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The survival of Salmonella in subtropical river water depends on genetic and metabolic reorganization for the expression of alternative metabolic pathways in response to starvation, which allows Sa...

Acute hepatopancreatic necrosis disease (AHPND) is a severe disease affecting recently stocked cultured shrimps. The disease is mainly caused by V. parahaemolyticus that harbors the pVA1 plasmid; this plasmid contains the pirA and pirB genes, which encode a delta-endotoxin. AHPND originated in China in 2009 and has since spread to several other Asian countries and recently to Latin America (2013). Many Asian strains have been sequenced, and their sequences are publicly accessible in scientific databases, but only four strains from Latin America have been reported. In this study, we analyzed nine pVA1-harboring V. parahaemolyticus sequences from strains isolated in Mexico along with the 38 previously available pVA1-harboring V. parahaemolyticus sequences and the reference strain RIMD 2210633. The studied sequences were clustered into three phylogenetic clades (Latin American, Malaysian, and Cosmopolitan) through pangenomic and phylogenomic analysis. The nucleotide sequence alignment of the pVA1 plasmids harbored by the Asian and Latin American strains confirmed that the main structural difference in the plasmid between the Asian and Latin American strains is the absence of the Tn3 transposon in the Asian strains; in addition, some deletions in the pirAB region were found in two of the Latin American strains. Our study represents the most robust and inclusive phylogenomic analysis of pVA1-harboring V. parahaemolyticus conducted to date and provides insight into the epidemiology of AHPND. In addition, this study highlights that disease diagnosis through the detection of the pirA and pirB genes is an inadequate approach due to the instability of these genes.

Acute hepatopancreatic necrosis disease (AHPND) is a life-threatening disease to recently stocked shrimp. This disease is mainly caused by Vibrio parahaemolyticus and, to date, it has not been effectively controlled. Bacteriophages are a promising method to control bacterial diseases in aquaculture and multiple phages that infect Asian strains of V. parahaemolyticus have been described. However, few studies have characterized the bacteriophages that infect Latin American strains. Here, two lytic Vibrio phages (vB_VpaP_AL-1 and vB_VpaS_AL-2) were isolated from estuary water in Sinaloa, Mexico. The host ranges were tested using ten AHPND-causing strains isolated from Mexico and phage AL-1 was able to infect two strains while AL-2 infected four. One-step growth curve showed that AL-1 produced 85 PFU/cell and AL-2 produced 68 PFU/cell in 30 and 40 min, respectively. Both phages were able to tolerate temperatures ranging from 20 to 50 °C and pH values ranging from 4 to 10. Phages AL-1 and AL-2 have double-stranded DNA genomes of 42,854 bp and 58,457 bp, respectively. In total, 53 putative ORFs associated with the phage structure, packing, host lysis, DNA metabolism, and additional functions were predicted in the AL-1 genome, while 92 ORFs associated with the same functions as the AL-1 and 1 tRNA were predicted in the AL-2 genome. The lifecycle was classified as virulent for both phages. Morphology, phylogeny, and comparative genomic analyses assigned phage AL-1 as a new member of the genus Maculvirus in the Autographiviridae family, and phage AL-2 as a new member of the Siphoviridae family. These findings suggest that vB_VpaP_AL-1 and vB_VpaS_AL-2 are potential biocontrol agents against AHPND-causing V. parahaemolyticus from Mexico.

The survival of

Salmonella in the environment have evolved genetically to maintain a stable cell metabolism. Nevertheless, a lack of common nutrients (such as glucose) causes these strains to metabolize alternative carbon sources. In this study, 21 strains of Salmonella Oranienburg isolated from subtropical river water were evaluated to compare their adaptation and preconditioning abilities for the consumption of environmental carbon sources (ECS). The results obtained in this study attributed important biological characteristics to the adaptation of the metabolism of Salmonella strains to diverse ECS; these characteristics include but are not limited to variations in plasticity and natural preconditioning in closely related microorganisms, such as environmental isolates belonging to the serotype Oranienburg.

Foodborne illnesses, mainly bacteria, are a major cause of morbidity and mortality worldwide. Pathogenic bacteria are involved in almost every step within the fresh produce chain compromising the companies' food safety programs and generating an ascending number of foodborne outbreaks in various regions of the world. Recently, bacteriophages return to the status of biocontrol agents. These bacteria-killing viruses are able to reduce or eliminate pathogenic bacterial load from raw and ready to eat foods. Phages are efficient, strain specific, easy to isolate and manipulate, and for that reasons, they have been used in pre and post harvest processes alone or mixed with antimicrobial agents for biocontrolling pathogenic bacteria. In this review, we focused on the feasibility of using lytic bacteriophage on fresh fruits and vegetables industry, considering challenges and perspectives mainly at industrial production level (packinghouses, supermarkets), where high volume of phage preparations and consequently high costs may be required.

In this study, packinghouses spraying disinfection process was simulated to compare the effectiveness of aqueous chlorine dioxide and sodium hypochlorite in reducing Escherichia coli O157:H7 on bell pepper surface. Bell peppers inoculated with 7.74 Log10 CFU of E. coli O157:H7 were placed in a rotary mixer (60 rpm), and sprayed with chlorine dioxide and sodium hypochlorite. Chlorine dioxide at 5 mg L-1 during 20-30 s reduces from 7.74 Log10 CFU of E. coli O157:H7 adhered on bell pepper to undetectable numbers, while the exposition with chlorine dioxide at pH 6 and 8 during 10 s was necessary to reduce 4.25 and 3.90 Log10 CFU, respectively. On the other hand, sodium hypochlorite at 200 mg L-1 at pH 6 reduced 2.89, 3.25 and 3.54 Log10 CFU/bell pepper at 10, 20 and 30 s, respectively. Spraying disinfection process using aqueous chlorine dioxide was more effective than sodium hypochlorite applications, therefor chlorine dioxide is an effective alternative to reduce E. coli O157:H7 on bell pepper surface.

Potato peels (PP) contain several bioactive compounds. These compounds are known to provide human health benefits, including antioxidant and antimicrobial properties. In addition, these compounds could have effects on human enteric viruses that have not yet been reported. The objective of the present study was to evaluate the phenolic composition, antioxidant properties in the acidified ethanol extract (AEE) and water extract of PP, and the antiviral effects on the inhibition of Av-05 and MS2 bacteriophages, which were used as human enteric viral surrogates. The AEE showed the highest phenolic content and antioxidant activity. Chlorogenic and caffeic acids were the major phenolic acids. In vitro analysis indicated that PP had a strong antioxidant activity. A 3 h incubation with AEE at a concentration of 5 mg/ml was needed to reduce the PFU/ml (plaque-forming unit per unit volume) of Av-05 and MS2 by 2.8 and 3.9 log₁₀, respectively, in a dose-dependent manner. Our data suggest that PP has potential to be a source of natural antioxidants against enteric viruses.

The aim of this research was to evaluate the efficacy of bacteriophage Av-08 for reducing Salmonella Montevideo and Escherichia coli O157:H7 presence from chicken skin surface at 4C. Significant reduction (P

Con el fin de cuantificar a Escherichia coli y determinar la presencia de Salmonella, se tomaron y analizaron 51 muestras de agua y 23 muestras de suelo en cuatro regiones del Valle de Culiacán, Sinaloa, México. Además, se determinó el perfil de resistencia de Salmonella a ampicilina, ciprofloxacino, trimetoprim-sulfametoxazol, tetraciclina, estreptomicina y gentamicina, y de los últimos tres antimicrobianos en E. coli. En las muestras de suelo analizadas hubo ausencia de E. coli y Salmonella, mientras que en las de agua se logró aislar 20 cepas de Salmonella. La serotipificación mostró que 13 cepas de Salmonella fueron Typhimurium, 2 de Infantis, 1 de Anatum, 1 de Agona, 1 de Oranienburg, 1 de Minnesota y 1 de Give. De las muestras de agua analizadas 98 % estaban contaminadas con E. coli, con mínimos de 4 UFC/100 mL, máximos de 4.5 x 105 UFC/100 mL y un promedio de 1.6 x 104 UFC/100 mL, de las cuales se seleccionaron cepas para evaluar el perfil de resistencia a antimicrobianos. Todas las cepas de Salmonella fueron susceptibles a ampicilina, ciprofloxacino y trimetoprim-sulfametoxazol y 12 presentaron resistencia a tetraciclina. De las 46 cepas de E. coli analizadas, nueve fueron resistentes a tetraciclina, 38 fueron resistentes a estreptomicina y sólo una cepa fue resistente a gentamicina; mientras que 23 cepas presentaron resistencia intermedia. Todas las cepas de Salmonella tuvieron altos niveles de resistencia a CuSO4•5H2O en dosis entre 1200 y 1600 µg mL-1 del antimicrobiano. El presente estudio demostró la existencia de diversos serotipos de Salmonella y altos niveles de E. coli en agua del Valle de Culiacán.

Lytic bacteriophages have reemerged as an alternative for the control of pathogenic bacteria. However, the effective use of phage relies on appropriate genomic characterization. In this study, we report the genome of bacteriophage Av-05 and its sequence analysis, which has strong lytic activity against Escherichia coli O157:H7 strains and several Salmonella serotypes. The analysis revealed that the phage Av-05 genome consists of 120,938 bp, containing 209 putative open reading frames (ORFs) and 9 tRNAs.

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.