Enumeration Methods and Production of Enterotoxins in Food-Derived Staphylococcus aureus

2012 ◽  
Vol 95 (1) ◽  
pp. 105-110 ◽  
Author(s):  
Chi Zhang ◽  
Deqin Zhang ◽  
Jun Yang ◽  
Jungui Zhou ◽  
Qilong Hu ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Culture Media ◽  
Food Poisoning ◽  
Food Samples ◽  
Rabbit Plasma ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Significant Difference ◽  
Statistical Relationships ◽  
Selection Of

Abstract Staphylococcal food poisoning is one of the most common foodborne diseases worldwide; it results from the ingestion of staphylococcal enterotoxins (SEs) in food, mainly Staphylococcus aureus. This study investigated the statistical relationships among morphological enumerations of food-derived S. aureus and production of SEs using different methodologies. Food samples naturally contaminated with coagulase-positive S. aureus were submitted for enumeration on Baird-Parker (BP) agar, Rabbit Plasma Fibrinogen agar (RPFA), and PetrifilmTM Staph Express count system (STX), and the morphologically typical colonies were isolated for VIDAS and real-time (RT) PCR tests. RPFA and STX displayed better performance for the enumeration of SE-positive S. aureus when compared with BP, including higher frequencies of SE-positive isolates and better correlation indices between typical and SE-positive counts. Among all the evaluated culture media, no significant difference (P > 0.05) was shown on the frequencies of typical colonies that carried 11 individual se genes. In addition, results for SE identification between VIDAS and RT-PCR assay were unparalleled. These data will be valuable for the selection of methods for inspection of food-derived S. aureus.

Multiplex PCR Detection of Enterotoxin Genes in Aeromonas spp. from Suspect Food Samples in Northern Taiwan

2008 ◽  
Vol 71 (10) ◽  
pp. 2094-2099 ◽  
Author(s):  
YU-CHANG CHANG ◽  
JAN-YI WANG ◽  
AMMAIYAPPAN SELVAM ◽  
SHU-CHEN KAO ◽  
SHANG-SHYNG YANG ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Diarrheal Disease ◽  
Simultaneous Detection ◽  
Food Poisoning ◽  
Food Samples ◽  
Pcr Assay ◽  
Multiplex Pcr Assay ◽  
Enterotoxin Genes ◽  
Causative Agents ◽  
Northern Taiwan

Aeromonads possess an array of virulence factors and are causative agents of a number of human infections. Among them, genes of one cytotoxic (Act) and two cytotonic (Alt, Ast) enterotoxins are implicated in a human diarrheal disease. A rapid, specific, simultaneous detection of these enterotoxin genes in suspected food poisoning samples is not yet reported. Hence, a multiplex PCR assay was designed to amplify the cytotoxic (act), heat-labile cytotonic (alt), and heat-stable cytotonic (ast) enterotoxin genes of aeromonads. The PCR assay was tested with 133 Aeromonas spp. isolated from suspect food poisoning samples and retail samples of poultry and fish from wet markets in and around Taipei, Northern Taiwan. The Aeromonas spp. isolates were divided into six genotypes based on absence or presence of one or more enterotoxin genes. Of these 133 isolates, Aeromonas caviae (52.5%) and Aeromonas hydrophila (43.4%) were the most frequently isolated species from food poisoning samples and retail samples, respectively. Among the species, A. hydrophila had a significantly higher proportion for harboring three enterotoxin genes than had the others, whereas Aeromonas encheleia, considered a nonpathogen, was found harboring three enterotoxin genes. The multiplex PCR assays are rapid and specific, and provide a useful tool for the detection and genotyping of enterotoxin genes of aeromonads.

scholarly journals Molecular Detection of Staphylococcal Enterotoxins and mecA Genes Products in Selected Food Samples Collected from Different Areas in Khartoum State

2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Mohammed Yahya Ahmed ◽  
Hashim Abdalbagi Ali ◽  
Babbiker Mohammed Taher Gorish ◽  
Sara Omer Ali ◽  
Eman Saif Aldein Abdalrhim ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Meca Gene ◽  
Food Poisoning ◽  
Meat Products ◽  
Low Frequency ◽  
Raw Milk ◽  
Food Samples ◽  
Increased Risk ◽  
Enterotoxin B ◽  
Staphylococcus Aureus Enterotoxins

Staphylococcal food poisoning is an intoxication that results from the consumption of improperly prepared or stored foods containing sufficient amounts of one or more preformed S. aureus enterotoxins. Nowadays, many researchers worldwide noted an emergence of resistant strains such as Staphylococci particularly for the antibiotic methicillin. Therefore, this study was aimed to determine the existence of Staphylococcus aureus and its enterotoxins, mecA genes, in selected food samples. A total of 400 selected food samples were collected from different areas in Khartoum State. The selected foods included cheese, meat products, fish, and raw milk. One hundred samples from each type of food were cultivated, and the resultant growth yielded 137 (34.25%) S. aureus, 126 (31.5%) bacteria other than S. aureus, and 137 (34.25%) yielded no growth. Eighty-four of the 137 S. aureus isolates were randomly selected and tested for the presence of mecA and enterotoxin genes. The oxacillin sensitivity test showed that 15 (11%) of 137 S. aureus isolates were oxacillin resistant. The PCR assay showed that the mecA gene was detected in 15 of 84 (17%) S. aureus isolates. Simultaneously, only 2 (2.385%) out of 84 S. aureus isolates showed an enterotoxin B gene product. There was a relatively moderate prevalence of methicillin-resistant Staphylococcus aureus with very low frequency of enterotoxin B gene in different kinds of selected food samples collected from Khartoum State. These findings elucidate the increased risk on public in Khartoum being affected by Staphylococcal food poisoning upon consumption of dairy or meat products prepared in unhygienic conditions that could lead to intoxication by Staphylococcus aureus enterotoxins.

Cellular Compatibility of a New Tantalum-Niobium Scaffold Material

2020 ◽  
Author(s):  
Jianan Ouyang ◽  
Zhenhan Deng ◽  
Kang Chen ◽  
Jianyi Xiong ◽  
Ying Li ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Type I Collagen ◽  
Material Surface ◽  
Control Group ◽  
Type I ◽  
Scaffold Material ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Porous Tantalum ◽  
Significant Difference

Abstract [Objective] To determine the cellular compatibility of porous tantalum-niobium (Ta-Nb) material. [Method] Rabbit osteoblasts were co-cultured with porous Ta-Nb material. The cell proliferation was detected by CCK-8 method, and the cell adhesion was observed under scanning electron microscope (SEM). The expressions of type-I collagen and osteocalcin were detected by RT-PCR assay. [Results] CCK-8 detection indicated that the cell proliferation on the porous Ta-Nb material showed no difference from that of the control group (P>0.05). SEM revealed that a large amount of cells adhered onto the surface and in the pores of the material. The number of cells on the material surface increased obviously over time. RT-PCR assay showed that with the prolonging of the time of co-culture, the expression of type-I collagen was enhanced (P<0.05), while the osteocalcin expression exhibited no significant difference (P>0.05[Conclusion] Porous Ta-Nb scaffold material can be used to promote the adhesion, growth and differentiation of osteoblasts with satisfactory cellular compatibility.

scholarly journals TSST-1, enterotoxin and bacteriocin-like substance production by Staphylococcus aureus isolated from foods

2013 ◽  
Vol 65 (5) ◽  
pp. 1537-1544 ◽  
Author(s):  
S.A. Carvalho ◽  
L.S. Carmo ◽  
E.F. Abreu ◽  
R.S. Dias ◽  
A.C.M. Apolônio ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Bovine Milk ◽  
Food Poisoning ◽  
Brain Heart Infusion ◽  
Food Samples ◽  
Antagonistic Activity ◽  
Toxic Shock ◽  
Toxic Shock Syndrome Toxin ◽  
Reference Strains ◽  
Infusion Agar

The production of Toxic Shock Syndrome Toxin-1 (TSST-1), enterotoxins and bacteriocin-like substances was evaluated in 95 strains of Staphylococcus aureus recovered from raw bovine milk (n=31) and from food samples involved in staphylococcal food poisoning (n=64). Enterotoxigenicity tests with the membrane over agar associated to optimal sensibility plate assays were performed and showed that 96.77% of strains recovered from milk and 95.31% from food samples produced enterotoxins A, B, C, D or TSST-1. Reference strains S. epidermidis, Bacillus cereus, Listeria monocytogenes, Lactobacillus casei, Pseudomonas aeruginosa, S. aureus, Salmonella Typhimurium, Escherichia coli, Enterococcus faecalis and Bacteroides fragilis were used as indicator bacteria in the antagonistic assays, the first five being sensitive to antagonistic substances. Brain heart infusion agar, in pH values ranging from 5.0 to 7.0 in aerobic atmosphere showed to be the optimum condition for antagonistic activity as evaluated with the best producer strains against the most sensitive indicator bacterium, L. monocytogenes. Sensitivity to enzymes confirmed the proteinaceous nature of these substances. Neither bacteriophage activity nor fatty acids were detected and the antagonistic activity was not due to residual chloroform. Results did not establish a positive correlation between the bacteriocinogenic profile and toxigenicity in the tested S. aureus strains.

An Immunomagnetic PCR Signal Amplification Assay for Sensitive Detection of Staphylococcus aureus Enterotoxins in Foods

2009 ◽  
Vol 72 (12) ◽  
pp. 2538-2546 ◽  
Author(s):  
LEENALITHA PANNEERSEELAN ◽  
PETER M. MURIANA
Keyword(s):  
Staphylococcus Aureus ◽  
Signal Amplification ◽  
Magnetic Beads ◽  
Food Poisoning ◽  
High Sensitivity ◽  
The United States ◽  
Food Samples ◽  
Dna Oligonucleotides ◽  
Amplification Assay ◽  
Covalently Linked

Enterotoxigenic strains of Staphylococcus aureus produce a variety of heat-stable staphylococcal enterotoxins (SEs) that are a prevalent cause of food poisoning in the United States and other countries. Many immunological and biochemical assays often work well in buffer systems but are hindered when tested in the complex chemical environment of foods. To overcome these biases and improve the limits of detection, we implemented an immunomagnetic PCR signal amplification assay (iPCR-SA) for recovery and detection of SEA and SEB in foods. Anti-SEA or anti-SEB primary antibodies were coated onto COOH-modified magnetic beads using 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide reagent. Secondary antibodies were covalently linked to amino-modified reporter DNA oligonucleotides (563 bp) via the linker molecule succinimidyl-4[N-maleimidomethyl]-cyclohexane-1-carboxylate. An internal 159-bp portion of the reporter DNA retained by the captured toxin molecule was then amplified by real-time PCR. A semiautomated Bead Retriever proved extremely helpful in both the application of the conjugation chemistries and required washes and the recovery and washing of bead-conjugated toxin from tested food samples. The procedure was simple, and analyses were completed in 5 to 6 h. The assay was sufficiently robust that we were able to detect SEA and SEB in tryptic soy broth, milk, lemon cream pie, tuna salad, deli turkey, and ground turkey at levels as low as 7.5 fg/ml. SE was still detected at high sensitivity after heating in food samples for typical pasteurization or cooking regimens. Sensitivity was diminished only when samples were subjected to extreme heating.

scholarly journals Serological detection of enterotoxin in foods implicated in staphylococcal food poisoning

1972 ◽  
Vol 70 (4) ◽  
pp. 755-762 ◽  
Author(s):  
R. J. Gilbert ◽  
Antonnette A. Wieneke ◽  
Janice Lanser ◽  
Magda Šimkovičová
Keyword(s):  
Staphylococcus Aureus ◽  
Food Poisoning ◽  
Food Samples ◽  
Serological Detection

SUMMARYTwo methods are described for the extraction of enterotoxin from foods incriminated in incidents of staphylococcal food poisoning. Enterotoxin was detected serologically in 12 of 24 food samples from 20 separate incidents: eight samples contained enterotoxin A, three contained D and one both A and B. The amount of enterotoxin in nine foods, based on 100 % recovery, varied from 0·02 to 0·09 μg./g.Data are also given on the numbers ofStaphylococcus aureusisolated from samples of food from 39 food poisoning incidents. Colony counts varied between 7·5 × 105and 9 × 109/g. with a median value of 7 × 107/g.

Investigations on the Frequency of Norovirus Contamination of Ready-to-Eat Food Items in Istanbul, Turkey, by Using Real-Time Reverse Transcription PCR

2011 ◽  
Vol 74 (5) ◽  
pp. 840-843 ◽  
Author(s):  
AYSUN YILMAZ ◽  
KAMIL BOSTAN ◽  
EDA ALTAN ◽  
KARLO MURATOGLU ◽  
NURI TURAN ◽  
...  
Keyword(s):  
Real Time ◽  
Reverse Transcription ◽  
Epidemiological Studies ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Food Items ◽  
Reverse Transcription Pcr ◽  
Significant Difference ◽  
Tomato Sample ◽  
Pcr Assays

Investigation of norovirus (NoV) contamination of food items is important because many outbreaks occur after consumption of contaminated shellfish, vegetables, fruits, and water. The frequency of NoV contamination in food items has not previously been investigated in Turkey. The aim of this study was to investigate the frequency of human NoV genogroups (G) I and II in ready-to-eat tomatoes, parsley, green onion, lettuce, mixed salads, and cracked wheat balls. RNA was extracted with the RNeasy Mini Kit, and a real-time reverse transcription (RT) PCR assay was performed using primers specific for NoV GI and GII. Among the 525 samples analyzed, NoV GII was detected in 1 green onion sample and 1 tomato sample by both SYBR Green and TaqMan real-time RT-PCR assays; no GI virus was detected. The Enterobactericaeae and Escherichia coli levels in the NoV-positive green onion were 6.56 and 1.28 log CFU/g, and those in the tomato were 5.55 and 1.30 log CFU/g, respectively. No significant difference in the bacterial levels was found between the NoV-positive and NoV-negative samples. This study is the first in which NoV GII was found in ready-to-eat food collected from Istanbul, Turkey; thus, these foods may be considered a risk to human health. Epidemiological studies and measures to prevent NoV infection should be considered.

scholarly journals Molecular detection of Staphylococcal enterotoxins and mecA genes products in food samples collected from different areas in Khartoum state

2020 ◽  
Author(s):  
Mohammed Yahya ◽  
Hashim Abdalbagi Ali ◽  
Babbiker Mohammed Taher Gorish ◽  
Sara Omer Ali ◽  
Eman Saif Aldein Abdalrhim ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Meca Gene ◽  
Food Poisoning ◽  
Meat Products ◽  
Low Frequency ◽  
Raw Milk ◽  
Food Samples ◽  
Enterotoxin Genes ◽  
Resistant Strains ◽  
Enterotoxin B

Abstract Background Staphylococcal Food Poisoning is an intoxication that results from the consumption of improperly prepared or stored foods containing sufficient amounts of one or more preformed S. aureus enterotoxins. Now days many researchers worldwide noted an emerging of resistant strains Staphylococci especially for the antibiotic Methicillin. Therefore, this study was aimed to determine the existence of Staphylococcus aureus and its enterotoxins, mecA genes in food samples. Results A total of 400 samples were collected from different areas in Khartoum state. The type of foods included Cheese, Meat products, Fish and Raw milk, 100 samples for each. out of 400 samples cultivated 137 (34.25%) isolates were identified as S. aureus, 126 (31.5%) were identified as bacteria other than S. aureus and 137 (34.25%) were yield no growth. Of 137 S.aureus isolates, 84 were randomly selected and examined for the presence mecA and enterotoxin genes products. Oxacillin sensitivity test showed that 15(11%) of 137 S.aureus isolates were Oxacillin resistant. The PCR assay showed that mecA gene was detected in 15 of 84 (17%) S. aureus isolates. While only 2 (2.385%) out of 84 S. aureus isolates were show an enterotoxin B gene product. Conclusion There was a relatively moderate prevalence of Methicillin resistant staphylococcus aureus with very low frequency of enterotoxin B gene in different kinds of food samples which collected from Khartoum state. These findings highlight the high potential risk for consumers of meat and dairy products especially in the absence of strict hygienic and preventive measures to avoid Staphylococcus aureus enterotoxins production in foods.

scholarly journals Molecular Characteristics of Staphylococcus aureus From Food Samples and Food Poisoning Outbreaks in Shijiazhuang, China

2021 ◽  
Vol 12 ◽  
Author(s):  
Guoping Lv ◽  
Ruiping Jiang ◽  
Han Zhang ◽  
Lei Wang ◽  
Lijie Li ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Potential Risk ◽  
Food Poisoning ◽  
Opportunistic Pathogen ◽  
Food Samples ◽  
Staphylococcal Enterotoxin ◽  
Molecular Characteristics ◽  
Resistant Gene ◽  
Human Infections ◽  
The Relationship

As an opportunistic pathogen worldwide, Staphylococcus aureus can cause food poisoning and human infections. This study investigated the sequence typing, the penicillin (blaZ) and methicillin (mec) resistance profiles of S. aureus from food samples and food poisoning outbreaks in Shijiazhuang City, and the staphylococcal enterotoxin (SE) types of the S. aureus isolates from food poisoning. A total of 138 foodborne S. aureus isolates were distributed into 8 clonal complexes (CCs) and 12 singletons. CC1, CC5, CC8, CC15, CC97, CC59, CC398, CC88, and CC7 were the predominant CCs of foodborne S. aureus isolates. Moreover, CC59, CC15, and CC5 were the most prevalent CCs in food poisoning outbreaks. SEE was the most commonly detected SE in food poisoning isolates. One hundred thirty-three S. aureus isolates harbored the penicillin-resistant gene blaZ, and nine isolates carried the mec gene. The present study further explained the relationship between S. aureus and foods and food poisoning and indicated the potential risk of S. aureus infection.

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