N. E. Martínez-Gonzáles

Comparison of rinsing and sanitizing procedures for reducing bacterial pathogens on fresh cantaloupes and bell peppers

Increased consumption of fruits and vegetables is linked to health benefits but also to an increase in the number of outbreaks of foodborne illness. To determine the effectiveness of different sanitizing treatments for reducing bacterial pathogens on fresh produce, fresh cantaloupes and bell peppers were harvested and inoculated with suspensions of Salmonella Typhimurium and Escherichia coli O157:H7. The inoculated fruits were treated with water wash alone or were washed and then waxed or rinsed with 200 mg/liter hypochlorite, 10% Ca(OH)2, or 2% lactic acid solutions applied by dipping for 15 s or spraying for 15 s. Preliminary experiments with chlorine treatments indicated that spraying with a 200, 600, or 1,000 mg/liter hypochlorite solution reduced populations of both pathogens by 2.1 to 2.6 and 1.5 to 2.1 log CFU for Salmonella Typhimurium and E. coli O157:H7, respectively. In general, no differences were observed between chlorine solutions without pH adjustment (pH 9.2) and those with pH adjusted to 6.0. When different wash regimes were applied to inoculated cantaloupes or bell peppers, water wash alone produced significantly lower counts of both pathogens on bell peppers in comparison to untreated controls. However, this reduction was not observed on cantaloupes, indicating a possible surface effect. Application of 2% L-lactic acid by spray was the treatment that resulted in the lowest bacterial counts on both cantaloupes and bell peppers. This treatment did not produce any deleterious change in the sensorial characteristics of the products tested. None of the pathogens studied was able to grow during refrigerated storage (5 degrees C for cantaloupes and 10 degrees C for bell peppers), although numbers close to the detection limit of the counting method were found in randomly tested individual samples at days 14 and 28 of storage, indicating that these pathogens can survive for long periods on the produce surface. These results indicate that selected produce commodities could be sanitized at the packing facility. However, these interventions should not be applied as a replacement for but only as a complement to good hygiene practices.

Internalization of Bacterial Pathogens in Tomatoes and Their Control by Selected Chemicals

The effect of different washing or sanitizing agents was compared for preventing or reducing surface and internal contamination of tomatoes by Salmonella Typhimurium and Escherichia coli O157:H7. The tomatoes were inoculated by dipping them in a bacterial suspension containing approximately 6.0 log CFU/ml of each pathogen and then rinsing them with tap water, hypochlorite solution (250 mg/liter), or lactic acid solution (2%, wt/vol). All treatments were applied by dipping or spraying, and solutions were applied at 5, 25, 35, and 55 degrees C. With the exception of the lactic acid dip at 5 degrees C, all treatments reduced both pathogens on the surfaces of the tomatoes by at least 2.9 cycles. No significantly different results were obtained (P > 0.05) with the dipping and spraying techniques. For internalized pathogens, the mean counts for tomatoes treated with water alone or with chlorine ranged from 0.8 to 2.1 log CFU/g. In contrast, after lactic acid spray treatment, all core samples of tomatoes tested negative for Salmonella Typhimurium and, except for one sample with a low but detectable count, all samples tested negative for E. coli O157:H7 with a plate count method. When the absence of pathogens was verified by an enrichment method, Salmonella was not recovered from any samples, whereas two of four samples tested positive for E. coli O157:H7 even though the counts were negative. Few cells of internalized pathogens were able to survive in the center of the tomato during storage at room temperature (25 to 28 degrees C). The average superficial pH of tomatoes treated with tap water, chlorine, or lactic acid was 4.9 to 5.2, 4.1 to 4.3, and 2.5, respectively (P < 0.05), whereas no differences were observed in the internal pH (3.6 to 3.7) of the tomatoes treated with different sanitizers. The general practice in the tomato industry is to wash the tomatoes in chlorinated water. However, chlorine is rapidly degraded by organic matter usually present in produce. Therefore, lactic acid sprays may be a more effective alternative for decontaminating tomato surfaces. The use of warm (55 degrees C) sprays could reduce pathogen internalization during washing.

Comparison of different washing treatments for reducing pathogens on orange surfaces and for preventing the transfer of bacterial pathogens to fresh-squeezed orange juice.

The objectives of this study were to compare the effectiveness of various washing treatments for reducing Escherichia coli O157:H7, Salmonella sp., and Listeria monocytogenes populations on orange surfaces and to measure the effect of some of these treatments in preventing the transfer of pathogens during juice extraction. Orange surfaces inoculated with L. monocytogenes or a mixture of E. coli O157:H7 and Salmonella Typhimurium were washed by water spray and then sprayed with or dipped in water at 80°C for 1 min, 70% ethanol for 15, 30, or 45 s or 1, 2, or 4 min, 2 or 4% lactic acid solution at 55°C for 15, 30, or 45 s or 1, 2, or 4 min, or 200 mg/liter hypochlorite at pH 6.5 or 10 for 15 s. The surviving populations of these pathogens on the oranges were enumerated after each treatment. In a further stage, the ability of these pathogens to be transferred to the juice during extraction was tested. Juice was obtained from inoculated oranges that were subjected to selected treatments using chlorine, lactic acid, ethanol, and hot water as described above, and then bacterial counts in orange juice were determined. The application of these treatments reduced the populations of pathogens on orange surfaces by 1.9 to >4.9 log, 1.9 to >4.6 log, and 1.4 to 3.1 log cycles for E. coli O157:H7, Salmonella Typhimurium, and L. monocytogenes, respectively. The treatments using hot water or lactic acid showed greater reductions than other treatments. The time, antimicrobial concentration, and form of application affected the bacterial reduction. All treatments resulted in undetectable counts in the juice. Nevertheless, pathogens were recovered by the enrichment-plating method. Treatment of oranges before juice extraction may reduce the risk associated with consuming orange juice.

Validation of a washing and sanitizing procedure for cantaloupes at a mexican packing facility.

In-plant validation of an alternative washing and sanitizing method was conducted at a cantaloupe packing operation in Mexico. This method consisted of a spray water wash followed by spraying warm (55 to 60 degrees C) 2% (L)-lactic acid solution and was compared with the existing method of spray washing the melons with tap water followed by immersion in a chlorinated water tank. Surface samples (100 cm(2)) were collected from 160 melons subjected to each processing method and tested for counts of aerobic bacteria, coliforms, and Escherichia coli. The aerobic plate counts from cantaloupes washed in the dump tank ranged from 3.6 to 5.2 log CFU/cm(2) and were significantly higher (P < 0.05) than those from melons treated with the alternative spray method, which ranged from 1.8 to 2.6 log CFU/cm(2). Coliform counts for cantaloupes treated in the dump tank were 0.2 to 2.2 log CFU/cm(2) and were below the detection level (-6.0 log CFU/cm(2)) on cantaloupes treated by the spray method. Growth of E. coli was observed in 2.5% of the samples of cantaloupes treated in the dump tank and in none of the samples of cantaloupes treated by lactic acid spray (P < 0.05). These results support the elimination of dump tanks in cantaloupe packing operations established by the Mexican government for certification of firms exporting cantaloupes to the United States. When a sanitizer is to be applied to the product, lactic acid seems to be a viable option, at least for products such as cantaloupes whose quality is not affected by an acid wash.

Incidence of Vibrio cholerae in Fresh Fish and Ceviche in Guadalajara, Mexico

The incidence of Vibrio cholerae O1 and non-Ol was determined in fresh fish and ceviche, a marinated raw fish dish ready for consumption. Fresh red snapper ( Lutjanus purpureous ) and mackerel ( Scomberomorus sierra ) were obtained from distribution centers, and ceviche from street vendors and small open restaurants in Guadalajara, Mexico. In addition to V. cholerae , the fish samples were tested for aerobic plate count (APC), total volatile nitrogen (TVN), trimethylamine (TMA), and the ceviche for APC, coliforms, and pH. V. cholerae O1 and non-O1 was isolated from 10% and 26% ofthe fish respectively. The mean data for the fish samples were in the region of: APC, 106 CFU/g of fish; more than 25 mg of TVN per 100 g of fish, but less than 5 mg of nitrogen as TMA per 100 g. Eleven percent of the ceviche obtained from street vendors and 6% obtained from restaurants were positive for V. cholerae O1. The mean APC and coliform counts were 6.6 and 4.8 log CFU/g of fish respectively, and the pH of the ceviche ranged from 3.0 to 4.5. All the strains of V. cholerae O1 isolated during this study were identified as biotype El Tor, serotypes Inaba and Ogawa. For both fresh fish and ceviche, the frequency of isolation of V. cholerae was highest during the summer months.

Survival of vibrio cholerae O1 in ceviche and its reduction by heat pretreatment of raw ingredients

The survival of Vibrio cholerae O1 serotypes Inaba and Ogawa was determined in ceviche prepared from inoculated ground fish. Ground mackerel purchased from a seafood distribution center was inoculated with V. cholerae and stored at 8 or 20 degrees C. Counts of V. cholerae decreased in 2.6 to 2.7 log10 CFU/g during 96 h of storage at 8 degrees C or 2.5 to 2.6 log10 CFU/g during 24 h at 20 degrees C. Survival studies indicated that serotype Inaba decreased its number following a linear or retarded trend, whereas serotype Ogawa followed an accelerated death trend. No effect of the initial level of inoculum was observed. Odor scores of ceviche indicated that this food became marginally acceptable within as little as 48 h of storage at 8 degrees C or 3 h at 20 degrees C and were related to total volatile nitrogen values but not to aerobic plate counts, pH, or coliform counts. A heat pretreatment that consisted of stirring 100 g of inoculated ground fish into 40 ml of boiling water produced an 8-log reduction of V. cholerae within 3 min without affecting the color, odor, or flavor of ceviche prepared with such pretreated fish. According to this study, V. cholerae present in contaminated ceviche will likely survive longer than the shelf life of this food. Preheating the ground raw fish used for preparing ceviche for 3 min should effectively eliminate V. cholerae O1, providing science-based conditions for implementing a critical control point if a hazard analysis critical control point plan were to be developed for preparation of ceviche.

Use of a novel medium, the Polymyxin Ceftazidime Oxford Medium, for isolation of Listeria monocytogenes from raw or non-pasteurized foods.

Polymyxin Ceftazidime Oxford Medium (PCOM), a novel selective and differential plating medium for Listeria monocytogenes was compared with Modified Oxford Agar (MOX) for efficacy to isolate L. monocytogenes and other Listeria spp. naturally present in non-pasteurized Mexican-style cheese (n = 50), non-pasteurized fresh squeezed orange juice (n = 50), raw beef chunks (n = 36), and fresh cabbage (n = 125). Samples were collected from retail markets and farms in Mexico and tested following the US Department of Agriculture enrichment technique. Listeria spp. were isolated from 23.4% of analyzed samples, and from those, 75.0% corresponded to raw beef chunks, 38.0% to non-pasteurized Mexican-style cheese, and 30.0% to fresh squeezed orange juice. No Listeria spp. were isolated from fresh cabbage samples. L. monocytogenes was recovered from 15.3% of food samples analyzed. Non-pasteurized Mexican-style cheese showed the highest proportion of L. monocytogenes positive samples (36.0%), followed by orange juice (26.0%) and raw beef (25.0%). The frequency of isolation of Listeria spp. and L. monocytogenes was not different (P > 0.05) between PCOM and MOX. The advantages of using PCOM when comparing to MOX, include the easier way to identify Listeria species, the lower cost per plate and the availability of its ingredients for Latin-American countries.

Microbiological Quality and Incidence of Salmonella on Cherry Tomatoes at Retail in Querétaro, México

Multiple outbreaks related to Salmonella in tomatoes require an evaluation of the risk associated with cherry tomatoes due to the increase in its production, consumption, and marketing in Mexicos central region. The purpose of this study was to determine the microbial quality of cherry tomatoes obtained from two retail sale points (supermarkets and local markets). Cherry tomato samples (333) were collected from four supermarkets and from four local markets, and the contents of aerobic plate count, molds and yeasts, total coliforms, and Escherichia coli were quantified; the presence of Salmonella was simultaneously determined. The median values of the microbial populations were obtained, and the data were analyzed per the sampling site by using the Wilcoxon and Kruskal-Wallis tests. The median of aerobic plate count content in tomatoes obtained from supermarkets ranged between 2.2 and 4.4 log CFU/g, and in markets from 2.9 to 4.8 log CFU/g. For molds and yeasts, the tomatoes from supermarkets (2.0 to 4.1 log CFU/g) and markets (1.5 to 4.5 log CFU/g) showed similar contents. Regardless of the sampling site, the values of total coliforms were very low, ranging from 1.0 to 1.8 log CFU/g. E. coli was detected in 5.4 and 20.1% of samples from supermarkets and markets, respectively; in both sites, the content was low (0.3 to 5.8 most probable number per g). The incidence of Salmonella was 14.1% in supermarkets and 7.8% in local markets. The results obtained from this investigation highlight the elevated risk for consumer health associated with the ingestion of cherry tomatoes.

Safety of fresh-squeezed juices

Abstract Outbreaks of foodborne illness resulting from consumption of contaminated fresh juices continue to occur worldwide. Between 1974 and 2014, fresh juices were involved in at least 48 foodborne illness outbreaks involving bacteria (Salmonella, Shigella flexneri, Shiga-toxin-producing Escherichia coli including groups O157 and O111 STEC, and enterotoxigenic E. coli), viruses (norovirus genotype II, hepatitis A virus), or protozoa (Cryptosporidium and Trypanosoma cruzi) as etiologic agents. Most of these outbreaks were linked to fresh-squeezed orange or apple juice, while outbreaks caused by T. cruzi were linked to fresh juice made from various exotic fruits. The sources of pathogens causing outbreaks in fresh-squeezed juice include animal feces contaminating the raw fruits or vegetables used for juice making, contaminated water used in the process, unsanitary equipment and utensils used in juice extraction and serving, and food handlers. These pathogens may be internalized into the fruits used to obtain juice, and survive despite the natural acidity of some fruit juices. Therefore, more effective strategies for reducing pathogens potentially present in fresh-squeezed juices should be applied. These strategies include the use of nondamaged raw materials, implementation of Good Manufacturing Practices during processing, and the implementation of an HACCP plan when possible.

Effect of the Use of a Neutralizing Step after Antimicrobial Application on Microbial Counts during Challenge Studies for Orange Disinfection

The effects of using a neutralizer after applying antimicrobial treatments and the effect of time lapse between treatment application and subsequent recovery and enumeration of Escherichia coli O157:H7 and Salmonella were investigated in Valencia oranges. Inoculated oranges surfaces were washed with distilled water for 15 s and then sprayed with a solution containing 200 mg/liter sodium hypochlorite (pH 6.5) for 15 s; they were then dipped in L-lactic acid (2.0% at 55°C) for 1 min or in distilled water at 80°C for 1 min. Posttreatment, oranges were divided into two groups. In the first group, oranges were dipped in neutralization treatment: 270 ml of buffered peptone water for 2 min for lactic acid-treated oranges, 270 ml of Dey-Engley broth for 2 min for chlorine-treated oranges, or 3.7 liters of tap water (25°C) for 10 s for hot water-treated oranges. The second group of treated oranges was not subjected to any neutralizer. All oranges then were kept at room temperature (average 26.2°C) and sampled at 0, 7.5, and 15 min for enumeration of surviving Salmonella and E. coli O157:H7. The orange surface (30 cm(2)) was excised for pathogen enumeration. The presence of free chlorine and changes in pH and temperature on the orange surface were determined in uninoculated, treated oranges. Free chlorine was detected on oranges after treatment; the change in temperature of orange surfaces was greater during treatment with hot water than with lactic acid. Nevertheless, pathogen enumeration did not show any impact of neutralizer use on the residual activity of antimicrobials or any impact of the time elapsed between antimicrobial treatment and recovery of bacterial pathogens from inoculated oranges (P ≥ 0.05). The results of this study indicate that the lack of a neutralizing step before enumeration of pathogens is not likely to affect the accuracy of results during challenge studies to test pathogen reduction strategies on oranges.