Immunomagnetic separation and conventional culture procedure for detection of naturally occurring Salmonella in raw pork sausages and chicken meat.

PubMed

Ripabelli, G; Sammarco, M L; Ruberto, A; Iannitto, G; Grasso, G M

1997-06-01

The aim of the study was to compare immunomagnetic separation (IMS) and conventional selective enrichment procedures using selenite cystine broth (SC) and Rappaport-Vassiliadis broth (RV) in 137 naturally contaminated food samples (69 raw pork sausages and 68 chicken meat). The utilization of SC or IMS appeared to be the most appropriate enrichment procedure: 15 out of 18 Salmonella-positive samples (83.3%) were detected by SC and 12 (66.7%) by IMS; RV yielded only seven positive isolations (38.9%). However, RV yielded the highest count of Salmonella colonies per plate and the lowest interference by competing organisms. IMS could become a reliable alternative to standard enrichment procedures and a combined IMS and selective enrichment broth could increase the chance of Salmonella recovery.

Evaluation of a newly developed triple buffered peptone broth for detection of Salmonella in broiler feed

USDA-ARS?s Scientific Manuscript database

Lactose broth (LB) and buffered peptone (BP) are used as pre-enrichment media to recover Salmonella from feed. Bacterial utilization of feed carbohydrates results in the production of acidic byproducts causing a drop in the media pH which can injure or kill Salmonella and yield false negative resul...

Visual and efficient immunosensor technique for advancing biomedical applications of quantum dots on Salmonella detection and isolation

NASA Astrophysics Data System (ADS)

Tang, Feng; Pang, Dai-Wen; Chen, Zhi; Shao, Jian-Bo; Xiong, Ling-Hong; Xiang, Yan-Ping; Xiong, Yan; Wu, Kai; Ai, Hong-Wu; Zhang, Hui; Zheng, Xiao-Li; Lv, Jing-Rui; Liu, Wei-Yong; Hu, Hong-Bing; Mei, Hong; Zhang, Zhen; Sun, Hong; Xiang, Yun; Sun, Zi-Yong

2016-02-01

It is a great challenge in nanotechnology for fluorescent nanobioprobes to be applied to visually detect and directly isolate pathogens in situ. A novel and visual immunosensor technique for efficient detection and isolation of Salmonella was established here by applying fluorescent nanobioprobes on a specially-designed cellulose-based swab (a solid-phase enrichment system). The selective and chromogenic medium used on this swab can achieve the ultrasensitive amplification of target bacteria and form chromogenic colonies in situ based on a simple biochemical reaction. More importantly, because this swab can serve as an attachment site for the targeted pathogens to immobilize and immunologically capture nanobioprobes, our mAb-conjugated QD bioprobes were successfully applied on the solid-phase enrichment system to capture the fluorescence of targeted colonies under a designed excitation light instrument based on blue light-emitting diodes combined with stereomicroscopy or laser scanning confocal microscopy. Compared with the traditional methods using 4-7 days to isolate Salmonella from the bacterial mixture, this method took only 2 days to do this, and the process of initial screening and preliminary diagnosis can be completed in only one and a half days. Furthermore, the limit of detection can reach as low as 101 cells per mL Salmonella on the background of 105 cells per mL non-Salmonella (Escherichia coli, Proteus mirabilis or Citrobacter freundii, respectively) in experimental samples, and even in human anal ones. The visual and efficient immunosensor technique may be proved to be a favorable alternative for screening and isolating Salmonella in a large number of samples related to public health surveillance.It is a great challenge in nanotechnology for fluorescent nanobioprobes to be applied to visually detect and directly isolate pathogens in situ. A novel and visual immunosensor technique for efficient detection and isolation of Salmonella was established here by applying fluorescent nanobioprobes on a specially-designed cellulose-based swab (a solid-phase enrichment system). The selective and chromogenic medium used on this swab can achieve the ultrasensitive amplification of target bacteria and form chromogenic colonies in situ based on a simple biochemical reaction. More importantly, because this swab can serve as an attachment site for the targeted pathogens to immobilize and immunologically capture nanobioprobes, our mAb-conjugated QD bioprobes were successfully applied on the solid-phase enrichment system to capture the fluorescence of targeted colonies under a designed excitation light instrument based on blue light-emitting diodes combined with stereomicroscopy or laser scanning confocal microscopy. Compared with the traditional methods using 4-7 days to isolate Salmonella from the bacterial mixture, this method took only 2 days to do this, and the process of initial screening and preliminary diagnosis can be completed in only one and a half days. Furthermore, the limit of detection can reach as low as 101 cells per mL Salmonella on the background of 105 cells per mL non-Salmonella (Escherichia coli, Proteus mirabilis or Citrobacter freundii, respectively) in experimental samples, and even in human anal ones. The visual and efficient immunosensor technique may be proved to be a favorable alternative for screening and isolating Salmonella in a large number of samples related to public health surveillance. Electronic supplementary information (ESI) available: One additional figure (Fig. S1), two additional tables (Tables S1 and S2) and additional information. See DOI: 10.1039/c5nr07424j

Detection of Salmonella enterica Serovar Montevideo and Newport in Free-ranging Sea Turtles and Beach Sand in the Caribbean and Persistence in Sand and Seawater Microcosms.

PubMed

Ives, A-K; Antaki, E; Stewart, K; Francis, S; Jay-Russell, M T; Sithole, F; Kearney, M T; Griffin, M J; Soto, E

2017-09-01

Salmonellae are Gram-negative zoonotic bacteria that are frequently part of the normal reptilian gastrointestinal flora. The main objective of this project was to estimate the prevalence of non-typhoidal Salmonella enterica in the nesting and foraging populations of sea turtles on St. Kitts and in sand from known nesting beaches. Results suggest a higher prevalence of Salmonella in nesting leatherback sea turtles compared with foraging green and hawksbill sea turtles. Salmonella was cultured from 2/9 and identified by molecular diagnostic methods in 3/9 leatherback sea turtle samples. Salmonella DNA was detected in one hawksbill turtle, but viable isolates were not recovered from any hawksbill sea turtles. No Salmonella was detected in green sea turtles. In samples collected from nesting beaches, Salmonella was only recovered from a single dry sand sample. All recovered isolates were positive for the wzx gene, consistent with the O:7 serogroup. Further serotyping characterized serovars Montevideo and Newport present in cloacal and sand samples. Repetitive-element palindromic PCR (rep-PCR) fingerprint analysis and pulsed-field gel electrophoresis of the 2014 isolates from turtles and sand as well as archived Salmonella isolates recovered from leatherback sea turtles in 2012 and 2013, identified two distinct genotypes and four different pulsotypes, respectively. The genotyping and serotyping were directly correlated. To determine the persistence of representative strains of each serotype/genotype in these environments, laboratory-controlled microcosm studies were performed in water and sand (dry and wet) incubated at 25 or 35°C. Isolates persisted for at least 32 days in most microcosms, although there were significant decreases in culturable bacteria in several microcosms, with the greatest reduction in dry sand incubated at 35°C. This information provides a better understanding of the epizootiology of Salmonella in free-ranging marine reptiles and the potential public health risks associated with human interactions with these animals in the Caribbean. © 2016 Blackwell Verlag GmbH.

Occurrence, genetic characterization and antimicrobial resistance of Salmonella isolated from chicken meat and giblets.

PubMed

Abd-Elghany, S M; Sallam, K I; Abd-Elkhalek, A; Tamura, T

2015-04-01

SUMMARY This study was undertaken to survey the presence of Salmonella in 200 chicken samples collected from Mansoura, Egypt. Salmonella was detected in 16% (8/50), 28% (14/50), 32% (16/50) and 60% (30/50) of whole chicken carcasses, drumsticks, livers and gizzards, respectively, with an overall prevalence of 34% (68/200) among all samples. One hundred and sixty-six isolates were identified biochemically as Salmonella, and confirmed genetically by PCR, based on the presence of invA and stn genes. The spvC gene, however, was detected in only 25.3% (42/166) of the isolates. Isolates were serotyped as Salmonella Enteritidis (37.3%), S. Typhimurium (30.1%), S. Kentucky (10.8%), S. Muenster (8.4%), S. Virchow (4.8%), S. Anatum (4.8%), S. Haifa (1.2%), and four were non-typable. Antimicrobial susceptibility tests of the Salmonella isolates revealed that 100% were resistant to each of erythromycin, penicillin, and amoxicillin, while 98.8%, 96.4%, 95.2%, and 91.6% were resistant to nalidixic acid, sulphamethoxazole, oxytetracycline, and ampicillin, respectively. Multidrug resistance was evident for 92.8% of the isolates. The high contamination level of chicken meat with multidrug-resistant Salmonella can constitute a problem for public health.

A Novel Chromogenic Ester Agar Medium for Detection of Salmonellae

PubMed Central

Cooke, Venitia M.; Miles, R. J.; Price, R. G.; Richardson, A. C.

1999-01-01

A novel agar medium, chromogenic Salmonella esterase (CSE) agar, for the differentiation of salmonellae is described. The agar contains peptones and nutrient extracts together with the following (grams per liter unless otherwise specified): 4-[2-(4-octanoyloxy-3,5-dimethoxyphenyl)-vinyl]-quinolinium-1-(propan-3-yl carboxylic acid) bromide (SLPA-octanoate; bromide form), 0.3223; lactose, 14.65; trisodium citrate dihydrate, 0.5; Tween 20, 3.0; ethyl 4-dimethylaminobenzoate, 0.035% (wt/vol), novobiocin, 70 mg liter−1. The key component of the medium is SLPA-octanoate, a newly synthesized ester formed from a C8 fatty acid and a phenolic chromophore. In CSE agar, the ester is hydrolyzed by Salmonella spp. to yield a brightly colored phenol which remains tightly bound within colonies. After 24 h of incubation at 37 or 42°C, colonies of typical Salmonella spp. were burgundy colored on a transparent yellow background, whereas non-Salmonella spp. were white, cream, yellow or transparent. CSE agar was evaluated by using a panel of strains including a high proportion of Salmonella and non-Salmonella strains giving atypical reactions on other differential agars. The sensitivity (93.1%) of CSE agar for non-typhi salmonellae compared favorably with those of Rambach (82.8%), xylose-lysine-deoxycholate (XLD; 91.4%), Hektoen-enteric (89.7%), and SM ID (91.4%) agars. The specificity (93.9%) was also comparable to those of other Salmonella media (SM ID agar, 95.9%; Rambach agar, 91.8%; XLD agar, 91.8%; Hektoen-enteric agar, 87.8%). Strains of Citrobacter freundii and Proteus spp. giving false-positive reactions with other media gave a negative color reaction on CSE agar. CSE agar enabled the detection of >30 Salmonella serotypes, including agona, anatum, enteritidis, hadar, heidelberg, infantis, montevideo, thompson, typhimurium, and virchow, which accounted for 91.8% of the salmonella isolates recorded by the Public Health Laboratory Service (Colindale, London, England) for 1997. PMID:9925620

Surveillance for human Salmonella infections in the United States.

PubMed

Swaminathan, Bala; Barrett, Timothy J; Fields, Patricia

2006-01-01

Surveillance for human Salmonella infections plays a critical role in understanding and controlling foodborne illness due to Salmonella. Along with its public health partners, the Centers for Disease Control and Prevention (CDC) has several surveillance systems that collect information on Salmonella infections in the United States. The National Salmonella Surveillance System, begun in 1962, receives reports of laboratory-confirmed Salmonella infections through state public health laboratories. Salmonella outbreaks are reported by state and local health departments through the Foodborne Disease Outbreak Reporting System, which became a Web-based, electronic system (eFORS) in 2001. PulseNet facilitates the detection of clusters of Salmonella infections through standardized molecular subtyping (DNA "fingerprinting") of isolates and maintenance of "fingerprint" databases. The National Antimicrobial Resistance Monitoring System for Enteric Bacteria (NARMS) monitors antimicrobial resistance in Salmonella by susceptibility testing of every 20th Salmonella isolate received by state and local public health laboratories. FootNet is an active surveillance system that monitors Salmonella infections in sentinel areas, providing population-based estimates of infection rates. Efforts are underway to electronically link all of the Salmonella surveillance systems at CDC to facilitate optimum use of available data and minimize duplication.

Paradigm Diagnostics Salmonella Indicator Broth (PDX-SIB) for detection of Salmonella on selected environmental surfaces.

PubMed

Olstein, Alan; Griffith, Leena; Feirtag, Joellen; Pearson, Nicole

2013-01-01

The Paradigm Diagnostics Salmonella Indicator Broth (PDX-SIB) is intended as a single-step selective enrichment indicator broth to be used as a simple screening test for the presence of Salmonella spp. in environmental samples. This method permits the end user to avoid multistep sample processing to identify presumptively positive samples, as exemplified by standard U.S. reference methods. PDX-SIB permits the outgrowth of Salmonella while inhibiting the growth of competitive Gram-negative and -positive microflora. Growth of Salmonella-positive cultures results in a visual color change of the medium from purple to yellow when the sample is grown at 37 +/- 1 degree C. Performance of PDX-SIB has been evaluated in five different categories: inclusivity-exclusivity, methods comparison, ruggedness, lot-to-lot variability, and shelf stability. The inclusivity panel included 100 different Salmonella serovars, 98 of which were SIB-positive during the 30 to 48 h incubation period. The exclusivity panel included 33 different non-Salmonella microorganisms, 31 of which were SIB-negative during the incubation period. Methods comparison studies included four different surfaces: S. Newport on plastic, S. Anatum on sealed concrete, S. Abaetetuba on ceramic tile, and S. Typhimurium in the presence of 1 log excess of Citrobacter freundii. Results of the methods comparison studies demonstrated no statistical difference between the SIB method and the U.S. Food and Drug Administration-Bacteriological Analytical Manual reference method, as measured by the Mantel-Haenszel Chi-square test. Ruggedness studies demonstrated little variation in test results when SIB incubation temperatures were varied over a 34-40 degrees C range. Lot-to-lot consistency results suggest no detectable differences in manufactured goods using two reference Salmonella serovars and one non-Salmonella microorganism.

Relationship between Salmonella infection, shedding and serology in fattening pigs in low-moderate prevalence areas.

PubMed

San Román, B; Garrido, V; Sánchez, S; Martínez-Ballesteros, I; Garaizar, J; Mainar-Jaime, R C; Migura-Garcia, L; Grilló, M J

2018-04-27

Salmonella is a major foodborne pathogen causing important zoonosis worldwide. Pigs asymptomatically infected in mesenteric lymph nodes (MLN) can be intermittent shedders of the pathogen through faeces, being considered a major source of human infections. European baseline studies of fattening pig salmonellosis are based on Salmonella detection in MLN. This work studies the relationship between Salmonella infection in MLN and intestinal content (IC) shedding at slaughter and the relationship between the presence of the pathogen and the serologic status at farm level. Mean Salmonella prevalence in the selected pigs (vertically integrated production system of Navarra, Spain) was 7.2% in MLN, 8.4% in IC and 9.6% in serum samples. In this low-moderate prevalence context, poor concordance was found between MLN infection and shedding at slaughter and between bacteriology and serology. In fact, most of shedders were found uninfected in MLN (83%) or carrying different Salmonella strains in MLN and in IC (90%). The most prevalent Salmonellae were Typhimurium resistant to ACSSuT ± Nx or ASSuT antibiotic families, more frequently found invading the MLN (70%) than in IC (33.9%). Multivariable analysis revealed that risk factors associated with the presence of Salmonella in MLN or in IC were different, mainly related either to good hygiene practices or to water and feed control, respectively. Overall, in this prevalence context, detection of Salmonella in MLN is an unreliable predictor of faecal shedding at abattoir, indicating that subclinical infections in fattening pigs MLN could have limited relevance in the IC shedding. © 2018 Blackwell Verlag GmbH.

Study of the cross-contamination and survival of Salmonella in fresh apples.

PubMed

Perez-Rodriguez, F; Begum, M; Johannessen, G S

2014-08-01

The present work aimed at studying the cross contamination of apples by Salmonella during the processing of commercial fresh apples and its survival capacity on apple at room temperature. For the first study, the typical process of fresh apples was simulated at laboratory scale in which an apple that was artificially contaminated by Salmonella at different concentration levels (8, 6 and 5 log cfu/apple) was introduced in one batch and processed including a simulated transport/washing step and drying step using sponges to simulate the porous material used in the industry. Results indicated that at 8 log cfu/apple, 50% fresh apples were contaminated after processing, with all analysed environmental samples being positive for the pathogen, consisting of washing water and sponges. However, at lower inoculum levels (5-6 log cfu/apple) no cross contamination was detected in apples, and only environmental samples showed contamination by Salmonella after processing including both water and sponges. Experiments on the survival of Salmonella on apple showed that the pathogen was capable to survive for 12 days, only showing a significant drop at the end of the experiment. Finally, two-class attribute sampling plans were assessed as tool to detect Salmonella in different contamination scenarios in fresh apple. This analysis indicated that with the highest inoculum level, a total of 16 apples would be needed to reach 95% of detecting Salmonella (i.e. lot rejection). In turn, when low levels were assessed (5-6 log cfu/apple), a large number of apples (n=1021) would have to be sampled to obtain the same confidence level (95%). If the environment is sampled (i.e. water and sponges), a lower number of samples would be needed to detect the pathogen. However, the feasibility of environmental sampling has not been assessed from a practical point of view. Overall, the results in this study evidenced that cross contamination by Salmonella might occur during processing of fresh apples and subsequently, the pathogen might survive for a noticeable period of time. Copyright © 2014 Elsevier B.V. All rights reserved.

Evaluation of the Thermo Scientific SureTect Salmonella species assay. AOAC Performance Tested Method 051303.

PubMed

Cloke, Jonathan; Clark, Dorn; Radcliff, Roy; Leon-Velarde, Carlos; Larson, Nathan; Dave, Keron; Evans, Katharine; Crabtree, David; Hughes, Annette; Simpson, Helen; Holopainen, Jani; Wickstrand, Nina; Kauppinen, Mikko

2014-01-01

The Thermo Scientific SureTect Salmonella species Assay is a new real-time PCR assay for the detection of Salmonellae in food and environmental samples. This validation study was conducted using the AOAC Research Institute (RI) Performance Tested Methods program to validate the SureTect Salmonella species Assay in comparison to the reference method detailed in International Organization for Standardization 6579:2002 in a variety of food matrixes, namely, raw ground beef, raw chicken breast, raw ground pork, fresh bagged lettuce, pork frankfurters, nonfat dried milk powder, cooked peeled shrimp, pasteurized liquid whole egg, ready-to-eat meal containing beef, and stainless steel surface samples. With the exception of liquid whole egg and fresh bagged lettuce, which were tested in-house, all matrixes were tested by Marshfield Food Safety, Marshfield, WI, on behalf of Thermo Fisher Scientific. In addition, three matrixes (pork frankfurters, lettuce, and stainless steel surface samples) were analyzed independently as part of the AOAC-RI-controlled laboratory study by the University of Guelph, Canada. No significant difference by probability of detection or McNemars Chi-squared statistical analysis was found between the candidate or reference methods for any of the food matrixes or environmental surface samples tested during the validation study. Inclusivity and exclusivity testing was conducted with 117 and 36 isolates, respectively, which demonstrated that the SureTect Salmonella species Assay was able to detect all the major groups of Salmonella enterica subspecies enterica (e.g., Typhimurium) and the less common subspecies of S. enterica (e.g., arizoniae) and the rarely encountered S. bongori. None of the exclusivity isolates analyzed were detected by the SureTect Salmonella species Assay. Ruggedness testing was conducted to evaluate the performance of the assay with specific method deviations outside of the recommended parameters open to variation (enrichment time and temperature, and lysis temperature), which demonstrated that the assay gave reliable performance. Accelerated stability testing was additionally conducted, validating the assay shelf life.

Evaluation of the Thermo Scientific™ SureTect™ Salmonella species Assay.

PubMed

Cloke, Jonathan; Clark, Dorn; Radcliff, Roy; Leon-Velarde, Carlos; Larson, Nathan; Dave, Keron; Evans, Katharine; Crabtree, David; Hughes, Annette; Simpson, Helen; Holopainen, Jani; Wickstrand, Nina; Kauppinen, Mikko

2014-03-01

The Thermo Scientific™ SureTect™ Salmonella species Assay is a new real-time PCR assay for the detection of Salmonellae in food and environmental samples. This validation study was conducted using the AOAC Research Institute (RI) Performance Tested MethodsSM program to validate the SureTect Salmonella species Assay in comparison to the reference method detailed in International Organization for Standardization 6579:2002 in a variety of food matrixes, namely, raw ground beef, raw chicken breast, raw ground pork, fresh bagged lettuce, pork frankfurters, nonfat dried milk powder, cooked peeled shrimp, pasteurized liquid whole egg, ready-to-eat meal containing beef, and stainless steel surface samples. With the exception of liquid whole egg and fresh bagged lettuce, which were tested in-house, all matrixes were tested by Marshfield Food Safety, Marshfield, WI, on behalf of Thermo Fisher Scientific. In addition, three matrixes (pork frankfurters, lettuce, and stainless steel surface samples) were analyzed independently as part of the AOAC-RI-controlled laboratory study by the University of Guelph, Canada. No significant difference by probability of detection or McNemars Chi-squared statistical analysis was found between the candidate or reference methods for any of the food matrixes or environmental surface samples tested during the validation study. Inclusivity and exclusivity testing was conducted with 117 and 36 isolates, respectively, which demonstrated that the SureTect Salmonella species Assay was able to detect all the major groups of Salmonella enterica subspecies enterica (e.g., Typhimurium) and the less common subspecies of S. enterica (e.g., arizoniae) and the rarely encountered S. bongori. None of the exclusivity isolates analyzed were detected by the SureTect Salmonella species Assay. Ruggedness testing was conducted to evaluate the performance of the assay with specific method deviations outside of the recommended parameters open to variation (enrichment time and temperature, and lysis temperature), which demonstrated that the assay gave reliable performance. Accelerated stability testing was additionally conducted, validating the assay shelf life.

Isolation of Salmonella enterica serovar Enteritidis from houseflies (Musca domestica) found in rooms containing Salmonella serovar Enteritidis-challenged hens.

PubMed

Holt, Peter S; Geden, Christopher J; Moore, Randle W; Gast, Richard K

2007-10-01

Houseflies (Musca domestica) released into rooms containing hens challenged with Salmonella enterica serovar Enteritidis (Salmonella serovar Enteritidis) rapidly became contaminated with Salmonella serovar Enteritidis. Forty to 50% of the flies were contaminated at 48 h, and the percentage increased to 50 to 70% at 4 and 7 days postexposure and then decreased to 30% at day 15. Initial attempts at recovering surface organisms for culture using an aqueous rinse were largely unsuccessful, while cultures of internal contents readily recovered Salmonella serovar Enteritidis. However, when 0.5% detergent was incorporated into the rinse, high recovery levels of bacteria were observed from both external and internal culture regimens, indicating equal distribution of the organism on and in the fly and a tighter interaction of the organism with the host than previously thought. Salmonella serovar Enteritidis was isolated routinely from the fly gut, on rare occasions from the crop, and never from the salivary gland. Feeding contaminated flies to hens resulted in gut colonization of a third of the birds, but release of contaminated flies in a room containing previously unchallenged hens failed to result in colonization of any of the subject birds. These results indicate that flies exposed to an environment containing Salmonella serovar Enteritidis can become colonized with the organism and might serve as a source for transmission of Salmonella serovar Enteritidis within a flock situation.

Isolation of Salmonella enterica Serovar Enteritidis from Houseflies (Musca domestica) Found in Rooms Containing Salmonella Serovar Enteritidis-Challenged Hens▿

PubMed Central

Holt, Peter S.; Geden, Christopher J.; Moore, Randle W.; Gast, Richard K.

2007-01-01

Houseflies (Musca domestica) released into rooms containing hens challenged with Salmonella enterica serovar Enteritidis (Salmonella serovar Enteritidis) rapidly became contaminated with Salmonella serovar Enteritidis. Forty to 50% of the flies were contaminated at 48 h, and the percentage increased to 50 to 70% at 4 and 7 days postexposure and then decreased to 30% at day 15. Initial attempts at recovering surface organisms for culture using an aqueous rinse were largely unsuccessful, while cultures of internal contents readily recovered Salmonella serovar Enteritidis. However, when 0.5% detergent was incorporated into the rinse, high recovery levels of bacteria were observed from both external and internal culture regimens, indicating equal distribution of the organism on and in the fly and a tighter interaction of the organism with the host than previously thought. Salmonella serovar Enteritidis was isolated routinely from the fly gut, on rare occasions from the crop, and never from the salivary gland. Feeding contaminated flies to hens resulted in gut colonization of a third of the birds, but release of contaminated flies in a room containing previously unchallenged hens failed to result in colonization of any of the subject birds. These results indicate that flies exposed to an environment containing Salmonella serovar Enteritidis can become colonized with the organism and might serve as a source for transmission of Salmonella serovar Enteritidis within a flock situation. PMID:17675422

Occurrence of Campylobacter and Salmonella in ducks and duck eggs in Selangor, Malaysia.

PubMed

Nor Faiza, S; Saleha, A A; Jalila, A; Fauziah, N

2013-03-01

The importance of Campylobacter and Salmonella as foodborne pathogens is well recognised globally. A recent work in Penang found ducks in commercial farms were infected with these organisms. The aim of the study was to detect the presence of Campylobacter and Salmonella in ducks and Salmonella in duck eggs in farms in a small part of Selangor. Cloacal swabs were obtained from 75 ducks and 30 duck eggs from three farms. The isolation and identification of Campylobacter and Salmonella were done using conventional methods. Twelve percent of Campylobacter and 16.0% of Salmonella were isolated from the ducks sampled. Salmonella was absent on and in eggs. Campylobacter isolates consisted of 22% Campylobacter jejuni and the remaining was Campylobacter coli. Three Salmonella serovars identified were Salmonella Agona, S. Braenderup and S. Corvallis. The presence of Campylobacter and Salmonella in ducks may cause contamination of the meat during processing and handling which can constitute public health hazard. Moreover, the farm workers may be exposed to the organisms through contact with the infected animals.

Rapid colorimetric sensing platform for the detection of Listeria monocytogenes foodborne pathogen.

PubMed

Alhogail, Sahar; Suaifan, Ghadeer A R Y; Zourob, Mohammed

2016-12-15

Listeria monocytogenes is a serious cause of human foodborne infections worldwide, which needs spending billions of dollars for inspection of bacterial contamination in food every year. Therefore, there is an urgent need for rapid, in-field and cost effective detection techniques. In this study, rapid, low-cost and simple colorimetric assay was developed using magnetic nanoparticles for the detection of listeria bacteria. The protease from the listeria bacteria was detected using D-amino acid substrate. D-amino acid substrate was linked to the carboxylic acid on the magnetic nanoparticles using EDC/NHS chemistry. The cysteine residue at the C-terminal of the substrate was used for the self-assembled monolayer formation on the gold sensor surface, which in turn the black magnetic nanobeads will mask the golden color. The color will change from black to golden color upon the cleavage of the specific peptide sequence by the Listeria protease. The sensor was tested with serial dilutions of Listeria bacteria. It was found that the appearance of the gold surface area is proportional to the bacterial concentrations in CFU/ml. The lowest detection limit of the developed sensor for Listeria was found to be 2.17×10(2) colony forming unit/ml (CFU/ml). The specificity of the biosensor was tested against four different foodborne associated bacteria (Escherichia coli, Salmonella, Shigella flexnerii and Staphylococcus aureus). Finally, the sensor was tested with artificially spiked whole milk and ground meat spiked with listeria. Copyright © 2016 Elsevier B.V. All rights reserved.

Effect of Low Dose of Fumonisins on Pig Health: Immune Status, Intestinal Microbiota and Sensitivity to Salmonella

PubMed Central

Burel, Christine; Tanguy, Mael; Guerre, Philippe; Boilletot, Eric; Cariolet, Roland; Queguiner, Marilyne; Postollec, Gilbert; Pinton, Philippe; Salvat, Gilles; Oswald, Isabelle P.; Fravalo, Philippe

2013-01-01

The objective of this study was to measure the effects of chronic exposure to fumonisins via the ingestion of feed containing naturally contaminated corn in growing pigs infected or not with Salmonella spp. This exposure to a moderate dietary concentration of fumonisins (11.8 ppm) was sufficient to induce a biological effect in pigs (Sa/So ratio), but no mortality or pathology was observed over 63 days of exposure. No mortality or related clinical signs, even in cases of inoculation with Salmonella (5 × 104 CFU), were observed either. Fumonisins, at these concentrations, did not affect the ability of lymphocytes to proliferate in the presence of mitogens, but after seven days post-inoculation they led to inhibition of the ability of specific Salmonella lymphocytes to proliferate following exposure to a specific Salmonella antigen. However, the ingestion of fumonisins had no impact on Salmonella translocation or seroconversion in inoculated pigs. The inoculation of Salmonella did not affect faecal microbiota profiles, but exposure to moderate concentrations of fumonisins transiently affected the digestive microbiota balance. In cases of co-infection with fumonisins and Salmonella, the microbiota profiles were rapidly and clearly modified as early as 48 h post-Salmonella inoculation. Therefore under these experimental conditions, exposure to an average concentration of fumonisins in naturally contaminated feed had no effect on pig health but did affect the digestive microbiota balance, with Salmonella exposure amplifying this phenomenon. PMID:23612754

Postmortem photonic imaging of lux-modified Salmonella typhimurium within the gastrointestinal tract of swine following oral inoculation in vivo

USDA-ARS?s Scientific Manuscript database

The study objective was to monitor Salmonella progression by photonic detection through segments of the gastrointestinal tract following oral inoculation. Pigs (~ 80 kg) were inoculated orally with 3.1 or 4.1×10*10 colony forming units (cfu) of Salmonella typhimurium transformed with plasmid pAK1-lu...

Biomolecular Architectures Molecular Biology

DTIC Science & Technology

2013-08-31

when the Salmonella beacon (13 nM) was tested in the presence of 800 ng bacterial genomic DNA in chicken broth (33%) (data not shown). Since it was...bacterium, Bacillus thuringiensis, transgenic tobacco containing the transgene, Bt cry1Ac, the Gram-negative bacterium, Salmonella Typhimurium, and the Gram... Salmonella Typhimurium, and the Gram-positive bacterium, Listeria monocytogenes, were monitored for detection by coupling molecular beacon (MB

[Detection of Salmonella and Mycobacterium species in seagulls captured in Talcahuano, Chile].

PubMed

López-Martín, Juana; Junod, Tania; Riquelme, Fredy; Contreras, Cecilia; González-Acuña, Daniel

2011-11-01

Salmonella can be isolated from the feces of seagulls. Therefore these birds can be a vector for dissemination of this pathogen. To evaluate the possible role of gulls as vectors of two important human and animal pathogens (My-cobacteria and Salmonella). One hundred twenty three Kelp gull (Larus dominicanus) and 60 Franklin gulls (Leucophaeus pipixcan) captured off the coast of the seaport of Talcahuano, were analyzed. Using traditional microbiological methods, the presence of Mycobacteria in cloacal swabs and feet lavages, was analyzed in both types of gulls. To detect the presence of Salmonella, feces, fecal and tracheal swabs, and feet lavage were analyzed from Franklin gulls. Feces, feet lavage, intestine, spleen, liver, kidney and lung, were examined in Kelp gulls. All Mycobacteria cultures were negative. Salmonella enterica cultures were positive in 25 % of Kelp gulls and 6.7 % of Franklin gulls. Four serovars were identified by serotyping. Enteritidis and Senfteberg serovars were found in both types of gulls. Anatum and Infantis serovars were found only in Kelp gulls. Feces of gulls captured during the winter had the highest yield of positive cultures (36.1%). Seagulls are an important Salmonella vector in Chile.

Simultaneous detection of Staphylococcus aureus and Salmonella typhimurium using multicolor time-resolved fluorescence nanoparticles as labels.

PubMed

Wang, Xiaole; Huang, Yukun; Wu, Shijia; Duan, Nuo; Xu, Baocai; Wang, Zhouping

2016-11-21

Foodborne illnesses caused by Staphylococcus aureus and Salmonella typhimurium are common public health issues worldwide, affecting both developing and developed countries. In this study, aptamers labeled with multicolor lanthanide-doped time-resolved fluorescence (TRFL) nanoparticles were used as signal probes, and immobilized by Fe 3 O 4 magnetic nanoparticles were used as the capture probes. The signal probes were bonded onto the captured bacteria by the recognition of aptamer to form the sandwich-type complex. Under the optimal conditions, TRFL intensity at 544nm was used to quantify S. typhimurium (y=10,213×-12,208.92, R 2 =0.9922) and TRFL intensity at 615nm for S. aureus (y=4803.20×-1933.87, R 2 =0.9982) in the range of 10 2 -10 5 CFU/ml. Due to the magnetic separation and concentration of Fe 3 O 4 nanoparticles, detection limits of the developed method were found to be 15, 20CFU/ml for S. typhimurium and S. aureus, respectively. The application of this bioassay in milk was also investigated, and results were consistent with those of plate-counting method. Therefore, this simple and rapid method owns a great potential in the application for the multiplex analysis in food safety. Copyright © 2016 Elsevier B.V. All rights reserved.

Prevalence and characterization of Salmonella isolated from chicken meat in Turkey.

PubMed

Siriken, Belgin; Türk, Haldun; Yildirim, Tuba; Durupinar, Belma; Erol, Irfan

2015-05-01

This study was conducted in a Turkish province to investigate the presence of Salmonella spp. in 150 chicken meat samples using 2 phenotyping techniques: classic culture technique (CCT) and immunomagnetic separation (IMS). For the confirmation of the isolates at molecular levels, invA gene was detected in these isolates. The presence of invA, class 1 (Cls1) integrons, and integrase (Int1) genes was demonstrated by PCR assay; and the resistance of the isolated Salmonella spp. strains to antibiotics was determined by disk diffusion test. All the cultural and PCR results were evaluated together; Salmonella spp. were detected in a total of 64 (42.66%) chicken meat samples. Contamination rate was higher in carcasses (53.33%, n = 75) than in meat pieces (32%, n = 75). When results of standard culture were compared with IMS technique, IMS (n = 54) showed a clear superiority over the CCT (n = 38). A very high resistance rate (≥ 89.28%) to vancomycin, tetracycline, streptomycin, or nalidixic acid was found. Trimethoprim-sulfamethoxazole resistance was present in 32.14%. Relatively lower incidence of resistance (≤ 8.33%) to gentamicin, chloramphenicol, ampicillin, and ceftriaxone was observed. Concurrent resistance to at least 4 antibiotics was detected in 92.85% of the isolates. Cls1 integrons and Int1 were positive in 80.95% and 95.23% of the isolates, respectively. However, Int1 alone was detected in 15.47% (n = 13). In conclusion, the high prevalence of Salmonella spp. in chicken meat may pose a potential public health risk, and the presence of antibiotic-resistant Salmonella spp. isolate together with Cls1 integron and/or integrase might play an important role in horizontal antibiotic gene transfer. © 2015 Institute of Food Technologists®

Evaluation of the ability of four ESBL-screening media to detect ESBL-producing Salmonella and Shigella.

PubMed

Sturød, Kjersti; Dahle, Ulf R; Berg, Einar Sverre; Steinbakk, Martin; Wester, Astrid L

2014-09-04

The aim of this study was to compare the ability of four commercially available media for screening extended-spectrum beta-lactamase (ESBL) to detect and identify ESBL-producing Salmonella and Shigella in fecal samples. A total of 71 Salmonella- and 21 Shigella-isolates producing ESBL(A) and/or AmpC, were received at Norwegian Institute of Public Health between 2005 and 2012. The 92 isolates were mixed with fecal specimens and tested on four ESBL screening media; ChromID ESBL (BioMèrieux), Brilliance ESBL (Oxoid), BLSE agar (AES Chemunex) and CHROMagar ESBL (CHROMagar). The BLSE agar is a biplate consisting of two different agars. Brilliance and CHROMagar are supposed to suppress growth of AmpC-producing bacteria while ChromID and BLSE agar are intended to detect both ESBL(A) and AmpC. The total sensitivity (ESBL(A)+AmpC) with 95% confidence intervals after 24 hours of incubation were as follows: ChromID: 95% (90.4-99.6), Brilliance: 93% (87.6-98.4), BLSE agar (Drigalski): 99% (96.9-100), BLSE agar (MacConkey): 99% (96.9-100) and CHROMagar: 85% (77.5-92.5). The BLSE agar identified Salmonella and Shigella isolates as lactose-negative. The other agars based on chromogenic technology displayed Salmonella and Shigella flexneri isolates with colorless colonies (as expected). Shigella sonnei produced pink colonies, similar to the morphology described for E. coli. All four agar media were reliable in screening fecal samples for ESBL(A)-producing Salmonella and Shigella. However, only ChromID and BLSE agar gave reliable detection of AmpC-producing isolates. Identification of different bacterial species based on colony colour alone was not accurate for any of the four agars.

Odorant binding protein based biomimetic sensors for detection of alcohols associated with Salmonella contamination in packaged beef.

PubMed

Sankaran, Sindhuja; Panigrahi, Suranjan; Mallik, Sanku

2011-03-15

Detection of food-borne bacteria present in the food products is critical to prevent the spread of infectious diseases. Intelligent quality sensors are being developed for detecting bacterial pathogens such as Salmonella in beef. One of our research thrusts was to develop novel sensing materials sensitive to specific indicator alcohols at low concentrations. Present work focuses on developing olfactory sensors mimicking insect odorant binding protein to detect alcohols in low concentrations at room temperature. A quartz crystal microbalance (QCM) based sensor in conjunction with synthetic peptide was developed to detect volatile organic compounds indicative to Salmonella contamination in packaged beef. The peptide sequence used as sensing materials was derived from the amino acids sequence of Drosophila odorant binding protein, LUSH. The sensors were used to detect alcohols: 3-methyl-1-butanol and 1-hexanol. The sensors were sensitive to alcohols with estimated lower detection limits of <5 ppm. Thus, the LUSH-derived QCM sensors exhibited potential to detect alcohols at low ppm concentrations. Copyright © 2011. Published by Elsevier B.V.

Presence and Persistence of Salmonella in Water: The Impact on Microbial Quality of Water and Food Safety.

PubMed

Liu, Huanli; Whitehouse, Chris A; Li, Baoguang

2018-01-01

Salmonella ranks high among the pathogens causing foodborne disease outbreaks. According to the Centers for Disease Control and Prevention, Salmonella contributed to about 53.4% of all foodborne disease outbreaks from 2006 to 2017, and approximately 32.7% of these foodborne Salmonella outbreaks were associated with consumption of produce. Trace-back investigations have suggested that irrigation water may be a source of Salmonella contamination of produce and a vehicle for transmission. Presence and persistence of Salmonella have been reported in surface waters such as rivers, lakes, and ponds, while ground water in general offers better microbial quality for irrigation. To date, culture methods are still the gold standard for detection, isolation and identification of Salmonella in foods and water. In addition to culture, other methods for the detection of Salmonella in water include most probable number, immunoassay, and PCR. The U.S. Food and Drug Administration (FDA) issued the Produce Safety Rule (PSR) in January 2013 based on the Food Safety Modernization Act (FSMA), which calls for more efforts toward enhancing and improving approaches for the prevention of foodborne outbreaks. In the PSR, agricultural water is defined as water used for in a way that is intended to, or likely to, contact covered produce, such as spray, wash, or irrigation. In summary, Salmonella is frequently present in surface water, an important source of water for irrigation. An increasing evidence indicates irrigation water as a source (or a vehicle) for transmission of Salmonella . This pathogen can survive in aquatic environments by a number of mechanisms, including entry into the viable but nonculturable (VBNC) state and/or residing within free-living protozoa. As such, assurance of microbial quality of irrigation water is critical to curtail the produce-related foodborne outbreaks and thus enhance the food safety. In this review, we will discuss the presence and persistence of Salmonella in water and the mechanisms Salmonella uses to persist in the aquatic environment, particularly irrigation water, to better understand the impact on the microbial quality of water and food safety due to the presence of Salmonella in the water environment.

Presence and Persistence of Salmonella in Water: The Impact on Microbial Quality of Water and Food Safety

PubMed Central

Liu, Huanli; Whitehouse, Chris A.; Li, Baoguang

2018-01-01

Salmonella ranks high among the pathogens causing foodborne disease outbreaks. According to the Centers for Disease Control and Prevention, Salmonella contributed to about 53.4% of all foodborne disease outbreaks from 2006 to 2017, and approximately 32.7% of these foodborne Salmonella outbreaks were associated with consumption of produce. Trace-back investigations have suggested that irrigation water may be a source of Salmonella contamination of produce and a vehicle for transmission. Presence and persistence of Salmonella have been reported in surface waters such as rivers, lakes, and ponds, while ground water in general offers better microbial quality for irrigation. To date, culture methods are still the gold standard for detection, isolation and identification of Salmonella in foods and water. In addition to culture, other methods for the detection of Salmonella in water include most probable number, immunoassay, and PCR. The U.S. Food and Drug Administration (FDA) issued the Produce Safety Rule (PSR) in January 2013 based on the Food Safety Modernization Act (FSMA), which calls for more efforts toward enhancing and improving approaches for the prevention of foodborne outbreaks. In the PSR, agricultural water is defined as water used for in a way that is intended to, or likely to, contact covered produce, such as spray, wash, or irrigation. In summary, Salmonella is frequently present in surface water, an important source of water for irrigation. An increasing evidence indicates irrigation water as a source (or a vehicle) for transmission of Salmonella. This pathogen can survive in aquatic environments by a number of mechanisms, including entry into the viable but nonculturable (VBNC) state and/or residing within free-living protozoa. As such, assurance of microbial quality of irrigation water is critical to curtail the produce-related foodborne outbreaks and thus enhance the food safety. In this review, we will discuss the presence and persistence of Salmonella in water and the mechanisms Salmonella uses to persist in the aquatic environment, particularly irrigation water, to better understand the impact on the microbial quality of water and food safety due to the presence of Salmonella in the water environment. PMID:29900166

Susceptibility of Salmonella enterica Isolates from Tomato Farm Environments to Fatty Acids Naturally Found on Tomato Fruit.

PubMed

Dev Kumar, Govindaraj; Micallef, Shirley A

2017-05-01

Salmonella enterica subsp. enterica can colonize tomato fruit as it interacts with fruit surface compounds. The exometabolome of tomato fruit contains a mixture of compounds, including fatty acids, which could affect Salmonella fitness. Fatty acids detected in fruit exudates were investigated for Salmonella inhibition. Pelargonic, lauric, myristic, palmitic, margaric, stearic, and oleic acids were suspended in water dissolved in dimethyl sulfoxide (DMSO) or emulsified in water and quillaja saponin to assess how bioavailability impacted Salmonella growth. The minimum inhibitory concentrations of fatty acids were determined using a resazurin assay. Quillaja saponin emulsion and DMSO solution of pelargonic acid were inhibitory to Salmonella at 31.25 mM. Lauric and myristic acid emulsions inhibited growth at 1 M concentrations in quillaja emulsions and 62.5 mM in DMSO. Lauric and myristic acids significantly affected growth of Salmonella Newport, Javiana, and Typhimurium (p ≤ 0.05). Growth curve analysis using the Baranyi model revealed reduced maxima populations for all treatments (p ≤ 0.001) and shorter lag phase durations for Salmonella Newport with lauric acid (p < 0.01) and Salmonella Javiana with lauric (p < 0.001) and myristic (p < 0.001) acids. Salmonella Newport and Javiana exhibited an accelerated growth rate with lauric acid (p < 0.001) as a result of early stationary phase transition (shorter log phase). In myristic acid-amended media, Salmonella Javiana also displayed a faster growth rate (p < 0.001). Pelargonic acid (31.25 mM) treatment of Salmonella cells resulted in a drop in culturable cells to below detection in an hour. Microscopic analysis with Cyto-dye and propidium iodide of bacterial cells treated with pelargonic acid indicated a mixture of live and dead cells, with cell lysis of some cells. A subset of cells exhibited elongation-possibly indicating filament formation, a known antibiotic stress response. The results suggest that fatty acids present in tomato fruit surface exudates may exert a restrictive effect on Salmonella growth on fruit.

Longitudinal Study of Distributions of Similar Antimicrobial-Resistant Salmonella Serovars in Pigs and Their Environment in Two Distinct Swine Production Systems

PubMed Central

Keelara, Shivaramu; Scott, H. Morgan; Morrow, William M.; Gebreyes, Wondwossen A.; Correa, Maria; Nayak, Rajesh; Stefanova, Rossina

2013-01-01

The aim of this longitudinal study was to determine and compare the prevalences and genotypic profiles of antimicrobial-resistant (AR) Salmonella isolates from pigs reared in antimicrobial-free (ABF) and conventional production systems at farm, at slaughter, and in their environment. We collected 2,889 pig fecal and 2,122 environmental (feed, water, soil, lagoon, truck, and floor swabs) samples from 10 conventional and eight ABF longitudinal cohorts at different stages of production (farrowing, nursery, finishing) and slaughter (postevisceration, postchill, and mesenteric lymph nodes [MLN]). In addition, we collected 1,363 carcass swabs and 205 lairage and truck samples at slaughter. A total of 1,090 Salmonella isolates were recovered from the samples; these were isolated with a significantly higher prevalence in conventionally reared pigs (4.0%; n = 66) and their environment (11.7%; n = 156) than in ABF pigs (0.2%; n = 2) and their environment (0.6%; n = 5) (P < 0.001). Salmonella was isolated from all stages at slaughter, including the postchill step, in the two production systems. Salmonella prevalence was significantly higher in MLN extracted from conventional carcasses than those extracted from ABF carcasses (P < 0.001). We identified a total of 24 different serotypes, with Salmonella enterica serovar Typhimurium, Salmonella enterica serovar Anatum, Salmonella enterica serovar Infantis, and Salmonella enterica serovar Derby being predominant. The highest frequencies of antimicrobial resistance (AR) were exhibited to tetracycline (71%), sulfisoxazole (42%), and streptomycin (17%). Multidrug resistance (resistance to ≥3 antimicrobials; MDR) was detected in 27% (n = 254) of the Salmonella isolates from the conventional system. Our study reports a low prevalence of Salmonella in both production systems in pigs on farms, while a higher prevalence was detected among the carcasses at slaughter. The dynamics of Salmonella prevalence in pigs and carcasses were reciprocated in the farm and slaughter environment, clearly indicating an exchange of this pathogen between the pigs and their surroundings. Furthermore, the phenotypic and genotypic fingerprint profile results underscore the potential role played by environmental factors in dissemination of AR Salmonella to pigs. PMID:23793629

The occurrence of Salmonella in raw and ready-to-eat bean sprouts and sprouted seeds on retail sale in England and Northern Ireland.

PubMed

Sadler-Reeves, L; Aird, H; de Pinna, E; Elviss, N; Fox, A; Kaye, M; Jorgensen, F; Lane, C; Willis, C; McLauchlin, J

2016-02-01

A total of 554 samples of bean sprouts or other sprouted seeds were collected at retail sale and submitted to nine Official Control Laboratories in England and Northern Ireland during January to March 2011. Samples (100 g) were tested for the presence of Salmonella using the EN ISO 6579:2002 method. Products labelled as ready-to-eat comprised 23% of the samples and 61% were labelled as raw or to-cook: the remaining 12% had no indication if the food was intended as ready-to-eat or ready-to-cook, and 4% were not recorded. Salmonella spp. were detected from four samples of mung-bean sprouts (0·7% of all the 554 samples) and all four isolates were confirmed as Salmonella enterica serovar Abaetetuba (11 : k : 1,5). Two of the samples where Salmonella was detected were sold as ready-to-eat (labelled 'rinse and serve' only): The remaining two were from samples labelled as ready-to-cook. Consumption of sprouted seeds have been associated with infections from a range of foodborne pathogens, particularly Salmonella and shigatoxin producing Escherichia coli (STEC). However, there is limited data (including that from EU monitoring) on foodborne pathogens in samples of this food type which are not associated with outbreaks of infection. Out of 554 raw and ready-to-eat bean sprouts and sprouted seeds sampled at retail, Salmonella spp. was detected from four samples. This study illustrated the potential of this product to be contaminated with a human pathogen and the importance of considering the intended use and preparation of specific food in assessing microbiological risks. © 2015 The Society for Applied Microbiology.

Use of high-throughput mass spectrometry to elucidate host pathogen interactions in Salmonella

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rodland, Karin D.; Adkins, Joshua N.; Ansong, Charles

Capabilities in mass spectrometry are evolving rapidly, with recent improvements in sensitivity, data analysis, and most important, from the standpoint of this review, much higher throughput allowing analysis of many samples in a single day. This short review describes how these improvements in mass spectrometry can be used to dissect host-pathogen interactions using Salmonella as a model system. This approach enabled direct identification of the majority of annotated Salmonella proteins, quantitation of expression changes under various in vitro growth conditions, and new insights into virulence and expression of Salmonella proteins within host cell cells. One of the most significant findingsmore » is that a very high percentage of the all annotated genes (>20%) in Salmonella are regulated post-transcriptionally. In addition, new and unexpected interactions have been identified for several Salmonella virulence regulators that involve protein-protein interactions, suggesting additional functions of these regulators in coordinating virulence expression. Overall high throughput mass spectrometry provides a new view of pathogen-host interactions emphasizing the protein products and defining how protein interactions determine the outcome of infection.« less

Use of Glycerol as an Optical Clearing Agent for Enhancing Photonic Transference and Detection of Salmonella typhimurium through Porcine Skin

USDA-ARS?s Scientific Manuscript database

The objective of this study was to evaluate glycerol (GLY) and GLY + dimethyl sulfoxide (DMSO) to increase photonic detection of transformed Salmonella typhimurium (S. typh-lux) through porcine skin. Skin was placed on 96-well plates containing S. typh-lux, imaged (5 min) using a CCD camera, and the...

Use of Glycerol as an Optical Clearing Agent for Enhancing Photonic Transference and Detection of Salmonella typhimurium Through Porcine Skin

USDA-ARS?s Scientific Manuscript database

The objective of this study was to evaluate glycerol (GLY) and GLY + dimethyl sulfoxide (DMSO) to increase photonic detection of transformed Salmonella typhimurium (S. typh-lux) through porcine skin. Skin was placed on 96-well plates containing S. typh-lux, imaged (5 min) using a CCD camera, and the...

Rapid detection of food pathogens using RNA aptamers-immobilized slide.

PubMed

Maeng, Jin-Soo; Kim, Namsoo; Kim, Chong-Tai; Han, Seung Ryul; Lee, Young Ju; Lee, Seong-Wook; Lee, Myung-Hyun; Cho, Yong-Jin

2012-07-01

The purpose of this study was to develop a simple and rapid detection system for foodborne bacteria, which consisted of an optical microscope and its slide chip with artificial antibodies, or RNA aptamers. From an RNA pool, three each RNA aptamers were built by the method of SELEX (systematic evolution of ligands by exponential enrichment) for components of cell wall, LPS (lipopolysaccharide) from E. coli O157:H7, teichoic acid from Staphylococcus aureus and a cell membrane protein of OmpC from Salmonella typhimurium, respectively. These aptamers were hybridized with thiol-conjugated 16 dT-linker molecules in order to be immobilized on silver surface which was, in advance, fabricated on glass slide, using a spin-coating method. To confirm that each aptamers retained its specific binding activities to their antigenic live bacteria, microscopic view of bound cells immobilized on silver film were observed. Furthermore, we observed the fluorescence-emitting bacteria-aptamer complex immobilized on silver film after adding RNA aptamers hybridized with fluorophore, FAM-conjugated 16 dT-linker molecules. As a result, the RNA aptamers-immobilized slide system developed in this study was a useful new tool to rapidly monitor individual food pathogens.

Antimicrobial effect of the Tunisian Nana variety Punica granatum L. extracts against Salmonella enterica (serovars Kentucky and Enteritidis) isolated from chicken meat and phenolic composition of its peel extract.

PubMed

Wafa, Ben Ajmia; Makni, Mohamed; Ammar, Sonda; Khannous, Lamia; Hassana, Amal Ben; Bouaziz, Mohamed; Es-Safi, Nour Eddine; Gdoura, Radhouane

2017-01-16

Punica granatum L. is widely recognized for its potency against a broad spectrum of bacterial pathogens. The purpose of this study was to explore the inhibitory and the bactericidal activities of Punica granatum against Salmonella strains. The effect of extracts obtained from different parts (peels, seeds, juice and flowers) of pomegranate and using different solvents against Salmonella enterica serovars Kentucky and Enteritidis isolated from chicken meat was thus investigated. Salmonella strains were identified with the standard API-20E system and confirmed by real time PCR. The obtained results showed that the highest antibacterial activity against Salmonella strains was observed with the peels ethanolic extract giving MIC values ranging from 10.75 to 12.5mg/mL. The ethanolic extract of P. granatum Nana peels at 0.8 and 1.6mg/g significantly inhibited the growth of Salmonella Kentucky in chicken meat stored at 4°C. The phenolic composition of the ethanolic peel extract was explored by HPLC coupled to both DAD and ESI/TOF-MS detections. The obtained results allowed the detection of 21 phytochemical compounds among which various phenolic compounds have been identified on the basis of their UV and MS spectra as well as with literature data. Among the detected compounds, anthocyanins, ellagitannins, ellagic acid derivatives and flavanols were further characterized through MS-MS analysis. Our results showed thus that the Tunisian variety Nana pomegranate constitutes a good source of bioactive compounds with potent antimicrobial activity on the growth of Salmonella strains suggesting that the studied pomegranate cultivar could be a natural remedy to minimize the emergence of Salmonella enterica strains which is often involved in food borne illness. Copyright © 2016 Elsevier B.V. All rights reserved.

Multi-laboratory validation study of multilocus variable-number tandem repeat analysis (MLVA) for Salmonella enterica serovar Enteritidis, 2015.

PubMed

Peters, Tansy; Bertrand, Sophie; Björkman, Jonas T; Brandal, Lin T; Brown, Derek J; Erdõsi, Tímea; Heck, Max; Ibrahem, Salha; Johansson, Karin; Kornschober, Christian; Kotila, Saara M; Le Hello, Simon; Lienemann, Taru; Mattheus, Wesley; Nielsen, Eva Møller; Ragimbeau, Catherine; Rumore, Jillian; Sabol, Ashley; Torpdahl, Mia; Trees, Eija; Tuohy, Alma; de Pinna, Elizabeth

2017-03-02

Multilocus variable-number tandem repeat analysis (MLVA) is a rapid and reproducible typing method that is an important tool for investigation, as well as detection, of national and multinational outbreaks of a range of food-borne pathogens. Salmonella enterica serovar Enteritidis is the most common Salmonella serovar associated with human salmonellosis in the European Union/European Economic Area and North America. Fourteen laboratories from 13 countries in Europe and North America participated in a validation study for MLVA of S. Enteritidis targeting five loci. Following normalisation of fragment sizes using a set of reference strains, a blinded set of 24 strains with known allele sizes was analysed by each participant. The S. Enteritidis 5-loci MLVA protocol was shown to produce internationally comparable results as more than 90% of the participants reported less than 5% discrepant MLVA profiles. All 14 participating laboratories performed well, even those where experience with this typing method was limited. The raw fragment length data were consistent throughout, and the inter-laboratory validation helped to standardise the conversion of raw data to repeat numbers with at least two countries updating their internal procedures. However, differences in assigned MLVA profiles remain between well-established protocols and should be taken into account when exchanging data. This article is copyright of The Authors, 2017.

Seven Salmonella Typhimurium Outbreaks in Australia Linked by Trace-Back and Whole Genome Sequencing.

PubMed

Ford, Laura; Wang, Qinning; Stafford, Russell; Ressler, Kelly-Anne; Norton, Sophie; Shadbolt, Craig; Hope, Kirsty; Franklin, Neil; Krsteski, Radomir; Carswell, Adrienne; Carter, Glen P; Seemann, Torsten; Howard, Peter; Valcanis, Mary; Castillo, Cristina Fabiola Sotomayor; Bates, John; Glass, Kathryn; Williamson, Deborah A; Sintchenko, Vitali; Howden, Benjamin P; Kirk, Martyn D

2018-05-01

Salmonella Typhimurium is a common cause of foodborne illness in Australia. We report on seven outbreaks of Salmonella Typhimurium multilocus variable-number tandem-repeat analysis (MLVA) 03-26-13-08-523 (European convention 2-24-12-7-0212) in three Australian states and territories investigated between November 2015 and March 2016. We identified a common egg grading facility in five of the outbreaks. While no Salmonella Typhimurium was detected at the grading facility and eggs could not be traced back to a particular farm, whole genome sequencing (WGS) of isolates from cases from all seven outbreaks indicated a common source. WGS was able to provide higher discriminatory power than MLVA and will likely link more Salmonella Typhimurium cases between states and territories in the future. National harmonization of Salmonella surveillance is important for effective implementation of WGS for Salmonella outbreak investigations.

40 CFR 503.8 - Sampling and analysis.

Code of Federal Regulations, 2013 CFR

2013-07-01

...-00000-1). (5) Salmonella sp. bacteria. Part 9260 D., “Standard Methods for the Examination of Water and... 20005; or Kenner, B.A. and H.P. Clark, “Detection and enumeration of Salmonella and Pseudomonas...

40 CFR 503.8 - Sampling and analysis.

Code of Federal Regulations, 2012 CFR

2012-07-01

...-00000-1). (5) Salmonella sp. bacteria. Part 9260 D., “Standard Methods for the Examination of Water and... 20005; or Kenner, B.A. and H.P. Clark, “Detection and enumeration of Salmonella and Pseudomonas...

[New methodological approaches to the detection and quantitative registration of Salmonella in the study of water objects].

PubMed

Aleshnia, V V; Panasovets, O P; Zhuravlev, P V; Sukhanova, S M; Golubenko, I A; Nedachin, A E; Talaeva, Iu G; Artemova, T Z; Gipp, E K; Butorina, N N; Zagaĭnova, A V; Shvager, M M; Mitrofanova, T V

2011-01-01

The paper gives data on the use of techniques to detect and register Salmonella in the water objects, by applying a new liquid nutrient medium. Experimental and field studies have shown its advantage over the accumulation media widely used in practical healthcare. It has been ascertained that the nutrient medium not only accumulates biomass, but also provides the restoration of the biological properties of uncultivated Salmonella species. The use of the nutrient medium at practical laboratories makes it possible to unify guidelines for the examination of water objects with varying degrees of biological pollution and to obtain the comparable results of analyses.

Prevalence and molecular profiles of Salmonella collected at a commercial turkey processing plant.

PubMed

Nde, Chantal W; Sherwood, Julie S; Doetkott, Curt; Logue, Catherine M

2006-08-01

In this study, whole carcasses were sampled at eight stages on a turkey-processing line and Salmonella prevalence was determined using enrichment techniques. Recovered Salmonella was further characterized using serotyping and the molecular profiles were determined using pulsed-field gel electrophoresis (PFGE). Prevalence data showed that contamination rates varied along the line and were greatest after defeathering and after chilling. Analysis of contamination in relation to serotypes and PFGE profiles found that on some visits the same serotype was present all along the processing line while on other days, additional serotypes were recovered that were not detected earlier on the line, suggesting that the birds harbored more than one serotype of Salmonella or there was cross-contamination occurring during processing. Overall, this study found fluctuations in Salmonella prevalence along a turkey-processing line. Following washing, Salmonella prevalence was significantly reduced, suggesting that washing is critical for Salmonella control in turkey processing and has significant application for controlling Salmonella at the postdefeathering and prechill stages where prevalence increased.

Structural and functional studies of a 50 kDa antigenic protein from Salmonella enterica serovar Typhi.

PubMed

Choong, Yee Siew; Lim, Theam Soon; Chew, Ai Lan; Aziah, Ismail; Ismail, Asma

2011-04-01

The high typhoid incidence rate in developing and under-developed countries emphasizes the need for a rapid, affordable and accessible diagnostic test for effective therapy and disease management. TYPHIDOT®, a rapid dot enzyme immunoassay test for typhoid, was developed from the discovery of a ∼50 kDa protein specific for Salmonella enterica serovar Typhi. However, the structure of this antigen remains unknown till today. Studies on the structure of this antigen are important to elucidate its function, which will in turn increase the efficiency of the development and improvement of the typhoid detection test. This paper described the predictive structure and function of the antigenically specific protein. The homology modeling approach was employed to construct the three-dimensional structure of the antigen. The built structure possesses the features of TolC-like outer membrane protein. Molecular docking simulation was also performed to further probe the functionality of the antigen. Docking results showed that hexamminecobalt, Co(NH(3))(6)(3+), as an inhibitor of TolC protein, formed favorable hydrogen bonds with D368 and D371 of the antigen. The single point (D368A, D371A) and double point (D368A and D371A) mutations of the antigen showed a decrease (single point mutation) and loss (double point mutations) of binding affinity towards hexamminecobalt. The architecture features of the built model and the docking simulation reinforced and supported that this antigen is indeed the variant of outer membrane protein, TolC. As channel proteins are important for the virulence and survival of bacteria, therefore this ∼50 kDa channel protein is a good specific target for typhoid detection test. Copyright © 2011 Elsevier Inc. All rights reserved.

Genomes of Salmonella with diverse patterns of antibiotic resistance (AR) revealed the dynamics of AR gene organization and detected resistance gene families found in Salmonella

USDA-ARS?s Scientific Manuscript database

We produced and assembled high quality draft genomes (~100X coverage) for 305 Salmonella from a diverse a group of over 100 serovars and diverse sources. Of these isolates, 119 were selected to capture a wide variety of different AR patterns. In our subsequent analyses we included 285 additional pub...

Postmortem Photonic Imaging of Lux-Modified Salmonella Typhimuium Within the Gastrointestinal Tract of Swine Following Oral Inoculation In Vivo

USDA-ARS?s Scientific Manuscript database

The study objective was to monitor Salmonella progression by photonic detection through segments of the gastrointestinal tract after oral inoculation. Pigs (~80 kg) were inoculated orally with 3.1 or 4.1 x 1010 cfu of Salmonella Typhimurium transformed with plasmid pAK1-lux for a 6-h (n = 6) or 12-h...

Effect of strategic administration of an encapsulated blend of formic acid, citric acid, and essential oils on Salmonella carriage, seroprevalence, and growth of finishing pigs.

PubMed

Walia, Kavita; Argüello, Hector; Lynch, Helen; Leonard, Finola C; Grant, Jim; Yearsley, Dermot; Kelly, Sinead; Duffy, Geraldine; Gardiner, Gillian E; Lawlor, Peadar G

2017-02-01

Controlling Salmonella at farm level can act as the first line of defence in reducing salmonellosis from pork. This study investigated the efficacy of an encapsulated blend of formic acid, citric acid, and essential oils (FormaXOL™) administered to finisher pigs for 28days prior to slaughter in controlling Salmonella shedding on a commercial farm with a history of high Salmonella seroprevalence. Fourteen pens of 8-10 pigs/pen were randomly assigned to a control (finisher diet without additive) or a treatment group (the same diet with 4kg/t of FormaXOL™) for 28 days. Faeces were collected from each pig on days 0, 14, and 28, while on day 29 blood, caecal digesta and ileocaecal-mesenteric lymph nodes were collected at slaughter. Pigs were weighed at the start and end of the trial, feed intake was recorded, and carcass quality parameters were recorded at slaughter. On day 14, Salmonella shedding was reduced in the treatment compared to the control group (27.9% versus 51.7% probability of detecting Salmonella in faeces, respectively; p=0.001). However, on day 28, no reduction was observed (20.6% versus 35.9% probability of detecting Salmonella in faeces, respectively; p=0.07). Interestingly, Salmonella shedding rates in the treated pigs remained stable throughout the trial compared to the control group. This suggests that the feed additive prevented additional pigs from acquiring the Salmonella infection. A lower Salmonella seroprevalence was detected at slaughter in the treatment compared to the control group using the 40% optical density cut-off (64.5% versus 88.5%, respectively; p=0.01). However, no significant differences in Salmonella recovery rates were observed in the caecal digesta or lymph nodes between treated and control groups. Treated pigs had a lower feed intake than pigs fed the control diet (p=0.001); however, average daily gain and feed conversion efficiency were not affected by treatment (p=0.45 and 0.55, respectively). Consequently, supplementing the diet with FormaXOL™ for 28days increased the feed cost per kg of live-weight gain by €0.08. Overall, results suggest that strategic administration of an encapsulated blend of formic acid, citric acid, and essential oils, to finishing pigs for 28days prior to slaughter has potential to prevent increased Salmonella shedding at certain time points as well as seroprevalence. However, this additive did not lower intestinal carriage, nor did it reduce seroprevalence to below the cut-off used for the high Salmonella risk category in Ireland (50%) or improve growth performance. Copyright © 2016 Elsevier B.V. All rights reserved.

Salmonella in beef and produce from honduras.

PubMed

Maradiaga, Martha; Miller, Mark F; Thompson, Leslie; Pond, Ansen; Gragg, Sara E; Echeverry, Alejandro; Garcia, Lyda G; Loneragan, Guy H; Brashears, Mindy M

2015-03-01

Salmonella continues to cause a considerable number of foodborne illnesses worldwide. The sources of outbreaks include contaminated meat and produce. The purpose of this study was to establish an initial investigation of the burden of Salmonella in produce and beef from Honduras by sampling retail markets and abattoirs. Retail produce samples (cantaloupes, cilantro, cucumbers, leafy greens, peppers, and tomatoes; n = 573) were purchased in three major cities of Honduras, and retail whole-muscle beef (n = 555) samples were also purchased in four major cities. Additionally, both hide and beef carcass (n = 141) samples were collected from two Honduran abattoirs. Whole-muscle beef samples were obtained using a sponge hydrated with buffered peptone water, and 10 ml of the buffered peptone water rinsate of each produce sample was collected with a dry sponge and placed in a bag to be transported back to the United States. Salmonella was detected using a commercially available, closeplatform PCR system, and positive samples were subjected to culture on selective media to obtain isolates. Overall, the prevalence of Salmonella-positive samples, based on PCR detection in Honduras (n = 555) retail beef was 10.1% (95% confidence interval = 7.8, 12.9), whereas 7.8% (n = 141) of beef carcass and hides samples were positive in both beef plants. The overall Salmonella prevalence for all produce samples (n = 573) collected was 2.1% (95% confidence interval = 1.2, 3.6). The most common serotypes identified in Honduras were Salmonella Typhimurium followed by Derby. These results provide an indication of Salmonella contamination of beef and produce in Honduras. Developing a Salmonella baseline for Latin America through an initial investigation like the one presented here contributes to a broader global understanding of the potential exposure through food, thus providing insight into the needs for control strategies.

Growth and survival of foodborne pathogens in beer.

PubMed

Menz, Garry; Aldred, Peter; Vriesekoop, Frank

2011-10-01

This work aimed to assess the growth and survival of four foodborne pathogens (Escherichia coli O157:H7, Salmonella Typhimurium, Listeria monocytogenes, and Staphylococcus aureus) in beer. The effects of ethanol, pH, and storage temperature were investigated for the gram-negative pathogens (E. coli O157:H7 and Salmonella Typhimurium), whereas the presence of hops ensured that the gram-positive pathogens (L. monocytogenes and S. aureus) were rapidly inactivated in alcohol-free beer. The pathogens E. coli O157:H7 and Salmonella Typhimurium could not grow in the mid-strength or full-strength beers, although they could survive for more than 30 days in the mid-strength beer when held at 4°C. These pathogens grew rapidly in the alcohol-free beer; however, growth was prevented when the pH of the alcohol-free beer was lowered from the "as received" value of 4.3 to 4.0. Pathogen survival in all beers was prolonged at lowered storage temperatures.

Salmonella Typhimurium induces SPI-1 and SPI-2 regulated and strain dependent downregulation of MHC II expression on porcine alveolar macrophages

PubMed Central

2012-01-01

Foodborne salmonellosis is one of the most important bacterial zoonotic diseases worldwide. Salmonella Typhimurium is the serovar most frequently isolated from persistently infected slaughter pigs in Europe. Circumvention of the host’s immune system by Salmonella might contribute to persistent infection of pigs. In the present study, we found that Salmonella Typhimurium strain 112910a specifically downregulated MHC II, but not MHC I, expression on porcine alveolar macrophages in a Salmonella pathogenicity island (SPI)-1 and SPI-2 dependent way. Salmonella induced downregulation of MHC II expression and intracellular proliferation of Salmonella in macrophages were significantly impaired after opsonization with Salmonella specific antibodies prior to inoculation. Furthermore, the capacity to downregulate MHC II expression on macrophages differed significantly among Salmonella strains, independently of strain specific differences in invasion capacity, Salmonella induced cytotoxicity and altered macrophage activation status. The fact that strain specific differences in MHC II downregulation did not correlate with the extent of in vitro SPI-1 or SPI-2 gene expression indicates that other factors are involved in MHC II downregulation as well. Since Salmonella strain dependent interference with the pig’s immune response through downregulation of MHC II expression might indicate that certain Salmonella strains are more likely to escape serological detection, our findings are of major interest for Salmonella monitoring programs primarily based on serology. PMID:22694285

A label-free ultrasensitive fluorescence detection of viable Salmonella enteritidis using enzyme-induced cascade two-stage toehold strand-displacement-driven assembly of G-quadruplex DNA.

PubMed

Zhang, Peng; Liu, Hui; Ma, Suzhen; Men, Shuai; Li, Qingzhou; Yang, Xin; Wang, Hongning; Zhang, Anyun

2016-06-15

The harm of Salmonella enteritidis (S. enteritidis ) to public health mainly by contaminating fresh food and water emphasizes the urgent need for rapid detection techniques to help control the spread of the pathogen. In this assay, an newly designed capture probe complex that contained specific S. enteritidis-aptamer and hybridized signal target sequence was used for viable S. enteritidis recognition directly. In the presence of the target S. enteritidis, single-stranded target sequences were liberated and initiated the replication-cleavage reaction, producing numerous G-quadruplex structures with a linker on the 3'-end. And then, the sensing system took innovative advantage of quadratic linker-induced strand-displacement for the first time to release target sequence in succession, leading to the cyclic reuse of the target sequences and cascade signal amplification, thereby achieving the successive production of G-quadruplex structures. The fluorescent dye, N-Methyl mesoporphyrin IX, binded to these G-quadruplex structures and generated significantly enhanced fluorescent signals to achieve highly sensitive detection of S. enteritidis down to 60 CFU/mL with a linear range from 10(2) to 10(7)CFU/mL. By coupling the cascade two-stage target sequences-recyclable toehold strand-displacement with aptamer-based target recognition successfully, it is the first report on a novel non-label, modification-free and DNA extraction-free ultrasensitive fluorescence biosensor for detecting viable S. enteritidis directly, which can discriminate from dead S. enteritidis. Copyright © 2016 Elsevier B.V. All rights reserved.

Immunosensor based on electrodeposition of gold-nanoparticles and ionic liquid composite for detection of Salmonella pullorum.

PubMed

Wang, Dan; Dou, Wenchao; Zhao, Guangying; Chen, Yan

2014-11-01

In order to increase the reproducibility and stability of electrochemical immunosensor, which is a key issue for its application and popularization, an accurate and stable immunosensor for rapid detection of Salmonella pullorum (S. pullorum) was proposed in this study. The immunosensor was fabricated by modifying Screen-printed Carbon Electrode (SPCE) with electrodeposited gold nanoparticles (AuNPs), HRP-labeled anti-S. pullorum and ionic liquids (ILs) (AuNP/HRP/IL). AuNPs are electrodeposited on the working electrode surface to increase the amount of antibodies that bind to the electrode and then modified with ILs to protect the antibodies from being inactivated in the test environment and maintain their biological activity and the stability of the detection electrode. The electrochemical characteristics of the stepwise modified electrodes and the detection of S. pullorum were investigated by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). As shown in the results of the experiments, AuNPs with unique electrochemical properties as well as biocompatibility characteristics have been proven to be able to strengthen the antibody combination effectively and to increase the electrochemical response signal. In addition, a crucial assessment regarding implementation of stability and reproducibility analysis of a range of immunosensors is provided. We found that application of AuNPs/ILs in the immune modified electrodes showed obvious improvement when compared with other groups. Given their high levels of reproducibility, stability, target specificity and sensitivity, AuNPs and ILs were considered to be excellent elements for electrode modification. Copyright © 2014 Elsevier B.V. All rights reserved.

Enteric Pathogens Associated with Childhood Diarrhea in Tripoli-Libya

PubMed Central

Rahouma, Amal; Klena, John D.; Krema, Zaineb; Abobker, Abdalwahed A.; Treesh, Khalid; Franka, Ezzedin; Abusnena, Omar; Shaheen, Hind I.; El Mohammady, Hanan; Abudher, Abdulhafid; Ghenghesh, Khalifa Sifaw

2011-01-01

Stool samples from children < 5 years of age with diarrhea (N = 239) were examined for enteric pathogens using a combination of culture, enzyme-immunoassay, and polymerase chain reaction methods. Pathogens were detected in 122 (51%) stool samples; single pathogens were detected in 37.2% and co-pathogens in 13.8% of samples. Norovirus, rotavirus, and diarrheagenic Escherichia coli (DEC) were the most frequently detected pathogens (15.5%, 13.4%, and 11.2%, respectively); Salmonella, adenovirus, and Aeromonas were detected less frequently (7.9%, 7.1%, and 4.2%). The most commonly detected DEC was enteroaggregative E. coli (5.4%). Resistance to ≥ 3 antimicrobials was observed in 60% (18/30) of the bacterial pathogens. Salmonella resistance to ciprofloxacin (63.1%) has become a concern. Enteric viral pathogens were the most significant causative agents of childhood diarrhea in Tripoli. Bacterial pathogens were also important contributors to pediatric diarrhea. The emergence of ciprofloxacin-resistant Salmonella represents a serious health problem that must be addressed by Libyan health authorities PMID:21633024

Non-protein coding RNA genes as the novel diagnostic markers for the discrimination of Salmonella species using PCR.

PubMed

Nithya, Ravichantar; Ahmed, Siti Aminah; Hoe, Chee-Hock; Gopinath, Subash C B; Citartan, Marimuthu; Chinni, Suresh V; Lee, Li Pin; Rozhdestvensky, Timofey S; Tang, Thean-Hock

2015-01-01

Salmonellosis, a communicable disease caused by members of the Salmonella species, transmitted to humans through contaminated food or water. It is of paramount importance, to generate accurate detection methods for discriminating the various Salmonella species that cause severe infection in humans, including S. Typhi and S. Paratyphi A. Here, we formulated a strategy of detection and differentiation of salmonellosis by a multiplex polymerase chain reaction assay using S. Typhi non-protein coding RNA (sRNA) genes. With the designed sequences that specifically detect sRNA genes from S. Typhi and S. Paratyphi A, a detection limit of up to 10 pg was achieved. Moreover, in a stool-seeding experiment with S. Typhi and S. Paratyphi A, we have attained a respective detection limit of 15 and 1.5 CFU/mL. The designed strategy using sRNA genes shown here is comparatively sensitive and specific, suitable for clinical diagnosis and disease surveillance, and sRNAs represent an excellent molecular target for infectious disease.

Streptomycin Induced Stress Response in Salmonella enterica Serovar Typhimurium Shows Distinct Colony Scatter Signature

PubMed Central

Singh, Atul K.; Drolia, Rishi; Bai, Xingjian; Bhunia, Arun K.

2015-01-01

We investigated the streptomycin-induced stress response in Salmonella enterica serovars with a laser optical sensor, BARDOT (bacterial rapid detection using optical scattering technology). Initially, the top 20 S. enterica serovars were screened for their response to streptomycin at 100 μg/mL. All, but four S. enterica serovars were resistant to streptomycin. The MIC of streptomycin-sensitive serovars (Enteritidis, Muenchen, Mississippi, and Schwarzengrund) varied from 12.5 to 50 μg/mL, while streptomycin-resistant serovar (Typhimurium) from 125–250 μg/mL. Two streptomycin-sensitive serovars (Enteritidis and Mississippi) were grown on brain heart infusion (BHI) agar plates containing sub-inhibitory concentration of streptomycin (1.25–5 μg/mL) and a streptomycin-resistant serovar (Typhimurium) was grown on BHI containing 25–50 μg/mL of streptomycin and the colonies (1.2 ± 0.1 mm diameter) were scanned using BARDOT. Data show substantial qualitative and quantitative differences in the colony scatter patterns of Salmonella grown in the presence of streptomycin than the colonies grown in absence of antibiotic. Mass-spectrometry identified overexpression of chaperonin GroEL, which possibly contributed to the observed differences in the colony scatter patterns. Quantitative RT-PCR and immunoassay confirmed streptomycin-induced GroEL expression while, aminoglycoside adenylyltransferase (aadA), aminoglycoside efflux pump (aep), multidrug resistance subunit acrA, and ribosomal protein S12 (rpsL), involved in streptomycin resistance, were unaltered. The study highlights suitability of the BARDOT as a non-invasive, label-free tool for investigating stress response in Salmonella in conjunction with the molecular and immunoassay methods. PMID:26252374

Assessment of the microscreen phage-induction assay for screening hazardous wastes (1989)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Houk, V.S.; DeMarini, D.M.

1989-01-01

The Microscreen phage-induction assay, which quantitatively measures the induction of prophage Lambda in Escherichia coli WP2s(Lambda), was used to test 14 crude (unfractionated) hazardous industrial-waste samples for genotoxic activity in the presence and absence of metabolic activation. Eleven of the 14 wastes induced prophage, and induction was observed at concentrations as low as 0.4 picograms per ml. Comparisons of the mutagenic activity of these waste samples in Salmonella and their ability to induce prophage Lambda indicate that the phage-induction assay was a more-sensitive indicator of genetic damage for this group of wastes. All but one of the wastes that weremore » mutagenic to Salmonella were detected by the phage-induction assay, and 5 wastes not mutagenic to Salmonella were genetically active in the phage assay. The enhanced ability of the phage-induction assay to detect genotoxic activity may be related to the constituents comprising these waste samples. Partial chemical characterizations of the wastes showed high concentrations of carcinogenic metals, solvents, and chlorinated compounds, most of which are detected poorly by the Salmonella assay.« less

Risk Factors for Salmonella, Shiga Toxin-Producing Escherichia coli and Campylobacter Occurrence in Primary Production of Leafy Greens and Strawberries

PubMed Central

Ceuppens, Siele; Johannessen, Gro S.; Allende, Ana; Tondo, Eduardo César; El-Tahan, Fouad; Sampers, Imca; Jacxsens, Liesbeth; Uyttendaele, Mieke

2015-01-01

The microbiological sanitary quality and safety of leafy greens and strawberries were assessed in the primary production in Belgium, Brazil, Egypt, Norway and Spain by enumeration of Escherichia coli and detection of Salmonella, Shiga toxin-producing E. coli (STEC) and Campylobacter. Water samples were more prone to containing pathogens (54 positives out of 950 analyses) than soil (16/1186) and produce on the field (18/977 for leafy greens and 5/402 for strawberries). The prevalence of pathogens also varied markedly according to the sampling region. Flooding of fields increased the risk considerably, with odds ratio (OR) 10.9 for Salmonella and 7.0 for STEC. A significant association between elevated numbers of generic E. coli and detection of pathogens (OR of 2.3 for STEC and 2.7 for Salmonella) was established. Generic E. coli was found to be a suitable index organism for Salmonella and STEC, but to a lesser extent for Campylobacter. Guidelines on frequency of sampling and threshold values for E. coli in irrigation water may differ from region to region. PMID:26295251

[Etiological surveillance and analysis of infectious diarrhea in Beijing in year 2010].

PubMed

Huang, Fang; Deng, Ying; Qu, Mei; Liu, Gui-Rong; Liu, Yuan; Zhang, Xin; Li, Jie; Yan, Han-Qiu; Gao, Zhi-Yong; Liu, Bai-Wei; Li, Xi-Tai; Li, Xin-Yu

2011-09-01

To explore the pathogenic form, epidemic features and serotype distribution of the pathogenic bacteria causing infectious diarrhea in Beijing. A total of 2118 samples of rectal swabs and stool specimens of diarrheal patients were collected from 6 surveillant intestinal tract clinics during the period between April and October, 2010. Enteric multiple pathogens including Vibrio cholerae, Vibrio parahaemolyticus, Salmonella, Shigella and diarrheagenic Escherichia coli were detected by the isolation culture, biochemical identification and serotyping methods. The population distribution, temporal distribution and serotype distribution of the above pathogenic bacteria were analyzed by descriptive statistical methods. 478 strains isolated from the total 2118 specimens were positive for pathogen detection, accounting to 22.6%. Among the 478 strains of pathogenic bacteria, Shigella accounting for 40.8% (195/478) was the most frequent pathogen, followed by Vibrio parahaemolyticus accouting for 23.8% (114/478), Salmonella accounting for 19.0% (91/478) and diarrheagenic Escherichia coli accounting for 4.8% (23/478). Enteric pathogenic bacteria spread mainly among adults aging between 20 and 39; and the distribution was different among different age groups, while the highest detected rate was in 30 - 39 age group, accounting for 27.2% (92/338). The detected rate of pathogenic bacteria showed evident seasonal variations, with a peak from July to October, whose detected rates were 23.5% (114/486), 32.8% (176/536), 36.1% (90/249) and 25.9% (29/112) respectively. The detected rates in other months were all under 16.0%. Shigella Sonnei was the dominant serotype, accounting for 83.1% (162/195). O3:K6 was the dominant serotype among Vibrio parahaemolyticus, accounting for 63.2% (72/114). Salmonella Enteritidis and Salmonella Typhimurium were dominant serotypes among Salmonella, accounting for 13.2% (12/91) and 12.1% (11/91) separately. Enterpathogenic Escherichia coli and enterotoxigenic Escherichia coli were the dominant serotypes among Diarrheagenic Escherichia coli, accounting for 69.6% (16/23) and 30.4% (7/23) respectively. The three main pathogenic bacteria causing infectious diarrhea in Beijing are Shigella, Vibrio parahaemolyticus, Salmonella; and there are obvious changes in the serotype distribution of Shigella and Samonella compared to previous years.

Determination of foodborne pathogenic bacteria by multiplex PCR-microchip capillary electrophoresis with genetic algorithm-support vector regression optimization.

PubMed

Li, Yongxin; Li, Yuanqian; Zheng, Bo; Qu, Lingli; Li, Can

2009-06-08

A rapid and sensitive method based on microchip capillary electrophoresis with condition optimization of genetic algorithm-support vector regression (GA-SVR) was developed and applied to simultaneous analysis of multiplex PCR products of four foodborne pathogenic bacteria. Four pairs of oligonucleotide primers were designed to exclusively amplify the targeted gene of Vibrio parahemolyticus, Salmonella, Escherichia coli (E. coli) O157:H7, Shigella and the quadruplex PCR parameters were optimized. At the same time, GA-SVR was employed to optimize the separation conditions of DNA fragments in microchip capillary electrophoresis. The proposed method was applied to simultaneously detect the multiplex PCR products of four foodborne pathogenic bacteria under the optimal conditions within 8 min. The levels of detection were as low as 1.2 x 10(2) CFU mL(-1) of Vibrio parahemolyticus, 2.9 x 10(2) CFU mL(-1) of Salmonella, 8.7 x 10(1) CFU mL(-1) of E. coli O157:H7 and 5.2 x 10(1) CFU mL(-1) of Shigella, respectively. The relative standard deviation of migration time was in the range of 0.74-2.09%. The results demonstrated that the good resolution and less analytical time were achieved due to the application of the multivariate strategy. This study offers an efficient alternative to routine foodborne pathogenic bacteria detection in a fast, reliable, and sensitive way.

Computational determination of the effects of virulent Escherichia coli and salmonella bacteriophages on human gut.

PubMed

Mostafa, Marwa Mostafa; Nassef, Mohammad; Badr, Amr

2016-10-01

Salmonella and Escherichia coli are different types of bacteria that cause food poisoning in humans. In the elderly, infants and people with chronic conditions, it is very dangerous if Salmonella or E. coli gets into the bloodstream and then they must be treated by phage therapy. Treating Salmonella and E. coli by phage therapy affects the gut flora. This research paper presents a system for detecting the effects of virulent E. coli and Salmonella bacteriophages on human gut. A method based on Domain-Domain Interactions (DDIs) model is implemented in the proposed system to determine the interactions between the proteins of human gut bacteria and the proteins of bacteriophages that infect virulent E. coli and Salmonella. The system helps gastroenterologists to realize the effect of injecting bacteriophages that infect virulent E. coli and Salmonella on the human gut. By testing the system over Enterobacteria phage 933W, Enterobacteria phage VT2-Sa and Enterobacteria phage P22, it resulted in four interactions between the proteins of the bacteriophages that infect E. coli O157:H7, E. coli O104:H4 and Salmonella typhimurium and the proteins of human gut bacterium strains. Several effects were detected such as: antibacterial activity against a number of bacterial species in human gut, regulation of cellular differentiation and organogenesis during gut, lung, and heart development, ammonia assimilation in bacteria, yeasts, and plants, energizing defense system and its function in the detoxification of lipopolysaccharide, and in the prevention of bacterial translocation in human gut. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Detection and identification of intestinal pathogenic bacteria by hybridization to oligonucleotide microarrays

PubMed Central

Jin, Lian-Qun; Li, Jun-Wen; Wang, Sheng-Qi; Chao, Fu-Huan; Wang, Xin-Wei; Yuan, Zheng-Quan

2005-01-01

AIM: To detect the common intestinal pathogenic bacteria quickly and accurately. METHODS: A rapid (<3 h) experimental procedure was set up based upon the gene chip technology. Target genes were amplified and hybridized by oligonucleotide microarrays. RESULTS: One hundred and seventy strains of bacteria in pure culture belonging to 11 genera were successfully discriminated under comparatively same conditions, and a series of specific hybridization maps corresponding to each kind of bacteria were obtained. When this method was applied to 26 divided cultures, 25 (96.2%) were identified. CONCLUSION: Salmonella sp., Escherichia coli, Shigella sp., Listeria monocytogenes, Vibrio parahaemolyticus, Staphylococcus aureus, Proteus sp., Bacillus cereus, Vibrio cholerae, Enterococcus faecalis, Yersinia enterocolitica, and Campylobacter jejuni can be detected and identified by our microarrays. The accuracy, range, and discrimination power of this assay can be continually improved by adding further oligonucleotides to the arrays without any significant increase of complexity or cost. PMID:16437687

Quartz-Crystal Microbalance (QCM) for Public Health: An Overview of Its Applications.

PubMed

Bragazzi, Nicola Luigi; Amicizia, Daniela; Panatto, Donatella; Tramalloni, Daniela; Valle, Ivana; Gasparini, Roberto

2015-01-01

Nanobiotechnologies, from the convergence of nanotechnology and molecular biology and postgenomics medicine, play a major role in the field of public health. This overview summarizes the potentiality of piezoelectric sensors, and in particular, of quartz-crystal microbalance (QCM), a physical nanogram-sensitive device. QCM enables the rapid, real time, on-site detection of pathogens with an enormous burden in public health, such as influenza and other respiratory viruses, hepatitis B virus (HBV), and drug-resistant bacteria, among others. Further, it allows to detect food allergens, food-borne pathogens, such as Escherichia coli and Salmonella typhimurium, and food chemical contaminants, as well as water-borne microorganisms and environmental contaminants. Moreover, QCM holds promises in early cancer detection and screening of new antiblastic drugs. Applications for monitoring biohazards, for assuring homeland security, and preventing bioterrorism are also discussed. © 2015 Elsevier Inc. All rights reserved.

Characterization and Antimicrobial Resistance of Salmonella Typhimurium Isolates from Clinically Diseased Pigs in Korea.

PubMed

Oh, Sang-Ik; Kim, Jong Wan; Chae, Myeongju; Jung, Ji-A; So, Byungjae; Kim, Bumseok; Kim, Ha-Young

2016-11-01

This study investigated the prevalence of Salmonella enterica serovar and antimicrobial resistance in Salmonella Typhimurium isolates from clinically diseased pigs collected from 2008 to 2014 in Korea. Isolates were also characterized according to the presence of antimicrobial resistance genes and pulsed-field gel electrophoresis patterns. Among 94 Salmonella isolates, 81 (86.2%) were identified as being of the Salmonella Typhimurium serotype, followed by Salmonella Derby (6 of 94, 6.4%), Salmonella 4,[5],12:i:- (4 of 94, 4.3%), Salmonella Enteritidis (2 of 94, 2.1%), and Salmonella Brandenburg (1 of 94, 1.1%). The majority of Salmonella Typhimurium isolates were resistant to tetracycline (92.6%), followed by streptomycin (88.9%) and ampicillin (80.2%). Overall, 96.3% of Salmonella Typhimurium isolates showed multidrug-resistant phenotypes and commonly harbored the resistance genes bla TEM (64.9%), flo (32.8%), aadA (55.3%), strA (58.5%), strB (58.5%), sulII (53.2%), and tetA (61.7%). The pulsed-field gel electrophoresis analysis of 45 Salmonella Typhimurium isolates from individual farms revealed 27 distinct patterns that formed one major and two minor clusters in the dendrogram analysis, suggesting that most of the isolates (91.1%) from diseased pigs were genetically related. These findings can assist veterinarians in the selection of appropriate antimicrobial agents to combat Salmonella Typhimurium infections in pigs. Furthermore, they highlight the importance of continuous surveillance of antimicrobial resistance and genetic status in Salmonella Typhimurium for the detection of emerging resistance trends.

Effect of drinking-water administration of experimental chlorate ion preparations on Salmonella enterica serovar Typhimurium colonization in weaned and finished pigs.

PubMed

Anderson, R C; Hume, M E; Genovese, K J; Callaway, T R; Jung, Y S; Edrington, T S; Poole, T L; Harvey, R B; Bischoff, K M; Nisbet, D J

2004-04-01

Foodborne disease caused by Salmonella is of public health and economic significance. In order to assess the practical effectiveness of a new intervention strategy, experimental chlorate preparations (ECP) were administered via the drinking water to weaned and finished pigs that had been orally challenged the previous day with 10(9)-10(10) colony-forming units of Salmonella serovar Typhimurium. After 24 or 36 h ad libitum access to 0X, 1X or 2X ECP treatment (where X is the concentration estimated to deliver a minimal daily effective dose), the pigs were euthanized and gut contents and lymph tissue collected at necropsy were cultured for the challenge Salmonella. Drinking water administration of ECP effectively reduced (p < 0.05) caecal Salmonella concentrations and, with the weaned pigs, tended (p < or = 0.10) to reduce rectal Salmonella concentrations. No negative effects of ECP treatment on water intake and animal wellbeing were observed and only marginal effects on gut fermentation characteristics occurred. The bactericidal effect of administering ECP in drinking water was relatively rapid, with reductions in caecal Salmonella concentrations occurring within 24 h. These results suggest that ECP administered to pigs just days before slaughter may reduce gut concentrations of Salmonella; however, the impacts of such reductions on slaughter hygiene have yet to be determined.

A biosensor platform for rapid detection of E. coli in drinking water.

PubMed

Hesari, Nikou; Alum, Absar; Elzein, Mohamad; Abbaszadegan, Morteza

2016-02-01

There remains a need for rapid, specific and sensitive assays for the detection of bacterial indicators for water quality monitoring. In this study, a strategy for rapid detection of Escherichia coli in drinking water has been developed. This strategy is based on the use of the substrate 4-methylumbelliferyl-β-d-glucuronide (MUG), which is hydrolyzed rapidly by the action of E. coli β-d-glucuronidase (GUD) enzyme to yield a fluorogenic 4-methylumbelliferone (4-MU) product that can be quantified and related to the number of E. coli cells present in water samples. In this study, the detection time required for the biosensor response ranged between 20 and 120 min, depending on the number of bacteria in the sample. This approach does not need extensive sample processing with a rapid detection capability. The specificity of the MUG substrate was examined in both, pure cultures of non-target bacterial genera such as Klebsiella, Salmonella, Enterobacter and Bacillus. Non-target substrates that included 4-methylumbelliferyl-β-d-galactopyranoside (MUGal) and l-leucine β-naphthylamide aminopeptidase (LLβ-N) were also investigated to identify nonspecific patterns of enzymatic activities in E. coli. GUD activity was found to be specific for E. coli and no further enzymatic activity was detected by other species. In addition, fluorescence assays were performed for the detection of E. coli to generate standard curves; and the sensitivity of the GUD enzymatic response was measured and repeatedly determined to be less than 10 E. coli cells in a reaction vial. The applicability of the method was tested by performing multiple fluorescence assays under pure and mixed bacterial flora in environmental samples. The results of this study showed that the fluorescence signals generated in samples using specific substrate molecules can be utilized to develop a bio-sensing platform for the detection of E. coli in drinking water. Furthermore, this system can be applied independently or in conjunction with other methods as a part of an array of biochemical assays in order to reliably detect E. coli in water. Copyright © 2015 Elsevier Inc. All rights reserved.

Complete genome sequence of a ciprofloxacin resistant Salmonella enterica subsp. enterica serovar Kentucky sequence of a ciprofloxacin strain, PU131, isolated from a human patient in Washington State.

USDA-ARS?s Scientific Manuscript database

A ciprofloxacin resistant (CipR) Salmonella enterica subsp. enterica serovar Kentucky ST198 has rapidly and extensively disseminated globally to become a major food-safety and public health concern. Here, we report a complete genome sequence of a CipR S. Kentucky ST198 strain PU131 isolated from a ...

Interdigitated microelectrode (IME) impedance sensor for the detection of viable Salmonella typhimurium.

PubMed

Yang, Liju; Li, Yanbin; Griffis, Carl L; Johnson, Michael G

2004-05-15

Interdigitated microelectrodes (IMEs) were used as impedance sensors for rapid detection of viable Salmonella typhimurium in a selective medium and milk samples. The impedance growth curves, impedance against bacterial growth time, were recorded at four frequencies (10Hz, 100Hz, 1kHz, and 10kHz) during the growth of S. typhimurium. The impedance did not change until the cell number reached 10(5)-10(6) CFUml(-1). The greatest change in impedance was observed at 10Hz. To better understand the mechanism of the IME impedance sensor, an equivalent electrical circuit, consisting of double layer capacitors, a dielectric capacitor, and a medium resistor, was introduced and used for interpreting the change in impedance during bacterial growth. Bacterial attachment to the electrode surface was observed with scanning electron microscopy, and it had effect on the impedance measurement. The detection time, t(D), defined as the time for the impedance to start change, was obtained from the impedance growth curve at 10Hz and had a linear relationship with the logarithmic value of the initial cell number of S. typhimurium in the medium and milk samples. The regression equations for the cell numbers between 4.8 and 5.4 x 10(5) CFUml(-1) were t(D) = -1.38 log N + 10.18 with R(2) = 0.99 in the pure medium and t(D) = -1.54 log N + 11.33 with R(2) = 0.98 in milk samples, respectively. The detection times for 4.8 and 5.4 x 10(5) CFUml(-1) initial cell numbers were 9.3 and 2.2 h, respectively, and the detection limit could be as low as 1 cell in a sample.

Rapid Sample Processing for Detection of Food-Borne Pathogens via Cross-Flow Microfiltration

PubMed Central

Li, Xuan; Ximenes, Eduardo; Amalaradjou, Mary Anne Roshni; Vibbert, Hunter B.; Foster, Kirk; Jones, Jim; Liu, Xingya; Bhunia, Arun K.

2013-01-01

This paper reports an approach to enable rapid concentration and recovery of bacterial cells from aqueous chicken homogenates as a preanalytical step of detection. This approach includes biochemical pretreatment and prefiltration of food samples and development of an automated cell concentration instrument based on cross-flow microfiltration. A polysulfone hollow-fiber membrane module having a nominal pore size of 0.2 μm constitutes the core of the cell concentration instrument. The aqueous chicken homogenate samples were circulated within the cross-flow system achieving 500- to 1,000-fold concentration of inoculated Salmonella enterica serovar Enteritidis and naturally occurring microbiota with 70% recovery of viable cells as determined by plate counting and quantitative PCR (qPCR) within 35 to 45 min. These steps enabled 10 CFU/ml microorganisms in chicken homogenates or 102 CFU/g chicken to be quantified. Cleaning and sterilizing the instrument and membrane module by stepwise hydraulic and chemical cleaning (sodium hydroxide and ethanol) enabled reuse of the membrane 15 times before replacement. This approach begins to address the critical need for the food industry for detecting food pathogens within 6 h or less. PMID:24014538

Imported edible leaves collected at retail sale in England during 2017 with an emphasis on betel and curry leaves: Microbiological quality with respect to Salmonella, Shiga-toxin-producing E. coli (STEC) and levels of Escherichia coli.

PubMed

McLauchlin, J; Aird, H; Charlett, A; Chattaway, M; Elviss, N; Hartman, H; Jenkins, C; Jørgensen, F; Larkin, L; Sadler-Reeves, L; Willis, C

2018-05-26

to investigate the microbiological quality of imported fresh leaves on retail-sale during 2017 with respect to Salmonella, Shiga-toxin-producing Escherichia coli (STEC) and levels of E.coli. 279 samples of imported edible leaves (69 banana, 77 betel, 118 curry and 15 other types) were tested. Salmonella spp. which were confirmed by whole genome sequencing and isolated from 44 samples, 26% from curry leaves, 14% from betel and 2.4% from all other leaf types: 80% of all samples contained ≥10 2 , 44% ≥10 3 and 22% ≥10 4 cfu of E.coli cfu/g. All samples where Salmonella were detected also yielded ≥20 cfu of E.coli/g. 54 samples were tested for STEC which was detected in 6 samples and isolated from three: one was identified as STEC O157:H7. this report further highlights an ongoing problem of Salmonella contamination of imported fresh edible leaves. amongst all food tested by Public Health England (approximately 11,000 per annum), curry leaves were the herb most commonly contaminated with Salmonella, and betel leaves were the most commonly contaminated ready-to-eat food. The high proportion with unsatisfactory E. coli levels and the detection of STEC suggests risks of contamination by multiple enteric pathogens. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Collaborative ring-trial of Dynabeads anti-Salmonella for immunomagnetic separation of stressed Salmonella cells from herbs and spices.

PubMed

Mansfield, L; Forsythe, S

1996-02-01

Eight laboratories participated in a Salmonella detection ring-trial which compared selective enrichment by conventional broths with immunomagnetic separation (IMS) using Dynabeads Anti-Salmonella. Laboratories analyzed six types of herbs and spices that were spiked with one of six freeze-dried Salmonella species. Each herb and spice analysis comprised of 12 samples (25 g each) which had been spiked at three different levels, plus a negative control and stored for one week prior to testing. Out of a total 468 samples analyzed, 195 (41.7%) were positive by both methods. Eighteen samples were positive only by IMS enrichment, in comparison with 19 positive samples by conventional enrichment broths and not IMS. These results confirm the potential use of IMS as an alternative to enrichment broths for Salmonella isolation.

Trisodium phosphate and sodium hypochlorite are more effective as antimicrobials against Campylobacter and Salmonella on duck as compared to chicken meat.

PubMed

Sarjit, Amreeta; Dykes, Gary A

2015-06-16

Little work has been reported on the use of commercial antimicrobials against foodborne pathogens on duck meat. We investigated the effectiveness of trisodium phosphate (TSP) and sodium hypochlorite (SH) as antimicrobial treatments against Campylobacter and Salmonella on duck meat under simulated commercial water chilling conditions. The results were compared to the same treatments on well-studied chicken meat. A six strain Campylobacter or Salmonella cocktail was inoculated (5 ml) at two dilution levels (10(4) and 10(8) cfu/ml) onto 25 g duck or chicken meat with skin and allowed to attach for 10 min. The meat was exposed to three concentrations of pH adjusted TSP (8, 10 and 12% (w/v), pH 11.5) or SH (40, 50 and 60 ppm, pH 5.5) in 30 ml water under simulated spin chiller conditions (4 °C, agitation) for 10 min. In a parallel experiment the meat was placed in the antimicrobial treatments before inoculation and bacterial cocktails were added to the meat after the antimicrobial solution was removed while all other parameters were maintained. Untreated controls and controls using water were included in all experiments. Bacterial numbers were determined on Campylobacter blood-free selective agar and Mueller Hinton agar or xylose deoxycholate agar and tryptone soya agar using the thin agar layer method for Campylobacter and Salmonella, respectively. All TSP concentrations significantly (p<0.05) reduced numbers of Campylobacter (~1.2-6.4 log cfu/cm(2)) and Salmonella (~0.4-6.6 log cfu/cm(2)) on both duck and chicken meat. On duck meat, numbers of Campylobacter were less than the limit of detection at higher concentrations of TSP and numbers of Salmonella were less than the limit of detection at all concentrations of TSP except one. On chicken meat, numbers of Campylobacter and Salmonella were less than the limit of detection only at the lower inoculum level and higher TSP concentrations. By contrast only some of the concentrations of SH significantly (p<0.05) reduced numbers of Campylobacter and Salmonella (~0.2-1.5 log cfu/cm(2)) on both duck and chicken meats. None of the SH treatments resulted in numbers of either pathogen being less than limit of detection. Results indicate that chicken meat has the ability to effectively protect Campylobacter and Salmonella against the impact of trisodium phosphate and sodium hypochlorite while duck meat does not. This study suggests that trisodium phosphate has a strong potential for application in a commercial poultry processing to reduce Campylobacter and Salmonella specifically on duck meat. Copyright © 2015 Elsevier B.V. All rights reserved.

The efficacy of different cleaning and disinfection procedures to reduce Salmonella and Enterobacteriaceae in the lairage environment of a pig abattoir.

PubMed

Walia, Kavita; Argüello, Hector; Lynch, Helen; Grant, Jim; Leonard, Finola C; Lawlor, Peadar G; Gardiner, Gillian E; Duffy, Geraldine

2017-04-04

This study investigated several cleaning and disinfection protocols for their ability to eliminate Salmonella and to reduce levels of Enterobacteriaceae, within the lairage pens of a commercial pig abattoir. Eight protocols were evaluated in each of 12 lairage pens at the end of the slaughtering day on 3 occasions (36 pens/protocol): (P1) high-pressure cold water wash (herein referred to as high-pressure wash); (P2) high-pressure wash followed by a quaternary ammonium compound (QAC)-based disinfectant without rinsing; (P3) high-pressure wash followed by a chlorocresol-based disinfectant without rinsing; (P4) high-pressure wash followed by a sodium hydroxide/sodium hypochlorite detergent with rinsing; (P5) P4 followed by P2; (P6) P4 followed by P3; (P7) P5 with drying for 24-48h; and (P8) P6 with drying for 24-48h. Two floor swabs and one wall swab were taken from each lairage pen before and after each protocol was applied, and examined for the presence of Salmonella and enumeration of Enterobacteriaceae. High-pressure washing alone (P1) did not reduce the prevalence of Salmonella in the lairage pens. When high-pressure washing, the probability of detecting Salmonella following application of the chlorocresol-based disinfectant (P3) was lower than with the QAC-based disinfectant, P2 (14.2% versus 34.0%, respectively; p<0.05). The probability of detecting Salmonella after the combined use of detergent and the chlorocresol-based disinfectant (P6) was also lower than application of detergent followed by the QAC-based disinfectant, P5 (2.2% versus 17.1%, respectively; p<0.05). Drying of pens (P7 and P8) greatly reduced the probability of detecting Salmonella. Only 3.8% of swabs were Salmonella-positive 48h after cleaning with detergent and the QAC-based disinfectant (P7); while an eradication of Salmonella was achieved 24h after cleaning with detergent and the chlorocresol-based disinfectant, P8. A reduction in Enterobacteriaceae counts to below the limit of detection (LOD; 10CFU/cm 2 ) was achieved following cleaning with detergent and disinfection with the chlorocresol-based disinfectant, regardless of drying (p<0.05), whereas, applying detergent and the QAC-based disinfectant (P7) did not reduce Enterobacteriaceae counts to below the LOD. Therefore ensuring that lairage pens are allowed to dry after intensive cleaning with detergent and a chlorocresol-based disinfectant is recommended as the most effective hygiene routine to eliminate Salmonella and reduce Enterobacteriaceae counts. Copyright © 2017 Elsevier B.V. All rights reserved.

Assessing the Impact of Manure Application in Commercial Swine Farms on the Transmission of Antimicrobial Resistant Salmonella in the Environment.

PubMed

Pornsukarom, Suchawan; Thakur, Siddhartha

2016-01-01

Land application of swine manure in commercial hog farms is an integral part of their waste management system which recycles the nutrients back to the soil. However, manure application can lead to the dissemination of bacterial pathogens in the environment and pose a serious public health threat. The aim of this study was to determine the dissemination of antimicrobial resistant Salmonella in the environment due to manure application in commercial swine farms in North Carolina (n = 6) and Iowa (n = 7), two leading pork producing states in the US. We collected manure and soil samples twice on day 0 (before and after manure application) from four distinct plots of lands (5 soil samples/plot) located at 20 feet away from each other in the field. Subsequent soil samples were collected again on days 7, 14, 21 from the same plots. A total of 1,300 soil samples (NC = 600; IA = 700) and 130 manure samples (NC = 60; IA = 70) were collected and analyzed in this study. The overall Salmonella prevalence was 13.22% (189/1,430), represented by 10.69% and 38.46% prevalence in soil and manure, respectively. The prevalence in NC (25.45%) was significantly higher than in IA (2.73%) (P<0.001) and a consistent decrease in Salmonella prevalence was detected from Day 0-Day 21 in all the farms that tested positive. Salmonella serotypes detected in NC were not detected in IA, thereby highlighting serotype association based on manure storage and soil application method used in the two regions. Antimicrobial susceptibility testing was done by the broth microdilution method to a panel of 15 antimicrobial drugs. A high frequency of isolates (58.73%) were multidrug resistant (resistance to three or more class of antimicrobials) and the most frequent resistance was detected against streptomycin (88.36%), sulfisoxazole (67.2%), and tetracycline (57.67%). Genotypic characterization by pulse field gel electrophoresis revealed clonally related Salmonella in both manure and soil at multiple time points in the positive farms. Our study highlights the potential role of swine manure application in the dissemination and persistence of antimicrobial resistant Salmonella in the environment.

Assessing the Impact of Manure Application in Commercial Swine Farms on the Transmission of Antimicrobial Resistant Salmonella in the Environment

PubMed Central

Pornsukarom, Suchawan; Thakur, Siddhartha

2016-01-01

Land application of swine manure in commercial hog farms is an integral part of their waste management system which recycles the nutrients back to the soil. However, manure application can lead to the dissemination of bacterial pathogens in the environment and pose a serious public health threat. The aim of this study was to determine the dissemination of antimicrobial resistant Salmonella in the environment due to manure application in commercial swine farms in North Carolina (n = 6) and Iowa (n = 7), two leading pork producing states in the US. We collected manure and soil samples twice on day 0 (before and after manure application) from four distinct plots of lands (5 soil samples/plot) located at 20 feet away from each other in the field. Subsequent soil samples were collected again on days 7, 14, 21 from the same plots. A total of 1,300 soil samples (NC = 600; IA = 700) and 130 manure samples (NC = 60; IA = 70) were collected and analyzed in this study. The overall Salmonella prevalence was 13.22% (189/1,430), represented by 10.69% and 38.46% prevalence in soil and manure, respectively. The prevalence in NC (25.45%) was significantly higher than in IA (2.73%) (P<0.001) and a consistent decrease in Salmonella prevalence was detected from Day 0-Day 21 in all the farms that tested positive. Salmonella serotypes detected in NC were not detected in IA, thereby highlighting serotype association based on manure storage and soil application method used in the two regions. Antimicrobial susceptibility testing was done by the broth microdilution method to a panel of 15 antimicrobial drugs. A high frequency of isolates (58.73%) were multidrug resistant (resistance to three or more class of antimicrobials) and the most frequent resistance was detected against streptomycin (88.36%), sulfisoxazole (67.2%), and tetracycline (57.67%). Genotypic characterization by pulse field gel electrophoresis revealed clonally related Salmonella in both manure and soil at multiple time points in the positive farms. Our study highlights the potential role of swine manure application in the dissemination and persistence of antimicrobial resistant Salmonella in the environment. PMID:27755598

Prevalence and antimicrobial susceptibilities of Vibrio, salmonella, and Aeromonas isolates from various uncooked seafoods in Thailand.

PubMed

Woodring, Joseph; Srijan, Apichai; Puripunyakom, Paksathorn; Oransathid, Wilawan; Wongstitwilairoong, Boonchai; Mason, Carl

2012-01-01

Uncooked seafood samples were collected from open markets and supermarkets in Bangkok, Thailand, and were examined for the presence of Vibrio, Salmonella, and Aeromonas species from January to February 2008. From 120 samples, 272 bacterial isolates were identified through biochemical testing. Of all sea bass, shrimp, oyster, and blood cockle samples (30 of each) that were processed for culture, 114 (95%) samples had at least one detectable isolate of Vibrio, Salmonella, or Aeromonas, leaving only 6 (5%) samples free of them. All oyster sample (100%) had at least one pathogen, followed by sea bass (97%), blood cockles (97%), and shrimp (90%). Overall, 111 (92%) of all samples had detectable Vibrio spp., 32 (27%) had detectable Aeromonas spp., and 25 (21%) had detectable Salmonella enterica. There was no overall difference between positive samples collected from fresh markets versus supermarkets (relative risk, 0.97; 95% CI, 0.89 to 1.05). Resistance to ampicillin among isolated pathogens was relatively high (56%), while resistance to 12 other antibiotics, including azithromycin, ciprofloxacin, and trimethoprim-sulfamethoxazole, was relatively low (0, 0, and 3%, respectively). Study results indicate that uncooked seafood in Bangkok, Thailand, commonly harbors enteric pathogens and that consumption of uncooked seafood should be avoided to reduce foodborne illnesses.

Droplet-based immunoassay on a 'sticky' nanofibrous surface for multiplexed and dual detection of bacteria using smartphones.

PubMed

Nicolini, Ariana M; Fronczek, Christopher F; Yoon, Jeong-Yeol

2015-05-15

We have developed a rapid, sensitive, and specific droplet-based immunoassay for the detection of Escherichia coli and Salmonella within a single-pipetted sample. Polycaprolactone (PCL) electrospun fibers on indium-tin-oxide (ITO) glass provide a sufficient surface to render a non-slip droplet condition, and while the PCL fibers lend a local hydrophilicity (contact angle θ=74°) for sufficient sub-micron particle adhesion, air pockets within the fibers lend an apparent hydrophobicity. Overall, the contact angle of water on this electrospun surface is 119°, and the air pockets cause the droplet to be completely immobile and resistant to movement, protecting it from external vibration. By using both anti-E. coli conjugated, 510 nm diameter green fluorescent particles (480 nm excitation and 520 nm emission) and anti-Salmonella conjugated, 400 nm diameter red fluorescent particles (640 nm excitation and 690 nm emission), we can detect multiple targets in a single droplet. Using appropriate light sources guided by fiber optics, we determined a detection limit of 10(2) CFU mL(-1). Immunoagglutination can be observed under a fluorescence microscope. Fluorescence detection (at the emission wavelength) of immunoagglutination was maximum at 90° from the incident light, while light scattering (at the excitation wavelength) was still present and behaved similarly, indicating the ability of double detection, greatly improving credibility and reproducibility of the assay. A power function (light intensity) simulation of elastic Mie scatter confirmed that both fluorescence and light scattering were present. Due to the size of the fluorescent particles relative to their incident excitation wavelengths, Mie scatter conditions were observed, and fluorescence signals show a similar trend to light scattering signals. Smartphone detection was included for true portable detection, in which the high contact angle pinning of the droplet makes this format re-usable and re-configurable. Copyright © 2014 Elsevier B.V. All rights reserved.

Landscape and meteorological factors affecting prevalence of three food-borne pathogens in fruit and vegetable farms.

PubMed

Strawn, Laura K; Fortes, Esther D; Bihn, Elizabeth A; Nightingale, Kendra K; Gröhn, Yrjö T; Worobo, Randy W; Wiedmann, Martin; Bergholz, Peter W

2013-01-01

Produce-related outbreaks have been traced back to the preharvest environment. A longitudinal study was conducted on five farms in New York State to characterize the prevalence, persistence, and diversity of food-borne pathogens in fresh produce fields and to determine landscape and meteorological factors that predict their presence. Produce fields were sampled four times per year for 2 years. A total of 588 samples were analyzed for Listeria monocytogenes, Salmonella, and Shiga toxin-producing Escherichia coli (STEC). The prevalence measures of L. monocytogenes, Salmonella, and STEC were 15.0, 4.6, and 2.7%, respectively. L. monocytogenes and Salmonella were detected more frequently in water samples, while STEC was detected with equal frequency across all sample types (soil, water, feces, and drag swabs). L. monocytogenes sigB gene allelic types 57, 58, and 61 and Salmonella enterica serovar Cerro were repeatedly isolated from water samples. Soil available water storage (AWS), temperature, and proximity to three land cover classes (water, roads and urban development, and pasture/hay grass) influenced the likelihood of detecting L. monocytogenes. Drainage class, AWS, and precipitation were identified as important factors in Salmonella detection. This information was used in a geographic information system framework to hypothesize locations of environmental reservoirs where the prevalence of food-borne pathogens may be elevated. The map indicated that not all croplands are equally likely to contain environmental reservoirs of L. monocytogenes. These findings advance recommendations to minimize the risk of preharvest contamination by enhancing models of the environmental constraints on the survival and persistence of food-borne pathogens in fields.

Landscape and Meteorological Factors Affecting Prevalence of Three Food-Borne Pathogens in Fruit and Vegetable Farms

PubMed Central

Strawn, Laura K.; Fortes, Esther D.; Bihn, Elizabeth A.; Nightingale, Kendra K.; Gröhn, Yrjö T.; Worobo, Randy W.; Wiedmann, Martin

2013-01-01

Produce-related outbreaks have been traced back to the preharvest environment. A longitudinal study was conducted on five farms in New York State to characterize the prevalence, persistence, and diversity of food-borne pathogens in fresh produce fields and to determine landscape and meteorological factors that predict their presence. Produce fields were sampled four times per year for 2 years. A total of 588 samples were analyzed for Listeria monocytogenes, Salmonella, and Shiga toxin-producing Escherichia coli (STEC). The prevalence measures of L. monocytogenes, Salmonella, and STEC were 15.0, 4.6, and 2.7%, respectively. L. monocytogenes and Salmonella were detected more frequently in water samples, while STEC was detected with equal frequency across all sample types (soil, water, feces, and drag swabs). L. monocytogenes sigB gene allelic types 57, 58, and 61 and Salmonella enterica serovar Cerro were repeatedly isolated from water samples. Soil available water storage (AWS), temperature, and proximity to three land cover classes (water, roads and urban development, and pasture/hay grass) influenced the likelihood of detecting L. monocytogenes. Drainage class, AWS, and precipitation were identified as important factors in Salmonella detection. This information was used in a geographic information system framework to hypothesize locations of environmental reservoirs where the prevalence of food-borne pathogens may be elevated. The map indicated that not all croplands are equally likely to contain environmental reservoirs of L. monocytogenes. These findings advance recommendations to minimize the risk of preharvest contamination by enhancing models of the environmental constraints on the survival and persistence of food-borne pathogens in fields. PMID:23144137

A longitudinal observational study of Salmonella shedding patterns by commercial turkeys during rearing and fattening, showing limitations of some control measures.

PubMed

Morris, V K; Carrique-Mas, J J; Mueller-Doblies, D; Davies, R H; Wales, A D; Allen, V M

2015-01-01

1. The onset and progression of Salmonella infections was investigated in commercial turkey flocks from placement at 1 d old until slaughter in "brood and move" systems using a longitudinal observational approach based on faeces and environmental sampling with subsequent culture of Salmonella. 2. Persistent Salmonella Newport contamination was found within rearing houses and on their external concrete aprons after cleaning and disinfection between crops of heavily shedding young birds. 3. Salmonella shedding was often detected by 5 d of age and the frequency of positive samples peaked at 14-35 d. Thereafter Salmonella isolations declined, especially in the later (fattening) stages. Samples were still Salmonella-positive at low prevalence in half of the intensively sampled houses at slaughter age. 4. A number of management interventions to combat Salmonella infection of flocks, including sourcing policy, competitive exclusion cultures and cleaning and disinfection, were inadequate to prevent flock infection, although improved disinfection on one unit was associated with a delay in the onset of flock infection.

Fate of Salmonella Typhimurium in laboratory-scale drinking water biofilms.

PubMed

Schaefer, L M; Brözel, V S; Venter, S N

2013-12-01

Investigations were carried out to evaluate and quantify colonization of laboratory-scale drinking water biofilms by a chromosomally green fluorescent protein (gfp)-tagged strain of Salmonella Typhimurium. Gfp encodes the green fluorescent protein and thus allows in situ detection of undisturbed cells and is ideally suited for monitoring Salmonella in biofilms. The fate and persistence of non-typhoidal Salmonella in simulated drinking water biofilms was investigated. The ability of Salmonella to form biofilms in monoculture and the fate and persistence of Salmonella in a mixed aquatic biofilm was examined. In monoculture S. Typhimurium formed loosely structured biofilms. Salmonella colonized established multi-species drinking water biofilms within 24 hours, forming micro-colonies within the biofilm. S. Typhimurium was also released at high levels from the drinking water-associated biofilm into the water passing through the system. This indicated that Salmonella could enter into, survive and grow within, and be released from a drinking water biofilm. The ability of Salmonella to survive and persist in a drinking water biofilm, and be released at high levels into the flow for recolonization elsewhere, indicates the potential for a persistent health risk to consumers once a network becomes contaminated with this bacterium.

Immunity to Intracellular Salmonella Depends on Surface-associated Antigens

PubMed Central

Claudi, Beatrice; Mazé, Alain; Schemmer, Anne K.; Kirchhoff, Dennis; Schmidt, Alexander; Burton, Neil; Bumann, Dirk

2012-01-01

Invasive Salmonella infection is an important health problem that is worsening because of rising antimicrobial resistance and changing Salmonella serovar spectrum. Novel vaccines with broad serovar coverage are needed, but suitable protective antigens remain largely unknown. Here, we tested 37 broadly conserved Salmonella antigens in a mouse typhoid fever model, and identified antigen candidates that conferred partial protection against lethal disease. Antigen properties such as high in vivo abundance or immunodominance in convalescent individuals were not required for protectivity, but all promising antigen candidates were associated with the Salmonella surface. Surprisingly, this was not due to superior immunogenicity of surface antigens compared to internal antigens as had been suggested by previous studies and novel findings for CD4 T cell responses to model antigens. Confocal microscopy of infected tissues revealed that many live Salmonella resided alone in infected host macrophages with no damaged Salmonella releasing internal antigens in their vicinity. In the absence of accessible internal antigens, detection of these infected cells might require CD4 T cell recognition of Salmonella surface-associated antigens that could be processed and presented even from intact Salmonella. In conclusion, our findings might pave the way for development of an efficacious Salmonella vaccine with broad serovar coverage, and suggest a similar crucial role of surface antigens for immunity to both extracellular and intracellular pathogens. PMID:23093937

Prevalence of Salmonella enterica, Listeria monocytogenes, and pathogenic Escherichia coli in bulk tank milk and milk filters from US dairy operations in the National Animal Health Monitoring System Dairy 2014 study.

PubMed

Sonnier, Jakeitha L; Karns, Jeffrey S; Lombard, Jason E; Kopral, Christine A; Haley, Bradd J; Kim, Seon-Woo; Van Kessel, Jo Ann S

2018-03-01

The dairy farm environment is a well-documented reservoir for zoonotic pathogens such as Salmonella enterica, Shiga-toxigenic Escherichia coli, and Listeria monocytogenes, and humans may be exposed to these pathogens via consumption of unpasteurized milk and dairy products. As part of the National Animal Health Monitoring System Dairy 2014 study, bulk tank milk (BTM, n = 234) and milk filters (n = 254) were collected from a total of 234 dairy operations in 17 major dairy states and analyzed for the presence of these pathogens. The invA gene was detected in samples from 18.5% of operations and Salmonella enterica was isolated from 18.0% of operations. Salmonella Dublin was detected in 0.7% of operations. Sixteen Salmonella serotypes were isolated, and the most common serotypes were Cerro, Montevideo, and Newport. Representative Salmonella isolates (n = 137) were tested against a panel of 14 antimicrobials. Most (85%) were pansusceptible; the remaining were resistant to 1 to 9 antimicrobials, and within the resistant strains the most common profile was resistance to ampicillin/clavulanic acid, ampicillin, cefoxitin, ceftiofur, ceftriaxone, chloramphenicol, streptomycin, sulfisoxazole, and tetracycline. Listeria spp. were isolated from 19.9% of operations, and L. monocytogenes was isolated from 3.0% of operations. Serogroups 1/2a and 1/2b were the most common, followed by 4b and 4a. One or more E. coli virulence genes were detected in the BTM from 30.5% of operations and in the filters from 75.3% of operations. A combination of stx 2 , eaeA, and γ-tir genes was detected in the BTM from 0.5% of operations and in the filters from 6.6% of operations. The results of this study indicate an appreciable prevalence of bacterial pathogens in BTM and filters, including serovars known to infect humans. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Prevalence and antibiotic resistance of Salmonella spp. in meat products, meat preparations and minced meat

NASA Astrophysics Data System (ADS)

Rašeta, M.; Mrdović, B.; Janković, V.; Bečkei, Z.; Lakićević, B.; Vidanović, D.; Polaček, V.

2017-09-01

This study aimed to determine Salmonella spp. prevalence in meat products, meat preparations and minced meat. Over a period of three years, a total of 300 samples were taken (100 RTE meat products, 100 meat preparations and 100 minced meat) and examined for the presence of Salmonella spp. Sampling was carried out at the warehouses of the food manufacturers. Salmonella spp. were not detected in RTE meat products, while 7% of semi-finished meat products (fresh sausages, grill meat formed and unformed) contained Salmonella, as did 18% of minced meats (minced pork II category, minced beef II category, mixed minced meat). The 25 Salmonella isolates obtained were examined for antibiotic resistance by the disk diffusion test, according to the NCCLS and CLSI guidelines. Isolates showed resistance to ampicillin and nalidixic acid (80%), tetracycline (72%), cefotaxime/clavulanic acid (48%), but not to gentamicin (8%) or trimethoprim/sulfamethoxazole (0%).

Salmonella serotypes and their antimicrobial susceptibility in apparently healthy dogs in Addis Ababa, Ethiopia.

PubMed

Kiflu, Bitsu; Alemayehu, Haile; Abdurahaman, Mukarim; Negash, Yohannes; Eguale, Tadesse

2017-05-19

The close bond between pet animals and family members poses risk of infection with zoonotic bacterial pathogens such as Salmonella. No data is available on occurrence of Salmonella in dogs in Ethiopia. The aim of this study was therefore to determine the prevalence, serotype distribution and antimicrobial resistance of Salmonella from feces of apparently healthy dogs in Addis Ababa, Ethiopia. Of the total 360 dogs examined, 42 (11.7%; 95% Confidence limit of 8.5%-15.4%) were positive for Salmonella. Fourteen serotypes were detected and the predominant ones were S. Bronx (n = 7; 16.7%), S. Newport (n = 6; 14.3%), followed by S. Typhimurium, S. Indiana, S. Kentucky, S. Saintpaul and S. Virchow (n = 4; 9.5%) each. Salmonella infection status was significantly associated with history of symptom of diarrhea during the past 60 days (OR = 3.78; CI = 1.76-8.13; p = 0). Highest resistance rates were found for oxytetracycline (59.5%), neomycin (50%), streptomycin (38.1%), cephalothin (33.3%), doxycycline (30.9%), ampicillin (30.9%) and amoxicillin + clavulanic acid (26.2%). Thirty eight (90.5%) of the isolates were resistant or intermediately resistant to at least one of the 16 antimicrobials tested. Resistance to two or more antimicrobials was detected in 30 (71.4%) of the isolates. Resistance to three or more antimicrobials was detected in 19 (45.2%) of the isolates. This study demonstrated high carriage rate of Salmonella serotypes known for causing human salmonellosis and large proportion of them were resistant to antimicrobials used in public and veterinary medicine for management of various bacterial infections, suggesting the possible risk of infection of human population in close contact with these dogs by drug resistant pathogens. Therefore, it is vital to work on raising public awareness on zoonotic canine diseases prevention measures and good hygienic practices.

Comparison of four chromogenic media and Hektoen agar for detection and presumptive identification of Salmonella strains in human stools.

PubMed

Perez, J M; Cavalli, P; Roure, C; Renac, R; Gille, Y; Freydiere, A M

2003-03-01

Several chromogenic media have been developed to enhance the specificity of Salmonella detection. We compared the performance of four commercial chromogenic media-namely, ABC medium (Lab M. Ltd., Bury, United Kingdom), COMPASS Salmonella agar (Biokar Diagnostics, Beauvais, France), CHROMagar Salmonella agar (CHROMagar Company, Paris, France), and SM ID agar (bioMerieux, Marcy l'Etoile, France)-with conventional Hektoen medium. Nine hundred sixteen stool samples from inpatients at three hospitals were cultured, in parallel, on the five media, both by direct inoculation and after selective enrichment in selenite broth. Sixty-four Salmonella strains with 12 serotypes were isolated on at least one medium. After 48 h of incubation, sensitivity before and after enrichment was 62.5 and 89.1% with ABC medium, 77.1 and 93.8% with COMPASS agar, 66.7 and 89.1% with CHROMagar, 68.8 and 85.9% with SM ID agar, and 85.4 and 98.4% with Hektoen agar, respectively. Broth enrichment and prolonged incubation (48 versus 24 h) increased the sensitivity of all five media. Only one strain was not isolated on Hektoen agar. The number of false-positive isolates was higher with all five media after enrichment in selenite broth and after incubation for 48 h compared to 24 h. The specificity of the four chromogenic media was better than 91% after incubation for 24 h (77.7% with Hektoen agar) and better than 84% after incubation for 48 h (74.8% with Hektoen agar). This higher specificity reduces the need for confirmatory tests, thereby cutting technical time and reagent requirements. Both COMPASS agar and CHROMagar Salmonella, which after simple additional tests showed close efficiencies (96 and 97%, respectively), can be recommended as single-plate media of choice for the detection and presumptive identification of salmonellae in stools.

The relationship between the numbers of Salmonella Enteritidis, Salmonella Heidelberg, or Salmonella Hadar colonizing reproductive tissues of experimentally infected laying hens and deposition inside eggs.

PubMed

Gast, Richard K; Guraya, Rupa; Guard, Jean; Holt, Peter S

2011-06-01

Contamination of eggs by Salmonella Enteritidis has been a prominent cause of human illness for several decades and is the focus of a recently implemented national regulatory plan for egg-producing flocks in the United States. Salmonella Heidelberg has also been identified as an egg-transmitted pathogen. The deposition of Salmonella strains inside eggs is a consequence of reproductive tract colonization in infected laying hens, but prior research has not determined the relationship between the numbers of Salmonella that colonize reproductive organs and the associated frequency of egg contamination. In the present study, groups of laying hens in two trials were experimentally infected with large oral doses of strains of Salmonella Enteritidis (phage type 13a), Salmonella Heidelberg, or Salmonella Hadar. Reproductive tissues of selected hens were cultured to detect and enumerate Salmonella at 5 days postinoculation, and the interior contents of eggs laid between 6 and 25 days postinoculation were tested for contamination. Significantly more internally contaminated eggs were laid by hens infected with Salmonella Enteritidis (3.58%) than with strains of either Salmonella Heidelberg (0.47%) or Salmonella Hadar (0%). However, no significant differences were observed between Salmonella strains in either isolation frequency or the number of colony-forming units (CFU) isolated from ovaries or oviducts. Salmonella isolation frequencies ranged from 20.8% to 41.7% for ovaries and from 8.3% to 33.3% for oviducts. Mean Salmonella colonization levels ranged from 0.10 to 0.51 log CFU/g for ovaries and from 0.25 to 0.46 log CFU/g for oviducts. Although parallel rank-orders were observed for Salmonella enumeration (in both ovaries and oviducts) and egg contamination frequency, a statistically significant relationship could not be established between these two parameters of infection.

Rapid detection of malignant bio-species using digital holographic pattern recognition and nano-photonics

NASA Astrophysics Data System (ADS)

Sarkisov, Sergey S.; Kukhtareva, Tatiana; Kukhtarev, Nickolai V.; Curley, Michael J.; Edwards, Vernessa; Creer, Marylyn

2013-03-01

There is a great need for rapid detection of bio-hazardous species particularly in applications to food safety and biodefense. It has been recently demonstrated that the colonies of various bio-species could be rapidly detected using culture-specific and reproducible patterns generated by scattered non-coherent light. However, the method heavily relies on a digital pattern recognition algorithm, which is rather complex, requires substantial computational power and is prone to ambiguities due to shift, scale, or orientation mismatch between the analyzed pattern and the reference from the library. The improvement could be made, if, in addition to the intensity of the scattered optical wave, its phase would be also simultaneously recorded and used for the digital holographic pattern recognition. In this feasibility study the research team recorded digital Gabor-type (in-line) holograms of colonies of micro-organisms, such as Salmonella with a laser diode as a low-coherence light source and a lensless high-resolution (2.0x2.0 micron pixel pitch) digital image sensor. The colonies were grown in conventional Petri dishes using standard methods. The digitally recorded holograms were used for computational reconstruction of the amplitude and phase information of the optical wave diffracted on the colonies. Besides, the pattern recognition of the colony fragments using the cross-correlation between the digital hologram was also implemented. The colonies of mold fungi Altenaria sp, Rhizophus, sp, and Aspergillus sp have been also generating nano-colloidal silver during their growth in specially prepared matrices. The silver-specific plasmonic optical extinction peak at 410-nm was also used for rapid detection and growth monitoring of the fungi colonies.

Use of amplicon sequencing to improve sensitivity in PCR-based detection of microbial pathogen in environmental samples.

PubMed

Saingam, Prakit; Li, Bo; Yan, Tao

2018-06-01

DNA-based molecular detection of microbial pathogens in complex environments is still plagued by sensitivity, specificity and robustness issues. We propose to address these issues by viewing them as inadvertent consequences of requiring specific and adequate amplification (SAA) of target DNA molecules by current PCR methods. Using the invA gene of Salmonella as the model system, we investigated if next generation sequencing (NGS) can be used to directly detect target sequences in false-negative PCR reaction (PCR-NGS) in order to remove the SAA requirement from PCR. False-negative PCR and qPCR reactions were first created using serial dilutions of laboratory-prepared Salmonella genomic DNA and then analyzed directly by NGS. Target invA sequences were detected in all false-negative PCR and qPCR reactions, which lowered the method detection limits near the theoretical minimum of single gene copy detection. The capability of the PCR-NGS approach in correcting false negativity was further tested and confirmed under more environmentally relevant conditions using Salmonella-spiked stream water and sediment samples. Finally, the PCR-NGS approach was applied to ten urban stream water samples and detected invA sequences in eight samples that would be otherwise deemed Salmonella negative. Analysis of the non-target sequences in the false-negative reactions helped to identify primer dime-like short sequences as the main cause of the false negativity. Together, the results demonstrated that the PCR-NGS approach can significantly improve method sensitivity, correct false-negative detections, and enable sequence-based analysis for failure diagnostics in complex environmental samples. Copyright © 2018 Elsevier B.V. All rights reserved.

Detection of Escherichia coli, Salmonella species, and Vibrio cholerae in tap water and bottled drinking water in Isfahan, Iran.

PubMed

Momtaz, Hassan; Dehkordi, Farhad Safarpoor; Rahimi, Ebrahim; Asgarifar, Amin

2013-06-07

The quality of drinking water has an important role in human infection and disease. This study was aimed at comparing polymerase chain reaction and culture in detecting Escherichia coli, Salmonella species and Vibrio cholera in tape water and bottled drinking water in various seasons in Isfahan province, Iran. A total of 448 water samples from tap water and bottled mineral water were taken over 6 months, from July 2010 to December 2010, and after filtration, samples were examined by culture and polymerase chain reaction methods for detection of Escherichia coli, Salmonella species, and Vibrio cholerae. The culture method showed that 34 (7.58%), 4 (0.89%) and 3 (0.66%) of all 448 water samples were positive for Escherichia coli, Salmonella species, and Vibrio cholera, respectively. The uidA gene from Escherichia coli, IpaB gene from Salmonella species, and epsM gene from Vibrio cholera were detected in 38 (26.38%), 5 (3.47%), and 3 (2.08%) of 144 tap-water samples, respectively. Escherichia coli was detected in 8 (2.63%) of 304 samples of bottled drinking water from 5 companies. The water of southern part of Isfahan and company 5 had the highest prevalence of bacteria. The Escherichia coli water contamination was significantly higher (P < 0.05) in the hot seasons (July-August) than cold (November-December) seasons and in company 5 than other companies. There were significant differences (P < 0.05) for the prevalence of bacteria between the tap waters of southern part and tap waters of central part of Isfahan. This study showed that the polymerase chain reaction assays can be an extremely accurate, fast, safe, sensitive and specific approach to monitor drinking water quality from purification facilities and bottled water companies. Also, our study confirmed the presence of Escherichia coli, Salmonella species, and Vibrio cholerae as water-borne pathogens in tap water and bottled drinking water of Isfahan, Iran. The present study showed the important public health problem in Isfahan, Iran.

Detection of Escherichia coli, Salmonella species, and Vibrio cholerae in tap water and bottled drinking water in Isfahan, Iran

PubMed Central

2013-01-01

Background The quality of drinking water has an important role in human infection and disease. This study was aimed at comparing polymerase chain reaction and culture in detecting Escherichia coli, Salmonella species and Vibrio cholera in tape water and bottled drinking water in various seasons in Isfahan province, Iran. Methods A total of 448 water samples from tap water and bottled mineral water were taken over 6 months, from July 2010 to December 2010, and after filtration, samples were examined by culture and polymerase chain reaction methods for detection of Escherichia coli, Salmonella species, and Vibrio cholerae. Results The culture method showed that 34 (7.58%), 4 (0.89%) and 3 (0.66%) of all 448 water samples were positive for Escherichia coli, Salmonella species, and Vibrio cholera, respectively. The uidA gene from Escherichia coli, IpaB gene from Salmonella species, and epsM gene from Vibrio cholera were detected in 38 (26.38%), 5 (3.47%), and 3 (2.08%) of 144 tap-water samples, respectively. Escherichia coli was detected in 8 (2.63%) of 304 samples of bottled drinking water from 5 companies. The water of southern part of Isfahan and company 5 had the highest prevalence of bacteria. The Escherichia coli water contamination was significantly higher (P < 0.05) in the hot seasons (July-August) than cold (November-December) seasons and in company 5 than other companies. There were significant differences (P < 0.05) for the prevalence of bacteria between the tap waters of southern part and tap waters of central part of Isfahan. Conclusions This study showed that the polymerase chain reaction assays can be an extremely accurate, fast, safe, sensitive and specific approach to monitor drinking water quality from purification facilities and bottled water companies. Also, our study confirmed the presence of Escherichia coli, Salmonella species, and Vibrio cholerae as water-borne pathogens in tap water and bottled drinking water of Isfahan, Iran. The present study showed the important public health problem in Isfahan, Iran. PMID:23742181

Molecular diagnosis of Salmonella typhi and its virulence in suspected typhoid blood samples through nested multiplex PCR.

PubMed

Prabagaran, Solai Ramatchandirane; Kalaiselvi, Vellingiri; Chandramouleeswaran, Naganathan; Deepthi, Krishnan Nair Geetha; Brahmadathan, Kootallur Narayanan; Mani, Mariappa

2017-08-01

A nested multiplex polymerase chain reaction (PCR) based diagnosis was developed for the detection of virulent Salmonella typhi in the blood specimens from patients suspected for typhoid fever. After the Widal test, two pairs of primers were used for the detection of flagellin gene (fliC) of S. typhi. Among them, those positive for fliC alone were subjected to identification of genes in Via B operon of Salmonella Pathogenesity Island (SPI-7) where four primer pairs were used to detect tviA and tviB genes. Among 250 blood samples tested, 115 were positive by fliC PCR; 22 of these were negative for tviA and tviB. Hence, the method described here can be used to diagnose the incidence of Vi-negative serovar typhi especially in endemic regions where the Vi vaccine is administered. Copyright © 2017 Elsevier B.V. All rights reserved.

SAS molecular tests Salmonella detection kit. Performance tested method 021202.

PubMed

Bapanpally, Chandra; Montier, Laura; Khan, Shah; Kasra, Akif; Brunelle, Sharon L

2014-01-01

The SAS Molecular tests Salmonella Detection method, a Loop-mediated Isothermal Amplification method, performed as well as or better than the U.S. Department of Agriculture-Food Safety Inspection Service Microbiology Laboratory Guidebook and the U.S. Food and Drug Administration Bacteriological Analytical Manual reference methods for ground beef, beef trim, ground turkey, chicken carcass rinses, bagged mixed lettuce, and fresh spinach. The ground beef (30% fat, 25 g test portion), poultry matrixes and leafy greens were validated in a 6-7 h enrichment, and ground beef (30% fat, 375 g composite test portion) and beef trim (375 g composite test portion) were validated in a 16-20 h enrichment. The method performance for meat and leafy green matrixes was shown to be acceptable under conditions of co-enrichment with Escherichia coli 0157. Thus, after a short 6-7 h co-enrichment step, ground beef, beef trim, lettuce, and spinach can be tested for both Salmonella and E. coli O157. Inclusivity and exclusivity testing revealed no false negatives and no false positives among the 100 Salmonella serovars and 30 non-Salmonella species examined. The method was shown to be robust when enrichment time, DNA extract hold time, and DNA volume were varied.

Salmonella Detection and Aerobic Colony Count in Deep-Frozen Carcasses of House Sparrow (Passer Domesticus) and Starling (Sturnus Vulgaris) Intended for Human Consumption

PubMed Central

De Cesare, Alessandra; Braggio, Simonetta; Manfreda, Gerardo

2014-01-01

Wild birds are potential vehicles of zoonotic pathogen transmission to humans. The zoonotic concern increases for small wild birds like house sparrows (Passer domesticus) and starlings (Sturnus vulgaris) which are hunted in developing countries and commercialised in Italy for human consumption. From June to October 2011, 330 house sparrows and 140 starlings were hunted and slaughtered. Deep-frozen carcasses were transported to Italy and stored for 6-8 months at -18°C. Aerobic colony count and Salmonella detection in carcasses were assessed following standard microbiological methods (ISO 4833:2003 and ISO 6579:2004, respectively). Carcasses of house sparrows showed higher levels of aerobic bacteria in comparison to starling carcasses (5.7 vs 3.2 log10 CFU/g). Moreover, 7 out of 11 lots of carcasses of house sparrows were positive for Salmonella. Among the 18 isolates of Salmonella, 14 were S. Typhimurium, 2 were S. Enteritidis, and 2 were not distinguishable. All of them were susceptible to antibiotics. All tested carcasses of starling were Salmonella negative. Deep-freezing was not efficient as a decontamination technique on carcasses of house sparrows. PMID:27800336

[Active etiological surveillance for foodborne diseases in Guangdong province, 2013-2014].

PubMed

Ke, B X; He, D M; Tan, H L; Zeng, H H; Yang, T; Li, B S; Liang, Y H; Lu, L L; Liang, J H; Huang, Q; Ke, C W

2016-10-10

Objective: To study the infection status, serotypes, drug resistance and molecular characteristics of Salmonella, Shigella, Vibrio parahemolyticus , enterotoxigenic Escherichia ( E. ) coli (ETEC), pathogenic E. coli (EPEC), Shiga Toxin producing E. coli (STEC) and Enteroinvasive E. coli (EIEC) collected from diarrhea patients in Guangdong. Methods: The strains of Salmonella, Shigella, V. parahemolyticus and 4 kinds of E. coli isolated from foodborne diseases surveillance during 2013-2014 were collected to conduct serotyping, drug resistance test and pulsed-field gel electrophoresis (PFGE). Results: A total of 3 372 stains of pathogens were isolated from 57 834 stool samples during 2013-2014, the overall positive rate was 5.83 % and the positive rate of Salmonella was highest, followed by that of V. parahemolyticus , 4 kinds of E. coli and Shigella . And 3 213 strains of Salmonella were divided into 143 serotypes. The most prevalent serotypes were Salmonella typhimurium , 4, 5, 12: i:-, Enteritidis , Stanley and Derby . Salmonella was sensitive to cephalosporin and fluoroquinolones, and showed significant differences in drug resistance rate among different serotypes. In top 10 common serotypes, S. enteritidis and S. derby were most resistant to cephalosporin and ciprofloxacin respectively. PFGE was performed for 2 289 strains of Salmonella , showing distribution diversity and significant fingerprint polymorphisms. The 85 strains of V. parahemolyticus were divided into 10 serotypes, O3∶K6 (61.18 % ) was the most common serotype, followed by O4∶K8. The results showed that the carrying rate of virulence genes tdh (81.18 % ) was high, while the carrying rate of trh was low (7.06 % ), and there were 10 strains carrying no the two kinds of virulence genes. The sensitive rate of V. parahemolyticus to imipenem, nalidixic acid, SMZ-TMP, chloramphenicol and tetracycline were more than 95 % . Thirteen strains of Shigella were detected, including 9 strains of Shigella sonnei , 3 strains of Shigella flexneri and 1 strains of Shigella bogdii . The strains all showed sensitivity to ceftazidime, ciprofloxacin and chloramphenicol (76.92 % ). There were 86 strains of E. coli detected, including 29 strains of ETEC (33.72 % ), 27 strains of EPEC (31.39 % ), 27 strains of STEC (31.39 % ) and 3 strains of EIEC (3.48 % ). Conclusions: In the active etiological surveillance for foodborne diseases in Guangdong during 2013-2014, the detection rate of Salmonella was highest (5.57 % ), followed by that of V. parahemolyticus , 4 kinds of E. coli and Shigella . Salmonella , V. parahemolyticus and Shigella were sensitive to cephalosporin and fluoroquinolones. Clustered cases of Salmonella infection were found in the surveillance, but no outbreaks occurred.

Evaluation of the addition of organic acids in the feed and/or water for broilers and the subsequent recovery of Salmonella Typhimurium from litter and ceca.

PubMed

Bourassa, D V; Wilson, K M; Ritz, C R; Kiepper, B K; Buhr, R J

2018-01-01

Three separate broiler Salmonella Typhimurium challenge experiments were conducted evaluating efficacy of formic and propionic acid feed supplements to suppress environmental and cecal Salmonella Typhimurium prevalence. In experiment 1, broilers were provided feed with 1 kg/ton formic acid or 5 kg/ton propionic acid feed additives or a basal control diet. At the day of placement, half of the pens were inoculated with seeder chicks orally challenged with a marker strain of Salmonella Typhimurium and to yield challenged and adjacent nonchallenged pens. No differences in weekly litter samples or cecal Salmonella prevalence at 3 or 6 wk among feeding treatments were detected. In experiment 2, treatments were: 2 kg/ton propionic acid in feed, 1.0 mL/L formic acid in water, both propionic acid in feed and formic acid in water, and a basal control. Every pen was challenged with seeder chicks inoculated with Salmonella Typhimurium. By 6 wk all pens maintained detectable litter Salmonella, and broilers provided both propionic acid in feed and formic acid in water had the lowest cecal recovery (35%), compared to the control (60%). In experiment 3, treatments were: formic acid at 4 or 6 kg/ton from wk 0 to 6 or for only the last wk, propionic acid at 5 or 10 kg/ton for only the last wk, and a basal control. Each pen was challenged with Salmonella Typhimurium inoculated seeder chicks. By 6 wk, broilers fed formic acid (4 kg/ton) for the entire growout had no Salmonella-positive ceca (0/30). All treatments that provided acid supplemented feed for only the last wk had 3-13% Salmonella-positive ceca. These experiments indicate that adding formic acid to broiler feed appears to prevent Salmonella colonization from challenge pens entering into the adjacent nonchallenge pens. Feeding formic acid (4 kg/ton) for 6 wk resulted in no recovery of Salmonella from ceca compared to the control prevalence of 17%. Published by Oxford University Press on behalf of Poultry Science Association 2017. This work is written by (a) US Government employee(s) and is in the public domain in the US.

Transmission and control of Salmonella in the pig feed chain: a conceptual model.

PubMed

Binter, Claudia; Straver, Judith Maria; Häggblom, Per; Bruggeman, Geert; Lindqvist, Per-Anders; Zentek, Jürgen; Andersson, Mats Gunnar

2011-03-01

Infected breeder pigs and contaminated feed represent potential sources of Salmonella introduction to fattening pig herds and may thereby cause human infections acquired via consumption of contaminated pork. Modelling approaches such as quantitative microbial risk assessment could improve the design of strategies for control and tracing of Salmonella in the feed chain. However, the construction of such models requires a thorough understanding of the dynamics of the feed chain, including production processes, microbial processes and transport logistics. The present article illustrates a conceptual model of Salmonella in the pig feed chain and explores the possibilities for quantitative modelling including identifying major gaps in data. Information was collected from peer-reviewed scientific journals, official documents and reports and by means of interviews with experts from authorities and the feed industry. Data on prevalence of Salmonella in different parts of the feed chain are difficult to compare as observed prevalence may be biased by variations in sampling procedures as well as limitations of the detection methods. There are almost no data on numbers of Salmonella in commodities of the feed chain, which often makes it difficult to evaluate risks, intervention strategies and sampling plans in a quantitative manner. Tracing the source of Salmonella contamination is hampered by the risk of cross-contamination as well as various mixing and partitioning events along the supply chain, which sometimes makes it impossible to trace the origin of a lot back to a batch or producer. Available information points to contaminated feed materials, animal vectors and persistent contamination of production environments as important sources of Salmonella in feed production. Technological procedures such as hydrothermal or acid treatment can be used to control Salmonella in feed production. However, a large fraction of pig feed is produced without decontamination procedures. Prevention of recontamination and control of moisture throughout the chain are thus critical factors for controlling Salmonella in feed production. To verify successful control it is necessary to have monitoring strategies able to detect low levels of Salmonella heterogeneously distributed in large volumes of feed and feed material in bulk. Experience from monitoring programs and research investigations indicates that sampling of dust and sweepings from control points along the production line is an efficient strategy to gain an indication of Salmonella contamination. Copyright © 2010 Elsevier B.V. All rights reserved.

Climate Patterns Governing the Presence and Permanence of Salmonellae in Coastal Areas of Bahia de Todos Santos, Mexico▿

PubMed Central

Simental, Lourdes; Martinez-Urtaza, Jaime

2008-01-01

Despite the importance of salmonellae as one of the major causes of food-borne infections worldwide, data regarding the presence of these organisms in the environment are limited. We investigated the presence of Salmonella spp. in Bahia de Todos Santos (Baja California, Mexico) and evaluated the environmental factors that affect the occurrence of Salmonella spp. in this arid region. A total of 1,331 samples collected from 21 sites along the coast during a period of 3 years were analyzed for Salmonella spp. Geographical and seasonal distribution of Salmonella spp. was evaluated in association with environmental parameters and with human infections in the area. The incidence of Salmonella bacteria throughout the study was 4.8%, with the highest incidence detected in wastewater (16.2%), followed by stream water (10.6%), mollusks (7.4%), and seawater (2.3%). Twenty different serotypes were identified among the 64 Salmonella isolates. The dominant serotype was Typhimurium (23.4%), followed by Vejle (6.2%). The presence of Salmonella spp. in coastal areas was mostly confined to rainy periods and areas of stream discharges, and runoff was identified as the predominant factor influencing the transport of Salmonella bacteria from source points to the sea via streams. Isolation of Salmonella spp. was negatively and significantly associated with temperature, probably because of the effect of solar radiation in the decline of permanence of Salmonella bacteria. Conversely, human infections prevailed during the warmest months and were negatively correlated with the presence of Salmonella spp. in the marine environment. PMID:18708509

Free-Living Turtles Are a Reservoir for Salmonella but Not for Campylobacter

PubMed Central

Marin, Clara; Ingresa-Capaccioni, Sofia; González-Bodi, Sara; Marco-Jiménez, Francisco; Vega, Santiago

2013-01-01

Different studies have reported the prevalence of Salmonella in turtles and its role in reptile-associated salmonellosis in humans, but there is a lack of scientific literature related with the epidemiology of Campylobacter in turtles. The aim of this study was to evaluate the prevalence of Campylobacter and Salmonella in free-living native (Emys orbicularis, n=83) and exotic ( Trachemys scripta elegans, n=117) turtles from 11 natural ponds in Eastern Spain. In addition, different types of samples (cloacal swabs, intestinal content and water from Turtle containers) were compared. Regardless of the turtle species, natural ponds where individuals were captured and the type of sample taken, Campylobacter was not detected. Salmonella was isolated in similar proportions in native (8.0±3.1%) and exotic (15.0±3.3%) turtles (p=0.189). The prevalence of Salmonella positive turtles was associated with the natural ponds where animals were captured. Captured turtles from 8 of the 11 natural ponds were positive, ranged between 3.0±3.1% and 60.0±11.0%. Serotyping revealed 8 different serovars among four Salmonella enterica subspecies: S. enterica subsp. enterica (n = 21), S. enterica subsp. salamae (n = 2), S. enterica subsp. diarizonae (n = 3), and S. enterica subsp. houtenae (n = 1). Two serovars were predominant: S. Thompson (n=16) and S . typhimurium (n=3). In addition, there was an effect of sample type on Salmonella detection. The highest isolation of Salmonella was obtained from intestinal content samples (12.0±3.0%), while lower percentages were found for water from the containers and cloacal swabs (8.0±2.5% and 3.0±1.5%, respectively). Our results imply that free-living turtles are a risk factor for Salmonella transmission, but do not seem to be a reservoir for Campylobacter . We therefore rule out turtles as a risk factor for human campylobacteriosis. Nevertheless, further studies should be undertaken in other countries to confirm these results. PMID:23951312

The occurrence of Salmonella spp. in duck eggs on sale at retail or from catering in England.

PubMed

Owen, M; Jorgensen, F; Willis, C; McLauchlin, J; Elviss, N; Aird, H; Fox, A; Kaye, M; Lane, C; de Pinna, E

2016-11-01

Since 2010, human salmonellosis outbreaks in the UK have been detected as associated with the consumption of duck eggs. Little data are available on the rate of occurrence of Salmonella in duck eggs. The aim of this study was to investigate the occurrence of Salmonella spp. in duck eggs on sale and from catering in England during 2011, particularly those from small-scale production. All samples were collected independently of human salmonellosis outbreak investigations. Composite samples of 6-10 eggs (shells and contents were examined separately) were examined for the presence of Salmonella spp. using the ISO 6579:2002 method. Salmonella spp. was recovered from two of 145 samples (1·4%). In one sample, Salmonella Typhimurium DT 8 was isolated from the shells while Salm. Typhimurium DT 8 and Salm. Typhimurium DT30 were isolated from the contents. Salmonella Typhimurium DT8 was isolated from the egg shells only in the second contaminated sample. This study provides baseline data for risk assessors, regulators and the food industry and may be helpful in communicating risks associated with the consumption of this product as well as evaluating risk management options to control food safety including vaccination of ducks. Human salmonellosis outbreaks in England and Northern Ireland due to Salmonella enterica serovar Typhimurium definitive phage type (DT) 8 have been identified as associated with the consumption of duck eggs since 2010. This study has shown that Salmonella spp. was detected in 1·4% of ducks egg samples providing baseline data for risk assessors, regulators and the food industry. This may be helpful in communicating risks associated with the consumption of this product as well as evaluating risk management options to control food safety including vaccination of ducks. © 2016 Crown copyright. Letters in Applied Microbiology © 2016 The Society for Applied Microbiology.

A new protocol to detect multiple foodborne pathogens with PCR dipstick DNA chromatography after a six-hour enrichment culture in a broad-range food pathogen enrichment broth.

PubMed

Hayashi, Masahiro; Natori, Tatsuya; Kubota-Hayashi, Sayoko; Miyata, Machiko; Ohkusu, Kiyofumi; Kawamoto, Keiko; Kurazono, Hisao; Makino, Souichi; Ezaki, Takayuki

2013-01-01

A quick foodborne pathogen screening method after six-hour enrichment culture with a broad-range food pathogen enrichment broth is described. Pathogenic factors of Salmonella enterica, Shigella spp., enteroinvasive Escherichia coli, and enterohemorrhagic E. coli are amplified with a cocktail primer and rapid polymerase chain reaction (PCR), which finishes amplification in 30 min. The PCR amplicon was differentiated with a dipstick DNA chromatography assay in 5-10 min. Starting from a four- to six-hour enrichment culture, this assay was finished within 45 min. Detection sensitivity of this protocol was less than 2.5 CFU/25 g for S. enterica and 3.3 CFU/25 g for enterohemorrhagic E. coli in spiked ground meat experiments.

EVALUATION OF A DNA PROBE TEST KIT FOR DETECTION OF SALMONELLAE IN BIOSOLIDS

EPA Science Inventory

Aims: Current United States regulations (40 CFR 503) for "Class A" biosolids requires use of multiple-tube fermentation techniques for fecal coliform or multiple tube enrichment techniques for Salmonella spp. followed by isolation and biochemical and serological confirmation. T...

Serotype determination of Salmonella by xTAG assay.

PubMed

Zheng, Zhibei; Zheng, Wei; Wang, Haoqiu; Pan, Jincao; Pu, Xiaoying

2017-10-01

Currently, no protocols or commercial kits are available to determine the serotypes of Salmonella by using Luminex MAGPIX®. In this study, an xTAG assay for serotype determination of Salmonella suitable for Luminex MAGPIX® is described and 228 Salmonella isolates were serotype determined by this xTAG assay. The xTAG assay consists of two steps: 1) Multiplex PCR to amplify simultaneously O, H and Vi antigen genes of Salmonella, and 2) Magplex-TAG™ microsphere hybridization to identify accurately the specific PCR products of different antigens. Compared with the serotyping results of traditional serum agglutination test, the sensitivity and specificity of the xTAG assay were 95.1% and 100%, respectively. The agreement rate of these two assays was 95.2%. Compared with Luminex xMAP® Salmonella Serotyping Assay (SSA) kit, the advantages of this xTAG assay are: First, the magnetic beads make it applicable to both the Luminex®100/200™ and MAGPIX® systems. Second, only primers rather than both primers and probes are needed in the xTAG assay, and the process of coupling antigen-specific oligonucleotide probes to beads is circumvented, which make the xTAG assay convenient to be utilized by other laboratories. The xTAG assay may serve as a rapid alternative or complementary method for traditional Salmonella serotyping tests, especially for laboratories that utilize the MAGPIX® systems. Copyright © 2017 Elsevier B.V. All rights reserved.

Visualization of gold and platinum nanoparticles interacting with Salmonella Enteritidis and Listeria monocytogenes

PubMed Central

Sawosz, Ewa; Chwalibog, André; Szeliga, Jacek; Sawosz, Filip; Grodzik, Marta; Rupiewicz, Marlena; Niemiec, Tomasz; Kacprzyk, Katarzyna

2010-01-01

Purpose Rapid development of nanotechnology has recently brought significant attention to the extraordinary biological features of nanomaterials. The objective of the present investigation was to evaluate morphological characteristics of the assembles of gold and platinum nanoparticles (nano-Au and nano-Pt respectively), with Salmonella Enteritidis (Gram-negative) and Listeria monocytogenes (Gram-positive), to reveal possibilities of constructing bacteria-nanoparticle vehicles. Methods Hydrocolloids of nano-Au or nano-Pt were added to two bacteria suspensions in the following order: nano-Au + Salmonella Enteritidis; nano-Au + Listeria monocytogenes; nano-Pt + Salmonella Enteritidis; nano-Pt + Listeria monocytogenes. Samples were inspected by transmission electron microscope. Results Visualization of morphological interaction between nano-Au and Salmonella Enteritidis and Listeria monocytogenes, showed that nano-Au were aggregated within flagella or biofilm network and did not penetrate the bacterial cell. The analysis of morphological effects of interaction of nano-Pt with bacteria revealed that nano-Pt entered cells of Listeria monocytogenes and were removed from the cells. In the case of Salmonella Enteritidis, nano-Pt were seen inside bacteria cells, probably bound to DNA and partly left bacterial cells. After washing and centrifugation, some of the nano-Pt-DNA complexes were observed within Salmonella Enteritidis. Conclusion The results indicate that the bacteria could be used as a vehicle to deliver nano-Pt to specific points in the body. PMID:20856838

A serological survey for pathogens in old fancy chicken breeds in central and eastern part of The Netherlands.

PubMed

de Wit, J J; van Eck, J H; Crooijmans, R P; Pijpers, A

2004-05-15

To get an impression of the presence of pathogens in multi-aged flocks of old fancy chicken breeds in the Netherlands, plasma samples originating from 24 flocks were examined for antibodies against 17 chicken pathogens. These flocks were housed mainly in the centre and east of the Netherlands, regions with a high poultry density. The owners of the tested flocks showed their chicken at national and international poultry exhibitions. Antibodies against Avian Influenza, Egg Drop Syndrome '76 virus, Pox virus, Salmonella pullorum/gallinarum, Salmonella Enteritidis or Salmonella Typhimurium were not detected. However, antibodies against other Salmonella species, Mycoplasma gallisepticum, infectious bursal disease virus, infectious bronchitis virus, avian encephalomyelitis virus, chicken anaemia virus, infectious laryngotracheitis virus, and avian leukosis virus, subgroups A and B, and subgroup J were detected in a varying proportion of the flocks. This study shows that antibodies against many chicken pathogens are present among the flocks of old fancy chicken breeds that are exhibited at international poultry exhibitions.

An outbreak of salmonella chester infection in Canada: rare serotype, uncommon exposure, and unusual population demographic facilitate rapid identification of food vehicle.

PubMed

Taylor, John; Galanis, Eleni; Wilcott, Lynn; Hoang, Linda; Stone, Jason; Ekkert, Judi; Quibell, Doug; Huddleston, Mark; McCormick, Rachel; Whitfield, Yvonne; Adhikari, Bijay; Grant, Christopher C R; Sharma, Davendra

2012-04-01

Salmonella Chester infection has rarely been reported in the literature. In 2010, 33 case patients were reported in 2 months in four Canadian provinces. We conducted an outbreak investigation in collaboration with public health agencies, food safety specialists, regulatory agencies, grocery store chains, and the product distributor. We used case patient interviews, customer loyalty cards, and microbiological testing of clinical and food samples to identify nationally distributed head cheese as the food vehicle responsible for the outbreak. The rare serotype, a limited affected demographic group, and an uncommon exposure led to the rapid identification of the source. Control measures were implemented within 9 days of notification of the outbreak.

Survey of Salmonella contamination in chicken layer farms in three Caribbean countries.

PubMed

Adesiyun, Abiodun; Webb, Lloyd; Musai, Lisa; Louison, Bowen; Joseph, George; Stewart-Johnson, Alva; Samlal, Sannandan; Rodrigo, Shelly

2014-09-01

This study was conducted to investigate the demography, management, and production practices on layer chicken farms in Trinidad and Tobago, Grenada, and St. Lucia and the frequency of risk factors for Salmonella infection. The frequency of isolation of Salmonella from the layer farm environment, eggs, feeds, hatchery, and imported day-old chicks was determined using standard methods. Of the eight risk factors (farm size, age group of layers, source of day-old chicks, vaccination, sanitation practices, biosecurity measures, presence of pests, and previous disease outbreaks) for Salmonella infection investigated, farm size was the only risk factor significantly associated (P = 0.031) with the prevalence of Salmonella; 77.8% of large farms were positive for this pathogen compared with 33.3 and 26.1% of medium and small farms, respectively. The overall isolation rate of Salmonella from 35 layer farms was 40.0%. Salmonella was isolated at a significantly higher rate (P < 0.05) from farm environments than from the cloacae. Only in Trinidad and Tobago did feeds (6.5% of samples) and pooled egg contents (12.5% of samples) yield Salmonella; however, all egg samples from hotels, hatcheries, and airports in this country were negative. Salmonella Anatum, Salmonella group C, and Salmonella Kentucky were the predominant serotypes in Trinidad and Tobago, Grenada, and St. Lucia, respectively. Although Salmonella infections were found in layer birds sampled, table eggs appear to pose minimal risk to consumers. However, the detection of Salmonella -contaminated farm environments and feeds cannot be ignored. Only 2.9% of the isolates belonged to Salmonella Enteritidis, a finding that may reflect the impact of changes in farm management and poultry production in the region.

Prevalence and antimicrobial susceptibility of Salmonella isolated from a variety of raw meat sausages in Gaborone (Botswana) retail stores.

PubMed

Samaxa, Ronald Gaelekolwe; Matsheka, Maitshwarelo Ignatius; Mpoloka, Sununguko Wata; Gashe, Berhanu Abegaz

2012-04-01

The objective of the study was to provide baseline data on the prevalence and antimicrobial susceptibility of Salmonella in different types of raw meat sausages directly accessible to the consumers in Gaborone, Botswana. A total of 300 raw sausages comprising 79 beef, 78 pork, 72 chicken, and 71 mutton samples were concurrently analyzed for the presence of Salmonella using a conventional culture method and a validated PCR method. The PCR assay results were in full concordance with those of the conventional culture method for the detection of Salmonella. Sixty-five (21.7%) of 300 samples were positive for Salmonella by both the conventional culture method and PCR assay. Even though more chicken samples contained Salmonella than did any other sausage type, the difference in the presence of Salmonella among the four sausages types was not significant. Eleven serotypes were identified, and Salmonella enterica subsp. salamae II was most prevalent in all the sausage types. Beef sausages generally had higher mesophilic bacterial counts than did the other three sausage types. However, higher microbial counts were not reflective of the presence of salmonellae. Susceptibility of the Salmonella enterica serotypes to 20 antimicrobial agents was determined, and Salmonella Muenchen was resistant to the widest array of agents and was mostly isolated from chicken sausages. Regardless of the meat of origin, all 65 Salmonella isolates were resistant to at least four antimicrobial agents: amikacin, gentamicin, cefuroxime, and tombramycin. This resistance profile group was the most common in all four sausage types, comprising 90% of all Salmonella isolates from beef, 71% from pork, 63% from mutton, and 35% from chicken. These results suggest that raw sausages pose a risk of transmitting multidrug-resistant Salmonella isolates to consumers.

Development of a Salmonella screening tool for consumer complaint-based foodborne illness surveillance systems.

PubMed

Li, John; Maclehose, Rich; Smith, Kirk; Kaehler, Dawn; Hedberg, Craig

2011-01-01

Foodborne illness surveillance based on consumer complaints detects outbreaks by finding common exposures among callers, but this process is often difficult. Laboratory testing of ill callers could also help identify potential outbreaks. However, collection of stool samples from all callers is not feasible. Methods to help screen calls for etiology are needed to increase the efficiency of complaint surveillance systems and increase the likelihood of detecting foodborne outbreaks caused by Salmonella. Data from the Minnesota Department of Health foodborne illness surveillance database (2000 to 2008) were analyzed. Complaints with identified etiologies were examined to create a predictive model for Salmonella. Bootstrap methods were used to internally validate the model. Seventy-one percent of complaints in the foodborne illness database with known etiologies were due to norovirus. The predictive model had a good discriminatory ability to identify Salmonella calls. Three cutoffs for the predictive model were tested: one that maximized sensitivity, one that maximized specificity, and one that maximized predictive ability, providing sensitivities and specificities of 32 and 96%, 100 and 54%, and 89 and 72%, respectively. Development of a predictive model for Salmonella could help screen calls for etiology. The cutoff that provided the best predictive ability for Salmonella corresponded to a caller reporting diarrhea and fever with no vomiting, and five or fewer people ill. Screening calls for etiology would help identify complaints for further follow-up and result in identifying Salmonella cases that would otherwise go unconfirmed; in turn, this could lead to the identification of more outbreaks.

Microbiological analysis of pre-packed sweet basil (Ocimum basilicum) and coriander (Coriandrum sativum) leaves for the presence of Salmonella spp. and Shiga toxin-producing E. coli.

PubMed

Delbeke, Stefanie; Ceuppens, Siele; Jacxsens, Liesbeth; Uyttendaele, Mieke

2015-09-02

Enteric pathogens, such as Salmonella spp. and pathogenic Escherichia coli, have been detected and associated with food borne outbreaks from (imported) fresh leafy herbs. Screening on imported herbs from South East Asian countries has been described. However, limited information on prevalence of these pathogens is available from other sourcing regions. Therefore, fresh pre-packed basil and coriander leaves from a Belgian trading company were investigated for the presence of Salmonella spp., Shiga toxin-producing E. coli (STEC), generic E. coli and coliforms. In total 592 samples were collected originating from Belgium, Israel and Cyprus during 2013-2014. Multiplex PCR followed by further culture confirmation was used for the detection of Salmonella spp. and STEC, whereas the Petrifilm Select E. coli and VRBL-agar were used, respectively, for the enumeration of E. coli and coliforms. Salmonella was detected in 10 out of 592 samples (25g) (1.7%; 5 from basil and 5 from coriander), of which two samples were sourced from Israel and eight from Cyprus. The presence of STEC was suspected in 11 out of 592 samples (25g) (1.9%; 3 basil and 8 coriander), due to the detection of stx and eae genes, of which one sample originated from Belgium, four from Israel and six from Cyprus. No STEC was isolated by culture techniques, but in three samples a serotype (O26, O103 or O111) with its most likely associated eae-variant (β or θ) was detected by PCR. Generic E. coli was enumerated in 108 out of 592 samples, whereby 55, 32 and 13 samples respectively between 10-100, 100-1000 and 1000-10,000cfu/g and 8 samples exceeding 10,000cfu/g. Coliforms were enumerated in all herb samples at variable levels ranging from 1.6 to 7.5logcfu/g. Further statistics indicate that the E. coli class (categorized by level) was significantly correlated with the presence of Salmonella (p<0.001) or STEC (p=0.019), while coliform counts were significant correlated with Salmonella (p<0.001), but not with STEC (p=0.405). Generic E. coli class is a better indicator for the presence of enteric pathogens than coliforms on fresh herbs, but the relationship between E. coli and Salmonella or STEC was not strong enough to provide a threshold value for E. coli to assure food safety (i.e. no pathogens present). Results indicate that fresh leafy herbs like basil and coriander sourced from different cultivation regions, may contain enteric pathogens and potentially pose a risk for human health. Copyright © 2015 Elsevier B.V. All rights reserved.

Gold nanoparticles enhanced SERS aptasensor for the simultaneous detection of Salmonella typhimurium and Staphylococcus aureus.

PubMed

Zhang, Hui; Ma, Xiaoyuan; Liu, Ying; Duan, Nuo; Wu, Shijia; Wang, Zhouping; Xu, Baocai

2015-12-15

Salmonella typhimurium and Staphylococcus aureus are most common causes of food-associated disease. A Raman based biosensor was developed for S. typhimurium and S. aureus detection simultaneously. The biosensor was based on nanoparticles enhanced Raman intensity and the specific recognition of aptamer. The Raman signal probe and the capture probe are built. Gold nanoparticles (GNPs) modified with Raman molecules (Mercaptobenzoic acid and 5,5'-Dithiobis(2-nitrobenzoic acid)) and aptamer are used as the signal probe for S. typhimurium and S. aureus, respectively. Fe3O4 magnetic gold nanoparticles (MGNPs) immobilized with both aptamer of S. typhimurium and S. aureus are used as the capture probe. When S. typhimurium and S. aureus are added in the reaction system, the capture probe will capture the target bacteria through the specific binding effect of aptamer. And then the signal probe will be connected to the bacteria also by the effect of aptamer to form the sandwich like detection structure. The Raman intensified spectrum was measured to quantify S. typhimurium and S. aureus. Under optimal conditions, the SERS intensity of MBA at 1582 cm(-1) are used to measure S. typhimurium (y=186.4762+704.8571x, R(2)=0.9921) and the SERS intensity of DNTB at 1333 cm(-1) are used to measure S. aureus (y=135.2381+211.4286x, R(2)=0.9946) in the range of 10(2)-10(7) cfu mL(-1). The LOD is 35 cfu mL(-1) for S. aureus and 15 cfu mL(-1) for S. typhimurium. This method is simple and rapid, results in high sensitivity and specificity, and can be used to detect actual samples. Copyright © 2015 Elsevier B.V. All rights reserved.

A label-free fluorescent direct detection of live Salmonella typhimurium using cascade triple trigger sequences-regenerated strand displacement amplification and hairpin template-generated-scaffolded silver nanoclusters.

PubMed

Zhang, Peng; Liu, Hui; Li, Xiaocheng; Ma, Suzhen; Men, Shuai; Wei, Heng; Cui, Jingjing; Wang, Hongning

2017-01-15

The harm of Salmonella typhimurium (S. typhimurium) to public health mainly by the consumption of contaminated agricultural products or water stresses an urgent need for rapid detection methods to help control the spread of S. typhimurium. In this work, an intelligently designed sensor system took creative advantage of triple trigger sequences-regenerated strand displacement amplification and self-protective hairpin template-generated-scaffolded silver nanoclusters (AgNCs) for the first time. In the presence of live S. typhimurium, single-stranded trigger sequences were released from aptamer-trigger sequences complex, initiating a branch migration to open the hairpin template I containing complementary scaffolds of AgNCs. Then the first strand displacement amplification was induced to produce numerous scaffolds of AgNCs and reporter strands which initiated a branch migration to open the hairpin template II containing complementary scaffolds of AgNCs. Then the second strand displacement amplification was induced to generate numerous scaffolds of AgNCs and trigger sequences which initiated the third branch migration and strand displacement amplification to produce numerous scaffolds of AgNCs and reporter strands in succession. Cyclically, the reproduction of the trigger sequences and cascade successive production of scaffolds were achieved successfully, forming highly fluorescent AgNCs, thus providing significantly enhanced fluorescent signals to achieve ultrasensitive detection of live S. typhimurium down to 50 CFU/mL with a linear range from 10 2 to 10 7 CFU/mL. It is the first report on a fluorescent biosensor for detecting viable S. typhimurium directly, which can distinguish from heat denatured S. typhimurium. And it develops a new strategy to generate the DNA-scaffolds for forming AgNCs. Copyright © 2016 Elsevier B.V. All rights reserved.

Cold plasma rapid decontamination of food contact surfaces contaminated with Salmonella biofilms

USDA-ARS?s Scientific Manuscript database

Cross-contamination of fresh produce and other foods from persistent pathogen reservoirs is a known risk factor in processing environments. Industry requires a rapid, waterless, zero-contact, chemical-free method for removing pathogens from food-contact surfaces. Cold plasma was tested for its abili...

Rapid pasteurization of shell eggs using RF

USDA-ARS?s Scientific Manuscript database

A novel method for rapidly pasteurizing eggs in the shell could enhance the safety of the United States’ food supply. Current federal regulations do not require eggs sold in stores to be pasteurized, yet these eggs are often consumed raw or undercooked and cause untold cases of salmonella illness ea...

[Prevalence and antimicrobial susceptibility of Salmonella isolated from broiler whole production process in four provinces of China].

PubMed

Li, W W; Bai, L; Zhang, X L; Xu, X J; Tang, Z; Bi, Z W; Guo, Y C

2018-04-06

Objective: To determine the prevalence and antimicrobial susceptibility of Salmonella isolated from broiler production process in 4 provinces of China. Methods: Using convenience sampling method, 238 sample sites from broiler whole production process were chosen in Henan, Jiangsu, Heilongjiang and Shandong provinces in 2012. A total of 11 592 samples were collected and detected to analyze prevalence baseline, including 2 090 samples from breeding chicken farms and hatcheries, 1 421 samples from broiler farms, 5 610 samples from slaughterhouses and 2 471 samples from distribution and retail stores. All Salmonella strains were isolated through selective enrichment, and were serotyped according to Kauffmann-White scheme. The antimicrobial susceptibilities of selected Salmonella strains were determined by the broth microdilution method and fourteen antimicrobial agents were examined. Results: During incubation course, the average prevalence of Salmonella was 5.5% in feces of breeding hens, feces of chicks, and hatching eggs, 123 Salmonella strains were isolated. During cultivation course, the prevalence of Salmonella was 8.0% in feces from broiler farms, soil, feed, and workers, 114 Salmonella strains were isolated. During slaughter course, the prevalence of Salmonella was 24.9% in swabs pre-slaughter, dressed broiler carcasses, pre-cooled broiler carcasses, water from precooling pool, cutter and chipping boards, frozen chicken portions, and workers, 1 438 Salmonella strains were isolated. During distribution and sale course, the prevalence of Salmonella was 20.9% in transport carts, frozen chicken portions, retail chicken portions and workers, 551 Salmonella strains were isolated. The dominant Salmonella serotypes were Salmonella Enteritidis ( n= 1 229) and Salmonella Indiana ( n= 621). Among 1 231 examined strains, 97.2% Salmonella isolates were resistant to at least one antimicrobial, 69.9% Salmonella strains were multi-drug resistant isolates. Conclusion: Our findings indicated that Salmonella contamination was common and serious in commercial broiler whole production process in China, especially in the course of defeathering, precooling and selling. The environment of broiler farm is the important source of Salmonella contamination. Additionally, antibiotic resistance of Salmonella was serious for common antimicrobials and multi-drug resistant strains existed widespread, which can pose potential risk on public health and clinical therapy.

Multidrug-resistant typhoid fever with neurologic findings on the Malawi-Mozambique border.

PubMed

Lutterloh, Emily; Likaka, Andrew; Sejvar, James; Manda, Robert; Naiene, Jeremias; Monroe, Stephan S; Khaila, Tadala; Chilima, Benson; Mallewa, Macpherson; Kampondeni, Sam D; Lowther, Sara A; Capewell, Linda; Date, Kashmira; Townes, David; Redwood, Yanique; Schier, Joshua G; Nygren, Benjamin; Tippett Barr, Beth; Demby, Austin; Phiri, Abel; Lungu, Rudia; Kaphiyo, James; Humphrys, Michael; Talkington, Deborah; Joyce, Kevin; Stockman, Lauren J; Armstrong, Gregory L; Mintz, Eric

2012-04-01

Salmonella enterica serovar Typhi causes an estimated 22 million cases of typhoid fever and 216 000 deaths annually worldwide. We investigated an outbreak of unexplained febrile illnesses with neurologic findings, determined to be typhoid fever, along the Malawi-Mozambique border. The investigation included active surveillance, interviews, examinations of ill and convalescent persons, medical chart reviews, and laboratory testing. Classification as a suspected case required fever and ≥1 other finding (eg, headache or abdominal pain); a probable case required fever and a positive rapid immunoglobulin M antibody test for typhoid (TUBEX TF); a confirmed case required isolation of Salmonella Typhi from blood or stool. Isolates underwent antimicrobial susceptibility testing and subtyping by pulsed-field gel electrophoresis (PFGE). We identified 303 cases from 18 villages with onset during March-November 2009; 214 were suspected, 43 were probable, and 46 were confirmed cases. Forty patients presented with focal neurologic abnormalities, including a constellation of upper motor neuron signs (n = 19), ataxia (n = 22), and parkinsonism (n = 8). Eleven patients died. All 42 isolates tested were resistant to ampicillin, chloramphenicol, and trimethoprim-sulfamethoxazole; 4 were also resistant to nalidixic acid. Thirty-five of 42 isolates were indistinguishable by PFGE. The unusual neurologic manifestations posed a diagnostic challenge that was resolved through rapid typhoid antibody testing in the field and subsequent blood culture confirmation in the Malawi national reference laboratory. Extending laboratory diagnostic capacity, including blood culture, to populations at risk for typhoid fever in Africa will improve outbreak detection, response, and clinical treatment.

Evaluation of aqueous and alcohol-based quaternary ammonium sanitizers for inactivating Salmonella spp., Escherichia coli O157:H7, and Listeria monocytogenes on peanut and pistachio shells.

PubMed

McEgan, Rachel; Danyluk, Michelle D

2015-05-01

This study evaluated the efficacy of aqueous (aQUAT) and isopropyl alcohol-based quaternary ammonium (ipQUAT) sanitizers for reducing Salmonella spp., Escherichia coli O157:H7, or Listeria monocytogenes populations on peanut and pistachio shell pieces. Inoculated nutshells were mixed with QUAT sanitizers, water, or 70% ethanol and enumerated immediately or after incubation at 30 °C for 48 h. None of the treatments had any immediate effect on Salmonella or E. coli O157:H7 populations on the peanut or pistachio shells. L. monocytogenes populations declined immediately on the peanut and pistachio shells treated with aQUAT or ipQUAT. After incubation, Salmonella and E. coli O157:H7 populations increased significantly on the water- or aQUAT-treated peanut and pistachio shells. L. monocytogenes populations also increased significantly on the water- or aQUAT-treated peanut shells, but levels did not change on the water-treated pistachio shells and levels were just above the limit of detection on the aQUAT-treated pistachio shells. After treatment with ipQUAT and 48-h incubation, Salmonella and E. coli O157:H7 populations decreased to or below the limit of detection on both shell types; L. monocytogenes populations remained at or below the limit of detection on both shell types. Copyright © 2014 Elsevier Ltd. All rights reserved.

Sensitivity of the ISO 6579:2002/Amd 1:2007 Standard Method for Detection of Salmonella spp. on Mesenteric Lymph Nodes from Slaughter Pigs

PubMed Central

Mainar-Jaime, R. C.; Andrés, S.; Vico, J. P.; San Román, B.; Garrido, V.

2013-01-01

The ISO 6579:2002/Amd 1:2007 (ISO) standard has been the bacteriological standard method used in the European Union for the detection of Salmonella spp. in pig mesenteric lymph nodes (MLN), but there are no published estimates of the diagnostic sensitivity (Se) of the method in this matrix. Here, the Se of the ISO (SeISO) was estimated on 675 samples selected from two populations with different Salmonella prevalences (14 farms with a ≥20% prevalence and 13 farms with a <20% prevalence) and through the use of latent-class models in concert with Bayesian inference, assuming 100% ISO specificity, and an invA-based PCR as the second diagnostic method. The SeISO was estimated to be close to 87%, while the sensitivity of the PCR reached up to 83.6% and its specificity was 97.4%. Interestingly, the bacteriological reanalysis of 33 potential false-negative (PCR-positive) samples allowed isolation of 19 (57.5%) new Salmonella strains, improving the overall diagnostic accuracy of the bacteriology. Considering the usual limitations of bacteriology regarding Se, these results support the adequacy of the ISO for the detection of Salmonella spp. from MLN and also that of the PCR-based method as an alternative or complementary (screening) test for the diagnosis of pig salmonellosis, particularly considering the cost and time benefits of the molecular procedure. PMID:23100334

Multiplex real-time PCR and culture methods for detection of Shiga toxin-producing Escherichia coli and Salmonella Thompson in strawberries, a lettuce mix and basil.

PubMed

Delbeke, S; Ceuppens, S; Holvoet, K; Samuels, E; Sampers, I; Uyttendaele, M

2015-01-16

An appropriate approach of high throughput multi-screening was verified for Shiga toxin-producing Escherichia coli (STEC) and Salmonella spp. in strawberries, lettuce and basil. Sample replicates were inoculated with STEC O157 or O26 and Salmonella Thompson (ca. 10-70, 100-700 and 1000-7000 cfu/25 g) and analysed after 1 and 5 days of storage (strawberries and lettuce at 7 °C and basil at 10 °C). After 18-24 h of enrichment at 37 °C in buffered peptone water, detection was performed using the GeneDisc multiplex PCR (stx1, stx2, eae and iroB genes) and selective culture media for isolation of STEC (with immunomagnetic separation (IMS)) and Salmonella spp. in parallel. After 1 day, the pathogenic strains were recovered from all samples for all inoculum levels, whereas reduced detection rates of STEC O157 and S. Thompson were observed after 5 days of storage in case of strawberries, in particular for the lowest inoculums level, suggesting superior survival potential for STEC O26. Overall, this study indicates the ability of PCR based screening methods for reproducible multi-detection of low numbers (10-70 cfu/25 g) of STEC and Salmonella in this type of foods. However, for the basil samples, PCR needed twofold dilution of the DNA extract to overcome inhibition. It was noted that on several occasions growth of competitive microbiota obstructed finding presumptive colonies on the selective agar media, whereas the use of an additional agar medium such as CHROMagar STEC (without IMS) improved recovery rate of STEC. Copyright © 2014 Elsevier B.V. All rights reserved.

Validation of the ANSR Salmonella method for detection of Salmonella spp. in selected foods and environmental samples.

PubMed

Mozola, Mark; Norton, Paul; Alles, Susan; Gray, R Lucas; Tolan, Jerry; Caballero, Oscar; Pinkava, Lisa; Hosking, Edan; Luplow, Karen; Rice, Jennifer

2013-01-01

ANSR Salmonella is a new molecular diagnostic assay for detection of Salmonella spp. in foods and environmental samples. The test is based on the nicking enzyme amplification reaction (NEAR) isothermal nucleic acid amplification technology. The assay platform features simple instrumentation, minimal labor, and, following a single-step 10-24 h enrichment (depending on sample type), an extremely short assay time of 30 min, including sample preparation. Detection is real-time using fluorescent molecular beacon probes. Inclusivity testing was performed using a panel of 113 strains of S. enterica and S. bongori, representing 109 serovars and all genetic subgroups. With the single exception of the rare serovar S. Weslaco, all serovars and genetic subgroups were detected. Exclusivity testing of 38 non-salmonellae, mostly Enterobacteriaceae, yielded no evidence of cross-reactivity. In comparative testing of chicken carcass rinse, raw ground turkey, raw ground beef, hot dogs, and oat cereal, there were no statistically significant differences in the number of positive results obtained with the ANSR and the U.S. Department of Agriculture-Food Safety and Inspection Service or U.S. Food and Drug Administration/Bacteriological Analytical Manual reference culture methods. In testing of swab or sponge samples from five different environmental surfaces, four trials showed no statistically significant differences in the number of positive results by the ANSR and the U.S. Food and Drug Administration/ Bacteriological Analytical Manual reference methods; in the trial with stainless steel surface, there were significantly more positive results by the ANSR method. Ruggedness experiments showed a high degree of assay robustness when deviations in reagent volumes and incubation times were introduced.

Estimation of the sensitivity of various environmental sampling methods for detection of Salmonella in duck flocks.

PubMed

Arnold, Mark E; Mueller-Doblies, Doris; Gosling, Rebecca J; Martelli, Francesca; Davies, Robert H

2015-01-01

Reports of Salmonella in ducks in the UK currently rely upon voluntary submissions from the industry, and as there is no harmonized statutory monitoring and control programme, it is difficult to compare data from different years in order to evaluate any trends in Salmonella prevalence in relation to sampling methodology. Therefore, the aim of this project was to assess the sensitivity of a selection of environmental sampling methods, including the sampling of faeces, dust and water troughs or bowls for the detection of Salmonella in duck flocks, and a range of sampling methods were applied to 67 duck flocks. Bayesian methods in the absence of a gold standard were used to provide estimates of the sensitivity of each of the sampling methods relative to the within-flock prevalence. There was a large influence of the within-flock prevalence on the sensitivity of all sample types, with sensitivity reducing as the within-flock prevalence reduced. Boot swabs (individual and pool of four), swabs of faecally contaminated areas and whole house hand-held fabric swabs showed the overall highest sensitivity for low-prevalence flocks and are recommended for use to detect Salmonella in duck flocks. The sample type with the highest proportion positive was a pool of four hair nets used as boot swabs, but this was not the most sensitive sample for low-prevalence flocks. All the environmental sampling types (faeces swabs, litter pinches, drag swabs, water trough samples and dust) had higher sensitivity than individual faeces sampling. None of the methods consistently identified all the positive flocks, and at least 10 samples would be required for even the most sensitive method (pool of four boot swabs) to detect a 5% prevalence. The sampling of dust had a low sensitivity and is not recommended for ducks.

Resistance to antimicrobial agents among Salmonella isolates recovered from layer farms and eggs in the Caribbean region.

PubMed

Adesiyun, Abiodun; Webb, Lloyd; Musai, Lisa; Louison, Bowen; Joseph, George; Stewart-Johnson, Alva; Samlal, Sannandan; Rodrigo, Shelly

2014-12-01

This investigation determined the frequency of resistance of 84 isolates of Salmonella comprising 14 serotypes recovered from layer farms in three Caribbean countries (Trinidad and Tobago, Grenada, and St. Lucia) to eight antimicrobial agents, using the disc diffusion method. Resistance among isolates of Salmonella was related to the country of recovery, type of sample, size of layer farms, and isolate serotype. Overall, all (100.0%) of the isolates exhibited resistance to one or more of seven antimicrobial agents tested, and all were susceptible to chloramphenicol. The resistance detected ranged from 11.9% to sulphamethoxazole-trimethoprim (SXT) to 100.0% to erythromycin. The difference was, however, not statistically significant (P = 0.23). Across countries, for types of samples that yielded Salmonella, significant differences in frequency of resistance were detected only to SXT (P = 0.002) in Trinidad and Tobago and to gentamycin (P = 0.027) in St. Lucia. For the three countries, the frequency of resistance to antimicrobial agents was significantly different for ampicillin (P = 0.001) and SXT (P = 0.032). A total of 83 (98.8%) of the 84 isolates exhibited 39 multidrug resistance patterns. Farm size significantly (P = 0.032) affected the frequency of resistance to kanamycin across the countries. Overall, among the 14 serotypes of Salmonella tested, significant (P < 0.05) differences in frequency of resistance were detected to kanamycin, ampicillin, and SXT. Results suggest that the relatively high frequency of resistance to six of the antimicrobial agents (erythromycin, streptomycin, gentamycin, kanamycin, ampicillin, and tetracycline) tested and the multidrug resistance detected may pose prophylactic and therapeutic concerns for chicken layer farms in the three countries studied.

Cross-sectional survey of indicator and pathogenic bacteria on vegetables sold from Asian vendors at farmers' markets in northern California.

PubMed

Pan, Fengguang; Li, Xunde; Carabez, Jennifer; Ragosta, Guy; Fernandez, Kristine L; Wang, Elaine; Thiptara, Anyarat; Antaki, Elizabeth; Atwill, Edward R

2015-03-01

A cross-sectional survey was conducted during summer 2013 to determine the occurrence of Escherichia coli, fecal coliforms (FCs), E. coli O157:H7, and Salmonella on raw vegetable commodities common to Asian cuisine from 21 vendors or farmers at six farmers' markets in northern California. Based on 242 samples from six commodities (basil, yardlong beans, bitter squash, okra, squash stems and leaves, cilantro), 100% of samples had detectable FCs and 20% had detectable E. coli. The mean concentrations were 0.67 log CFU/g and 1.26 log CFU per bundle for E. coli and 4.00 log CFU/g and 6.26 log CFU per bundle for FCs. Vegetables irrigated with ground versus surface water contained lower concentrations of FCs, but this difference was not observed for E. coli. Yardlong beans, bitter squash, and okra had lower levels of FCs compared with basil, cilantro, and squash stems and leaves. Sixteen (6.6%) samples had detectable levels of Salmonella serovars (Newport, Enteritidis, Agona, and Worthington), with the majority of positives found in cilantro and squash stems and leaves. There was a twofold higher probability of Salmonella contamination in samples from growers or vendors who stated that they used organic farming practices compared with samples from those using conventional farming practices. Lastly, the concentrations of FC and E. coli bacteria were significantly associated with Salmonella contamination: for each additional 100 CFU/g or bundle, the probability of Salmonella contamination increased by ∼15 and ∼30%, respectively. None of the samples had detectable E. coli O157:H7.

Irrigation water quality and the benefits of implementing good agricultural practices during tomato (Lycopersicum esculentum) production.

PubMed

Estrada-Acosta, M; Jiménez, M; Chaidez, C; León-Félix, J; Castro-Del Campo, N

2014-07-01

The implementation of good agricultural practices (GAP) from irrigation water to the tomato packaging process enhances the safety of fresh produce and its value throughout the food chain. The aim of the present study was to show that fresh produce farms that apply and enforce GAP could reduce the presence of Salmonella in finished produce. Samples were collected biweekly from six packing houses from the central region of Sinaloa, México, for the isolation of Salmonella spp by the ISO 6579:2002 method, and the isolated strains were serotyped and genotyped by the Kauffmman-White scheme and pulsed field gel electrophoresis (PFGE), respectively. Salmonella strains were detected in 13 (36.1 %) irrigation water samples, while only two tomato samples were positive (5.5 %). Eight different serotypes were identified in irrigation water, and Salmonella Oranienburg (34 %) was the most prevalent; however, only Salmonella Agona and Salmonella Weltevreden were present on tomatoes. Salmonella Oranienburg was the most widely dispersed and variable serotype, with 10 different PFGE profiles. Salmonella Weltevreden was isolated from both types of samples, albeit with distinct genetic profiles, implying that the sources of contamination differ. These results confirm the utility of implementing good agricultural practices to reduce Salmonella contamination in irrigation water and the packaging process.

Ten years experience of Salmonella infections in Cambridge, UK.

PubMed

Matheson, Nicholas; Kingsley, Robert A; Sturgess, Katherine; Aliyu, Sani H; Wain, John; Dougan, Gordon; Cooke, Fiona J

2010-01-01

Review of all Salmonella infections diagnosed in the Cambridge area over 10 years. All Salmonella enterica isolated in the Clinical Microbiology Laboratory, Addenbrooke's Hospital between 1.1.1999 and 31.12.2008 were included. Patient demographics, serotype and additional relevant details (travel history, resistance-type, phage-type) were recorded. 1003 episodes of Salmonella gastroenteritis were confirmed by stool culture, representing 88 serotypes. Serotypes Enteritidis (59%), Typhimurium (4.7%), Virchow (2.6%), Newport (1.8%) and Braenderup (1.7%) were the 5 most common isolates. There were an additional 37 invasive Salmonella infections (32 blood cultures, 4 tissue samples, 1 CSF). 13/15 patients with Salmonella Typhi or Salmonella Paratyphi isolated from blood or faeces with an available travel history had returned from the Indian subcontinent. 8/10 S. Typhi or Paratyphi isolates tested had reduced susceptibility to fluoroquinolones (MIC > or = 0.125 mg/L). 7/21 patients with non-typhoidal Salmonella bacteraemia were known to be immunosuppressed. This study describes Salmonella serotypes circulating within a defined geographical area over a decade. Prospective molecular analysis of isolates of S. enterica by multi-locus sequence typing (MLST) and single nucleotide polymorphism (SNP) detection will determine the geo-phylogenetic relationship of isolates within our region. 2009 The British Infection Society. Published by Elsevier Ltd. All rights reserved.

A Multiplex RT-PCR Assay for S. aureus, L. monocytogenes, and Salmonella spp. Detection in Raw Milk with Pre-enrichment.

PubMed

Ding, Tian; Suo, Yuanjie; Zhang, Zhaohuan; Liu, Donghong; Ye, Xingqian; Chen, Shiguo; Zhao, Yong

2017-01-01

This study firstly developed a multiplex real-time PCR (RT-PCR) technique combined with a pre-enrichment step to simultaneously detect Staphylococcus aureus ( S. aureus ), Listeria monocytogenes ( L. monocytogenes ) and Salmonella spp. in raw milk and the dairy farm environment (feces, soil, feed, water) in one reaction. Brain heart infusion (BHI) broth was selected for the enrichment step to increase the density of the target bacteria by using an incubation of 4 h before multiplex RT-PCR. The results showed that the detection limit of the multiplex real-time assay was approximately 10 2 CFU/mL for pure cultures and artificially contaminated milk without enrichment, while 12, 14, and 10 CFU/25 mL, respectively, for S. aureus, L. monocytogenes , and Salmonella spp. after pre-enrichment. The newly developed multiplex RT-PCR assay was applied to 46 dairy farm environmental samples and raw milk samples covering a wide variety of sample types. The results demonstrated that the multiplex RT-PCR assay coupled with the BHI enrichment broth was suitable for the simultaneous screening of S. aureus, L. monocytogenes , and Salmonella spp. in the pasture environment and in raw milk. The multiplex RT-PCR assay clearly and successfully shortened the total detection time and reduced labor compared to conventional culture-based methods for testing natural samples.

A Multiplex RT-PCR Assay for S. aureus, L. monocytogenes, and Salmonella spp. Detection in Raw Milk with Pre-enrichment

PubMed Central

Ding, Tian; Suo, Yuanjie; Zhang, Zhaohuan; Liu, Donghong; Ye, Xingqian; Chen, Shiguo; Zhao, Yong

2017-01-01

This study firstly developed a multiplex real-time PCR (RT-PCR) technique combined with a pre-enrichment step to simultaneously detect Staphylococcus aureus (S. aureus), Listeria monocytogenes (L. monocytogenes) and Salmonella spp. in raw milk and the dairy farm environment (feces, soil, feed, water) in one reaction. Brain heart infusion (BHI) broth was selected for the enrichment step to increase the density of the target bacteria by using an incubation of 4 h before multiplex RT-PCR. The results showed that the detection limit of the multiplex real-time assay was approximately 102 CFU/mL for pure cultures and artificially contaminated milk without enrichment, while 12, 14, and 10 CFU/25 mL, respectively, for S. aureus, L. monocytogenes, and Salmonella spp. after pre-enrichment. The newly developed multiplex RT-PCR assay was applied to 46 dairy farm environmental samples and raw milk samples covering a wide variety of sample types. The results demonstrated that the multiplex RT-PCR assay coupled with the BHI enrichment broth was suitable for the simultaneous screening of S. aureus, L. monocytogenes, and Salmonella spp. in the pasture environment and in raw milk. The multiplex RT-PCR assay clearly and successfully shortened the total detection time and reduced labor compared to conventional culture-based methods for testing natural samples. PMID:28620364

Protective action of Lactobacillus kefir carrying S-layer protein against Salmonella enterica serovar Enteritidis.

PubMed

Golowczyc, M A; Mobili, P; Garrote, G L; Abraham, A G; De Antoni, G L

2007-09-30

Eight Lactobacillus kefir strains isolated from different kefir grains were tested for their ability to antagonize Salmonella enterica serovar Enteritidis (Salmonella enteritidis) interaction with epithelial cells. L. kefir surface properties such as autoaggregation and coaggregation with Salmonella and adhesion to Caco-2/TC-7 cells were evaluated. L. kefir strains showed significantly different adhesion capacities, six strains were able to autoaggregate and four strains coaggregated with Salmonella. Coincubation of Salmonella with coaggregating L. kefir strains significantly decreased its capacity to adhere to and to invade Caco-2/TC-7 cells. This was not observed with non coaggregating L. kefir strains. Spent culture supernatants of L. kefir contain significant amounts of S-layer proteins. Salmonella pretreated with spent culture supernatants (pH 4.5-4.7) from all tested L. kefir strains showed a significant decrease in association and invasion to Caco-2/TC-7 cells. Artificially acidified MRS containing lactic acid to a final concentration and pH equivalent to lactobacilli spent culture supernatants did not show any protective action. Pretreatment of this pathogen with spent culture supernatants reduced microvilli disorganization produced by Salmonella. In addition, Salmonella pretreated with S-layer proteins extracted from coaggregating and non coaggregating L. kefir strains were unable to invade Caco-2/TC-7 cells. After treatment, L. kefir S-layer protein was detected associated with Salmonella, suggesting a protective role of this protein on association and invasion.

Disinfectant susceptibility of different Salmonella serotypes isolated from chicken and egg production chains.

PubMed

Long, M; Lai, H; Deng, W; Zhou, K; Li, B; Liu, S; Fan, L; Wang, H; Zou, L

2016-09-01

The study aimed to serotype the Salmonella isolates recovered from chicken and egg production chains, and to investigate the disinfectant resistance phenotypes and genotypes of these isolates. The Salmonella isolates were serotyped, and the minimal inhibitory concentrations (MICs) of disinfectants were determined. Results showed that the Salmonella isolates recovered from both chains were diverse, and the serotypes in each part of the production chain and between the two production chains were significantly different. In the chicken production chain, 19 different serotypes were recovered, while only five serotypes were found in the egg production chain. The isolates showed a high susceptibility to didecyldimethylammonium bromide (DDAB) but a low susceptibility to benzalkonium chloride (BC), benzalkonium bromide (BAB) and chlorhexidine (CHX). Salmonella Enteritidis and Salmonella Typhimurium were more resistant to BC and BAB. The qacEΔ1 and qacF resistance genes were detected in 26·7 and 7·7% of the isolates respectively. The qacEΔ1 gene was frequently found in Salmonella Derby and Salm. Enteritidis (P < 0·05). Our findings indicated that Salmonella was commonly present in both chains, and could serve as a critical vector in spreading disinfectant resistance associated with different serotypes. This study first demonstrated disinfectant resistance phenotypes and genotypes of serotyped Salmonella. The study highlights the need for monitoring the disinfectant resistance varied in different Salmonella serotypes. © 2016 The Society for Applied Microbiology.

Sampling naturally contaminated broiler carcasses for Salmonella by three different methods

USDA-ARS?s Scientific Manuscript database

Postchill neck skin (NS) maceration and whole carcass rinsing (WCR) are frequently used methods to detect salmonellae from commercially processed broilers. These are practical, nondestructive methods, but they are insensitive and may result in frequent false negatives (20 to 40%). NS samples only ...

Accelerating sample preparation through enzyme-assisted microfiltration of Salmonella in chicken extract

USDA-ARS?s Scientific Manuscript database

Microfiltration of chicken extracts has the potential to significantly decrease the time required to detect Salmonella, as long as the extract can be efficiently filtered and the pathogenic microorganisms kept in a viable state during this process. We present conditions that enable microfiltration ...

Effect Of Colony Numbers Selected From Plating Media On Salmonella Serogroup Detection From Naturally Contaminated Chicken Carcasses

USDA-ARS?s Scientific Manuscript database

Introduction: Attribution studies are being undertaken to link Salmonella serotypes associated with human illness to food sources. However, studies have demonstrated that culture techniques often influence the sensitivity and specificity associated with bacterial recovery. Purpose: This study ...

Investigations of Salmonella enterica serovar newport infections of oysters by using immunohistochemistry and knockout mutagenesis.

PubMed

Morrison, Christopher M; Dial, Sharon M; Day, William A; Joens, Lynn A

2012-04-01

The consumption of raw oysters is an important risk factor in the acquisition of food-borne disease, with Salmonella being one of a number of pathogens that have been found in market oysters. Previous work by our lab found that Salmonella was capable of surviving in oysters for over 2 months under laboratory conditions, and this study sought to further investigate Salmonella's tissue affinity and mechanism of persistence within the oysters. Immunohistochemistry was used to show that Salmonella was capable of breaching the epithelial barriers, infecting the deeper connective tissues of the oysters, and evading destruction by the oysters' phagocytic hemocytes. To further investigate the mechanism of these infections, genes vital to the function of Salmonella's two main type III secretion systems were disrupted and the survivability of these knockout mutants within oysters was assayed. When the Salmonella pathogenicity island 1 and 2 mutant strains were exposed to oysters, there were no detectable deficiencies in their abilities to survive, suggesting that Salmonella's long-term infection of oysters does not rely upon these two important pathogenicity islands and must be due to some other, currently unknown, mechanism.