The microbiology of minimally processed fresh fruits and vegetables

Minimally processed fresh (MPF) fruits and vegetables are good media for growth of microorganisms. They have been involved in outbreaks because of the consumption of products contaminated by pathogens. They are also sensitive to various spoilage microorganisms such as pectinolytic bacteria, saprophytic Gram‐negative bacteria, lactic acid bacteria, and yeasts. Contamination of MPF fruits and vegetables occurs at every stage of the food chain, from cultivation to processing. Polluted environments during cultivation or poor hygienic conditions in processing increase the risk of contamination with foodborne pathogens. Although MPF fruits and vegetables may harbor psychrotrophic microorganisms such as fluorescent pseudomonads or Listeria monocytogenes, good control of refrigeration temperature limits growth of spoilage and pathogenic microorganisms. Modified atmospheres are often efficient to maintain or improve visual and organoleptic quality of MPF fruits and vegetables, but their effects on microorganisms are inconsistent. Chemical disinfection can partially reduce the initial bacterial contamination; irradiation seems to be more efficient. The applications of legislations and quality assurance systems to control contamination, survival, and growth of foodborne pathogens in MPF fruits and vegetables are discussed.     

Ultrasound decontamination of minimally processed fruits and vegetables

The effectiveness of power ultrasound for the microbial decontamination of minimally processed fruits and vegetables was studied. Reductions in Salmonella typhimurium attached to iceberg lettuce obtained by cleaning with water, chlorinated water, ultrasound with water and ultrasound with chlorinated water were 0.7, 1.7, 1.5 and 2.7 logs, respectively, for small-scale (2 L) trials. The cleaning action of cavitation appears to remove cells attached to the surface of fresh produce, rendering the pathogens more susceptible to the sanitizer. For large-scale (40 L) trials, the addition of chlorine to water in the tank gave a systematic difference in Escherichia coli decontamination efficiency. However, the frequency of ultrasound treatment (25, 32–40, 62–70 kHz) had no significant effect on decontamination efficiency ( P  > 0.69). With the potentially high capital expenditure together with the expensive process of optimization and water treatment, it is unlikely that the fresh produce industry would be willing to take up this technology. Furthermore, the additional one log reduction achieved by applying ultrasound to a chlorinated water wash does not completely eliminate the risk of pathogens on fresh produce.     

The microbiological safety of minimally processed vegetables

Summary Demand for fresh, convenient, minimally processed vegetables has led to an increase in the quantity and variety of products available to the consumer. Modified atmosphere packaging, in combination with refrigeration, is increasingly being employed as a mild preservation technique to ensure quality and storage-life. The fresh nature of these products, together with the mild processing techniques and subsequent storage conditions, have presented indigenous and pathogenic microorganisms with new ecosystems and potential infection vehicles; a number of outbreaks of foodborne disease being attributed to their consumption. Psychrotrophic pathogens and pathogens which are capable of maintaining an infectious potential under mild preservation regimes are of particular concern; Listeria monocytogenes , Aeromonas hydrophila and Clostridium botulinum being amongst the most notable. This review describes the processing, packaging and storage procedures involved in the production of minimally processed vegetables, and details their impact upon the survival and growth of associated pathogens. Gaps in our current understanding of the consequences of this novel technology for microbiological safety are highlighted.     

Decontamination methods to prolong the shelf-life of minimally processed vegetables, state-of-the-art

Minimally processed vegetables (MPV) are any fresh vegetables that have been physically altered from their original form, but remains in a fresh state. Microorganisms present in MPV can cause foodborne illnesses or spoilage; hence, decontamination of MPV can produce more stable products. The present review examines the difficulties to decontaminate and prolong the shelf-life of MPV, evaluating the current way of data analysis and interpretation. It addresses the different aspects of the problem of the accessibility of sanitizers to microorganisms (irregularities of produce surface, injuries, internalization, attachment, and biofilms). It also includes a critical exposition of the methodological problems to estimate the prolongation of shelf-life due to a decontamination method, namely: the variability among samples, the reproducibility of the results, and the interpolation when lacking some crucial data. Furthermore, it revises the difficulties to control the microbial loads of decontaminated MPV during storage (the enhanced growth rate of microorganisms in decontaminated MPV, the patterns of microbial growth in non decontaminated and decontaminated MPV, and the role of temperature in keeping the decontamination effect).     

冷却猪肉中腐败微生物鉴定及其消长规律的研究

目的对冷却猪肉中腐败微生物进行鉴定,研究其在0～4℃条件下贮藏时的消长规律。方法采用选择性培养基对冷却猪肉中的腐败微生物进行分离培养,利用Biolog微生物自动鉴定系统对菌株进行鉴定。结果共鉴定出11株具有代表性的细菌:肠杆菌4株,假单胞菌1株,热杀索丝菌1株,不动杆菌1株,乳酸菌2株,葡萄球菌2株。冷却猪肉中腐败微生物初始菌相结构为:热杀索丝菌54.9%,肠杆菌科8.7%,假单胞菌属3.6%,乳酸菌属29.5%,葡萄球菌/微球菌0.6%,霉菌/酵母菌2.7%。在0～4℃条件下贮藏时,热杀索丝菌、肠杆菌科和假单胞菌属是冷却猪肉中的优势腐败菌,假单胞菌属和肠杆菌科在菌相结构中的比例增长最高,特别是假单胞菌属的数量增长最快。结论鉴定出了冷却猪肉中的主要腐败微生物,确定了其初始菌相和优势腐败菌。     

Potential applications of plant based derivatives as fat replacers,antioxidants and antimicrobials in fresh and processed meat products

Growing concern about diet and health has led to development of healthier food products. In general consumer perception towards the intake of meat and meat products is unhealthy because it may increase the risk of diseases like cardiovascular diseases, obesity and cancer, because of its high fat content (especially saturated fat) and added synthetic antioxidants and antimicrobials. Addition of plant derivatives having antioxidant components including vitamins A, C and E, minerals, polyphenols, flavanoids and terpenoids in meat products may decrease the risk of several degenerative diseases. To change consumer attitudes towards meat consumption, the meat industry is undergoing major transformations by addition of nonmeat ingredients as animal fat replacers, natural antioxidants and antimicrobials, preferably derived from plant sources.     

Comparison of four commercial DNA extraction kits for PCR detection of Listeria monocytogenes, Salmonella, Escherichia coli O157:H7, and Staphylococcus aureus in fresh, minimally processed vegetables

Four commercial DNA extraction methods, PrepMan Ultra (Applied Biosystems), InstaGene Matrix (BioRad), DNeasy Tissue kit (Qiagen), and UltraClean (MoBio), were tested for PCR detection of Listeria monocytogenes, Escherichia coli O157: H7, Salmonella, and Staphylococcus aureus in fresh, minimally processed vegetables. For comparative purposes, sensitivity assays with specific PCRs were carried out after DNA extraction with the four methods in green pepper, broccoli, and onion artificially inoculated with the four pathogens separately. As confirmed by statistical analysis, the DNeasy Tissue kit rendered the highest sensitivity values in the three matrices assayed for Salmonella, L. monocytogenes, and E. coli O157:H7 and in onion for S. aureus. Despite being the most expensive of the methods compared, the DNeasy Tissue Kit can be successfully applied for any of the four most commonly studied pathogens, thus saving time and overall reducing the cost of the analysis.     

Susceptibility of minimally processed green pepper and cucumber to chilling injury as observed by apparent respiration rate

Apparent respiration rate and total microbial count were measured for intact and cut vegetables of green chilli pepper and cucumber stored at 5 and 10 °C. The minimally processed vegetables were more sensitive to chilling injury at 5 °C compared to the intact produce. Their response of increased respiration to chilling injury at 5 °C was noticed earlier than spoilage due to microbial growth, and determined the limit of shelf-life (4 days) in storage. Due to increased susceptibility to chilling injury when these products were stored at 5 °C, they had shelf-lives shorter than or equal to those at 10 °C.     

A multiplex RTi-PCR reaction for simultaneous detection of Escherichia coli O157:H7, Salmonella spp. and Staphylococcus aureus on fresh, minimally processed vegetables

In this work, a new multiplex single-tube real-time PCR approach is presented for the detection of Escherichia coli O157:H7, Salmonella spp. and Staphylococcus aureus, three of the more frequent food-borne bacterial pathogens that are usually investigated in a variety of food matrices. The study includes the design and specificity testing, of a new primer and probe specific for Salmonella spp. Reaction conditions were adjusted for the simultaneous amplification and detection of specific fragments in the beta-glucuronidase (uidA, E. coli) and Thermonulease (nuc, Sta. aureus) genes, and in the replication origin sequence (oriC, Salmonella spp.). Melting-curve analysis using a SYBR Green I RTi-PCR approach showed characteristic T(m) values demonstrating the specific and efficient amplification of the three fragments. Subsequently, a TaqMan RTi-PCR approach was settled, using FAM, NED and VIC fluorescently labelled specific probes for an automated detection. It was equally sensitive than uniplex RTi-PCR reactions in Sta. aureus and E. coli O157:H7, using same amounts of purified DNA, and allowed detection of 10 genome equivalents in the presence of 10(2) or 10(4) genome equivalents of the other two pathogens. Finally, it was tested in artificially inoculated fresh, minimally processed vegetables, revealing a sensitivity of 10(3)CFUg(-1) each of these pathogens in direct detection, following DNA extraction with DNeasy Tissue Kit (Qiagen). The multiplex RTi-PCR developed scored the sensitivity recognised for PCR in food and it allows a high-throughput and automation, thus it is promising as a rapid and cost-effective test for the food industry.