Shiga toxin-producing Escherichia coli (STEC) are zoonotic enteric pathogens associated with human gastroenteritis worldwide. Cattle and small ruminants are important animal reservoirs of STEC. The present study investigated animal reservoirs for STEC in small rural farms in the Culiacan Valley, an important agricultural region located in Northwest Mexico. A total of 240 fecal samples from domestic animals were collected from five sampling sites in the Culiacan Valley and were subjected to an enrichment protocol followed by either direct plating or immunomagnetic separation before plating on selective media. Serotype O157:H7 isolates with the virulence genes stx2, eae, and ehxA were identified in 40% (26/65) of the recovered isolates from cattle, sheep and chicken feces. Pulse-field gel electrophoresis (PFGE) analysis grouped most O157:H7 isolates into two clusters with 98.6% homology. The use of multiple-locus variable-number tandem repeat analysis (MLVA) differentiated isolates that were indistinguishable by PFGE. Analysis of the allelic diversity of MLVA loci suggested that the O157:H7 isolates from this region were highly related. In contrast to O157:H7 isolates, a greater genotypic diversity was observed in the non-O157 isolates, resulting in 23 PFGE types and 14 MLVA types. The relevant non-O157 serotypes O8:H19, O75:H8, O111:H8 and O146:H21 represented 35.4% (23/65) of the recovered isolates. In particular, 18.5% (12/65) of all the isolates were serotype O75:H8, which was the most variable serotype by both PFGE and MLVA. The non-O157 isolates were predominantly recovered from sheep and were identified to harbor either one or two stx genes. Most non-O157 isolates were ehxA-positive (86.5%, 32/37) but only 10.8% (4/37) harbored eae. These findings indicate that zoonotic STEC with genotypes associated with human illness are present in animals on small farms within rural communities in the Culiacan Valley and emphasize the need for the development of control measures to decrease risks associated with zoonotic STEC.

Escherichia coli O157:H7 has become a global public health and a food safety problem. Despite the implementation of control strategies that guarantee the safety in various products, outbreaks persist and new alternatives are necessary to reduce this pathogen along the food chain. Recently, our group isolated and characterised lytic bacteriophages against E. coli O157:H7 with potential to be used as biocontrol agents in food. To this end, phages need certain requirements to allow their manufacture and application. The aim of this study was to determine the physical stability and allergenic potential of free and microencapsulated (ME) bacteriophage cocktails against E. coli O157:H7. In vitro and in vivo studies were performed to determine phage survival under different pH, gastrointestinal conditions, temperature and UV light intensities. Results showed that the stability of ME phages was significantly (P<0.05) higher than free phages after ultraviolet irradiation, pH conditions between 3 to 7, and exposure to temperatures between at -80°C and 70°C. Both formulations were highly sensitive to very low pH in simulated gastric fluid, but stable in bile salts. In vivo studies in mice confirmed these phages passed through the gastrointestinal tract and were excreted in faeces. In silico, full-length alignment analysis showed that all phage proteins were negative for allergenic potential, but different predicting criteria classified seven phage proteins with a very low probability to be an allergen. In conclusion, these data demonstrated that microencapsulation provided a greater stability to phage formulation under stress conditions and assure a more suitable commercial formulation for the biological control of E. coli O157:H7.

Aims:  To identify and quantify the presence of Escherichia coli, Staphylococcus aureus, Salmonella, hepatitis A and norovirus in households and to assess the effect of chlorine and quaternary ammonium–based disinfectants following a prescribed use. Methods and Results:  Eleven sites distributed in kitchen, bathroom, pet and children′s areas of two groups of 30 homes each: (i) a nonprescribed disinfectant user group and (ii) a disinfectant protocol user group. During the 6‐week study, samples were collected once a week except for week one when sample collection occurred immediately before and after disinfectant application to evaluate the disinfectant protocol. The concentration and occurrence of bacteria were less in the households with prescribed use of disinfectants. The greatest reductions were for E. coli (99%) and Staph. aureus (99·9999%), respectively. Only two samples were positive for HAV, while norovirus was absent. Disinfection protocols resulted in a significant (P < 0·05) microbial reduction in all areas of the homes tested compared to homes not using a prescribed protocol. Conclusions:  The study suggests that disinfectant product application under specific protocol is necessary to achieve greater microbial reductions. Significance and Impact of the Study:  Prescribed protocols constitute an important tool to reduce the occurrence of potential disease‐causing micro‐organisms in households.

Four phages isolated from cattle and poultry feces were analyzed for their ability to lyse Salmonella serotypes and Escherichia coli O157:H7. The phage one-step growth curves, morphology, and genetic characteristics were determined. All phages showed a lytic effect on various Salmonella serotypes and E. coli O157:H7, which lysed at least 70% of the 234 strains tested. The phages had latent periods ranging from 10 to 15 min and generation times of 30 to 45 min, while burst size fluctuated between 154 and 426 PFU/cell. Phages morphology showed isometric and elongated heads and rigid contractile tails, consistent with morphology of the Myoviridae family. Phages' DNA dendrograms showed a distinctive RFLP when digested by HindIII and EcoRV, and SDS-PAGE profile showed distinctive proteins expression as well. In vitro phage challenge showed a total reduction of E. coli O157:H7, Salmonella Typhimurium and Saintpaul counts at 2 h, whereas for Salmonella Montevideo a reduction and retardation growth, at a multiplicity of infection (MOI) of 100, was observed; however, under a MOI of 10 000, no viable cells were detected after 4 h. The wide host ranges of these phages suggested they could be used for simultaneous biocontrol of some Salmonella serotypes and E. coli O157:H7.

The potential health effects of Salmonella found in wastewater residuals is dependent on the exposure of individuals to the organism. This paper provides a risk assessment for human infection from Salmonella due to direct contact with Class B biosolids, and from contact with Class A biosolids following regrowth of Salmonella. In addition, a risk assessment is provided for infection via airborne transport of bioaerosols from Class B biosolids and biosolids in which regrowth had occurred, to off-site communities. Results of the risk characterization imply that the risk of human infection from direct contact with Class B land applied residuals and subsequent ingestion is low. In contrast, the risk from direct contact with Class A residuals following regrowth is greater. Risks from airborne transport of Salmonella via bioaerosols away from a Class B land application site are also low. However, once again the risk from aerosols resulting from biosolids in which regrowth had occurred was greater. Based on these analyses, we conclude that it is highly unlikely that Salmonella infections will occur from land applied Class A or B residuals. However, risks become significant if Class A biosolids are stored anaerobically i.e. saturated, prior to land application.

Irrigation water can serve as a vehicle for transporting pathogenic microorganisms, and numerous cases of bacterial infections from consumption of irrigated fresh produce have been reported in recent years. Chlorine-based disinfectants applied when produce is packed are widely used to control microorganisms. When applied properly, the chlorine products are effective. However, hazardous disinfection breakdown products can be formed, and chlorine disinfectants have high oxidant activity that can affect produce quality and pose a risk to food handlers. Quaternary Ammonium Compounds (QACs) are a disinfectant alternative for the washing of fruits and vegetables. They can control a great number of microorganisms, have low toxicity when used at recommended doses, and are stable in storage. The purpose of this work was to assess the disinfectant activity of QACs against Escherichia coli and Staphylococcus aureus under worst-case and average-case turbidity conditions, (2 and 100 nephelometric units); two disinfectant concentrations (100 and 200 mg/L; and two contact times (30 and 120 seconds). Our research showed that QACs were effective against both bacteria. The percentage reduction of Escherichia coli was significantly higher in the less turbid solution (P = 0.027), while turbidity did not affect the reduction of Staphylococcus aureus (P > 0.05). E. coli was more resistant to QAC treatment than S. aureus. Based on the data obtained we can conclude that QACs could be an alternative in washing processes of fruits and vegetables.

The purpose of this study was to compare chlorinated and ozonated water in reducing Salmonella typhimurium inoculated onto fresh ripe tomatoes. Surface-inoculated tomatoes were immersed/sprayed with chlorinated (200 mg l(-1)) and ozonated water (1 and 2 mg l(-1)) under 2 and 100 nefelometric turbidity units (NTU). Contact times were 120 and 30 s for immersing and spraying applications, respectively. Immersing in chlorinated water and low turbidity resulted in the most effective application with 3.61 log(10) bacterial reduction, while 1 and 2 mg l(-1) of ozone reduced 2.32 and 2.53 log(10), respectively. High turbidity and chlorine reduced the bacterial counts by 3.39 log(10), while 1 and 2 mg l(-1) of ozonated water and low turbidity reduced the bacteria by 1.48 and 1.92 log(10), respectively. Spraying chlorinated water reduced bacteria by 3 log(10), and ozonated water at 1 and 2 mg l(-1) reduced counts by 1.84 and 2.40 log(10), respectively. No statistical differences were found between chlorine and ozone (2 mg l(-1)) during spraying applications (p < 0.05). The use of ozonated water both in immersing and spraying applications is suggested when water turbidity remains low.

The implementation of good agricultural practices (GAP) from irrigation water to the tomato packaging process enhances the safety of fresh produce and its value throughout the food chain. The aim of the present study was to show that fresh produce farms that apply and enforce GAP could reduce the presence of Salmonella in finished produce. Samples were collected biweekly from six packing houses from the central region of Sinaloa, México, for the isolation of Salmonella spp by the ISO 6579:2002 method, and the isolated strains were serotyped and genotyped by the Kauffmman-White scheme and pulsed field gel electrophoresis (PFGE), respectively. Salmonella strains were detected in 13 (36.1 %) irrigation water samples, while only two tomato samples were positive (5.5 %). Eight different serotypes were identified in irrigation water, and Salmonella Oranienburg (34 %) was the most prevalent; however, only Salmonella Agona and Salmonella Weltevreden were present on tomatoes. Salmonella Oranienburg was the most widely dispersed and variable serotype, with 10 different PFGE profiles. Salmonella Weltevreden was isolated from both types of samples, albeit with distinct genetic profiles, implying that the sources of contamination differ. These results confirm the utility of implementing good agricultural practices to reduce Salmonella contamination in irrigation water and the packaging process.