Toxin genes profiles and toxin production ability of Bacillus cereus isolated from clinical and food samples

Bacillus cereus can cause diarrheal and emetic type of food poisoning but little study has been done on the main toxins of food poisoning caused by B. cereus in Korea. The objective of this study is to characterize the toxin gene profiles and toxin-producing ability of 120 B. cereus isolates from clinical and food samples in Korea. The detection rate of nheABC, hblCDA, entFM, and cytK enterotoxin gene among all B. cereus strains was 94.2, 90.0, 65.8, and 52.5%, respectively. The ces gene encoding emetic toxin was not detected in all strains. Bacillus cereus strains carried at least 1 of the 8 enterotoxin genes were classified into 12 groups according to the presence or absence of 8 virulence genes. The 3 major patterns, I (nheABC, hblCDA, entFM, and cytK gene), II (nheABC, hblCDA and entFM gene), and VI (nheABC and hblCDA gene), accounted for 79.2% of all strains (95 out of 120 B. cereus isolates). Non-hemolytic enterotoxin (NHE) and hemolysin BL (HBL) enterotoxins were produced by 107 and 100 strains, respectively. Our finding revealed that NHE and HBL enterotoxins encoded by nhe and hbl genes were the major toxins among B. cereus tested in this study and enterotoxic type of B. cereus was predominant in Korea.     

Incidence and characterization of Bacillus cereus isolated from traditional fermented meals in Nigeria

The aim of this study was to examine the presence of Bacillus cereus in fermented meals used in food seasoning in Nigeria. The microbial profiles of iru and ogiri, two Nigerian fermented vegetable proteins, were examined for presence of B. cereus. In the 50 samples tested, B. cereus was detected in all the samples, with the level of detection ranging from log 6.3 to log 8.3 g(-1) sample. Phenotypic characteristics of the B. cereus isolates showed that all of them could not ferment many sugars, most especially mannitol, but they utilized propionate citrate as a source of carbon and grew anaerobically. The isolates do not produce gas from glucose but hydrolyzed starch, casein, and gelatin. API-50CHB combined with API-20E identified the isolates as B. cereus. The diarrheal enterotoxin was detected by a reversed passive latex agglutination test kit. Results showed no significant difference in toxin production between ogiri and iru B. cereus isolated from different sources; all the isolates also demonstrated positive hemolytic activity. The API-ZYM enzyme profile showed that the strains have poor hydrolytic enzyme potential; hence, their possible contributions to the fermentation of vegetable protein is doubtful. This study established the proliferation of B. cereus in fermented protein meal and determined the diarrheal toxin production potential of the organism.     

Enterotoxins and emetic toxins production by Bacillus cereus and other species of Bacillus isolated from Soumbala and Bikalga, African alkaline fermented food condiments

The ability of various species of Bacillus from fermented seeds of Parkia biglobosa known as African locust bean (Soumbala) and fermented seeds of Hibiscus sabdariffa (Bikalga) was investigated. The study included screening of the isolates by haemolysis on blood agar, detection of toxins in broth and during the fermentation of African locust bean using the Bacillus cereus Enterotoxin Reverse Passive Latex Agglutination test kit (BCET-RPLA) and the Bacillus Diarrhoeal Enterotoxin Visual Immunoassay (BDEVIA). Detection of genes encoding cytotoxin K (CytK), haemolysin BL (Hbl A, Hbl C, Hbl D), non-hemolytic enterotoxin (NheA, NheB, NheC) and EM1 specific of emetic toxin producers was also investigated using PCR with single pair and multiplex primers. Of 41 isolates, 29 Bacillus belonging to the species of B. cereus, Bacillus subtilis, Bacillus licheniformis and Bacillus pumilus showed haemolysis on blood agar. Using RPLA, enterotoxin production was detected for three isolates of B. cereus in broth and all B. cereus (9) in fermented seeds. Using BDEVIA, enterotoxin production was detected in broth as well as in fermented seeds for all B. cereus isolates. None of the isolates belonging to the other Bacillus species was able to produce enterotoxins either by RPLA or BDEVIA. Nhe genes were detected in all B. cereus while Hbl and CytK genes were detected respectively in five and six B. cereus strains. A weak presence of Hbl (A, D) and CytK genes was detected in two isolates of B. subtilis and one of B. licheniformis but results were inconsistent, especially for Hbl genes. The emetic specific gene fragment EM1 was not detected in any of the isolates studied.     

Genetic diversity, antimicrobial resistance, and toxigenic profiles of Bacillus cereus strains isolated from Sunsik

Bacillus cereus can cause emetic and diarrheal types of food poisoning, but little study has been done on the toxins and toxin-encoding genes of B. cereus strains isolated from Sunsik, a Korean ready-to-eat food prepared from grains, fruits, and vegetables. In this study, 39 unique B. cereus strains were isolated and identified from Sunsik samples, with an average contamination level of 10 to 200 CFU/g. The detection rates of the hblACD, cytK, and bceT genes among all the strains were 48.7, 66.7, and 87.1%, respectively. All 39 B. cereus strains carried nheABC and entFM genes, and 36 strains also had the ces gene, which encodes an emetic toxin. Nonhemolytic enterotoxin and hemolysin BL enterotoxin were produced by 39 and 26 strains, respectively. The strains were separated into 13 profiles based on the presence or absence of toxins and their genes, as determined by antibody tests and PCR analysis. Profile 1 was the largest group, comprising 30.7% (12 of 39) of the B. cereus strains tested; these strains harbored all toxins and their genes. The B. cereus strains were susceptible to most of the antibiotics tested but were highly resistant to b -lactam antibiotics. The repetitive element sequence polymorphism PCR fingerprints of the B. cereus strains were not influenced by the presence of toxin genes or antibiotic resistance profiles. Our results suggest that B. cereus strains from Sunsik could cause either the diarrheal or emetic types of food poisoning because all strains isolated contained at least one toxin and its gene, although the level of B. cereus contamination in Sunsik was low.     

Detection of Bacillus cereus in foods by colony hybridization using PCR-generated probe and characterization of isolates for toxins by PCR

Isolates of Bacillus cereus from traditional Indian foods were detected by colony hybridization using the PCR-generated phospholipase (PL-1) probe. In all, 29 isolates picked up by the probe were confirmed as B. cereus by conventional cultural and biochemical characteristics. All the isolates reacted positively in PCR with phospholipase (PL-1) primers. Among the native isolates, 11 of them showed the discontinuous pattern of haemolysin BL activity in gel diffusion assay. Though 14 isolates reacted positively in PCR with primers (Ha-1) specific to the B gene of haemolysin BL, only four of them showed both the presence of gene and haemolysin BL activity. More than 50% of the isolates indicated their potential enterotoxigenicity by reacting positively with primers specific for the BceT gene encoding for diarrhoeal enterotoxin. PCR with primers for different inverse (IS) repeat elements revealed that isolates carrying transposon IS 231-P 231-1 did not carry IS 240-P 240. Some of the isolates were devoid ofthese IS elements. The study demonstrated the potential of using of a PCR-generated labelled PL-1 probe for the direct detection of B. cereus in food samples and PCR for characterizing the toxigenic isolates.     

Incidence,Antibiotic Susceptibility,and Toxin Profiles of Bacillus cereus sensu lato Isolated from Korean Fermented Soybean Products

Korean fermented soybean products, such as doenjang, kochujang, ssamjang, and cho‐kochujang, can harbor foodborne pathogens such as Bacillus cereus sensu lato (B. cereus sensu lato). The aim of this study was to characterize the toxin gene profiles, biochemical characteristics, and antibiotic resistance patterns of B. cereus sensu lato strains isolated from Korean fermented soybean products. Eighty‐eight samples of Korean fermented soybean products purchased from retails in Seoul were tested. Thirteen of 26 doenjang samples, 13 of 23 kochujang samples, 16 of 30 ssamjang samples, and 5 of 9 cho‐kochujang samples were positive for B. cereus sensu lato strains. The contamination level of all positive samples did not exceed 4 log CFU/g of food (maximum levels of Korea Food Code). Eighty‐seven B. cereus sensu lato strains were isolated from 47 positive samples, and all isolates carried at least one enterotoxin gene. The detection rates of hblCDA, nheABC, cytK, and entFM enterotoxin genes among all isolates were 34.5%, 98.9%, 57.5%, and 100%, respectively. Fifteen strains (17.2%) harbored the emetic toxin gene. Most strains tested positive for salicin fermentation (62.1%), starch hydrolysis (66.7%), hemolysis (98.9%), motility test (100%), and lecithinase production (96.6%). The B. cereus sensu lato strains were highly resistant to β‐lactam antibiotics such as ampicillin, penicillin, cefepime, imipenem, and oxacillin. Although B. cereus sensu lato levels in Korean fermented soybean products did not exceed the maximum levels permitted in South Korea (<10⁴ CFU/g), these results indicate that the bacterial isolates have the potential to cause diarrheal or emetic gastrointestinal diseases.     

Impact of intestinal microbiota and gastrointestinal conditions on the in vitro survival and growth of Bacillus cereus

Ingestion of B. cereus can result in diarrhea, if these bacteria survive gastrointestinal passage and achieve growth and enterotoxin production in the small intestine. The gastrointestinal survival of vegetative cells and spores of the diarrheal food poisoning strain B. cereus NVH 1230-88 was investigated during in vitro batch experiments simulating the stomach, duodenum and ileum using simulation media and competing intestinal microbiota. All spores and approx. 30% of the vegetative B. cereus cells survived the 2 h incubation in gastric medium with pH 4.0. Sterile intestinal medium induced germination of spores and enabled outgrowth of vegetative cells to approx. 7 log CFU/mL. The behavior of B. cereus in the intestinal environment with competing intestinal bacteria was determined by their relative concentrations. Besides the numbers of intestinal bacteria, the nutrition and composition of the intestinal community were also very important for the growth inhibition of B. cereus.     

金黄色葡萄球菌食品分离株肠毒素基因分布及分型研究

目的 对2006-2009年上海市Staphylococcus aureus(S.aureus)食品分离株进行肠毒素基因检测和基因分型研究,以了解肠毒素基因的分布规律及S.aureus的流行特点.方法 利用PCR方法检测食品中金黄色葡萄球菌肠毒素基因,包括5种传统肠毒素基因(SEA-SEE)和4种新型肠毒素基因(SEG-SEJ);利用脉冲场凝胶电泳法对49株食品分离株进行基因分型.结果 本研究发现49株食品分离株中有19株含有肠毒素基因,16株含有传统肠毒素SEA和SEC,且SEC占93.8％,并检测到新型肠毒素SEG、SEI、SEJ和SEH.PFGE法基因分型显示5株菌不能被分型,其余44株可分为28个基因型,表现为基因型的多样性,且分离自不同时间的菌株具有相同的带型.结论 应加强食品中S.aureus的监测分析,为其引起的食物中毒的预防和控制提供科学依据.     

Production of Bacillus cereus emetic toxin (cereulide) in various foods

To determine the role of Bacillus cereus as a potential pathogen in food poisoning, the production of an emetic toxin (cereulide) by B. cereus was quantified in various food sources. The amount of emetic toxin in 13 of 14 food samples implicated in vomiting-type food poisoning cases ranged from 0.01 to 1.28 microg/g. A vomiting-type strain, B. cereus NC7401, was inoculated into various foods and incubated for 24 h at 20, 30, and 35 degrees C. In boiled rice, B. cereus rapidly increased to 10(7)-10(8) cfu/g and produced emetic toxin at both 30 and 35 degrees C. In farinaceous foods, the production of emetic toxin was as high as that in the food samples implicated in food poisoning. Low levels of emetic toxin were detectable in egg and meat and their products and a small quantity of toxin was detectable in liquid foods such as milk and soymilk when not aerated. Bacterial growth and toxin production was inhibited in foods cooked with vinegar, mayonnaise, and catsup, supposedly by the decreased pH of acetic acid. This is the first report that has quantified emetic toxin of B. cereus in various foods.     

Characteristics of enterotoxin H-producing Staphylococcus aureus isolated from clinical cases and properties of the enterotoxin productivity

Staphylococcal enterotoxin H (SEH) is predicted to be involved in staphylococcal food poisoning. To characterize SEH-producing Staphylococcus aureus isolates from staphylococcal food poisoning cases in Japan, we investigated the relationship between SEH production and coagulase serotype, which is an epidemiological marker, and compared the properties of SEH production with those of staphylococcal enterotoxins A (SEA) and B (SEB). SEH production was determined by a newly developed sandwich enzyme-linked immunosorbent assay. Eighty-six (59.7%) of 144 isolates from staphylococcal food poisoning cases produced SEH. Seventy-one of the SEH-producing isolates simultaneously produced SEA, SEB, or both. All SEH-producing isolates belonged to coagulase type VII, which was the predominant type, representing 99 (68.8%) of 144 isolates. The amount of SEH produced in brain heart infusion was almost the same as the amount of SEA and approximately 10-fold lower than that of SEB. SEH and SEA were produced mainly during the late exponential phase of growth, whereas SEB was produced mostly during the stationary phase. The production levels of SEH and SEA were gradually affected by decreases in water activity, but the production of SEB was greatly reduced under conditions of low water activity. These findings indicate that SEH-producing S. aureus isolates are of high prevalence in staphylococcal food poisoning cases. Given the unique epidemiological characteristic of these isolates, SEH and SEA probably are responsible for food poisoning.     

Enterotoxigenic and genetic profiles of Bacillus cereus strains of food origin in Brazil

In Brazil, the incidence of Bacillus cereus outbreaks is unknown, and there is little information about B. cereus occurrence in food. In addition, data on toxin production and genetic characterization of the B. cereus isolates cannot be found. This pathogen causes two distinct types of toxin-mediated foodborne illnesses known as diarrheal and emetic syndromes. Diarrheal syndrome has been linked to three different enterotoxins: two protein complexes, hemolysin BL (HBL) and nonhemolytic enterotoxin (NHE); and an enterotoxic protein, cytotoxin K (cytK). Emetic syndrome is related to cereulide, a toxin encoded by the ces gene. In this study, NHE and HBL production capacities of 155 strains of B. cereus isolated from Brazilian food products were evaluated with an immunoassay. Strains were also tested for the presence of the genes of the HBL and NHE complexes, cytK, cytK-1, cytK-2, and ces, using PCR. HBL was detected in 105 (67.7%) strains and NHE in 154 (99.4%) strains. All the strains harbored at least one gene of the NHE complex, while 96.1% of them were positive for at least one of those of the HBL complex. Genes cytK1 and ces were not detected. All strains showed toxigenic capacity and could represent a risk for consumers if good practices are not followed. This is the first report on toxigenic and genetic profiles of B. cereus strains isolated in Brazil.     

蜡样芽孢杆菌致吐毒素的研究现状

蜡样芽孢杆菌致吐毒素是热处理也不能灭活的毒素,没有方法可以去除食品中致吐毒素Cereulide。因此,探讨食品中哪些因素影响致吐毒素产生显得至关重要。本文对蜡样芽孢杆菌孢子、致吐毒素产生株的特点及致吐食源性中毒,分离、检测和定量食品中Cereulide的国内外新方法及影响致吐毒素产生的因素等进行了综述。     

MODELING OF THE GERMINATION OF SPORES FROM CLOSTRIDIUM PERFRINGENS FOOD POISONING ISOLATES

Clostridium perfringens type A isolates carrying a chromosomal gene for enterotoxin production (chromosomal c. perfringens enterotoxin [ C- cpe] strains) are frequent causative agents of food poisoning. Predictive models for their growth in meat products have been published; however, the development of germination models needs further research. In this study, the Weibull function was used to describe the germination of C- cpe food poisoning strains in phosphate and Tris–HCl buffer as affected by pH (5.8–8.5), germinant concentration (1–100 mM KCl) and germination temperature (23–60C). As indicated by estimators of model accuracy and bias, an empirical model for spore germination as a function of germination temperature only, predicted accurately the germination potential of 4 C- cpe food poisoning strains in buffer and in laboratory media in the 23–50C range.

PRACTICAL APPLICATIONS

Thermal processing models based on the inactivation of a heat-resistant spore ignore the potential germination and outgrowth of spore survivors as recommended by new food safety guidelines. Germination models for spore survivors to be used in conjunction with outgrowth models should be developed using as samples the product of interest because the bioavailability and concentration of germinants such as ions and free amino acids vary with the product and its manufacturing process. Another germination model application is in conjunction with new processing technologies such as high pressure processing (HPP). The inactivation of bacterial spores has been a major challenge to HPP process developers. Since germinated spores are sensitive to pressure, germination models would be useful to design combined strategies of food thermal treatment and HPP.     

即食米面制品中蜡样芽胞杆菌分离鉴定及毒力基因研究

为探究即食米面制品中蜡样芽胞杆菌的污染状况及食源性蜡样芽胞杆菌中毒力基因的分布规律,以餐饮服务场所采集245份即食米面制品为蜡样芽胞杆菌的污染调查材料,通过国标法及持家基因分离鉴定得到49株蜡样芽胞杆菌阳性株,并对其进行13种毒力基因的PCR检测。结果表明:蜡样芽胞杆菌的平均检出率为10.61%(26/245),阳性检出样品主要为盒饭及米饭,其蜡样芽胞杆菌平均检出水平为3032 CFU/g,其中以盒饭的检出程度最高。49株分离菌株均检出两种或两种以上的毒力基因,非溶血性肠毒素基因nhe和entFM检出率最高,分别达到100%及91.84%,是分离菌株的主要毒力基因;溶血素基因hblA、hblC、hblD检出率分别为20.41%、38.78%和40.82%。同时携带nhe三个基因又携带hblA、hblC和hblD基因的强毒株有7株,占14.29%,且这7株菌均含有entFM基因。呕吐型毒力基因ces、cer、EM1仅2株检出,检出率为4.08%。本研究对即食米面制品中蜡样芽胞杆菌的监控、预警及爆发引起食物中毒后追踪其感染源和传播途径及构建基因指纹图谱库和分子溯源平台具有指导意义。     

Enterotoxin production by Staphylococcus aureus isolated from mastitic cows

Staphylococcus aureus is an important cause of mastitis in cows. The ability of S. aureus strains to produce one or more enterotoxins in milk and dairy products is linked to staphylococcal food poisoning. To determine whether staphylococci causing bovine mastitis could cause human foodborne intoxication, the production of staphylococcal enterotoxins A through D (SEA, SEB, SEC, and SED) by 160 S. aureus isolates was evaluated with the use of a reverse passive latex agglutination enterotoxin kit. All S. aureus strains were isolated over a 9-month period from 2,343 routine submissions of a composite quarter collection of individual mastitic cows at 18 dairy farms in the San Joaquin Valley in California. Prior to enterotoxin detection, isolates were grown by a method that enhances the in vitro synthesis of enterotoxin. Twenty-two of 160 S. aureus isolates produced enterotoxin. Seven produced SEC, 12 produced SED, and 3 produced both SEC and SED. None of the isolates produced SEA or SEB.     

Prevalence of Bacillus cereus in dried milk products used by Chilean School Feeding Program

The prevalence of Bacillus cereus, in a total of 381 samples of dried milk products (milk with rice, milk substitute, milk powder, milk-cereal-rice, pudding milk, flan, and mousse) used by the Chilean School Feeding Program, was investigated. The potential of 94 selected isolates of B. cereus to produce diarrhoeal enterotoxin (by the BCET-RPLA test) in BHI culture, as well as the ability of enterotoxigenic-strains to grow at psychrotrophic temperatures were also verified. B. cereus was found in 175 of 381 of the samples analysed (45.9%), reaching levels from 3.0 to 10(4) spores g(-1). As expected, the higher prevalence and counts were observed in those products that contained whole rice, cereals and pulses extruded, and food additives. Of the 94 isolates of B. cereus tested for diarrhoeal enterotoxin production, 28 (29.8%) were positive, and none of these was able to grow at < or = 7 degrees C. The prevalence of B. cereus in dried milk products analysed was fairly high, although it was present in low number. However, as they were composed to a large extent of enterotoxigenic mesophilic strains, the potential risk for the safety of reconstituted products held at improper temperature should not be neglected.     

Functionality of selected strains of moulds and yeasts from Vietnamese rice wine starters

The role of starch-degrading mycelial fungi, and the alcohol production and ethanol tolerance of the yeasts isolated from selected Vietnamese traditional rice wine starters were examined, and optimum conditions for these essential steps in rice wine fermentation were determined. Of pure isolates from Vietnamese rice wine starters, mould strains identified as Amylomyces rouxii, Amylomyces aff. rouxii, Rhizopus oligosporus and Rhizopus oryzae, were superior in starch degradation, glucose production and amyloglucosidase activity during the saccharification of purple glutinous rice. A. rouxii was able to produce up to 25%w/w glucose with an amyloglucosidase activity up to 0.6 Ug(-1) of fermented moulded mass. Five yeast isolates identified as Saccharomyces cerevisiae were selected for their superior alcohol productivity. They were able to deplete a relatively high initial percentage of glucose (20% w/v), forming 8.8% w/v ethanol. The ethanol tolerance of S. cerevisiae in challenge tests was 9-10% w/v, and 13.4% w/v as measured in fed-batch fermentations. Optimum conditions for the saccharification were: incubation for 2 d at 34 degrees C, of steamed rice inoculated with 5 log cfu g(-1); for the alcoholic fermentation 4 d at 28.3 degrees C, of saccharified rice liquid inoculated with 5.5 log cfu mL(-1).     

Toxin profile, antibiotic resistance, and phenotypic and molecular characterization of Bacillus cereus in Sunsik

Sunsik, a ready-to-eat food in Korea, is comprised of various agricultural and marine products, and has been an important concern in Bacillus cereus food poisoning. The aim of this study was to investigate the toxin profiles, genotypic and phenotypic patterns as well as antibiotic resistance of B. cereus strains isolated from Sunsik. A subtyping method known as automated repetitive sequence-based PCR system (DiversiLab™) was used to assess the intraspecific biodiversity of these isolates. Thirty-five B. cereus strains were isolated from 100 commercial Sunsik samples, all of which harbored at least 1 enterotoxin gene. The detection rates of nheABC, hblCDA, cytK, and entFM enterotoxin gene among all isolates were 97%, 86%, 77%, and 100%, respectively. Most strains also produced corresponding enterotoxins such as HBL (83%) and NHE (94%). One strain (2.9%) carried the emetic toxin genes, including ces and EM1, and was positive for the HEp-2 cell emetic toxin assay. Most strains were positive for various biochemical tests such as salicin hydrolysis (86%), starch fermentation (89%), hemolysis (89%), motility test (100%) and lecithinase hydrolysis (89%). All isolates were susceptible to most antibiotics although they were highly resistant to β-lactam antibiotics. By using the automated rep-PCR system, all isolates were successfully differentiated, indicating the diversity of B. cereus strains present in Sunsik.     

一起由金黄色葡萄球菌引起食物中毒的实验室诊断

目的对一起食物中毒进行相关样品的实验室检测,针对分离病原菌进行分子分型溯源和耐药分析。方法对可疑食品(冰激凌、牛奶)样品、患者呕吐物和粪便标本使用国家标准或其他方法进行沙门菌、志贺菌、致泻大肠埃希菌、金黄色葡萄球菌、单核细胞增生李斯特菌、蜡样芽胞杆菌和诺如病毒检测。对检出的金黄色葡萄球菌进行肠毒素检测、脉冲场凝胶电泳(PFGE)分型和抗生素敏感性检测。结果从生产冰激凌剩余牛奶及冰激凌样品中检出金黄色葡萄球菌12株,患者呕吐物和粪便标本中检出4株,菌株均产生A+C+E混合型肠毒素,PFGE显示患者呕吐物、粪便、中毒同批冰激凌样品、生产冰激凌用牛奶中检出的金黄色葡萄球菌菌株同源性为100.0%;药敏结果显示,这16株菌株均对青霉素和红霉素耐药。结论本起事件是由冰激凌加工点使用了被金黄色葡萄球菌污染且产生大量肠毒素的牛奶生产冰激凌所致。建议监管部门加强对农村食品小作坊、牛奶供应站等生产经营场所的监管,预防类似事件的发生。     

Present state of knowledge on staphylococcal intoxication

Globally, staphylococcal intoxication remains a very common food poisoning. In this review, emphasis is being placed on epidemiological aspects of the problem and the effect of food environment on the survival and growth of staphylococci and production of enterotoxins. The high prevalence of staphylococci in raw foods of animal origin requires effective processing for safety. Man remains a major reservoir for post-process recontamination. The effect of the intrinsic characteristics of foods (pH, water activity, Eh, preservatives competing microbial flora, natural food) and extrinsic parameters of processing and storage (temperature, freezing, irradiation, dehydration, packaging, humidity) on staphylococcal survival, growth and enterotoxin production has been evaluated extensively. While staphylococci can be destroyed easily the enterotoxins can survive practically all food processing. While extreme levels of intrinsic variables can control enterotoxin production, yet the environment of most foods is conductive to staphylococcal growth. Rapid food cooling, refrigeration and consumer, food handler and processor education remain the key to staphylococcal food poisoning prevention.