Quantification of Salmonella transfer in cross-contamination scenarios found in chicken slaughterhouses.

Chicken are among the main reservoirs of Salmonella, and slaughterhouses have been identified as key sites for cross-contamination of this pathogen. This study aimed to quantify the transfer rate of Salmonella in different cross-contamination scenarios found in chicken slaughterhouses. To this end, a pool of Salmonella spp. Was inoculated onto chicken carcasses and thighs, reaching out concentrations of 2-5 log10 CFU/g. After inoculation, carcasses and thighs were used to reproduce four cross-contamination scenarios based on industrial reality as follows: 1. Transfer of Salmonella from chicken carcasses to stainless steel and polyethylene surfaces; 2. Transfer of Salmonella between hanging chicken carcasses; 3. Transfer of Salmonella from stainless steel surfaces to chicken carcasses, and 4. Transfer of Salmonella from thighs to stainless steel and polyethylene surfaces. The results showed that the transfer rates (TR) of Salmonella on the chicken carcass to stainless steel and polyethylene were 25.77 ± 22.63% and 24.71 ± 13.93%, respectively, while the TR between hanged chicken carcasses was 5.11 ± 1.71%. When sliding carcasses through a stainless steel ramp, 41.47 ± 1.32% of the Salmonella present on the ramp adhered to the chicken carcasses, and the greater transfer seems to be linked to the wet surfaces. The transfer rates from the thighs to the stainless steel and polyethylene were 1.81 ± 0.66% and 9.0 ± 1.34%, respectively. Cross-contamination occurred regardless of the sample weight, time of contact, and amount of inoculum.

Phenotypic and genotypic characterization of antibiotic resistance of Salmonella Heidelberg in the south of Brazil.

Salmonella is the main human pathogen present in the poultry chain. Salmonella Heidelberg is one of the most important serovars for public health since it has been frequently isolated in broiler chickens from different countries and may present multidrug resistance (MDR). This study was carried out with 130 S. Heidelberg isolates collected from pre-slaughter broiler farms in 2019 and 2020 in 18 cities from three Brazilian states to study relevant aspects regarding their genotypic and phenotypic resistance. The isolates were tested and identified using somatic and flagellar antiserum (0:4, H:2, and H:r), and an antimicrobial susceptibility test (AST) was performed against 11 antibiotics for veterinary use. The strains were typed by Enterobacterial Repetitive Intergenic Consensus (ERIC)-PCR, and representatives of the main clusters of the identified profiles were sequenced by Whole Genome Sequencing (WGS). AST results showed that all isolates were resistant to sulfonamide, 54 % (70/130) were resistant to amoxicillin, and only one was sensitive to tetracycline. Twelve isolates (15.4 %) were MDR. The dendrogram obtained from the ERIC-PCR showed that the strains were grouped into 27 clusters with similarity above 90 %, with some isolates showing 100 % similarity but with different phenotypic profiles of antimicrobial resistance. Identical strains collected on the same farm on other dates were identified, indicating that they were residents. WGS identified 66 antibiotic-resistance genes. The sul2 (present in all sequenced samples) and tet(A) genes were highlighted and validated in the experimental analysis. The fosA7 gene was also identified in all sequenced samples, but resistance was not observed in the phenotypic test, possibly due to the heteroresistance of the S. Heidelberg strains evaluated. Considering that chicken meat is one of the most consumed meats in the world, the data obtained in the present study can corroborate the mapping of the origin and trends of antimicrobial resistance.

Assessing the Listeria monocytogenes transference during mechanical slicing of mozzarella cheese.

Listeriosis is a foodborne disease caused by Listeria monocytogenes (LM) and has been linked to the consumption of sliced mozzarella cheese. This study aimed to assess the LM transference during mechanical slicing of mozzarella cheese and its growth during refrigerated storage. Mozzarella cheese was contaminated with LM and 100 slices containing approximately 5 log CFU/slice were produced. Next, 100 slices of non-contaminated cheese were sliced using a contaminated blade (3.67log CFU/10 cm2). LM was quantified on the blade and slices right after slicing and after storage at 10 °C for 10 and 15 days. Results demonstrated that increasing counts of LM were transferred to the blade, comparing the first and the fifth slices (2.71 and 3.22log CFU/10 cm2, respectively, p ≤ 0.05), however, transference stabilized after the 50th slice (3.75 CFU/10 cm2). The blade transferred 1.69-2.66 log CFU/g of LM to different slices. At the end of the storage at 10 °C by 10 and 15 days, LM counts increased to 1.51 and 1.69 log CFU/g, respectively, indicating that LM population can increase if the cheese is stored for a long time.

First Fatal Cases due to Escherichia coli O157 and Campylobacter jejuni subsp. jejuni Outbreak Occurred in Southern Brazil.

In December 2019, four children of the same school were hospitalized due to severe diarrhea, vomiting, and fever, and the mother of one child was diagnosed with hemolytic uremic syndrome (HUS). Escherichia coli O157 NM was isolated from the stool of one child, whereas Campylobacter jejuni isolates were found in feces, raw foods, environmental samples, and tap water. In addition, the same pulsed-field gel electrophoresis profile was identified in C. jejuni isolated from feces and tap water. One child died of renal failure and another due to respiratory failure. Based on symptoms and bacterial isolation, the deaths were assigned to E. coli O157 NM, but coinfection with C. jejuni may have contributed to the severity of symptoms. These were the first deaths assigned to E. coli O157 NM registered in Brazil.

Bactericidal effect of marinades on meats against different pathogens: a review.

Marinades are seasoned liquids used to improve tenderness, palatability, flavor, color and/or texture of different meats. In addition to contribute to the sensory characteristics, marinates can inactivate food microorganism as well. The purpose of this study was to assess the current state of knowledge regarding the effect of marinades on meats and important food pathogens. Using a systematic review of literature, different types of marinades were evaluated, identifying its ingredients, concentrations, temperature, marinating time and their effect on Salmonella, Escherichia coli, Listeria monocytogenes, Campylobacter and Vibrio. Findings demonstrated that the use of marinades on meats not only prevents the growth of pathogens but also inactivates food pathogens. Most marinades were able to reduce < 3 log CFU/g of pathogens, and Vibrio populations demonstrated the highest reductions (> 4 log CFU/g). The pH was the most pronounced parameter influencing the pathogens inactivation, however, ingredients and storage temperature also affected pathogen reduction in marinades.

Evaluation of PhageDX Salmonella Assay for Salmonella Detection in Hydroponic Curly Lettuce.

Lettuce is one of the most consumed leafy vegetables worldwide and has been involved in multiple foodborne outbreaks. Salmonella is one of the most prevalent etiological agents of foodborne disease (FBD) in lettuces, and its detection may take several days depending on the chosen method. This study evaluates a new rapid method that uses recombinant bacteriophages to detect Salmonella in hydroponic curly lettuce. First, the ability of the assay to detect six Salmonella serovars at three different concentrations (1, 10, and 100 CFU/well) was tested. Second, the detection of Salmonella was tested in lettuces using a cocktail of the same Salmonella serovars and concentrations after a 7 h enrichment. The results of these experiments showed that the detection limit was dependent on the serovar tested. Most serovars were detected in only 2 h when the concentration was 100 CFU/well. Salmonella was detected in 9 h (7 h enrichment + 2 h bioluminescence assay) in all lettuce samples with 10 CFU/25 g or more. Salmonella detection was not influenced by natural microbiota of lettuces. This study demonstrated that the phage assay was sensitive and faster than other detection methods, indicating that it is a better alternative for Salmonella detection on lettuces.

Modelling the growth of Staphylococcus aureus on cooked broccoli under isothermal conditions.

In this study, we developed predictive models describing the growth of Staphylococcus aureus on cooked broccoli florets. A pool of 3.5 log CFU/g of five S. aureus strains were inoculated on 10 g broccoli portions. The samples were then stored at 10, 20, 30 and 37 °C, and colonies were enumerated at different time intervals. Baranyi and Roberts model was fitted to the data using a Bayesian Adaptive Markov Chain Monte Carlo for estimation of the growth parameters. S. aureus showed low growth at 10 °C on broccoli samples and at 20-37 °C interval, Baranyi and Roberts model fitted well to the experimental data (R2>0.97). Estimated growth parameters were correlated with the possibility of toxin production and indicate the potential presence of these biological hazards on contaminated broccoli after heat treatment. Additionally, linear regression was performed for growth rate as storage temperature function. This secondary model followed a linear tendency with R2=0.997 and was compared with two tertiary models (ComBase Predictor and Pathogen Modeling Program) and literature data, demonstrating similar growth rate values of both. These results can be helpful for food services and managers to establish food safety standards for S. aureus growth on cooked broccoli.

Mathematical modelling and validation of Salmonella enterica growth in sushi exposed to different time-temperature scenarios found in real sushi establishments.

In this study mathematical models to predict Salmonella enterica growth in sushi at different temperatures were developed considering data obtained in 26 restaurants in Southern Brazil. The sushi type chosen to develop the models was the one that presented the highest total aerobic mesophilic counts among sushis collected in the establishments. Salmonella was inoculated (2-3 log UFC/g) in this sushi type prepared in the laboratory (pH 4.8; aw 0.98) and incubated under isothermal conditions at 7, 15, 20, 25 and 37 °C. Baranyi and Roberts model was used to describe Salmonella growth curves, generating R2 values of ≥0.98 and RMSE values of <0.24 log CFU/g/h for primary models. Ratkowsky's equation was used in secondary model, generating R2 of 0.99 and RMSE of 0.02 log CFU/g/h. The model validation was simulated under non-isothermal conditions, using the worst-case scenario that was built through data from the environmental conditions and data obtained from the restaurants. The non-isothermal conditions were performed at 36.3 °C for 6 h, 10 °C for 24 h and 29.5 °C for 6 h sequentially, reaching 6.7 log CFU/g of Salmonella and generating RMSE of 0.06 log CFU/g/h, Bias factor of 0.97 and Accuracy factor of 1.03. The negligible growth time (ς) for Salmonella, considering the average of higher distribution temperatures of chosen sushi type (approximately 18 °C), was 8.9 h. However, growth rates of total aerobic mesophilic demonstrated that at 15 °C and 20 °C, the lag phases were approximately 11 and 5 h respectively. Based on these results, we suggest for sushi distribution the use of temperatures of ≤15 °C for 6 h (maximum time of distribution allowed in Brazil) considering the Salmonella growth.

Food safety behavior and handling practices during purchase, preparation, storage and consumption of chicken meat and eggs.

The aim of this study was to assess the risk of occurrence of a foodborne outbreak and point practices determinant to achieve high Good Hygienic Practice level during handling practice from purchase to consumption of chicken meat and eggs. The risk behavior of respondents and the risk of the occurrence of a foodborne outbreak were measured using Weighted Harmonic Outbreak Prevention Index (WHOPI). WHOPI were not correlated to socioeconomic data and perception of risks. Different profiles of handling practices were identified inside each WHOPI level. Chicken meat defrost, time and temperature of egg cooking and the point of yolk were identified as the most important procedures responsible for the WHOPI level upgrades. The consumption of chicken meat and eggs were characterized as discrete distributions. The average consumption of chicken meat was 113.48 g/per day and eggs daily intake distribution was 0.92 units/day. Our results can be applied for future microbiological food safety risk assessments related to the consumption of chicken meat and eggs.

Salmonella spp. and Escherichia coli O157:H7 prevalence and levels on lettuce: A systematic review and meta-analysis.

Lettuce (Lactuca sativa), one of the most consumed leafy vegetables in the world, is frequently implicated with foodborne disease (FBD) outbreaks, with Salmonella spp. and Escherichia coli O157:H7 being the most common bacteria to cause this illness. Estimates of prevalence and levels of these pathogens on lettuce are scarce in developed or in developing countries, which hinders risk assessment attempts. In here, we present a systematic review and meta-analysis of reported prevalence and levels of Salmonella spp. and E. coli O157:H7 on lettuce using the worldwide available data. Literature was reviewed and examined the results for inclusion of articles in the meta-analysis. Data (prevalence and/or concentration of Salmonella spp. and E. coli O157:H7 on lettuce, sample characteristic, country of origin, and Salmonella identified serovars) were extracted, and meta-analysis was performed using Open Meta-Analyst, Task Order # 2 software. Although only one work reported the presence of E coli O157:H7 on lettuce, several reports indicated the presence of other, distinct enterohemorrhagic E. coli (EHEC) strains, with a mean prevalence of 0.041 (95% CI: 0.005-0.078) and concentration varying from <3.0 MPN/g to >1100 MPN/g. Furthermore, the mean prevalence of Salmonella spp. on lettuce was 0.041 (95% CI: 0.030-0.052), with reported concentrations varying between 0.054 ±â€¯0.058 CFU/g to 218.78 MPN/g. In addition, subgroup analysis of the presence of Salmonella spp. in lettuce revealed a mean prevalence of the bacteria of 0.028 (95% CI: 0.014-0.042) in developed nations and 0.064 (0.041-0.087) in developing nations, with reports varying from 0.001 in Japan to 0.5 in Burkina Faso. Despite a relatively low prevalence, consumption of lettuce is inherently risky because it usually is eaten raw, without thermal treatment to inactivate pathogens. This potential risk further supports performance of quantitative risk assessments to quantify the probability of FBD caused by Salmonella spp. and E. coli O157:H7 transmitted to lettuce.

Detection of virulence genes in Salmonella Heidelberg isolated from chicken carcasses.

During the last years, Brazilian government control programs have detected an increase of Salmonella Heidelberg in poultry slaughterhouses a condition that poses a threat to human health However, the reasons remain unclear. Differences in genetic virulence profiles may be a possible justification. In addition, effective control of Salmonella is related to an efficient epidemiological surveillance system through genotyping techniques. In this context, the aim of this study was the detection of 24 virulence-associated genes in 126 S. Heidelberg isolates. We classified the isolates into 56 different genetic profiles. None of the isolates presented all the virulence genes. The prevalence of these genes was high in all tested samples as the lowest number of genes detected in one isolate was 10/24. The lpfA and csgA (fimbriae), invA and sivH (TTSS), and msgA and tolC (intracellular survival) genes were present in 100% of the isolates analyzed. Genes encoding effector proteins were detected in the majority of SH isolates. No single isolate had the sefA gene. The pefA gene was found in only four isolates. We have also performed a screening of genes associated with iron metabolism: 88.9% of isolates had the iroN geneand 79.4% the sitC gene . Although all the isolates belong to the same serotype, several genotypic profiles were observed. These findings suggest that there is a diversity of S. Heidelberg isolates in poultry products. The fact that a single predominant profile was not found in this study indicates the presence of variable sources of contamination caused by SH. The detection of genetic profiles of Salmonella strains can be used to determine the virulence patterns of SH isolates.

Genome analysis reveals insights into high-resistance and virulence of Salmonella Enteritidis involved in foodborne outbreaks.

Salmonella enterica serovar Enteritidis strain SE86 has been associated with several foodborne diseases occurring in Southern Brazil, becoming and important causative agent of human salmonellosis. In this work, the complete genome of the bacterium Salmonella Enteritidis SE86 was sequenced using the Illumina MiSeq platform. An in silico analysis of the SE86 genome was performed in order to compare it with different Salmonella strains as well as to investigate the presence of stress-resistance and virulence genes. This strain showed a variety of genes that can be involved in antimicrobial and biocide resistance, acid and thermal resistance as well as virulence and adhesion. These genetic features could explain its increased resistance and the prevalence of this strain in foodborne outbreaks in Southern Brazil.

Distribuição de probabilidade da temperatura na cadeia de carne de frango brasileira: fonte para avaliação de risco / Probability distribution of temperatures in the Brazilian chicken meat chain: source of risk assessment

O presente estudo tem como objetivo descrever a distribuição de probabilidades da temperatura de cada etapa da cadeia produtiva de carne de frango resfriada produzida sob inspeção federal no Brasil. Os resultados gerados geram subsídio para realização de uma Análise de Risco de salmonelose pelo consumo de carne de frango no país. Foram compilados registros de temperatura e os dados foram ajustados às distribuições de probabilidade utilizando o software @Risk. Nas etapas de abatedouro-frigorífico, centro de distribuição, serviço de alimentação e supermercado a média de temperatura foi inferior a 5,5 ºC, demostrando sucesso na manutenção da cadeia de frio. A distribuição de probabilidade Logistic demonstrou melhor ajuste para descrever as etapas de processamento. Os dados de distribuição de probabilidade gerados poderão ser utilizados em modelos matemáticos e de distribuição para predizer o crescimento microbiano em alimentos expostos a diferentes temperaturas.

Avaliação do comportamento de risco de manipuladores de alimentos e hábito de consumo de carne de frango no Brasil / Evaluation of risk behavior of food manipulators and consumption habit of chicken meat in Brazil

O manipulador de alimentos possui grande impacto na ocorrência de doenças transmitidas por alimentos devido às más práticas de higiene e manipulação de alimentos. Assim, com o objetivo de avaliar os hábitos de consumo e caracterizar a manipulação de carne de frango no Brasil foi desenvolvido e aplicado um questionário. Foram obtidas e analisadas 1217 respostas. A maior parte dos entrevistados foi classificada com nível médio de boas práticas, independentemente de sua condição socioeconômica (Tukey-Test, p> 0,005). Em relação ao nível de conformidade dos parâmetros avaliados, observou-se que os respondentes cumpriam primeiramente conformidade em questões relacionadas a temperatura, seguido de binômio tempo-temperatura, tempo e, por fim, boas práticas. Os resultados desse estudo servem como suporte no emprego de ações de saúde coletiva, além de poderem ser aplicadas em ferramentas robustas de gestão da segurança de alimentos, como a Análise de Risco.

Assessment of Salmonella spp. and Escherichia coli O157:H7 growth on lettuce exposed to isothermal and non-isothermal conditions.

This study aimed to assess the growth of Salmonella and Escherichia coli O157:H7 on lettuce exposed to isothermal and non-isothermal conditions. Pathogens were inoculated on lettuce separately and stored under isothermal condition at 5 °C, 10 °C, 25 °C, 37 °C for both bacteria, at 40 °C for Salmonella and 42 °C for E. coli O157:H7. Growth curves were built by fitting the data to the Baranyi's DMFit, generating R2 values greater than 0.92 for primary models. Secondary models were fitted with Ratkowsky equations, generating R2 values higher than 0.91 and RMSE lower than 0.1. Experimental data showed that both bacteria could grow at all temperatures. Also, the growth of both pathogens under non-isothermal conditions was studied simulating temperatures found from harvest to supermarkets in Brazil. Models were analysed by R2, RMSE, bias factor (Bf) and accuracy factor (Af). Salmonella and E. coli O157:H7 were able to grow in this temperature profile and the models could predict the behavior of these microorganisms on lettuce under isothermal and non-isothermal conditions. Based on the results, a negligible growth time (ς) was proposed to provide the time which lettuce could be exposed to a specific temperature and do not present an expressive growth of bacteria. The ς was developed based on Baranyi's primary model equation and on growth potential concept. ς is the value of lag phase added of the time necessary to population grow 0.5 log CFU/g. The ς of lettuce exposed to 37 °C was 1.3 h, while at 5 °C was 3.3 days.

Prevalence of Salmonella spp. in poultry carcasses samples collected in slaughterhouses of Southern Brazil from 2006 to 2015.

INTRODUCTION: This study aimed to evaluate the prevalence of Salmonella on poultry carcasses produced in slaughterhouses of Southern Brazil participating of the Official Pathogen Reduction Program conducted by the Ministry of Agriculture, Livestock, and Supply. METHODOLOGY: From 2006 to 2015, 77,165 poultry carcasses were analyzed for presence/absence of Salmonella spp. and the results were statistically evaluated. RESULTS: Prevalence varied from 2.92% to 5.24%, with a mean percentage of 4.04%. The difference in prevalence numbers was not significant during all the period analyzed. Higher Salmonella prevalence has been reported worldwide, indicating the efficacy of Brazilian control measures implemented in the productive chain and the low risk associated to Brazilian poultry meat consumption. However, additional information about the acceptable and safe prevalence of Salmonella on poultry should be defined by risk analysis studies, considering the reality of Brazilian companies and scientific data. CONCLUSIONS: The results of the present study can be the first step for a national Risk Assessment and may contribute to improvements in self-controlling programs and with the current Brazilian poultry regulation.

Phenotypic and Molecular Characterization of Salmonella Enteritidis SE86 Isolated from Poultry and Salmonellosis Outbreaks.

Salmonella Enteritidis remains a standout among the leading causes of foodborne diseases worldwide. Previous studies have demonstrated that a unique clonal group of Salmonella Enteritidis, named SE86, is involved in foodborne outbreaks in southern Brazil and is frequently identified among strains isolated from poultry. The aim of this study was to determine the influence of the isolation source (food products involved in salmonellosis outbreaks and poultry sources) on the phenotypic and molecular characteristics of Salmonella Enteritidis SE86. A biofilm formation assay, antimicrobial susceptibility test, polymerase chain reaction identification of virulence-associated genes, and phage type 4 (PT4) assessment were performed to characterize Salmonella Enteritidis SE86. The human strains presented less antimicrobial resistance than the poultry strains. Resistance to some substances was related to the isolation source of the strain. Strains of the same clonal group presented different biofilm production abilities. Biofilm formation was independent of the isolation source at all temperatures. Temperature influenced biofilm formation only by the poultry strains. Most of the investigated genes presented a high frequency and a regular distribution, regardless of the isolation source. The spvB, spiA, pagC, sipB, prgH, spaN, sitC, and lpfC genes were associated with the avian strains, whereas iroN was associated with the strains isolated from food products involved in salmonellosis outbreaks. Most strains belonged to PT4. No relationship was found between biofilm production and antimicrobial resistance or between the virulence profile and biofilm production or antimicrobial resistance.

First Report of Human Gastroenteritis Caused by Escherichia coli O157:NM in Brazil.

In September 2005, the Sanitary Surveillance Service of Rio de Janeiro (SSS/RJ), Brazil, investigated a case of gastroenteritis involving a 13-year-old teenager hospitalized because of bloody diarrhea and severe abdominal pain. Owing to the severity of the symptoms, an epidemiological investigation was conducted in two states of Brazil. Escherichia coli O157:NM was isolated from stools and from a tomato and cheese salad prepared at the school canteen where the teenager attended. This is the first report of a human case of gastroenteritis related to E. coli O157:NM infection in Brazil.

Antimicrobial activity of free and liposome-encapsulated thymol and carvacrol against Salmonella and Staphylococcus aureus adhered to stainless steel.

Antimicrobial activity of thymol, carvacrol and thymol/carvacrol liposomes (TCL) was evaluated against two bacterial pools, each one consisting of four strains of Staphylococcus aureus or Salmonella enterica. TCL were prepared using thin-film hydration, showing 270.20nm average diameter (polydispersity index of 0.33) and zeta potential of +39.99mV. Minimum inhibitory concentration (MIC) of thymol, carvacrol and TCL against S. aureus pool was 0.662mg/ml, while MIC for Salmonella pool was 0.331mg/ml for thymol and carvacrol, and for TCL was 0.662mg/ml. Bacterial pools (8.0logCFU/ml), allowed in contact on stainless steel AISI 304 coupons in UHT skim milk for 15min, resulted in adhered populations of 5.6-6.1logCFU/cm2. Adhered S. aureus (±6.1logCFU/cm2) were inhibited after 1-min and 10-min treatments using thymol or carvacrol at MIC and 2.0 MIC. Reductions of 1.47-1.76logCFU/cm2 and 1.87-2.04logCFU/cm2 were obtained using 0.5 MIC of thymol and carvacrol, respectively. A 10-min contact with free (MIC and 2.0 MIC) and encapsulated (MIC) antimicrobials inhibited attached Salmonella (±6.0logCFU/cm2); however, after 1-min of contact, 2.0 MIC of thymol and carvacrol were not able to inactivate adhered Salmonella MIC of TCL inactivated S. aureus and Salmonella after 10min; however, after 1-min contact, adhered S. aureus and Salmonella populations were decreased in 1.62logCFU/cm2 and 2.01logCFU/cm2, respectively. Considering antimicrobial concentrations and contact times, thymol, carvacrol, and TCL could be employed in food-contact surfaces to prevent biofilm formation at early stages of bacterial attachment. Further investigations should be performed considering long-term antibacterial effects of TCL.

Microbial quality of irrigation water used in leafy green production in Southern Brazil and its relationship with produce safety.

Irrigation water has been recognized as an important microbial risk factor for fruits and vegetables in many production areas, but there is still a lack of information about how the microbiological quality of different irrigation water sources and climatic conditions influence the safety of vegetables produced in Brazil. This study evaluated the distribution of generic E. coli and the prevalence of E. coli O157:H7 in two different water sources (ponds and streams bordering farmlands and urban areas) used for irrigation and on commercially produced lettuces in Southern Brazil. We also evaluated the effect of agricultural factors and meteorological conditions in the potential contamination of water and produce samples. A longitudinal study was conducted on four farms during a year (July 2014 to August 2015). The results showed generic E. coli prevalence of 84.8% and 38.3% in irrigation water samples and on lettuces, respectively, indicating irrigation water as an important source of contamination of lettuces. No significant differences were detected in the counts of E. coli between the two different surface water sources. The climatic conditions, particularly rainfall and environmental temperature, have influenced the high concentration of E. coli. The highest loads of E. coli in irrigation water and on lettuces were found during the warmest time of the year. E. coli O157:H7 was detected by qualitative polymerase chain reaction (qPCR) in 13 water samples but only 4 were confirmed by isolation in culture media.