Log MPN Research Articles

Analysis of the drought effects on the physicochemical and bacteriological quality of the Inaouene River water (Taza, Morocco)

The Inaouene basin's waters are deeply affected by pollution, which undermines their quality. Indeed, due to the increasing population and industrial development of Taza city, the Inaouene River and its tributaries Larbaa and Lahdar are currently at risk of large quantities of urban discharge, exacerbated by successive drought periods. The objective of this study was to determine the pollution levels and assess the impact of drought on the water quality of the Inaouene River, based on physicochemical, and bacteriological analyses at seven stations over two hydrological cycles (June 2020 to May 2022). The results of physicochemical analyses during the first hydrological cycle revealed averages of 15.93 °C for Temperature, 1434.43 µS cm-1 for electrical conductivity (EC), 0.93 and 0.63 mg L-1 respectively for total phosphorus (TP) and orthophosphate (PO43-), 9 mg L-1 for NTK, 134.23 and 278.43 mg L-1 for BOD5 and COD respectively, 3.99 mg L-1 for Dissolved oxygen (DO) and 203.59 mg L-1 for sulfate (SO42-). For water bacteriological quality, the total coliforms (TC), Escherichia coli (E. coli), and intestinal enterococcus (IE) have shown averages of 3.45, 2.96, and 2.73 Log10 MPN per 100 ml respectively. During the second hydrological cycle, marked by low rainfall, a pronounced degradation in water quality has been observed, affecting all parameters. Notably, water quality went from average to poor or very poor according to the Moroccan surface water quality standards (MSWQS) and from very poor to unsuitable for consumption for the Water Quality Index (WQI) during the drought period. This study confirmed the exacerbation of deterioration in the water quality of the Inaouene River during the drought period, notably evidenced by heightened concentrations of organic matter (increases of 41.53 % and 37.08 % for BOD5 and COD, respectively), fecal contaminants (elevations of 11.48 %, 10.18 %, and 14.7 % for IE, TC, and E. coli, respectively), nutrients (increases of 38.83 % for TP, 48.67 % for PO43-, and 35.82 % for NTK), and mineral content (augmentations of 28.89 % for EC, and 31.73 % for sulfate). Additionally, this degradation in quality is reflected in the elevation of water temperature and the reduction in oxygenation (of 9.39 % for Temperature, and 46,26 % for DO).

Development of a depuration protocol for commercially important edible bivalve molluscs of India: Ensuring microbiological safety

We investigated the effect of depuration of three naturally contaminated commercially important tropical edible bivalve molluscs by varying temperature, salinity and body-size of animals harvested from Ashtamudi and Vembanad estuaries, India using a static depuration system to ensure microbiological food safety. Before depuration, the levels of faecal indicators and pathogens were above the acceptable limits for live consumption. The depuration water temperature had a significant effect on bacterial elimination. Log reduction of faecal coliforms (FC) and E. coli varied between room-temperature (RTDS) and low-temperature depuration system (LTDS) and it was in the range of 1.39–2.44 and 1.88–2.82 log MPN, respectively under RTDS and LTDS. The elimination of bacterial pathogens such as Vibrio and Salmonella spp. was rapid in RTDS compared to LTDS. The highest elimination of FC and E. coli (2.39 and 2.92 log) was at 35 psμ depuration and the lowest (0.87 and 1.65 log) at 15 psμ depuration. The reduction of FC and E. coli was higher in the medium-sized animals compared to the small animals. Based on the results it is recommended that depuration using the static system under room-temperature (30 ± 1 °C); a salinity range of (25–35 psμ) and using medium-size bivalves (clam>30 mm; mussel >45 mm, and oyster >65 mm length) as optimum conditions for producing microbiologically safe bivalves for live consumption within 36 h.

Dietary Inclusion of a Saccharomyces cerevisiae-Derived Postbiotic Is Associated with Lower Salmonella enterica Burden in Broiler Chickens on a Commercial Farm in Honduras.

Postbiotic feed additives may aid foodborne pathogen reduction during poultry rearing. The study objective was to evaluate a postbiotic additive in parallel to an industry control diet and the subsequent associated burden of Salmonella enterica on a single, commercial broiler farm in Honduras. Twelve houses were matched and assigned the standard diet (CON) or standard diet plus postbiotic (SCFP). New litter was placed in each house and retained across flock cycles with sampling prior to each chick placement and three consecutive rearing cycles. At ~33–34 days, 25 ceca were collected on-farm from each house, treatment, and cycle. Salmonella prevalence in litter for CON (30.6%) and SCFP (27.8%) were equivalent; however, Salmonella load within positive samples was lower (p = 0.04) for SCFP (3.81 log10 MPN/swab) compared to CON (5.53 log10 MPN/swab). Cecal prevalence of Salmonella was lower (p = 0.0006) in broilers fed SCFP (3.4%) compared to CON (12.2%). Salmonella load within positive ceca were numerically reduced (p = 0.121) by 1.45 log10 MPN/g for SCFP (2.41 log10 MPN/g) over CON (3.86 log10 MPN/g). Estimated burden was lower (p = 0.003) for SCFP flocks (3.80 log10 MPN) compared to CON (7.31 log10 MPN). These data demonstrate the preharvest intervention potential of postbiotics to reduce Salmonella enterica in broiler chickens.

Evaluation of bacterial contamination on three different surfaces within domestic refrigerators

The objectives of this study were to analyze the number of microorganisms, fungal composition and the correlation between bacterial enrichment and air quality on three internal surfaces (the inner wall, shelf, and basket) of domestic refrigerators. The results showed that the inner wall had a significantly lower number of coliforms (<italic>P</italic> &lt; 0.05), and the range was 0.2-2.5 log MPN cm⁻². The total bacterial counts and psychrophilic bacterial counts on three internal surfaces in the same refrigerator tended to be consistent. Moreover, the inner wall owned a simpler bacterial community structure. At the genus level of fungi, the dominant flora of both the inner wall and shelf were Saccharomyces spp. and Candida spp., while Saccharomyces spp., Candida spp. and Fistulina spp. took superiority in the basket. Specifically, Shannon index and Simpson index, which represent the bacterial community diversity, were the lowest on the wall, and 6 bacterial species on the inner wall had relative abundance higher than 0.5% of the total operational taxonomic units (OTUs), while for shelf and basket, there were 12 and 11 bacterial species respectively. Also, there was a significantly negative correlation in the basket between the chao1 index and PM2.5. This study could provide guidance for the sanitation and recommend an adequate package of foods stored in refrigerators.

Detection, quantification, and characterization of Salmonella enterica in mango, tomato, and raw chicken purchased in the central region of Mexico

To estimate human exposure to Salmonella enterica, it is essential to understand the pathogen distribution and characteristics. Prevalence and concentration of S. enterica were determined in mango, tomato, and raw chicken samples purchased in three states (Aguascalientes, Querétaro, and Guadalajara) located in the central region of Mexico during two seasons. In addition, S. enterica isolates were characterized by absence/presence of 13 virulence genes (chromosomal, prophage, and plasmid) and resistance to 14 antibiotics. A total of 300 samples of mango, 272 of tomato, and 354 of raw chicken were analyzed. The mean of the prevalence (24.9%) and concentration (-0.61 Log MPN/g) of S. enterica in chicken was higher than in mango (1.3%, -1.7 Log MPN/g) and tomato (1.1%, -1.7 Log MPN). Among S. enterica isolates (284), there were 7 different virulotypes, belonging 68.7% of isolates to V2; there was high variability in the presence of mobile genetic elements. The occurrence of specific mobile elements ranged from 81.4% to 11.3% among isolates. Among the isolates, 91.5% were resistant to at least one antibiotic with ampicillin being the most frequent; 54.9% of isolates were multidrug resistant. Data from this study can be used for quantitative microbial risk assessment of S. enterica related to mango, tomato, and raw chicken consumption in the central region of Mexico. PRACTICAL APPLICATION: Data on the prevalence and concentration of Salmonella enterica obtained in this study can be used to estimate the exposure assessment for the consumption of mango, tomato, and chicken in the central region of Mexico. In addition, the characteristics of the S. enterica isolates could be used to select representative strains for future studies to evaluate the intraspecies variability.

Hazard Identification Related to the Presence of Vibrio spp., Biogenic Amines, and Indole-Producing Bacteria in a Non-Filter Feeding Marine Gastropod (Tritiamutabilis) Commercialized on the Italian Market.

Tritia mutabilis is a carrion-feeder edible marine gastropod with an open circulatory system. Therefore, biological, and chemical contaminants associated with the feed can reach all body tissues. The aim of the present study was to investigate the possible association of these characteristics with some food safety hazards. Vibrio spp. load, and the prevalence of pathogenic V. parahaemolyticus, V. vulnificus, and V. cholerae, were investigated. Moreover, biogenic amines (BAs) and indole-producing bacteria (IPB), markers of seafood decomposition, were quantified for the first time in an edible carrion-feeder. Overall, 49 batches were analyzed (38 from retail, and 11 from primary production). The Vibrio spp. load resulted of 5.64 ± 0.69 log10 CFU g−1 at retail, and 5.27 ± 0.74 at harvest but all batches resulted negative for pathogenic Vibrio. Histamine, putrescine, cadaverine, and tyramine were detected both at harvest and at the retail level. Their sum (BAs Index) showed a mean value of 50.45 and 65.83 mg Kg−1 in batches at harvest and at retail, respectively. IPB were detected at harvest and upon refrigeration for three days (T1–T3). The mean load resulted in 2.52 ± 0.85 log10 MPN g−1 at T0, 3.31 ± 1.23 at T3 in batches immediately refrigerated, and 3.22 ± 1.18 at T3 in batches previously immersed in clean seawater. Our results contribute to identifying food-borne hazards for T. mutabilis that may be related to the retention of biogenic amines and indole-producing bacteria due to carrion feeding.

Monte Carlo Simulation Model for Predicting Salmonella Contamination of Chicken Liver as a Function of Serving Size for Use in Quantitative Microbial Risk Assessment

The first step in quantitative microbial risk assessment (QMRA) is to determine the distribution of pathogen contamination among servings of the food in question at some point in the farm-to-table chain. In the present study, the distribution of Salmonella contamination among servings of chicken liver for use in the QMRA was determined at meal preparation. Salmonella prevalence (P), most probable number (MPN, N), and serotype for different serving sizes were determined by use of a combination of five methods: (i) whole sample enrichment; (ii) quantitative PCR; (iii) culture isolation; (iv) serotyping; and (v) Monte Carlo simulation. Epidemiological data also were used to convert serotype data to virulence (V) values for use in the QMRA. A Monte Carlo simulation model based in Excel and simulated with @Risk predicted Salmonella P, N, serotype, and V as a function of a serving size of one (58 g) to eight (464 g) chicken livers. Salmonella P of chicken livers was 72.5% (58 of 80) per 58 g. Four Salmonella serotypes were isolated from chicken livers: (i) Infantis (P = 28%, V = 4.5); (ii) Enteritidis (P = 15%, V = 5); (iii) Typhimurium (P = 15%, V = 4.8); and (iv) Kentucky (P = 15%, V = 0.8). Salmonella N was 1.76 log MPN/58 g (median) with a range of 0 to 4.67 log MPN/58 g, and the median Salmonella N was not affected (P > 0.05) by serotype. The model predicted a nonlinear increase (P ≤ 0.05) of Salmonella P from 72.5%/58 g to 100%/464 g, a minimum N of 0 log MPN/58 g to 1.28 log MPN/464 g, and a median N from 1.76 log MPN/58 g to 3.22 log MPN/464 g. Regardless of serving size, predicted maximum N was 4.74 log MPN per serving, mean V was 3.9 per serving, and total N was 6.65 log MPN per lot (10,000 chicken livers). The data acquired and modeled in this study address an important data gap in the QMRA for Salmonella and whole chicken liver.

Microbiological Characteristics and Behavior of Staphylococcus aureus, Salmonella spp., Listeria monocytogenes and Staphylococcal Toxin during Making and Maturing Cotija Cheese

Cotija cheese is an artisanal matured Mexican cheese from unpasteurized milk. This work determined the microbiological characteristics and behavior of Staphylococcus aureus, Salmonella spp., Listeria monocytogenes and staphylococcal toxin during cheese elaboration and ripening. Sixteen 20-kg cheeses were used, eight inoculated with 6 log CFU/mL of each pathogen, and eight uninoculated, and samples were taken at each stage of the process. In the uninoculated samples, the survival of S. aureus and L. monocytogenes decreased after 30 days of ripening. The average counts of S. aureus in milk, curd, and serum were 7 log MPN /mL, and 8.7 log MPN /g in cheese, decreasing from day 15. Salmonella spp. counts (initial inoculum: 7.2 log MPN /mL) decreased after 24 h, and L. monocytogenes counts (8.7 log MPN/g at 24 h) decreased from day 15 in the cheese. Salmonella spp. and L. monocytogenes were not detected in any sample after 60 days of ripening, unlike S. aureus, which was detected at the end of the study. Lactic acid bacteria counts were 9 log CFU/mL in milk and whey and 7.2 log CFU/g in cheese. Pathogens behavior during the ripening process reduces the health risks by consuming products made with unpasteurized milk when subjected to ripening.

A Bayesian inference approach to quantify average pathogen loads in farmyard manure and slurry using open-source Irish datasets

Farm-to-fork quantitative microbial risk assessments (QMRA) typically start with a preliminary estimate of initial concentration (Cinitial) of microorganism loading at farm level, consisting of an initial estimate of prevalence (P) and the resulting pathogen levels in animal faeces. An average estimation of the initial concentration of pathogens can be achieved by combining P estimates in animal populations and the levels of pathogens in colonised animals' faeces and resulting cumulative levels in herd farmyard manure and slurry (FYM&S). In the present study, 14 years of data were collated and assessed using a Bayesian inference loop to assess the likely P of pathogens. In this regard, historical and current survey data exists on P estimates for a number of pathogens, including Cryptosporidium parvum, Mycobacterium avium subspecies paratuberculosis (MAP), Salmonella spp., Clostridium spp., Campylobacter spp., pathogenic E. coli, and Listeria monocytogenes in several species (cattle, pigs, and sheep) in Ireland. The results revealed that Cryptosporidium spp. has potentially the highest mean P (Pmean) (25.93%), followed by MAP (15.68%) and Campylobacter spp. (8.80%) for cattle. The Pmean of E. coli is highest (7.42%) in pigs, while the Pmean of Clostridium spp. in sheep was estimated to be 7.94%. Cinitial for Cryptosporidium spp., MAP., Salmonella spp., Clostridium spp., and Campylobacter spp. in cattle faeces were derived with an average of 2.69, 4.38, 4.24, 3.46, and 3.84 log10 MPN g −1, respectively. Average Cinitial of Cryptosporidium spp., Salmonella spp., Clostridium spp., and E. coli in pig slurry was estimated as 1.27, 3.12, 3.02, and 4.48 log10 MPN g −1, respectively. It was only possible to calculate the average Cinitial of Listeria monocytogenes in sheep manure as 1.86 log10 MPN g −1. This study creates a basis for future farm-to-fork risk assessment models to base initial pathogen loading values for animal faeces and enhance risk assessment efforts.

Microbiological Quality Assessment of Sashimi and Sushi Sold in the Federal District, Brazil

This study aimed to evaluate the microbiological quality of sashimi and sushi sold in the Federal District, Brazil. A total of 54 samples (27 sashimi and 27 sushi) were collected in 25 commercial establishments. Samples were tested for counts of mesophilic and psychrotrophic bacteria, determination of total and thermotolerant coliforms, Staphylococcus aureus counts, and presence of Salmonella. The results showed that 40.7% of sashimi (11/27) and 25.9% of sushi (7/27) samples were unfit for consumption according to Brazilian legislation. Salmonella (genetically confirmed by the invA gene’s presence) was present in 25.9% of the sashimi samples (7/27). Thermotolerant coliforms were positive in 92.5% of sashimi samples (25/27) and 74.0% of sushi samples (20/27), and one sashimi sample was unfit for consumption (≥ 2.0 log MPN g-1). S. aureus bacteria exceeded the acceptable limit of 3.0 log CFU g-1 in 14.9% of sashimi samples (4/27) and 25.9% of sushi samples (7/27). A high count of S. aureus bacteria in some sushi and sashimi samples indicates improper personal hygiene while processing these ready-to-eat foods. Salmonella spp. can occur by improper handling of these foods or by the contact of fish with waters contaminated with sewage. These results can pose a health risk to consumers and indicated the need to improve good hygienic practices in establishments selling sashimi and sushi in Federal District, Brazil.

The efficacy of X-ray doses on Vibrio vulnificus in pure culture and Vibrio parahaemolyticus in pure culture and inoculated farm raised live oysters (Crassostrea virginica) with different acceleration voltages

Vibrio vulnificus and Vibrio parahaemolyticus are halophilic bacteria and can cause illness associated with the consumption of oysters. In this study, we tested the hypothesis to see if the adjustment of the acceleration voltage can change the efficacy of X-ray doses on V. vulnificus and V. parahaemolyticus in pure culture and inoculated V. parahaemolyticus within farm-raised live oysters (Crassostrea virginica). By using a tailor-made X-ray irradiator (Kimtron 350), the acceleration voltage was adjusted to 50, 200 or 350 kV and irradiated on pure culture of V. vulnificus or V. parahaemolyticus and farm-raised live oysters accumulated with a three-strain mixture of V. parahaemolyticus. Inactivation of V. vulnificus and V. parahaemolyticus pure cultures without filter had a lower inactivation when compared with filtered X-ray. With a 1 mm aluminum filter, 250–300 Gy X-ray reduced more than 6 log CFU ml−1 at 350 kV acceleration voltage. When 350 kV X-ray was applied on live oysters with the aluminum filter, 6 log MPN g−1 reduction of V. parahaemolyticus was achieved with 1250 Gy. The V. parahaemolyticus was significantly (p < 0.05) reduced in whole shell oysters from 8.11 ± 0.19 to 6.11 ± 0.19, 3.97 ± 0.47, 4.17 ± 0.17 and 2.77 ± 0.28 log MPN g−1 after processing with 250, 500, 750 and 1000 Gy, respectively. These results showed that the pathogen inactivation can be improved by the optimization of energy level with the adjustment of irradiation filter and acceleration voltage.

Sources of food contamination in a closed hydroponic system.

This study investigated potential contamination sources in a commercial, closed hydroponic system. Water, substrate and lettuce (Lactuca sativa) samples were evaluated for microbiological indicator populations, including aerobic plate count (APC), coliform bacteria (CB) and yeast and mould (YM). Listeria spp. detection via cultural enrichment and agglutination was negative for all samples. Peat moss substrate (postharvest) had the highest counts for APC (6·8 log CFU per g), CB (4·5 log MPN per g) and YM (5·1 and 4·8 log CFU per g respectively). Roots embedded in plugs demonstrated counts for all populations nearly as high as the substrate. Among water samples, a seedling water reservoir housing germinated plants yielded the highest count for APC (5·1 log CFU per g) and CB (2·4 log MPN per g) likely due to the large numbers of plugs and their close proximity in the reservoir. Harvested lettuce leaves demonstrated higher APC (4·1 log CFU per g) than preharvest leaves (1·7 log CFU per g) due to the transfer of microbes from the root ball. These data suggest that substrates are a significant potential source of contamination in hydroponic systems and likely facilitate microbial transfer to harvested leaves. There is, therefore, the need to further investigate mitigation of potential contamination events. SIGNIFICANCE AND IMPACT OF THE STUDY: Hydroponic production is known to provide safe, clean produce. This study, however, suggests that the hydroponic substrate (peat moss plug) is a possible source of contamination in the hydroponic system. This finding is important as most harvested hydroponic lettuces are packaged and sold with substrate and root ball intact. This implies a high probability of microbial transfer from the root ball to edible harvested lettuce leaves.

First report ofEscherichia coliO157:H7 in ready‐to‐eat sushi

The aim of this study was to evaluate the microbiological quality of commercially prepared ready-to-eat (RTE) sushi by enumerating aerobic mesophilic bacteria (AMB) and thermotolerant coliforms (TC) and detecting Escherichia coli and Salmonella ssp. An isolate was identified as E. coli O157:H7 which was evaluated for its virulence and antimicrobial resistance profiling as well as its ability to form biofilms on stainless steel. There were four sampling events in seven establishments, totalling 28 pools of sushi samples. Mean AMB counts ranged between 5·2 and 7·7 log CFU per gram. The enumeration of TC varied between 2·1 and 2·7 log MPN per gram. Salmonella ssp. were not detected, and one sample was positive for E. coli and was identified as E. coli O157:H7. To the best of our knowledge, this is the first report of E. coli O157:H7 in sushi samples in the world literature. This isolate presented virulence genes stx1, stx2, eae and hlyA. It was also susceptible to 14 antimicrobials tested and had the ability to form biofilms on stainless steel. There is a need to improve the good hygiene practices adopted in establishments selling sushi in the city of Pelotas, Brazil. In addition, the isolated E. coli O157:H7 carries a range of important virulence genes being a potential risk to consumer health, as sushi is a RTE food. This isolate also presents biofilm formation ability, therefore, may trigger a constant source of contamination in the production line of this food. The increase in the consumption of sushi worldwide attracts attention regarding the microbiological point of view, since it is a ready-to-eat food. To our knowledge, this was the first time that E. coli O157:H7 was identified in sushi samples.

Assessing the relationship between organic farming practices and microbiological characteristics of organic lettuce varieties (Lactuca sativa L.) grown in Sao Paulo, Brazil.

This study aimed to gather information on farming practices employed in organic lettuce fields in Sao Paulo, Brazil and associate these practices with the microbiological characteristics of the products. Practices were surveyed using a questionnaire applied in ten farms, where 200 heads of lettuce were collected and submitted to enumeration of total coliforms and generic Escherichia coli and tested for Salmonella spp. using culture and molecular (qPCR) methods. Based on the responses, the farms could be clustered in two groups: group 1, comprised by six farms, where chicken manure was used as fertilizer in most of them and the composting process was not performed on site; and group 2, comprised by four farms, where other types of fertilizer were used, and the composting process was performed on site. Generic E. coli was detected in 56 (28%) samples, with an average of 1·1±0·7log MPN per g. Salmonella DNA was detected in two (1%) samples by qPCR. The prevalence and bacterial loads of generic E. coli, and the occurrence of Salmonella, even at low populations undetectable by conventional culture methods, highlight the need for control measures during farming practices to reduce microbial contamination and risks of foodborne illnesses. These measures include the use of properly composted manure and appropriate washing procedures for leafy vegetables before consumption. The obtained data contribute to a better understanding of the farming practices of organically grown lettuces in Sao Paulo, Brazil.

Developing, cross-validating and applying regression models to predict the concentrations of faecal indicator organisms in coastal waters under different environmental scenarios

This study developed, cross-validated and applied a regression-based model to predict concentrations of faecal indicator organisms (FIOs) under different environmental conditions in the North and South bays of Santa Catarina, South of Brazil. The model was developed using a database of FIO concentrations in seawater sampled at 50 sites and the validation was performed using a different database by comparing 288 pairs of measured and modelled results for 15 sites. The index of agreement between the model outputs and the FIO concentrations measured during the validation period was 66%; the mean average error was 0.43 log10 and the root mean square error was 0.58 log10 MPN.100mL−1. These validation results indicate that the model provides a fair representation of the FIO contamination in the bays for the meteorological conditions under which the model was trained. The simulation of different scenarios showed that under typical levels of resident human population in the catchments and median rainfall and solar radiation conditions, the median FIO concentration in the bays is 0.4 MPN.100mL−1. Under extreme meteorological conditions, the combined effect of high rainfall and low solar radiation increased FIO concentrations up to 5 log10 MPN.100mL−1. The simulated scenarios also show that increases in resident population during the summer tourist season and average rainfall concentrations do not increase median FIO concentrations in the bays relative to periods of time with average population, possibly because of higher bacterial die-off in the waters. The models can be an effective tool for management of human health risks in bathing and shellfish waters impacted by sewage pollution.

Survival and detection of Bacillus cereus in the presence of Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans after rechallenge in make-up removers.

Pathogenic contamination of cosmetics intended to be applied on or around the eye area, including make-up removers, may lead to severe eye infections. To assess the efficacy of antimicrobial preservatives in these products, we investigated the survival and detection of Bacillus cereus F 4227A spiked into make-up removers, alone and in the presence of other relevant micro-organisms. Four brands of make-up removers, A, B, C and D, were challenged three times (day 0, day 7 and day 14) using B. cereus, in pure and mixed cultures, at a final concentration of 5 log CFU per mL of Bacillus cereus or 6 log CFU per mL for other micro-organisms. Inoculated samples were diluted and spiral-plated after 30 min and 24 h of each challenge onto selective media for recovery of surviving micro-organisms: BACARA (B. cereus), MacConkey (E. coli), ChromID (P. aeruginosa), XLT4 (S. enteritidis), Baird Parker agar (Staph. aureus) and PDA+chlortetracycline HCL (C. albicans). The population of B. cereus spiked as a pure culture increased significantly from the first to the third challenge after 30-min exposure time, going from 0.73 to 2.59 in A, from 0.80 to 2.69 in B and from 0.80 to 1.67 log CFU per mL in C (P < 0.05). Likewise, the B. cereus population from the mixed cultures had a significantly higher survival count at the third challenge: from 0.12 log MPN per mL to 2.16 log CFU per mL in A, 0.57 to 2.27 log CFU per mL in B and from undetected (LOD = 0.48 log MPN) to 0.98 log CFU per mL in C, respectively. After challenges, Staph. aureus, C. albicans and P. aeruginosa increased in B; Staph. aureus and C. albicans in C; and E. coli and Staph. aureus in D. The growth of other bacteria types was unaffected by the number of challenges, but B. cereus population was detected with the third challenge. It is appropriate to assess the antimicrobial efficacy of preservatives using at least three challenges, especially for cosmetics that are subjected to repetitive contamination by users.

Dynamics of Salmonella Shedding and Welfare of Hens in Free-Range Egg Production Systems.

The current study investigated the effect of environmental stressors (i.e., weather changes) on Salmonella shedding in free-range production systems and the correlations with behavioral and physiological measures (i.e., fecal glucocorticoid metabolites). This involved longitudinal and point-in-time surveys of Salmonella shedding and environmental contamination on four commercial free-range layer farms. The shedding of Salmonella was variable across free-range farms and in different seasons. There was no significant effect of season on the Salmonella prevalence during this investigation. In this study, the combined Salmonella most probable number (MPN) counts in environmental (including feces, egg belt, dust, nest box, and ramp) samples were highest in samples collected during the summer season (4th sampling, performed in February). The predominant serovars isolated during this study were Salmonella enterica serovar Mbandaka and Salmonella enterica serovar Typhimurium phage types 135 and 135a. These two phage types were involved in several egg product-related Salmonella outbreaks in humans. Multilocus variable-number tandem-repeat analysis (MLVA) results indicated that MLVA types detected from human food poisoning cases exhibited MLVA patterns similar to the strains isolated during this study. All Salmonella isolates (n = 209) were tested for 15 different genes involved in adhesion, invasion, and survival of Salmonella spp. We also observed variations for sopA, ironA, and misL There were no positive correlations between fecal corticosterone metabolite (FCM) and Salmonella prevalence and/or shedding in feces. Also, there were no positive correlations between Salmonella prevalence and Salmonella count (log MPN) and any of the other welfare parameters.IMPORTANCE In this study, the welfare of laying hens and Salmonella shedding were compared over a prolonged period of time in field conditions. This study investigated the long-term shedding of Salmonella serovars in a free-range egg production system. Given that there is increasing demand for free-range eggs, it is essential to understand the risks associated with such a production system.

Microbiological quality of lettuce (Lactuca sativa) irrigated with wastewater in Addis Ababa, Ethiopia and effect of green salads washing methods

BackgroundIn Addis Ababa, where irrigation water for vegetable production is commonly derived from the highly polluted Akaki river, information on microbial contamination of water and irrigated vegetable is scanty. An assessment was done to determine the microbiological quality of irrigation water and lettuce harvested from 10 urban farming sites of Addis Ababa. The efficacy of 5 lettuce washing methods were also assessed. A total of 210 lettuce and 90 irrigation water samples were analyzed for faecal coliform and helminth eggs population levels.ResultsThe mean faecal coliform levels of irrigation water ranged from 4.29-5.61 log10 MPN 100 ml−1, while on lettuce, the concentrations ranged from 3.46-5.03 log10 MPN 100 g−1. Helminth eggs and larvae were detected in 80% of irrigation water and 61% of lettuce samples. Numbers ranged from 0.9-3.1 eggs 1000 ml−1 and 0.8-3.7 eggs 100 g−1 wet weight for irrigation water and lettuce, respectively. The helminth eggs identified included those of Ascaris lumbricoides, Hookworm, Enterobius vermicularis, Trichuris trichiura, Taenia and Strongloyides larvae. Ascaris lumbricoides and Hookworm were most prevalent in both irrigation water and lettuce samples. Compared with the WHO recommendations and international standards, the faecal coliform and helminth eggs levels in irrigation water and lettuce samples exceeded the recommended levels. Irrespective of the tested washing methods, faecal coliform and helminth eggs levels were somehow reduced. Among the washing methods, potable tap water washing - rinsing (2 min) followed by dipping in 15 000 ppm vinegar solution for a minute supported the highest faecal coliorm reduction of 1.7 log10 units, whereas lowest reduction of 0.8 log10 units was achieved for the same procedure without vinegar.ConclusionCompared with international standards, both faecal coliform and helminth eggs levels exceeded recommended thresholds in water and lettuce, but still in a potential risk range which can be easily mitigated if farmers and households are aware of the potential risk. Aside preventing occupational exposure, potential risk reduction programs should target households which have so far no guidance on how best to wash vegetables. The result of the present study suggest that the vinegar based washing methods are able to reduce faecal coliform towards low level while the physical washing with running water may help to substantially decrease potential risk of helminth parasitic infections.

Influence of cutting and deboning operations on the microbiological quality and shelf life of buffalo meat

Considering the specific biochemical composition of buffalo (Bubalus bubalis) meat (high iron content, high biological value proteins and essential fatty acids, low amounts of fat and cholesterol), we evaluated the influence of cutting and deboning operations on the microbiological quality and shelf-life of vacuum-packed buffalo meat stored under refrigeration. On the processing day, samples were collected from carcass, deboning room surfaces and meat cuts. Samples from meat cuts were evaluated weekly for two months. On the processing day, higher counts of Pseudomonas spp. were observed in samples from meat cuts compared with the hindquarters and the processing surfaces. For thermotolerant coliform scores, the averages were −0.5log MPN·cm−2, −0.4log MPN·cm−2 and 0.9log MPN·g−1, respectively. Higher counts of Pseudomonas spp. and LAB in meat cuts were observed on the processing day and after the first week of storage, respectively, remaining constant during shelf life. Listeria grayi was identified in two samples of hindquarters and meat cuts during storage. Listeria innocua was identified in one meat cut. In conclusion, cutting and deboning operations influence the microbiological quality and shelf life of vacuum-packed buffalo meat stored under refrigeration.

Microbiological Profile and Incidence of Salmonella and Listeria monocytogenes on Hydroponic Bell Peppers and Greenhouse Cultivation Environment

The aim of this study was to generate information regarding the microbiological profile, including Salmonella and Listeria monocytogenes incidence, of hydroponically grown bell peppers and materials associated with their production in greenhouses located in Mexico. Samples of coconut fiber (24), knives (30), drippers (20), conveyor belts (161), pepper transportation wagons (30), air (178), water (16), nutrient solution for plant irrigation (78), and bell pepper fruits (528) were collected during one cycle of production (2009 to 2010) for the quantification of microbial indicators (aerobic plate counts [APC], molds, coliforms, and Escherichia coli) and the detection of Salmonella and L. monocytogenes. With regard to surfaces (conveyor belts and wagons) and utensils (knives and drippers), the APC, coliform, and mold counts ranged from 3.0 to 6.0, from 1.4 to 6.3, and from 3.6 to 5.2 log CFU/100 cm(2) or per utensil, respectively. The air in the greenhouse contained low median levels of APC (1.2 to 1.4 log CFU/100 liters) and molds (2.2 to 2.5 log CFU/100 liters). The median content of APC and coliforms in water were 0.5 log CFU/ml and 0.3 log MPN/100 ml, respectively. The median content of coliforms in nutrient solution ranged from 1.8 to 2.4 log MPN/100 ml, and E. coli was detected in 18 samples (range, <0.3 to 1.2 log MPN/100 ml). On bell pepper analyzed during the study, populations (median) of APC, coliforms, and molds were 5.4, 3.6, and 5.8 log CFU per fruit, respectively; E. coli was detected in 5.1% of the samples (range, 0.23 to 1.4 log MPN per fruit). Salmonella was isolated from only one sample (1.6%) of conveyor belt located at the packing area and in four bell pepper samples (3%). L. monocytogenes was not detected. This information could help producers to establish effective control measures to prevent the presence of foodborne pathogens in bell peppers based on a scientific approach.