Presence and growth of Bacillus cereus in dehydrated potato flakes and hot-held, ready-to-eat potato products purchased in New Zealand

Potato products prepared from dehydrated potato flakes have been implicated in foodborne illness incidents involving Bacillus cereus intoxications. B. cereus can survive as spores in potato flakes and can germinate and multiply in the rehydrated product. This study assessed the frequency and concentration of B. cereus in dehydrated potato flakes and hot-held, ready-to-eat mashed potato products. Of 50 packets of potato flakes tested, eight contained greater than 100 CFU/g B. cereus (maximum 370 CFU/g). The temperature of the potato portion of 44 hot-held food products was measured immediately after purchase, and 86% were below the safe hot-holding temperature of 60 degrees C. The potato portions were subsequently tested for B. cereus. Only two of the potato portions contained B. cereus at greater than 100 CFU/g, a potato-topped pastry (1000 CFU/g) and a container of potato and gravy (120 CFU/g). To assess multiplication of B. cereus in this food, we held rehydrated potato flakes with naturally occurring B. cereus at 37, 42, and 50 degrees C and tested them over 6 h. By 6 h, the number of B. cereus in potato stored at 37 degrees C had exceeded 10(3) CFU/g, was greater than 10(4) CFU/g at 50 degrees C, and was close to 10(6) CFU/g at 42 degrees C. Growth data were compared to predictions from the U.S. Department of Agriculture Pathogen Modeling Program (PMP 7.0). The PMP predictions were found to simulate the measured growth better at 42 degrees C than at 37 degrees C. Hot-held potato products should be safe for consumption if held at 60 degrees C or above or discarded within 2 h.     

Prevalence and characterisation of Bacillus cereus in vacuum packed potato puree

Refrigerated processed foods of extended durability (REPFED) potato puree was analysed for Bacillus cereus contamination along the production line and during the product shelf‐life. Isolated B. cereus strains were tested for their psychrotrophic character and the ability to produce enterotoxins. Bacillus cereus contamination during four subsequent productions was in the range of 2.3–4.0 log cfu g^?1. Productions five and six were significantly less contaminated with B. cereus (≤1 log cfu g ^?1). All B. cereus isolates from the first four productions were able to grow at 7 ° and 10 °C, whereas the majority of the isolates from productions five and six did not. No B. cereus isolates grew at 4 °C. randomly amplified polymorphic DNA (RAPD) fingerprinting showed that the most of B. cereus contamination originated from one source. In total, 30.4% of isolates expressed enterotoxic character. The present study points out the necessity to prevent an ‘in house’ colonisation and contamination during food processing in order to accomplish the safety of REPFED throughout the shelf‐life. It also indicates the most critical steps in the production line of ready‐to‐eat potato puree and impact of failures regarding the food safety. The data provided can be used for risk assessment studies regarding B. cereus in REPFED.     

Presence of Bacillus cereus in street foods in Gaborone, Botswana

The purpose of this study was to evaluate the microbiological safety and quality of street foods sold in Gaborone, Botswana. A total of 148 point-of-sale composite street food samples were bought and analyzed between June 2001 and May 2002. The analysis focused on the level of contamination of various street foods with Bacillus cereus. The B. cereus (vegetative and spores), total spore, and total viable counts were determined on all the samples. Also B. cereus isolates from 444 individual point-of-sale food samples were characterized with respect to their biochemical profiles and enterotoxigenic properties. The B. cereus contamination rate for point-of-sale foods was 65%. The B. cereus counts ranged from not detectable to levels as high as 9.1 log CFU/g. Despite the high rate of contamination of some samples, generally, most samples had B. cereus counts of less than 4 log CFU/g; hence, they were of acceptable microbiological quality. Bacillus diarrheal enterotoxin was detected from 52 isolates from individual portions of meals using the B. cereus enterotoxin reversed passive latex agglutination kit. Results of the assay revealed that 59.6% of the B. cereus isolates were enterotoxigenic. Most of the enterotoxigenic isolates were obtained from vegetable samples.     

Detection of Bacillus cereus on selected retail chicken products

Samples from five chicken meat products, obtained at retail stores, were evaluated for the presence of Bacillus cereus. The products tested were as follows: breaded, fully cooked, frozen nuggets (NUGGETS); breaded, fully cooked, frozen tenders (TENDERS); fully cooked, frozen, white-meat fajita-style strips (STRIPS); raw, refrigerated, boneless, skinless, marinated breast fillets (FILLETS); and raw, refrigerated, cut-up, tray-pack bone-in parts (PARTS), either split breasts or thighs. Four packages of each item were obtained on three different days (n = 60). Frozen and refrigerated products were held overnight in their respective environments as appropriate; then packages were opened aseptically, and a total of 25 g of tissue was excised from multiple pieces within a package. The 25-g samples were enriched in 225 ml of Trypticase soy-polymixin broth for 18 to 24 h at 30 degrees C and then plated on mannitol-egg yolk-polymixin agar and incubated for 18 to 24 h at 30 degrees C. Colonies characteristic of B. cereus were chosen and replated for isolation on mannitol-egg yolk-polymixin agar. Suspect colonies were confirmed as Bacillus spp. by Gram stain, hemolysis on blood agar, and a biochemical test strip. Isolates were further confirmed as B. cereus using Bacteriological Analytical Manual procedures, including tests for motility, rhizoid growth, hemolysis, and protein toxin crystal production. B. cereus was detected in 27 of 60 total samples. By product, the prevalence levels were as follows: NUGGETS, 11 of 12 positive; TENDERS, 8 of 12 positive; STRIPS, 6 of 12 positive; FILLETS, 0 of 12 positive; and PARTS, 2 of 12 positive. Isolates were tested by PCR for presence of the toxin-encoding genes bceT, nheABC, hblACD, and cytK. Results indicate that B. cereus organisms were present on four of the five retail poultry products tested in this study, with the highest rates reported for the three fully cooked items, especially the two breaded products. All strains isolated contained the gene(s) for at least one of the toxins, although none of the strains contained the cytK gene.     

Bacillus cereus in personal care products: risk to consumers

Bacillus cereus is ubiquitous in nature and thus occurs naturally in a wide range of raw materials and foodstuffs. B. cereus spores are resistant to desiccation and heat and able to survive dry storage and cooking. Vegetative cells produce several toxins which on ingestion in sufficient numbers can cause vomiting and/or diarrhoea depending on the toxins produced. Gastrointestinal disease is commonly associated with reheated or inadequately cooked foods. In addition to being a rare cause of several acute infections (e.g. pneumonia and septicaemia), B. cereus can also cause localized infection of post‐surgical or trauma wounds and is a rare but significant pathogen of the eye where it may result in severe endophthalmitis often leading to loss of vision. Key risk factors in such cases are trauma to the eye and retained contaminated intraocular foreign bodies. In addition, rare cases of B. cereus‐associated keratitis (inflammation of the cornea) have been linked to contact lens use. Bacillus cereus is therefore a microbial contaminant that could adversely affect product safety of cosmetic and facial toiletries and pose a threat to the user if other key risk factors are also present. The infective dose in the human eye is unknown, but as few as 100 cfu has been reported to initiate infection in a susceptible animal model. However, we are not aware of any reports in the literature of B. cereus infections in any body site linked with use of personal care products. Low levels of B. cereus spores may on occasion be present in near‐eye cosmetics, and these products have been used by consumers for many years. In addition, exposure to B. cereus is more likely to occur through other routes (e.g. dustborne contamination) due to its ubiquity and resistance properties of spores. The organism has been recovered from the eyes of healthy individuals. Therefore, although there may be a perceived hazard, the risk of severe eye infections as a consequence of exposure through contaminated near‐eye cosmetics is judged to be vanishingly small. It is unlikely that more stringent microbiological standards for near‐eye cosmetics will have any impact on the risk of severe eye infections caused by B. cereus, as these are not linked to use of personal care products.     

Antimicrobial effect of nisin and carvacrol and competition between Bacillus cereus and Bacillus circulans in vacuum-packed potato puree

A study was undertaken to investigate the growth and competition between Bacillus cereus and B. circulans strains in both BHI and ready-to-eat potato puree. In addition, antimicrobial potential of supplemented nisin and carvacrol was evaluated against inoculated B. cereus and B. circulans strains. The accomplished growth inhibition was observed for both B. cereus and B. circulans, where B. circulans strains were more sensitive. MIC values were decreased by lowering the incubation temperature in separate applications of nisin and carvacrol, while the effect of combined application of nisin and carvacrol appeared to be more obvious at higher temperatures. The overall effect of interactions between spoilage and pathogenic Bacillus spp. was dependent on the psychrotrophic character of both cultures. The complete inhibition of B. circulans in the co-culture experiments corresponded to B. cereus population density of approximately 6 log units. Microbial challenge testing in potato puree showed that, at both 7 and 10 °C, B. cereus TZ415 and B. circulans 4.1 could grow to high counts when no antimicrobial substances (nisin and carvacrol) were applied, while no visible spoilage occurred. A more pronounced antimicrobial activity of nisin and nisin–carvacrol combination was observed in potato puree compared to the BHI medium.     

湿粉类制品蜡样芽胞杆菌污染风险及控制研究

目的 以广州市为例分析湿粉类制品蜡样芽胞杆菌污染情况和生产加工过程的风险点,提出相应的控制措施.方法 选择广州市范围生产湿粉类制品的生产单位,采集生产环节和销售环节样品及同批次产品生产使用的原料,对蜡样芽胞杆菌进行定量检测.结果 共采集32份出厂的湿粉类制品、139份市售湿粉类制品、32份原料大米、5份小麦淀粉、3份玉...     

Microbiological quality and incidence of Staphylococcus aureus and Bacillus cereus in vegetarian food products

A total of 320 samples of vegetarian food purchased from local markets were examined in order to determine the microbiological quality of these products. Aerobic count, psychrotrophic count, anaerobic count, E. Coli, coliforms, S. aureus, B. cereus, spores of Bacillus as well as enterotoxins produced by B. cereus andS. aureus were also evaluated. The incidence of E. coli and coliforms in these vegetarian food products were 28·1% and 32·5%, respectively, while 18·1% and 3·4% of the samples were found to contain S. aureus and B. cereus, respectively. The vegetarian food products investigated in the present study were divided into five groups, based on their major components. The samples made from soybeans showed the highest detection rate (74·5%) when S. aureus, B. cereus, E. coli and coliforms were used as indicators. S. aureus was detected in all the five groups, with the highest detection rate in cereal and Konjac group (23·8%), while B. cereus was detected only in soybean, cereal and Konjac groups with the incidence of 5·1%, 9·5% and 2·4%, respectively. Among the 38 strains of S. aureus isolates, 17 strains were observed to produce enterotoxins of types A, B, C and D. Whereas, among the seven strains of B. cereus isolates, one produced diarrheal toxin while four others killed adult ICR mice. The present results also revealed that vacuum-packed vegetarian foods have the lowest incidence of micro-organisms tested in this study.     

Prevalence and characterization of toxigenic Bacillus cereus group isolated from low-moisture food products

Food Science and Biotechnology - The present study was conducted to determine the occurrence of B. cereus group members in low-moisture food products by phenotypic and genetic assessment and to...     

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.     

Aroma of dehydrated pear products

Pear purees and cubes were dehydrated with sugars (sucrose and trehalose) addition. Freeze drying and foam-mat drying were used. Manual solid-phase microextraction (SPME) coupled with gas chromatography (GC-flame ionization detector (FID) and GC-MS) was applied to determine the changes in retention of aroma compounds in dehydrated pear purees and cubes. The best retention of aroma compounds in dehydrated pear purees was noticed in the case when freeze drying and trehalose addition were combined. In dehydrated pear cubes, previously dipped in trehalose solution, the highest aroma retention was also determined. This study showed possible application of trehalose as potentially beneficial food ingredient, with the aim to improve the quality of dehydrated fruit products, especially their aroma, and to produce superior dried fruit products or ingredients, which are widely used in food formulation.     

Bacillus cereus in free-stall bedding

To increase the understanding of how different factors affect the bacterial growth in deep sawdust beds for dairy cattle, the microbiological status of Bacillus cereus and coliforms in deep sawdust-bedded free stalls was investigated over two 14-d periods on one farm. High counts of B. cereus and coliforms were found in the entire beds. On average, 4.1 log₁₀B. cereus spores, 5.5 log₁₀B. cereus, and 6.7 log₁₀ coliforms per gram of bedding could be found in the upper layers of the sawdust likely to be in contact with the cows’ udders. The highest counts of B. cereus spores, B. cereus, and coliforms were found in the bedding before fresh bedding was added, and the lowest immediately afterwards. Different factors of importance for the growth of B. cereus in the bedding material were explored in laboratory tests. These were found to be the type of bedding, pH, and the type and availability of nutrients. Alternative bedding material such as peat and mixtures of peat and sawdust inhibited the bacterial growth of B. cereus. The extent of growth of B. cereus in the sawdust was increased in a dose-dependent manner by the availability of feces. Urine added to different bedding material raised the pH and also led to bacterial growth of B. cereus in the peat. In sawdust, a dry matter content greater than 70% was needed to lower the water activity to 0.95, which is needed to inhibit the growth of B. cereus. In an attempt to reduce the bacterial growth of B. cereus and coliforms in deep sawdust beds on the farm, the effect of giving bedding daily or a full replacement of the beds was studied. The spore count of B. cereus in the back part of the free stalls before fresh bedding was added was 0.9 log units lower in stalls given daily bedding than in stalls given bedding twice weekly. No effect on coliform counts was found. Replacement of the entire sawdust bedding had an effect for a short period, but by 1 to 2 mo after replacement, the counts of B. cereus spores in the beds had increased about 2 log units and were as high as they were before bed replacement. Therefore, free-stall management could, to a limited extent, reduce the content of B. cereus spores in the beds by daily bedding and entire bed replacement.     

蜡样芽胞杆菌生长特性及大蒜精油对其抑菌活性的研究

主要研究了37℃条件下3种蜡样芽胞杆菌在营养肉汤中的生长状况,建立了37℃下蜡样芽胞杆菌在营养肉汤中的Boltzmann牛长模犁,3条牛长曲线相关系数鼯均大于0．97;检测了不同培养时间蜡样芽胞杆菌的产芽胞情况,结果表明1号菌株和14号菌株较早产芽胞,培养相同时间,产芽胞数：1号菌株,4号菌株〉标准菌株;采用牛沣杯法和平板计数法研究了大蒜精油对蜡样芽胞杆菌的抑制效果,结果表明浓度为lO。的人蒜精油对3种蜡样芽胞杆菌都有很好的抑制效果,3种菌株中l号菌株最难抑制。     

Emetic toxin-producing strains of Bacillus cereus show distinct characteristics within the Bacillus cereus group

One hundred representative strains of Bacillus cereus were selected from a total collection of 372 B. cereus strains using two typing methods (RAPD and FT-IR) to investigate if emetic toxin-producing hazardous B. cereus strains possess characteristic growth and heat resistance profiles. The strains were classified into three groups: emetic toxin (cereulide)-producing strains (n=17), strains connected to diarrheal foodborne outbreaks (n=40) and food-environment strains (n=43), these latter not producing the emetic toxin. Our study revealed a shift in growth limits towards higher temperatures for the emetic strains, regardless of their origin. None of the emetic toxin-producing strains were able to grow below 10 degrees Celsius. In contrast, 11% (9 food-environment strains) out of the 83 non-emetic toxin-producing strains were able to grow at 4 degrees Celsius and 49% at 7 degrees Celsius (28 diarrheal and 13 food-environment strains). non-emetic toxin-producing strains. All emetic toxin-producing strains were able to grow at 48 degrees Celsius, but only 39% (16 diarrheal and 16 food-environment strains) of the non-emetic toxin-producing strains grew at this temperature. Spores from the emetic toxin-producing strains showed, on average, a higher heat resistance at 90 degrees Celsius and a lower germination, particularly at 7 degrees Celsius, than spores from the other strains. No difference between the three groups in their growth kinetics at 24 degrees Celsius, 37 degrees Celsius, and pH 5.0, 7.0, and 8.0 was observed. Our survey shows that emetic toxin-producing strains of B. cereus have distinct characteristics, which could have important implication for the risk assessment of the emetic type of B. cereus caused food poisoning. For instance, emetic strains still represent a special risk in heat-processed foods or preheated foods that are kept warm (in restaurants and cafeterias), but should not pose a risk in refrigerated foods.     

酱腌菜中蜡样芽孢杆菌的检测与分析

对来自超市的50个酱腌菜样品进行了蜡样芽孢杆菌的检测与分析。研究结果表明,各样品中都有蜡样芽胞杆菌检出,数量为2×102～2×104 CFU/g,其中糖醋渍菜中蜡样芽孢杆菌的检出率较高。本调查分析对我国酱腌菜中蜡样芽孢杆菌污染的溯源奠定基础。建议完善酱腌菜中蜡样芽孢杆菌的限量标准。     

Sterilization of dehydrated potato granules with propylene oxide

Summary Results were provided for propylene oxide cold sterilization of dehydrated mashed potato granules that contained 7% moisture and 0.044% chloride. Treatment with an optimum concentration of 0.1% w/w sterilant for 6 days at 22° C gave a bacterial count reduction from 3.4 × 10⁵ to less than 5 × 10³, a low glycol residue of 29 ppm, and 12 ppm of chlorohydrin, with the 1-chloro-2-propanol isomer constituting 94% of the total. This indicated that, in spite of the granule pH of 5.62, nucleophilic attack in dehydrated granules involves nonprotonated propylene oxide. Based on NMR analysis, evidence was given that such sterilization does not result in detectable etherification of starch, even though its content of 83% on a dry basis places it as the major potentially reactive constituent.

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Sterilisation von getrocknetem Kartoffelpulver mit Propylenoxyd

Zusammenfassung Propylenoxyd wurde zur Kaltsterilisation von Kartoffelpulver mit 7% Feuchtigkeit und einen Chloridgehalt von 0,044% verwendet. Bei einem optimalen Gehalt des Sterilisationsmittels von 0,1% wurde die Behandlung während 6 Tagen bei 22° C durchgeführt, was eine Reduktion des Keimgehaltes von 3,4 × 10⁵ auf weniger als 5 × 10³ bewirkte. Der Glykolrückstand war mit 29 ppm niedrig, ebenfalls das Chlorohydrin mit 12 ppm, wobei das 1-Chlor-2-propanol 94% der beiden Isomere ausmachte. Dies ergab einen Hinweis, daß im Pulver trotz des pH-Wertes 5,62 nicht protoniertes Propylenoxyd als Acceptor nucleophiler Übergänge dient. Aufgrund von NMR-Untersuchungen wurde gefunden, daß die obengenannte Behandlung keine nachweisbare Ätherbildung der Stärke bewirkte, obwohl ihr Gehalt 83% des Trockengewichtes beträgt und damit als potentieller Hauptreaktionspartner anzusehen wäre.

     

Analytical methods for Bacillus cereus and other Bacillus species

Bacillus cereus can give rise to two distinct forms of foodborne disease, the emetic and the diarrhoeal syndromes. The emetic syndrome is believed to be associated with an emetic toxin pre-formed in food. Cooked rice is the most common vehicle, and the symptoms are similar to those of Staphylococcus aureus intoxication. The diarrhoeal type is caused by an enterotoxin and the symptoms generally parallel those of the Clostridium perfringens food poisoning. The heat resistance of B. cereus spores and the non-fastidious nature of the organism facilitates its survival and/or growth in a wide variety of foods. This review describes analytical methods available for the isolation, identification, and enumeration of the organism, in addition to details about biological and immunological methods for toxin assay. Data are also presented concerning the incidence and epidemiology of B. cereus food poisoning around the world, and especially in Japan.