Circulación de genes de virulencia asociados a Escherichia coli diarrogénica en aguas residuales del Gran Área Metropolitana de Costa Rica / Circulation of diarrheagenic Escherichia coli associated virulence genes in Costa Rican Great Metropolitan Area wastewaters

Resumen Introducción: la enfermedad diarreica es una de las principales causas de muerte en niños menores de cinco años. Uno de los agentes etiológicos bacterianos más reconocido es Escherichia coli, que puede causar infecciones a través de diversos mecanismos asociados a la adquisición de genes de virulencia. Metodología: se recolectaron muestras de aguas residuales en los afluentes y los efluentes de cinco plantas de tratamiento de la Gran Área Metropolitana, durante tres días consecutivos, en los meses de marzo, mayo, octubre y diciembre del año 2013, a fin de cuantificar la concentración de E. coli, mediante la técnica de fermentación en tubos múltiples, y evaluar la presencia de los genes de virulencia aatA, bfpA, eaeA, ipaH, lt, st, stx1 y stx2, vinculados con patotipos de E. coli diarrogénica, con la técnica de PCR punto final. Resultados: el estudio reveló niveles de E. coli similares a los encontrados en afluentes de otras regiones del mundo, en cambio, los efluentes presentaron concentraciones superiores a los 4 Log10 NMP/100 mL. La circulación anual de genes de virulencia fue relacionada con los patotipos enterotoxigénica, productores de toxina Shiga/enterohemorrágica, enteroagregativa y enteropatógena. No se detectaron genes referentes a la variedad enteroinvasora. Además, se determinó que no todas las plantas de tratamiento remueven significativamente la carga de E. coli. Conclusiones: el flujo de E. coli diarrogénica es común en nuestro país y su diseminación en el ambiente es posible cuando los efluentes se descargan en cuerpos de aguas superficiales

Abstract Introduction. Diarrheic disease is one of the most important diseases causes in children under five years. One of the most recognized etiological agents is Escherichia coli. This bacterium has the capacity of causing infections by multiple mechanisms associated with the acquisition of different virulence genes. Methodology: water samples were collected from the affluents and effluents of five wastewater treatment plants located in the Costa Rican Great Metropolitan Area, during three sequential days, in March, May, October, and December from 2013. The aim was to quantify E. colis concentration by multiple tubes fermentation method and to evaluate the presence of aatA, bfpA, eaeA, ipaH, lt, st, stx1, and stx2 virulence genes, related with diarrheagenic E. coli, by PCR end-point method. Results: The study revealed that the levels of E. coli into the affluents are similar to those reported in other world regions. The effluents concentration presented higher than 4 Log10 NMP/100 mL level of E. coli. A year-round circulation of virulence genes was demonstrated, and it was associated with enterotoxigenic, Shiga-toxin producers/ enterohemorrhagic, enteroaggregative, and enteropathogenic pathotypes. Genes related to enteroinvasive variety did not were detected. Not all wastewater treatment plants removed E. coli efficiently. Conclusions: Diarrheagenic E. coli circulation is common in our county with the possibility of dissemination to the environment by effluents wastewater discharge in surface water bodies.

Antimicrobial susceptibility and serotypes of Neisseria meningitidis and Streptococcus pneumoniae in Sri Lanka: Experience from the National Reference Laboratory

Objective: To determine the antimicrobial susceptibility and serotypes of Neisseria (N.) meningitidis and Streptococcus (S.) pneumoniae in Sri Lankan patients. Methods: We retrospectively analyzed 11 blood culture specimens from suspected patients with invasive meningococcal disease and 26 S. pneumoniae clinical isolates. We tested 6 antimicrobials against N. meningitidis and 12 antimicrobials against S. pneumoniae. Meningococcal serogroup was determined by realtime PCR and Quellung serotyping was used for pneumococcal analysis. Results: N. meningitidis serogroup B was the most common in this study. Intermediate-susceptibility to penicillin was seen in 75.0% (6/8) of strains. Susceptibility to ciprofloxacin, levofloxacin and cotrimoxazole was 62.5% (5/8), 62.5% (5/8) and 87.5% (7/8), respectively. Excellent susceptibility was seen in cefotaxime and meropenem. In S. pneumoniae, the most common serotype was 19F in both invasive and non-invasive pneumococcal diseases. The majority of strains showed multidrug resistance. Penicillin non-susceptibility in non-meningeal strains were 13.6% and all meningeal strains were penicillin resistant. Erythromycin was highly resistant in both groups. Amoxicillin showed excellent susceptibility in non-invasive pneumococcal diseases strains. Linezolid, levofloxacin and vancomycin showed 100.0% susceptibility in all pneumococcal isolates. Conclusions: Implementation of vaccines should be considered, especially for children and high-risk populations. This may contribute to reducing pneumococcal and meningococcal invasive disease burden and help prevent emergence of antimicrobial resistant strains.

Staphylococcus aureus and Streptococcus agalactiae: prevalence, resistance to antimicrobials, and their relationship with the milk quality of dairy cattle herds in Minas Gerais state, Brazil / Staphylococcus aureus e Streptococcus agalactiae: prevalência, resistência aos antimicrobianos e suas relações com a qualidade do leite em rebanhos leiteiros de Minas Gerais, Brasil

Bovine mastitis is the most frequent disease worldwide in dairy herds, causing high economic losses to producers and industry, as well as having implications for public health due to the zoonotic potential of some agents involved in its etiology and the increased risk of antimicrobial residues in milk and its derivatives. Considering the multifactorial aspect of this disease, knowledge of the agents involved in its etiology and their antimicrobial susceptibility profiles is very important. This study was conducted with 306 dairy herds from the Campo das Vertentes region, located in the south of Minas Gerais state, whose owners were milk suppliers to a dairy in the same region. The study involved approximately 34,000 dairy cows and covered an area of approximately 12,564 km2. In these herds, prevalence rates of Staphylococcus aureus and Streptococcus agalactiae and their relationship with bulk milk somatic cell counts (BMSCC), total bacterial counts (TBC), and daily production were evaluated. In addition, analyses of resistance of these pathogens to the antimicrobials most commonly used in the treatment of mastitis in dairy herds were performed. Microbiological analyses of milk samples from collect from bulk milk tanks were performed aiming to evaluate the prevalence of S. aureus and S. agalactiae. For these proposes, the modified Baird-Parker Agar medium was used for detection of S. aureus and the modified Edwards Agar medium, enriched with 5% defibrinated sheep blood, was used for detection of S. agalactiae. The disc diffusion technique was applied to evaluate antimicrobial resistance. Results show high prevalence rates of S. aureus (70.3%) and S. agalactiae (67.0%) in the dairy farms studied, with 47.71% of the herds showing both pathogens. Associations between BMSCC and the presence of pathogens S. aureus and S. agalactiae and between TBC and the presence of S. agalactiae were observed, demonstrating the influence of these pathogens in milk quality. No variation was observed in the distribution of S. aureus and S. agalactiae in the different strata of daily production. High levels of resistance and multi-resistance were observed among the pathogens S. aureus and S. agalactiae. The results indicate the need for more effective control measures for mastitis caused by S. aureus and S. agalactiae in the dairy herds of the region studied and more judicious use of antimicrobials in order to reduce the problem of resistance to them.(AU)

A mastite bovina é a doença de maior frequência em rebanhos leiteiros em nível mundial, acarretando grandes prejuízos econômicos aos produtores e à indústria. Além disso, esta enfermidade tem implicações na saúde pública, devido ao potencial zoonótico de alguns agentes envolvidos em sua etiologia e por aumentar os riscos de resíduos de antimicrobianos no leite e derivados. Considerando o aspecto multifatorial da mastite bovina, o conhecimento dos agentes envolvidos em sua etiologia e os perfis de suscetibilidade aos antibióticos é de suma importância. O estudo envolveu 306 fazendas de leite da região de Campo das Vertentes, localizada no sul de Minas Gerais, cujos proprietários eram fornecedores de leite para um laticínio da região, totalizando aproximadamente 34.000 animais e abrangendo uma área aproximada 12.564 km2. Nestes rebanhos, avaliaram-se a prevalência de Staphylococcus aureus e Streptococcus agalactiae e a relação destes agentes com os índices de contagem de células somáticas do leite do tanque de expansão (CCSt), contagem bacteriana total (CBT) e produção diária. Analisou-se também a resistência destes patógenos aos antimicrobianos mais comumente utilizados no tratamento da mastite em rebanhos leiteiros. Análises microbiológicas de amostras de leite dos tanques de expansão foram realizadas para se determinar as prevalências dos patógenos S. aureus e S. agalactiae. Para a detecção de S. aureus, utilizou-se o meio seletivo Ágar Baird-Parker modificado e para a detecção de S. agalactiae, o meio seletivo Ágar Edwards modificado, enriquecido com 5% de sangue ovino desfibrinado. Foi utilizada a técnica de difusão em discos para a avaliação de resistência aos antimicrobianos. Os resultados apontaram altas prevalências de S. aureus (70,3%) e de S. agalactiae (67,0%), com 47,71% dos rebanhos examinados apresentando ambos os agentes. Verificaram-se associações entre a CCSt e a presença dos patógenos S. aureus e S. agalactiae, e também entre a CBT e a presença de S. agalactiae, demonstrando a interferência negativa destes patógenos nestes quesitos de qualidade. Não se observaram variações nas distribuições dos patógenos S. aureus e nem S. agalactiae em função da produção diária das propriedades estudadas. Níveis elevados de resistência e de multirresistência foram observados para ambos os agentes. Os resultados apontam a necessidade de medidas mais efetivas de controle para S. aureus e S. agalactiae nos rebanhos da região estudada e do uso mais criterioso dos antimicrobianos, visando minimizar o problema da resistência aos mesmos.(AU)

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.

Dominant bacterial diseases in the extensive and semi-intensive animal breeding and their treatment method by ethnoveterinary medicine in Benin.

In Africa, various medicines are used in animal diseases treatment. This study aims to identify the medicinal plants that are often used to cure bacterial diseases in domestic animals, in order to promote their rational and sustainable use. The work was performed by working with 787 breeders in the 8 agro-ecological zones of Benin, from June to December 2013. An ethnobotanical survey has identified 32 medicinal plants species among which Afzelia africana, Crossopteryx febrifuga, Albizia chevalieri, Khaya senegalensis, Mangifera indica and Entada africana are used for bovine pasteurellosis. The bronchitis is treated by 11 species of medicinal plants which include Allium sativum, Garcinia kola, Terminalia laxiflora, Zanthoxylum zanthoxyloides and Detarium microcarpum. For infectious coryza treatment, 8 plants have been identified among which, we have Securinega virosa, Capsicum annuum, Anacardium occidentale and Vitellaria paradoxa. About cutaneous streptothricosis, 3 plants (Zea mays, Piliostigma thonningii and Parkia biglobosa) were cited for its treatment. Contagious bovine peripneumonia is treated by 3 plants (Tamarindus indica, Oxythenanthera abyssinica and Khaya senegalensis). Avian salmonellosis is also treated by 3 plants (Bridelia ferruginea, Cissus quadrangularis and Ocimum gratissimum). Concerning Blackleg and enterotoxemia, they are respectively treated by Aganope stulhmannii and Ocimum gratissimum.

Roles of Mesenchymal Stem Cells (MSCs) in bacterial diseases / 동물의과학연구지

Mesenchymal stem cell (MSC) based cell therapy has emerged as a promising therapeutic approach for treatment of several degenerative, infectious and non-infectious diseases. Numerous studies have demonstrated the remarkable immunosuppressive and antibacterial effects of MSCs both in vitro and in vivo, in animal models and in humans. However, the antibacterial effects of MSCs rely heavily on their paracrine factors rather than direct cell-to-cell contact and the effect is specific to disease and site of infection or injury. Furthermore, recent studies have demonstrated the double-edged sword effect of MSCs in bacterial infectious diseases. Despite their inherent potential for repair of damaged tissues, immunosuppression, and alleviation of various autoimmune as well as infectious diseases, MSCs also play a critical role in promoting persistent bacterial infection and disease progression. Therapeutic administration of MSCs successfully inhibited the bacterial growth and enhances survival by improved clearance of pathogenic bacteria in sepsis and pneumonic conditions. However, due to their abnormal transformation, they assist in long lasting survival and persistent infection of Mycobacterium tuberculosis (M. tuberculosis) and may also be responsible for progression of gastric cancer. This review focuses on recent advances that have broadened our understanding of MSC based therapy for bacterial diseases and provides new insight into the possible therapeutic targets of fatal bacterial diseases.

Brucella canis infection in dogs attended in veterinary clinics from patos, Paraíba state, Brazil

To determine the frequency of anti-Brucella canis antibodies in dogs attended in veterinary clinics from Patos, Paraíba State, Brazil, as well as to identify risk factors and to isolate and identify the agent, 193 dogs were used. Agar gel immunodiffusion test (AGID) was used to detect B. canis antibodies in sera. Isolation of B. canis was carried out in blood and bone marrow from seropositive animals. Six animals tested seropositive in AGID, resulting in a frequency of 3.11 percent. B. canis was isolated from bone marrow of one seropositive animal, with confirmation by PCR. Lack of cleaning of the dog's environment was identified as risk factor (odds ratio = 7.91). This is the first report of isolation of B. canis in dogs from the Northeast region of Brazil.

Síndrome do abscesso pituitário em bezerros na Região Centro-Oeste / Pituitary abscess syndrome in calves from Mid-Western Brazil

A síndrome do abscesso pituitário é uma doença neurológica responsável por casos esporádicos e surtos, principalmente em bezerros, ocasionando alto índice de mortalidade. Descreve-se a ocorrência e os achados clínicos, laboratoriais e anátomo-patológicos em três bezerros com síndrome do abscesso pituitário no Centro-Oeste do Brasil. Os animais tinham 8-11 meses de idade e os sinais clínicos mais marcantes relacionaram-se aos sinais nervosos de origem cerebral e do tronco encefálico com evolução clínica de 7-20 dias. A hematologia revelou leucocitose por neutrofilia e hiperfibrinogenemia. A análise do líquido céfalo-raquidiano apresentou pleocitose neutrofílica. Arcanobacterium pyogenes foi isolado do líquido céfalo-raquidiano. Um dos bezerros apresentou recuperação após antibioticoterapia. A mortalidade foi de 66,6 por cento (2/3). Os achados de necropsia consistiram em um único abscesso de localização parapituitária ou situado no parênquima da glândula; um dos bezerros apresentou rinite necrosante e outro, broncopneumonia abscedativa. O exame histológico do sistema nervoso central revelou ausência quase completa do tecido hipofisário normal, devido à necrose extensa e infiltrado inflamatório neutrofílico difuso concomitante. Reitera-se a importância da realização de práticas de manejo adequadas a fim de reduzir a incidência de inúmeras enfermidades, principalmente em bezerros, dentre elas a síndrome do abscesso pituitário.

Pituitary abscess syndrome is a neurologic disease responsible for sporadic cases and outbreaks especially in calves leading to high mortality rates. This paper aimed to report the occurrence and the clinical, laboratorial and pathologic findings in three 8 to 11-month-old calves with pituitary abscess syndrome from Mid-Western Brazil. The most important clinical findings were nervous signs of cerebral and brainstem origin with clinical evolution of 7-20 days. Hematology revealed leucocytosis by neutrophilia and hyperfibrinogenemia. Cerebrospinal fluid analysis showed neutrophilic pleocytosis. Arcanobacterium pyogenes was isolated from cerebrospinal fluid. One calf recovered after antibiotic treatment. Mortality rate was 66.6 percent (2/3). Necropsy findings included single para-hypophyseal abscesses or located in the glandular parenchyma; one calf showed necrotizing rhinitis and another abscedative pneumonia. Histological exams of the central nervous system reveal complete absence of normal pituitary tissue due to the wide necrosis and neutrophilic inflammatory infiltrate. The authors reiterate the importance of adequate management practices to reduce incidence of several diseases especially in calves, including the pituitary abscess syndrome.

Bacteriophage: laboratorial diagnosis and phage therapy / Bacteriofagos: diagnóstico laboratorial e terapia fágica

Bacteriophages have been researched as a new alternative to antibiotics. These viruses inject their genetic material into bacteria and use their host machinery to multiply themselves. The research of bacteriophages in Brazil will certainly provide low-cost treatment of multidrug resistant bacteria, new microbiological diagnosis and advantages for the Brazilian food industry.

Bacteriófagos têm sido pesquisados como uma alternativa ao uso de antibióticos. Estes vírus infectam as bactérias e utilizam a maquinaria celular para multiplicar o próprio material genético. O estudo de bacteriófagos no Brasil levará ao desenvolvimento de tratamentos de baixo custo, novos testes diagnósticos e vantagens para a industria alimentícia.

Suscetibilidade à azitromicina de isolados bacterianos de processos infecciosos em cães e gatos / Susceptibility to azithromycin of bacteria isolated from infectious processes in dogs and cats

O presente estudo avaliou o perfil de suscetibilidade à azitromicina de patógenos bacterianos prevalentes em diferentes sítios infecciosos de animais de companhia. Adicionalmente, foram estudados o perfil de atividade in vitro de azitromicina contra esses patógenos e sua concentração inibitória mínima (CIM). Testes como a difusão em disco e a microdiluição em caldo detectaram resistência respectivamente em 48,6 por cento e 55 por cento dos isolados de Staphylococcus spp. e em 55,3 por cento e 72,7 por cento dos bastonetes Gram-negativos. A CIM50 para S. aureus foi 4,0mg/mL, para S. intermedius foi de 1,0mg/mL, para Staphylococcus spp. coagulase-negativas foi de e"512mg/mL e para bastonetes Gram-negativos foi de 256mg/mL. Quinze por cento (9/60) dos isolados oxacilina-resistente e multidroga-resistentes, mecA-positivos, de Staphylococcus spp. apresentaram também resistência à azitromicina. A disseminação de bactérias multidroga-resistentes aponta para a necessidade da avaliação da atividade antimicrobiana para selecionar o fármaco mais indicado e, assim, minimizar falhas terapêuticas na conduta clínica veterinária.

The susceptibility pattern to azithromycin of bacterial pathogens from various infectious sites, and the in vitro activity and minimum inhibitory concentration (MIC) of azithromycin were studied. Tests such as disc diffusion and broth microdilution detected respectively 48.6 percent and 55 percent of resistant Staphylococcus spp., and 55.3 percent and 72.7 percent resistant gram-negative rods. MIC50 for S. aureus was 4.0mg/mL, that for S. intermedius was 1.0mg/mL, for coagulase-negative Staphylococcus e"512mg/mL, and for gram-negative rods 256mg/mL. Fifteen percent (9/60) of oxacilin-resistant, multidrug-resistant and mecA-positive Staphylococcus spp. isolates were also azithromycin resistant. The dissemination of multidrug resistant bacteria points out to the need of antimicrobial evaluation activity in order to select the best indicated drug and thus minimizing therapeutic failures in veterinary practice.