Evaluation of real-time PCR coupled with immunomagnetic separation or centrifugation for the detection of healthy and sanitizer-injured Salmonella spp. on mung bean sprouts

Abstract 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 bothcost 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. protocol. The results of the method modification were compared to standard cultural reference methods for diced tomatoes, chocolate, and deli ham. All threematrixes 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 reliabilityas a rapid, simplified, method for the preparation of samples and subsequent detection of Salmonella in diced tomatoes, chocolate, and deliham.

Download Full-text

Efficacy of chlorine and peroxyacetic acid on reduction of natural microflora, Escherichia coli O157:H7, Listeria monocyotgenes and Salmonella spp. on mung bean sprouts

Food Microbiology ◽

10.1016/j.fm.2013.05.001 ◽

2013 ◽

Vol 36 (2) ◽

pp. 475-480 ◽

Cited By ~ 33

Author(s):

Shan Yu Neo ◽

Pei Yan Lim ◽

Li Kai Phua ◽

Gek Hoon Khoo ◽

Su-Jung Kim ◽

...

Keyword(s):

Escherichia Coli ◽

Mung Bean ◽

Escherichia Coli O157 ◽

Peroxyacetic Acid ◽

Salmonella Spp ◽

Mung Bean Sprouts ◽

Natural Microflora ◽

Bean Sprouts

Download Full-text

Combination of Immunomagnetic Separation with Real-Time PCR for Rapid Detection of Salmonella in Milk, Ground Beef, and Alfalfa Sprouts

Journal of Food Protection ◽

10.4315/0362-028x-68.3.557 ◽

2005 ◽

Vol 68 (3) ◽

pp. 557-561 ◽

Cited By ~ 43

Author(s):

BIRCE MERCANOGˇLU ◽

MANSEL W. GRIFFITHS

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Rapid Detection ◽

Ground Beef ◽

Immunomagnetic Separation ◽

Pcr Assay ◽

Salmonella Spp ◽

Pasteurized Milk ◽

Alfalfa Sprouts ◽

Sensitive Method

A rapid, specific, and sensitive method for detecting Salmonella spp. in pasteurized milk, ground beef, and alfalfa sprouts was developed. The method combined immunomagnetic separation with a real-time PCR assay based on the double-stranded DNA binding dye SYBR Green I. The primers used produced a product with a melting temperature of 87 ± 0.5°C during the PCR assay by amplifying a 284-bp sequence from the invasive gene (invA) of Salmonella. The method was successful in detecting 20 Salmonella strains, but the expected PCR product was not formed by any of 11 other bacterial strains. To test this combined method for the monitoring of Salmonella, Salmonella enterica serotype Newport was inoculated into 52 samples each of pasteurized milk, ground beef, and alfalfa sprouts. Following a 10-h nonselective enrichment step in buffered peptone water, cells were removed by immunomagnetic separation and DNA extracted using the High Pure PCR template preparation kit. The DNA produced was used as a template in the real-time PCR assay. When spiked pasteurized milk, ground beef, and alfalfa sprout samples were analyzed by this protocol, an initial inoculum of 1 CFU/ml, 25 CFU/25 g, and 1.5 CFU/25 g, respectively, was detectable within 13 h. These results indicate that the combination of immunomagnetic separation and real-time PCR assay was a highly specific and sensitive method for the rapid detection of Salmonella.

Download Full-text

A Simple Spectrofluorometric Method for the Determination of Total Auxins in Mung Bean Sprouts

Journal of Analytical Chemistry ◽

10.1134/s1061934820110155 ◽

2020 ◽

Vol 75 (11) ◽

pp. 1404-1407

Author(s):

Aziguli Yigaimu ◽

Jiahua Chang ◽

Amina Hoji ◽

Turghun Muhammad ◽

Burabiye Yakup ◽

...

Keyword(s):

Mung Bean ◽

Mung Bean Sprouts ◽

Spectrofluorometric Method ◽

Bean Sprouts

Download Full-text

Scanning electron microscopy of native biofilms on mung bean sprouts

Canadian Journal of Microbiology ◽

10.1139/w03-002 ◽

2003 ◽

Vol 49 (1) ◽

pp. 45-50 ◽

Cited By ~ 32

Author(s):

William F Fett ◽

Peter H Cooke

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Food Safety ◽

Vigna Radiata ◽

Mung Bean ◽

Fibrillar Material ◽

Mung Bean Sprouts ◽

Native Microflora ◽

Bean Sprouts ◽

Scanning Electron

Native biofilms present on the adaxial surface of cotyledons of mung bean sprouts (Vigna radiata) were studied by use of scanning electron microscopy. Biofilms were abundant on the cotyledon surfaces and were comprised of rod-shaped bacteria, cocci-shaped bacteria, or yeasts, often with one type of microbe predominant. In contrast to our earlier study of biofilms on green sprouts (alfalfa, clover, broccoli, and sunflower), yeast and cocci were abundant on mung bean. Filamentous fungi were not observed. Sheet-like or fibrillar material (presumably composed of secreted microbial polysaccharides, proteins, lipids, and nucleic acids) fully or partially covered the biofilms. Biofilms up to 5 mm in length were observed, and some biofilms were comprised of more than just a monolayer of microbial cells. Native biofilms on sprout surfaces undoubtedly play an important role in the ecology of plant epiphytic microbes and may also afford protected sites for plant and human bacterial pathogens.Key words: mung bean sprouts, biofilms, native microflora, scanning electron microscopy, food safety.

Download Full-text

Enzymes of Nucleic Acid Metabolism from Mung Bean Sprouts

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)96545-2 ◽

1966 ◽

Vol 241 (12) ◽

pp. 2876-2880 ◽

Cited By ~ 2

Author(s):

Hubert S. Loring ◽

J.E. McLennan ◽

Tom L. Walters

Keyword(s):

Nucleic Acid ◽

Mung Bean ◽

Acid Metabolism ◽

Nucleic Acid Metabolism ◽

Mung Bean Sprouts ◽

Bean Sprouts

Download Full-text

A Nuclease from Mung Bean Sprouts

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)93952-9 ◽

1962 ◽

Vol 237 (2) ◽

pp. 506-511 ◽

Cited By ~ 6

Author(s):

Shan-ching Sung ◽

M. Laskowski

Keyword(s):

Mung Bean ◽

Mung Bean Sprouts ◽

Bean Sprouts

Download Full-text

Effect of putative probiont Enterococcus hirae on the hematological parameters of juvenile African catfish, Clarias gariepinus (Burchell, 1822) during pre- and post-challenge against Aeromonas hydrophila

Malaysian Journal of Fundamental and Applied Sciences ◽

10.11113/mjfas.v14n3.1097 ◽

2018 ◽

Vol 14 (3) ◽

pp. 423-428 ◽

Cited By ~ 1

Author(s):

Nur Hidayahanum Hamid ◽

Hassan Mohd Daud ◽

Prapansak Srisapoome ◽

Hasliza Abu Hassim ◽

Md Sabri Mohd Yusoff ◽

...

Keyword(s):

Aeromonas Hydrophila ◽

Mung Bean ◽

Hematological Parameters ◽

Clarias Gariepinus ◽

African Catfish ◽

Post Challenge ◽

Control Groups ◽

Blood Samples ◽

Mung Bean Sprouts ◽

Bean Sprouts

Probiotics have been widely known to have the ability to improve the immune system of livestock and aquatic animal. The present study was carried out to evaluate the effect of dietary supplementation of two probiotic isolates of Enterococcus faecium on hematological parameters of juvenile African catfish, Clarias gariepinus during pre- and post-challenge with aquatic pathogen, Aeromonas hydrophila. The probiotics were previously isolated from vegetable wastes (mung bean sprouts, Vigna radiate and cucumber, Cucumis sativus) which have been fermented for 7 days. The experimental fish (270 tails) with an average weight of 5.13 ± 1.03 g were distributed and divided randomly into i) control (30 tails), fed with commercial diet ii) E1 (30 tails), fed with diets supplemented with 108 CFU/ml of E. faecium isolated from fermented cucumber, iii) E2 (30 tails), fed with 108 CFU/ml of E. faecium isolated from fermented mung bean sprouts. The feeding trial was conducted for 50 days. All experimental groups were then challenged with A. hydrophila (1.5 × 106 CFU/mL) via intraperitoneal injection on day 51. Prior to challenge, blood samples were collected from five fish randomly selected from each group on the day 51 (pre-challenge). After 72 hours of post-challenge, blood samples were again collected from five fish from each groups. The hematological parameters such as total erythrocyte count (RBC), total leucocyte count (WBC), packed cell volume (PCV), hemoglobin (Hb), the derived blood indices of mean corpuscular volume (MCV) and corpuscular hemoglobin concentration (MCHC) were examined. Hematological profiles of pre- and post-challenge infected juvenile catfish were compared with the control groups. The RBC, Hb, WBC, PCV, MCV and MCHC of fish fed with probiotics showed higher significant difference (P<0.05) as compared to control groups during pre- and post-challenge of pathogen. The high level of RBC and WBC during pre- and post-challenge showed the capability of the probiotics to improve the immune response of juvenile African catfish and thus increased the fish disease resistance against A. hydrophila infection. The result suggested that E. faecium could be used effectively as a probiotic in aquaculture.

Download Full-text