Screening, biochemical characterization and antibiotics resistance/susceptibility of bacteria isolated from native soil and water samples / Triagem, caracterização bioquímica e resistência/suscetibilidade a antibióticos de bactérias isoladas de amostras nativas de solo e água

The present study was conducted to isolate and characterize bacteria from water and soil sample taken from the Lahore Canal at different sites i.e. Mall Road, Mohlanwal and Khera site. Isolated bacterial strains were identified on the basis of morphological and biochemical tests. Identification was confirmed by culturing bacteria on selective media. Antibiotic resistance test was also performed to observe the resistance of bacteria against different antibiotics. Blood agar test was performed for identification of different pathogenic bacteria. The result revealed that water and soil samples of Lahore Canal Lahore from different sites were contaminated with Escherichia coli, Salmonella sp., Vibrio sp., Bacillus spp., Enterococcus sp. and Staphylococcus spp. Due to presence of these pathogens, this water is not suitable for any domestic and irrigation use. Study also revealed that water of the Lahore Canal is harmful for human health as it is contaminated with bacteria that can cause severe disease e.g., Escherichia coli can cause gastroenteritis, Bacillus spp. can cause nausea and vomiting, Enterococcus may infect urinary tract, Salmonella sp. is responsible for Bacteremia, Staphylococcus spp. can cause mild fever and Vibrio sp. can be the reason of cholera. Thus it is rendered unfit for any kind of human use even other than drinking like swimming, bathing, washing etc., until and unless some remedial measures are employed to eradicate pathogenic microorganisms by WASA and LWMS according to standards of WHO. Similarly, it is quite harmful, when and where ever it is used for irrigation without proper treatment.

O presente estudo foi realizado para isolar e caracterizar bactérias de amostras de água e solo retiradas do Canal Lahore, em Lahore, em diferentes locais, ou seja, Mall Road, Mohlanwal e Khera. As cepas bacterianas isoladas foram identificadas com base em testes morfológicos e bioquímicos. A identificação foi confirmada por cultura de bactérias em testes de meios seletivos. O teste de resistência aos antibióticos também foi realizado para observar a resistência das bactérias a diferentes antibióticos. Foi realizado o teste de ágar sangue para identificar diferentes bactérias patogênicas. O resultado revelou que amostras de água e solo do Canal Lahore, Lahore, de diferentes localidades estavam contaminadas com Escherichia coli, Salmonella sp., Vibrio sp., Bacillus spp., Enterococcus sp. e Staphylococcus spp. Por causa da presença desses patógenos, essa água não é adequada para qualquer uso doméstico e de irrigação. O estudo revelou que a água do Canal Lahore é prejudicial à saúde humana, pois está contaminada com bactérias que podem causar doenças graves, por exemplo: Escherichia coli pode ocasionar gastroenterite; Bacillus spp. pode causar náuseas e vômitos; Enterococcus sp. pode infectar o trato urinário; Salmonella sp. é responsável pela bacteremia; Staphylococcus spp. pode causar febre leve; e Vibrio sp. pode ser a razão da cólera. Assim, torna-se imprópria para uso humano, como natação, banho, lavagem etc., até que algumas medidas corretivas sejam empregadas para erradicar microrganismos patogênicos por WASA e LWMS de acordo com os padrões da OMS. Da mesma forma, é bastante prejudicial, quando usada para irrigação sem tratamento adequado.

Vibrio sp. ArtGut-C1, a polyhydroxybutyrate producer isolated from the gut of the aquaculture live diet Artemia (Crustacea)

BACKGROUND: Vibrio species display variable and plastic fitness strategies to survive and interact with multiple hosts, including marine aquaculture species that are severely affected by pathogenic Vibrios. The culturable Vibrio sp. strain ArtGut-C1, the focus of this study, provides new evidence of such phenotypic plasticity as it accumulates polyhydroxybutyrate (PHB), a biodegradable polymer with anti-pathogen activity, particularly in the marine larviculture phase. The strain was isolated from the gut of laboratory-reared Artemia individuals, the live diet and PHB carrier used in larviculture. Its main phenotypic properties, taxonomic status and genomic properties are reported based on the whole-genome sequencing. RESULTS: Vibrio sp. ArtGut-C1 yielded 72.6% PHB of cells' dry weight at 25 C. The genomic average nucleotide identity (ANI) shows it is closely related to V. diabolicus (ANI: 88.6%). Its genome contains 5,236,997- bp with 44.8% GC content, 3,710 protein-coding sequences, 96 RNA, 9 PHB genes functionally related to PHB metabolic pathways, and several genes linked to competing and colonizing abilities. CONCLUSIONS: This culturable PHB-accumulating Vibrio strain shows high genomic and phenotypic variability. It may be used as a natural pathogen biocontrol in the marine hatchery and as a potential cell factory for PHB production.

Miocarditis por gram negativos / Gram-Negative Myocarditis

Resumen Objetivo: describir las características epidemiológicas, clínicas y microbiológicas de los pacientes que cursan con miocarditis por Enterobacterias. Métodos: se realizó una revisión sistemática de la literatura, en la que se incluyeron Pubmed, Ovid, Scopus, SciELO y LILACS sin exclusión por tipo de idioma. La población objetivo de estudio fueron los pacientes con diagnóstico de infección bacteriana por bacilo gram negativo mediante cultivo, técnicas moleculares o histopatología, y quienes presentaban biopsia de miocardio o, en su defecto, resonancia magnética cardiaca con hallazgos sugestivos de miocarditis. Resultados: se encontraron 742 artículos, de los cuales se incluyeron 24; en estos se reportaron 27 pacientes. La edad promedio fue de 31 años. El 81% de los pacientes eran de sexo masculino. El síntoma principal fue diarrea (80%), seguido de fiebre (53%) y dolor torácico (38%). El 37% de los pacientes fallecieron. El hallazgo más común en el electrocardiograma fue la elevación del segmento ST (36,7%). En quienes se realizó ecocardiograma se encontraron anormalidades en 50% de los casos, siendo más frecuente la disminución en la fracción de eyección. El microorganismo más común fue el Campylobacter jejuni, seguido por Salmonela sp. Conclusiones: la miocarditis causada por enterobacterias es más frecuente en pacientes adultos jóvenes de sexo masculino. Los síntomas gastrointestinales suelen estar presentes al momento de la presentación clínica. El diagnóstico requiere de alta sospecha clínica teniendo en cuenta que las anormalidades eléctricas y en ecocardiograma no se encuentran en todos los pacientes.

Abstract Objective: To describe the epidemiological, clinical, and microbiological characteristics of patients with myocarditis due to Enterobacteria. Methods: A systematic review was carried out on the literature, which included Pubmed, Ovid, Scopus, SciELO, and LILACS, with no exclusions due to language. The target population of the study were patients with a diagnosis of bacterial infection due to gram negative bacillus by means of a culture, or using molecular or histopathology technique. They also had to have had a myocardial biopsy or, if not, a cardiac magnetic resonance scan with findings suggestive of myocarditis. Results: Out of a total of 742 articles found, 24 of these, in which 27 patients were described, were included. The mean age was 31 years, and 81% were male. The main symptom was diarrhoea (80%), followed by fever (53%), and chest pain (38%). More than one-third (37%) of the patients died. The most common finding on the electrocardiogram (ECG) was elevation of the ST segment (36.7%). Abnormalities were found in 50% of the cases, on whom a cardiac ultrasound was performed, with a decrease in the ejection fraction being the most common. The most common microorganism was Campylobacter jejuni, followed by Salmonella spp. Conclusions: Myocarditis caused by enterobacteria is most common in young male patients. The gastrointestinal symptoms are usually present from the clinical onset. The diagnosis requires a high clinical suspicion, taking into account that the abnormalities in the ECG and cardiac ultrasound are not found in all patients.

Actividad antitumoral de biopolímeros aislados de Vibrio sp. en cáncer de mama inducido en ratas / Antitumor activity of vibrio sp. isolated biopolymers in induced breast cancer in rats

RESUMEN Objetivo: Evaluar la actividad antitumoral del extracto crudo de biopolímeros aislados de la bacteria marina Vibrio sp. en cáncer de mama inducido por N-Methyl-N-nitrosourea (MNU) en ratas. Materiales y métodos: Se cultivó la bacteria marina Vibrio sp. durante siete días, luego se filtró, precipitó y concentró el sobrenadante crudo. Se administró una dosis única de MNU 50 mg/kg a 39 ratas Holtzman y fueron tratadas diariamente durante nueve semanas por vía oral: G1 (n=13): suero fisiológico 0,1 mL/100g; G2 (n=13): extracto crudo de biopolímeros de Vibrio sp. 20 mg/kg; G3 (n=13): tamoxifeno 100 mg/kg. El G4 (n=11) solo recibió suero fisiológico 0,1 mL/100g. Se valoró semanalmente el peso corporal y la aparición de tumores mamarios identificados mediante palpación; así como el examen histopatológico al final del tratamiento. Resultados: El 77% de las ratas del grupo G1 desarrollaron tumores a partir de la séptima semana en un promedio de 2,2 tumores por cada animal; en contraste al grupo tratado con el extracto crudo de biopolímeros y tamoxifeno; donde solo una rata (8%) en cada grupo desarrolló tumores y posterior a la semana nueve de la inducción (p=0,001). Los resultados histopatológicos sostienen que todos los tumores extirpados corresponden a adenocarcinoma ductal de mama con distintos patrones: sólido, papilar y quístico. Asimismo, se evidenciaron focos necróticos en el 30% de los tumores del grupo G1. Conclusión: El extracto crudo de biopolímeros aislados de Vibrio sp. presentan efecto antitumoral en cáncer de mama inducido en ratas.

ABSTRACT Objective: To evaluate the antitumor activity of the raw extract from biopolymers isolated from the Vibrio sp. marine bacteria in breast cancer induced by N-Methyl-N-nitrosourea (MNU) in rats. Materials and methods: The Vibrio sp. marine bacteria was cultured for seven days, then the raw supernatant was filtered, precipitated and concentrated. MNU was administered in a single dose of 50 mg/kg to 39 Holtzman rats and were daily treated for 9 weeks orally: G1 (n = 13): 0.1 mL/100 g of saline solution; G2 (n = 13): 20 mg/kg of raw extract from Vibrio sp. biopolymers; G3 (n = 13): 100 mg/kg of tamoxifen; G4 (n = 11) received no MNU and only 0.1 mL/100 g of saline solution. Body weight and the appearance of breast tumors identified by palpation were assessed weekly, as well as histopathological examination at the end of treatment. Results: Seventy-seven percent of the rats in the G1 group developed tumors from week 7 onwards in an average of 2.2 tumors per animal; in contrast to the group treated with the raw biopolymer extract and tamoxifen; where only one rat (8%) in each group developed tumors after week nine of induction (p = 0.001). The histopathological results support that all the removed tumors correspond to breast ductal adenocarcinoma with different patterns: solid, papillary and cystic. Likewise, necrotic foci were evidenced in 30% of the tumors of the G1 group. Conclusion: The raw extract of biopolymers isolated from Vibrio sp. present antitumor effect in breast cancer induced in rats.

Diagnóstico de la calidad físico química y biológica del agua en el Golfo de Fonseca, La Unión, El Salvador. Propuestas para el desarrollo sostenible: en asocio con el Centro de Educación e Instrucción Naval de El Salvador, CEIN-Base Naval de La Unión / Diagnosis of the physical, chemical and biological quality of water in the Gulf of Fonseca, La Unión, El Salvador. Proposals for sustainable development: in association with the Naval Education and Training Center of El Salvador, CEIN-La Unión Naval Base

Este estudio permitió conocer la calidad microbiológica del agua del Golfo de Fonseca, a través de un programa de monitoreo realizado durante 12 meses, en 9 puntos de muestreo distribuidos en 3 sectores. "A" estero La Manzanilla-Isla Perico, "B" Ciudad de La Unión ­ bocana río Goascorán y "C" estero El Tamarindo ­ Isla Meanguera. El propósito fue obtener registros de contaminación de aguas costeras para el país y el impacto que las aguas residuales producen en el ambiente costero y la pérdida de la calidad del agua para actividades tales como la maricultura y la recreación. Entre las bacterias encontradas se encuentran: Enterococcus, Pseudomonas, Vibrios y Heterótrofas. Dichas bacterias están asociadas con diferentes enfermedades para la salud humana, tal es el caso de las pseudomonas, las cuales están frecuentemente asociadas a infecciones de los tractos respiratorio y urinario. Los resultados obtenidos de Vibrio indican que el agua no es apta para uso en acuicultura, ni para uso recreativo, el agua se encuentra en el límite de referencia en cuanto al uso con propósito de producción acuícola y límite de referencia en cuanto al riesgo de consumo de productos pesqueros.

This study allowed to know the microbiological quality of the water of Golfo de Fonseca, through a monitoring program carried out during 12 months, in 9 sampling points distributed in 3 sectors. "A" La Manzanilla estuary- Isla Perico, "B" City of La Unión - Río Goascorán mouth and "C" El Tamarindo estuary - Isla Meanguera. The purpose was to obtain records of coastal water pollution for the country and the impact that wastewater produces on the coastal environment and the loss of water quality for activities such as mariculture and recreation. Among the bacteria found are: Enterococcus, Pseudomonas, Vibrios and Heterotrophs. These bacteria are associated with different diseases for human health, such is the case of pseudomonas, which are frequently associated with infections of the respiratory and urinary tracts. The results obtained from Vibrio indicate that the water is not suitable for use in aquaculture, nor for recreational use, the water is at the reference limit in terms of use for the purpose of aquaculture production and the reference limit regarding the risk of consumption. of fishery products.

Aislamiento y caracterización de una bacteria marina degradadora de testosterona: Vibrio sp. N3 / Isolation and characterization of a marine testosterone-degrading bacterium: Vibrio sp. N3

Steroids, including testosterone, estrone, 17β-estradiol, estriol and 17β-ethinyl estradiol, are harmful not only to the population dynamics of aquatic life forms but also to public health. In this study, a marine testosterone-degrading bacterium (strain N3) was isolated from Nanao Island in the South China Sea. In addition, the strain could also use 17β-estradiol (E2), 17β-ethinyl estradiol (EE2), estriol (E3) or cholesterol as a sole carbon source. According to the 16S rRNA gene sequence analysis, strain N3 was identified as Vibrio sp. Further characterization showed that the strain is aerobic, gram-negative, and mobile and exhibits resistance to ampicillin, carbenicillin, penicillin and spectinomycin. For enhancing its capacity of testosterone degradation, the Plackett-Burman factorial design and the central composite design were used to optimize the culture condition. Under optimal conditions, 92% of testosterone was degraded by Vibrio sp. N3 in 48 h.

Los esferoides-que incluyen la testosterona, la estrona, el 17 β-estradiol, el estriol y el 17 p-etinilestradiol-son nocivos no solo para la población dinámica de las formas de vida acuática, sino también para la salud pública. En este estudio se aisló una bacteria marina degradadora de testosterona de la isla de Nanao, en el Mar del Sur de China, a la que se denominó cepa N3. Se determinó que esta cepa también podría usar 17 β-estradiol (E2), 17 p-etinilestradiol (EE2), estriol (E3) o colesterol como únicas fuentes de carbono. De acuerdo con el análisis de la secuencia del gen 16S rRNA, la cepa N3 se identificó como Vibrio sp. La caracterización adicional mostró que dicha bacteria es un organismo aerobio, gram negativo y móvil, y que presenta resistencia a ampicilina, carbenicilina, penicilina y espectinomicina. Para optimizar la condición de cultivo en relación con su capacidad de degradar la testosterona, se utilizaron el diseño factorial Plackett-Burman y el diseno compuesto central. En condiciones óptimas, el 92% de la testosterona fue degradada por Vibrio sp. N3 en 48 h.

Bacteriophages in the control of pathogenic vibrios

Vibrios are common inhabitants of marine and estuarine environments. Some of them can be pathogenic to humans and/or marine animals using a broad repertory of virulence factors. Lately, several reports have indicated that the incidence of Vibrio infections in humans is rising and also in animals constitute a continuing threat for aquaculture. Moreover, the continuous use of antibiotics has been accompanied by an emergence of antibiotic resistance in Vibrio species, implying a necessity for efficient treatments. One promising alternative that emerges is the use of lytic bacteriophages; however, there are some drawbacks that should be overcome to make phage therapy a widely accepted method. In this work, we discuss about the major pathogenic Vibrio species and the progress, benefits and disadvantages that have been detected during the experimental use of bacteriophages to their control.

Incidencia de parásitos y bacterias del genero vibrio en el cultivo de camarón marino desarrollados en cooperativas camaroneras del municipio de Jiquilisco, Departamento de Usulután / Incidence of parasites and bacteria of the vibrio genus in the marine shrimp culture developed in shrimp cooperatives of the municipality of Jiquilisco, Department of Usulután

En El Salvador el 59.3% del camarón marino de la especie Litopenaeus vannamei se produce en la Bahía de Jiquilisco, departamento de Usulután, el cultivo tiene una duración de 60 a 90 días y el peso de cosecha oscila entre 7 y 12 gramos. El bajo crecimiento y las altas mortalidades son problemas que preocupan a este sector productivo. Por esta razón la Escuela Especializada en Ingeniería ITCA-FEPADE a través de la carrera de Gestión Integral de Recursos Marino Costeros, desarrolló una investigación aplicada orientada a identificar la existencia de parásitos y bacterias del género Vibrio en los cultivos de camarón marino que se desarrollan en la zona de Salinas del Potrero y El Zompopero, así como evaluar su incidencia en el cultivo. Con la investigación se detectó la presencia de cinco géneros de parásitos que están presentes en branquias, intestinos y urópodos en el cultivo de camarón marino en grado 1 y 2; ambas categorías se consideran las etapas iniciales de propagación de los parásitos. También se evidenció a través del análisis bacteriológico en el medio de cultivo TCBS, la presencia de bacterias del genero Vibrio en agua, sedimento y camarón.

In El Salvador, 59.3% of the marine shrimp of the Litopenaeus vannamei species is produced in the Bay of Jiquilisco, department of Usulután, the culture lasts from 60 to 90 days and the harvest weight ranges between 7 and 12 grams. Low growth and high mortality are problems that concern this productive sector. For this reason, Escuela Espcializada en Ingeniería ITCA-FEPADE , through the major of Gestión Integral de Recursos Marino Costeros, developed an applied research aimed at identifying the existence of parasites and bacteria of the genus Vibrio in marine shrimp cultures that develop in the area of ​​Salinas del Potrero and El Zompopero, as well as evaluating their impact on the crop. The investigation detected the presence of five types of parasites that are present in gills, intestines and uropods in marine shrimp culture in grade 1 and 2; Both categories are considered the initial stages of parasite spread. The presence of Vibrio bacteria in water, sediment and shrimp is also evidenced through bacteriological analysis in the TCBS culture medium.

Pathogenic Vibrio species isolated from estuarine environments (Ceará, Brazil) - antimicrobial resistance and virulence potential profiles

ABSTRACT Detection of virulent strains associated with aquatic environment is a current concern for the management and control of human and animal health. Thus, Vibrio diversity was investigated in four estuaries from state of Ceará (Pacoti, Choró, Pirangi and Jaguaribe) followed by antimicrobial susceptibility to different antimicrobials used in aquaculture and detection of main virulence factors to human health. Isolation and identification were performed on TCBS agar (selective medium) and dichotomous key based on biochemical characteristics, respectively. Nineteen strains of genus Vibrio were catalogued. Vibrio parahaemolyticus (Choró River) and V. alginolyticus (Pacoti River) were the most abundant species in the four estuaries. All strains were submitted to disk diffusion technique (15 antimicrobials were tested). Resistance was found to: penicillin (82%), ampicillin (54%), cephalotin (7%), aztreonan (1%), gentamicin, cefotaxime and ceftriaxone (0.5%). Five pathogenic strains were chosen to verification of virulence factors. Four estuaries showed a high abundance of species. High number of tested positive strains for virulence is concerning, since some of those strains are associated to human diseases, while others are known pathogens of aquatic organisms.

Diagnóstico e incidencia de enfermedades bacterianas y parasitarias que afectan el cultivo de camarón marino en estanques del sector El Zompopero y Salinas del Potrero, municipio de Jiquilísco, departamento de Usulután: en vínculo con Cooperativas de San Hilario / Diagnosis and incidence of bacterial and parasitic diseases that affect the cultivation of marine shrimp in ponds of El Zompopero and Salinas del Potrero sectors, municipality of Jiquilísco, department of Usulután: in connection with cooperatives of San Hilario

Las enfermedades bacterianas y parasitarias pueden llegar a comprometer los niveles de sobrevivencia de un cultivo de camarón marino. La frecuencia de aparición de camarones muertos en las orillas de los estanques o que aparecen en las atarrayas, son solo un signo o señal de que algo marcha mal en el estanque. Cuando los camarones están sometidos a factores estresantes se reducen su actividad limpiadora, no mudan y por tanto, son altamente susceptibles a la invasión de ectoparásitos. En grados de severidad 3 y 4 de la infestación por ectoparásitos en la superficie corporal del camarón, trae como consecuencia dificultades en la locomoción, la alimentación, la respiración, y finalmente sobreviene la muerte. Por otra parte, las enfermedades de origen bacteriano reportados en los sistemas de cultivo causadas por el género Vibrio, ha llegado a ser la enfermedad económicamente más importante en el cultivo de organismos marinos, afectando un gran número de especies; es catalogada como la causa de serias pérdidas económicas en la producción de camarón de cultivo en diversos países.

Bacterial and parasitic diseases can compromise the survival levels of a marine shrimp culture. The frequency of the appearance of dead shrimp on the banks of the ponds or that appear on the cast nets, are just a sign that something is wrong in the pond. When shrimp are subjected to stress, their cleaning activity is reduced, they do not molt and therefore are highly susceptible to invasion by ectoparasites. In degrees of severity 3 and 4, the infestation by ectoparasites on the body surface of the shrimp, results in difficulties in locomotion, feeding, breathing, and finally, death ensues. On the other hand, the diseases of bacterial origin reported in culture systems caused by the Vibrio genus have become the most economically important disease in the cultivation of marine organisms, affecting a large number of species; It is classified as the cause of serious economic losses in the production of farmed shrimp in various countries.

Tipificación molecular de aislados de vibrio cholerae O1 causantes de dos brotes de cólera en Venezuela / Molecular typing of vibrio cholerae O1 isolates causing two outbreaks of cholera in Venezuela

En Venezuela, en junio de 1996, se reportó que los casos de cólera eran causados por V. cholerae O1 serotipo Ogawa. A finales de 1998 se detectó un segundo brote de cólera causado por V. cholerae O1 serotipo Inaba resistente a la ampicilina y el trimetoprim-sulfametoxazol. Para estudiar las relaciones entre las cepas se examinaron veinticinco aislados de Vibrio cholerae O1 obtenidos desde 1996 a 2000 en Venezuela, para determinar la presencia de genes de virulencia y perfiles genómicos. Mediante la reacción en cadena de la polimerasa se determinó la presencia de genes de virulencia. Para determinar el perfil genómico de los aislamientos se utilizó ribotipificación y electroforesis en gel de campo pulsado (PFGE). Todos los aislados resultaron positivos para los genes ctxA, ctxB, zot y ace. El análisis RFLP de los genes RNAr mostró un único patrón de ribotipo V. El análisis de PFGE mostró una similitud de 91,5% independientemente del año o lugar de aislamiento, lo que indica la relación genómica entre los aislados. En conjunto, los datos sugieren que la cepa de V. cholerae O1 resistente a los antibióticos que apareció en 1998 surgió de la cepa epidémica anterior o de otro estrechamente relacionado con el clon anterior, con cambio de serotipo y ganancia de determinantes de resistencia a antibióticos. Es muy importante monitorear continuamente la aparición de la variantes porque mejorará la comprensión de la evolución de nuevos clones de V. cholerae

In Venezuela, cholera reported in June 1996 was caused by V. cholerae O1 serotype Ogawa. Second outbreak of cholera caused by V. cholerae O1 serotype Inaba, resistant to ampicillin and trimethoprim- Sulfamethoxazole, was notify at the end of 1998. Twenty-five isolates of Vibrio cholerae O1 obtained from 1996 to 2000 in Venezuela were examined to study the relationships between strains, presence of virulence genes and genomic profiles. Presence of virulence genes was detected by Polymerase Chain Reaction. Ribotyping and pulsed-field gel electrophoresis (PFGE) were used to determine the genomic profile of isolates. All isolates shown PCR product for ctxA, ctxB, zot and ace genes. RFLP analysis of rRNA gene showed one unique pattern from ribotype V. PFGE analysis revealed a similarity of 91.5%, regardless year or place of isolation, suggesting genomic relatedness among them. Overall, these data suggest that antibiotic resistant V. cholerae O1 El Tor strain that appeared in 1998 emerged from the previous epidemic strain or from another closely related to the previous clone. It is important the continuous monitor the emergence of variants because it will improve our understanding of the evolution of new clones V. cholerae

Isolamento de Vibrio spp de camarões comercializados in natura na cidade de São Gonçalo - RJ / Isolation of Vibrio spp from shrimp commercialized in natura in the city of Sao Gonçalo - RJ

Bactérias do gênero Vibrio fazem parte da microbiota de camarões, pois têm capacidade de associar-se à quitina presente no exoesqueleto destes animais e ao zooplancton, que por sua vez são consumidos por estes animais. O gênero contém pelo menos 12 espécies patogênicas, incluindo V. cholerae, responsável por várias pandemias de cólera. A contaminação humana acontece através do consumo de alimentos, principalmente de origem marinha, crus ou mal cozidos. Por se tratar de um tipo de pescado amplamente consumido pela população, este trabalho teve como objetivo investigar a presença de espécies de Vibrio em camarões comercializados in natura na cidade de São Gonçalo-RJ. Os camarões foram adquiridos em duas peixarias da cidade e caracterizados por metodologia convencional e molecular; cento e vinte e nove amostras testaram positivamente para as provas bioquímicas realizadas e, destas, cinquenta e duas testaram positivamente para os testes moleculares. Visando investigar a identidade das espécies de Vibrio, as amostras foram submetidas ao PCR multiplex para 4 espécies (V. cholerae, V. mimicus, V. parahaemolyticus, V. vulnificus). Doze isolados foram identificados como V. parahaemolyticus e 9 como V.cholerae não O1. Dentre os demais isolados, 31 demonstram se tratar de outras espécies de Vibrio spp. O sítio com o maior número de isolados foi a casca, seguida pelo hepatopâncreas/ hemolinfa. A ribotipagem por PCR das 21 cepas demonstrou claramente a separação das cepas de V.parahaemolyticus e V.cholerae. As cepas de V. cholerae e V. parahaemolyticus demonstraram alto índice de resistência a ampicilina (83,33%) e 100% de sensibilidade à nitroflurantoína e tetraciclina. Sete cepas (38,8%) apresentaram perfil de multirresistência a dois antimicrobianos. Nossos resultados demonstram a presença de espécies patogênicas de Vibrio em amostras de pescados amplamente consumidos pela população.

Análisis microbiológicodel pulpo rojo en puertos pesqueros de Campeche, México / Microbiological analysis of red octopus in fishing ports of Campeche, Mexico

Resumen: Objetivo: Estudiar la calidad microbiológica del pulpo rojo dado su importante impacto económico y social en la región sur-sureste de México. Material y métodos: Se tomaron muestras en diversas zonas de captura de la especie y se analizaron con pruebas bioquímicas descritas en las normas oficiales mexicanas. Se identificaron cepas pertenecientes al género Vibrio, Salmonella, coliformes fecales y E. coli O157:H7. Con el empleo del Sistema BAx, se logró la identificación de microorganismos a través de su ADN bacteriano. Los resultados obtenidos en los métodos bioquímicos y moleculares fueron contrastados. Resultados: El método estadístico de Bland-Altman indicó que ambas técnicas pueden usarse indistintamente. La prueba de McNemar demostró que ambos métodos cuentan con la misma eficacia para la identificación de patógenos (valor X2=0.5 ρ=0.4795). Conclusión: La calidad microbiológica del pulpo en la región sur-sureste de México es deficiente debido a la presencia de flora bacteriana patógena que podría representar un riesgo epidemiológico. Los índices establecidos por las normas sugieren la necesidad de aplicar técnicas de identificación eficaces y rápidas como el Sistema BAx. Este método alternativo de análisis puede coadyuvar a la implementación de estrategias efectivas que permitan cumplir con especificaciones mínimas sanitarias durante el procesamiento de los productos pesqueros, y así elevar los sistemas de control para disminuir los riesgos de brotes epidemiológicos en la región.

Abstract: Objective: In this work we studied the microbiological quality of the red octopus given its important economic and social impact on the region South-Southeast of Mexico. Materials and methods: Samples were taken in different areas of capture of the species and analyzed with biochemical tests described in the Mexican official standards, identifying strains belonging to the genus Vibrio, Salmonella and faecal coliforms, and E. coli O157: H7. We used the BAx System for the identification of microorganisms through their bacterial DNA. The results obtained in biochemical and molecular methods were confirmed. Results: Bland-Altman statistical method pointed out that both techniques can be used interchangeably. McNemar test showed that both methods have the same efficacy for the identification of pathogens (value X2=0.5 ρ=0.4795). Conclusion: The microbiological quality of the octopus in the South-Southeast region of Mexico is deficient due to the presence of pathogenic intestinal flora that might represent an epidemiological risk. The indexes established by the regulations suggest the need to apply effective and rapid identification technologies, such as the BAx System.This alternative method of analysis can contribute to the implementation of effective strategies that allow compliance with the minimal sanitary specifications during the processing of fishing products, thus strengthening the control systems to decrease the risks of epidemiological outbreaks in the region.

Molecular variations in Vibrio alginolyticus and V. harveyi in shrimp-farming systems upon stress

A study was performed to investigate the genomic variations in the shrimp farm isolates of Vibrio alginolyticus and V. harveyi when the isolates were subjected to environmental stress. Samples of shrimps, water and sediment were collected from Southern Indian coastal shrimp farms. Vibrio isolates were biochemically identified and confirmed using 16S rDNA and gyrB gene specific PCR. The bacterial strains were genotyped by PCR fingerprinting using GTG(5) and IS (Insertion Sequence) primers. Seven strains each of V. alginolyticus and V. harveyi were subjected to 10 passages through trypticase soya broth (TSB), which contained different NaCl concentrations (3, 6 and 8%) and trypticase soya agar (TSA). V. alginolyticus was also passaged through TSB with a 12% NaCl concentration. PCR fingerprinting, which was performed on the strains that were passaged through different salt concentrations, confirmed that V. alginolyticus and V. harveyi could affect the genomic variations, depending on the environmental conditions of the culture. The study highlights the complex genotypic variations that occur in Vibrio strains of tropical aquatic environment because of varied environmental conditions, which result in genetic divergence and/or probable convergence. Such genetic divergence and/or convergence can lead to the organismal adaptive variation, which results in their ability to cause a productive infection in aquatic organisms or generation of new strains.

Evaluación del riesgo microbiológico a Vibrio spp. en alimentos de origen marino en México / Microbial risk assessment of Vibrio spp. in seafood products in Mexico

Las enfermedades transmitidas por los alimentos son uno de los mayores problemas de salud pública que actualmente existen. La evaluación del riesgo microbiológico es un proceso utilizado para examinar los peligros ocultos en los alimentos, la probabilidad de exposición a éstos y su impacto en la salud pública. La evaluación del riesgo se realiza en cuatro fases: identificación del peligro, caracterización del peligro, evaluación de la exposición y caracterización del riesgo. De acuerdo con el proceso/resultado, las evaluaciones de riesgo microbiológico se clasifican en dos categorías: cualitativa y cuantitativa. La presente revisión pretende enmarcar la importancia de implementar estas evaluaciones en alimentos de origen marino que son consumidos crudos, fortaleciendo así el acceso a los alimentos inocuos y de buena calidad para beneficio del consumidor, y la necesidad de evaluaciones de riesgo microbiológico que hay en México.

Food-borne diseases are among the major public health problems that currently exist. Microbiological risk assessment is a process used to evaluate the hidden hazards in food, the likelihood of exposure to these hazards and their impact on public health. Risk assessment is performed in four steps: hazard identification, hazard characterization, assessment of exposure and risk characterization. According to the process/response microbial risk assessment is classified in two categories, qualitative and quantitative. The aim of this review is to underline the importance of implementing assessments in seafood that is usually consumed raw, strengthening access to good quality and safe food for the consumer's benefit and to stress the necessity of microbiological risks assessments in Mexico.

Occurrence of potentially pathogenic Vibrio in oysters (Crassostrea gigas) and waters from bivalve mollusk cultivations in the South Bay of Santa Catarina

Introduction This research aimed to identify and quantify potentially pathogenic Vibrio from different cultivations of bivalve shellfish in the State of Santa Catarina, Brazil, and water regions in the South Bay, as well as correlate the incidence of these microorganisms with the physicochemical parameters of marine waters. Methods Between October 2008 and March 2009, 60 oyster and seawater samples were collected from six regions of bivalve mollusk cultivation, and these samples were submitted for Vibrio counts. Results Twenty-nine (48.3%) oyster samples were revealed to be contaminated with one or more Vibrio species. The Vibrio parahaemolyticus and Vibrio vulnificus counts in the samples ranged from < 0.5 log10 Most Probable Number (MPN) g–1 to 2.3 log10 MPN g–1 oyster and from < 0.5 log10 MPN g–1 to 2.1 log10 MPN g–1 oyster, respectively. Of the 60 seawater samples analyzed, 44 (73.3%) showed signs of contamination with one or more vibrio species. The counts of V. parahaemolyticus and V. vulnificus in the samples ranged from < 0.3 log10 MPN·100mL–1 to 1.7 log10MPN·100mL–1 seawater and from < 0.3 log10 MPN·100mL–1 to 2.0 log10 MPN·100mL–1 seawater, respectively. A positive correlation between V. vulnificus counts and the seawater temperature as well as a negative correlation between the V. parahaemolyticus counts and salinity were observed. Conclusions The results suggest the need to implement strategies to prevent vibrio diseases from being transmitted by the consumption of contaminated bivalve shellfish. .

Development of duplex-PCR for identification of Aeromonas species

Introduction The number of reports of intestinal infections caused by Aeromonas spp. has increased significantly in recent years. In most clinical laboratories, identification of these bacteria is carried out by general phenotypic tests that sometimes do not accurately differentiate Aeromonas and Vibrio. Methods A duplex-polymerase chain reaction (PCR) was developed directed to 2 targets identifying Aeromonas spp. pathogenic to humans. Results The duplex-PCR results were reproducible and specific for Aeromonas spp. pathogenic to humans. Conclusions This method will allow differentiation between Vibrio and Aeromonas spp. in patients with in cholera-like symptoms and can also be used in water quality monitoring. .

Microbiota of Vibrio sp. in the hepatopancreas of cultured white pacific shrimp (Litopenaeus vannamei) / Vibrio sp., microbiota en el hepatopáncreas del camarón blanco del Pacífico (Litopenaeus vannamei)

Objective. The present study aimed to investigate the presence of vibrios in the hepatopancreas of cultured shrimp. Materials and methods. Vibrios from the hepatopancreas of fifteen samples of five specimens each, of apparently healthy Pacific white shrimp (Litopenaeus vannamei) were isolated, identified and quantified. Results. The vibrio density ranged from 430 to 2,400 MPN g-1 (rs MPN cm-1=-0.114; rs MPN g-1 = 0.211). Thirty isolations were obtained, most of which belonged to the species V. cholerae (n=11) and V. parahaemolyticus (n=7). Conclusions. The outcomes of the present study suggest that, even in the absence of symptoms of vibriosis, the microbiota of the hepatopancreas of cultured shrimp may include sucrose positive and negative vibrios.

Objetivo. El presente estudio tuvo como objetivo investigar la presencia de vibrios en el hepatopáncreas del camarón de cultivo. Material y métodos. En este estudio, fueron aislados, identificados y cuantificados los vibrios del hepatopáncreas de 75 camarones blancos del Pacífico (Litopenaeus vannamei), aparentemente sanos, oriundos de un cultivo en la región nordeste de Brasil. Quince muestras, cada una consta de cinco camarones, se pusieron a prueba. Resultados. La densidad de Vibrio varió de 430 a 2.400 NMP g-1 (rs NMP cm-1 = -0.114; rs NMP g-1 = 0.211). Treinta aislamientos fueron obtenidos, la mayoría de los cuales pertenecían a la especie V. cholerae (n=11) y V. parahaemolyticus (n=7). Conclusiones. Los hallazgos del presente estudio sugieren que, incluso en ausencia de síntomas de la vibriosis, la microbiota de lo hepatopáncreas del camarón de cultivo puede incluir vibrios sacarosa positivos y negativos.

Hemolytic and urease activities in vibrios isolated from fresh and frozen oysters

INTRODUCTION: The present study aimed to survey the Vibrio microbiota of oysters (Crassostrea rhizophorae) obtained from restaurants in Fortaleza, State of Ceará, Brazil, and to identify virulence factors. METHODS: The isolated vibrios were submitted to biochemical identification and were tested for hemolytic and urease activities. RESULTS: The isolated strains belonged to 13 species, with predominance of Vibrio mimicus. Of the strain isolates only from fresh samples, 20.5% and 2.8% showed hemolytic and urease activities, respectively. CONCLUSIONS: The findings support the little-publicized claim that Vibrio species other than V. parahaemolyticus and V. vulnificus can represent a health risk to public health.