Molecular characterization and pathology of field isolates of foot-and-mouth virus in Swiss albino mice / Caracterização molecular e patologia de isolados de campo do vírus da febre aftosa em camundongos albinos suíços

Foot-and-mouth disease is responsible for severe economic losses to the livestock industry of Pakistan. This study aimed to use Swiss albino mice as a cost-effective experimental animal model to study different immunological and histopathological aspects of FMDV instead of natural targeted species like cattle. After isolation of field isolates FMDV on BHK-21 cell line, biological titer of the virus and mice infectious dose50 was calculated. Virus was injected in 45 Swiss albino mice (group A) through intraperitoneal route. The gross, histopathological and immunopathological lesions in heart, trachea and lungs were recorded at different day's intervals. Histopathologically, the heart showed congestion, hemorrhages and necrosis of cardiac muscles. Trachea showed deciliated epithelium and lungs showed hemorrhages, bronchial edema and alveolar emphysema. Immunohistochemical studies revealed the presence of virus in cardiac muscles, tracheal and bronchial epithelium and alveolar lumen. The findings evoked a thought that laboratory animals could be an alternative to large animals to meet budget limitations for further research on foot-and-mouth-disease.

A febre aftosa (FMD) é responsável por graves perdas econômicas para a indústria pecuária do Paquistão. Este estudo teve como objetivo usar camundongos albinos suíços como um modelo animal experimental de baixo custo para estudar diferentes aspectos imunológicos e histopatológicos do FMDV em vez de espécies naturais como o gado. Após o isolamento dos isolados de campo do FMDV na linhagem celular BHK-21, calculou-se o título biológico do vírus e a dose infecciosa dos camundongos50. O vírus foi injetado em 45 camundongos albinos suíços (grupo A) por via intraperitoneal. As lesões macroscópicas, histopatológicas e imunopatológicas no coração, traqueia e pulmões foram registradas em diferentes intervalos de dias. Histopatologicamente, o coração apresentava congestão, hemorragias e necrose dos músculos cardíacos. A traqueia apresentava epitélio deciliado e os pulmões apresentavam hemorragias, edema brônquico e enfisema alveolar. Estudos imuno-histoquímicos revelaram a presença de vírus em músculos cardíacos, epitélio traqueal e orçamentárias para pesquisas adicionais sobre a febre aftosa

Co-existence of two Yersinia ruckeri biotypes and serotype O1a retrieved from rainbow trout (Oncorhynchus mykiss) farmed in Puno, Peru.

Yersinia ruckeri causes important economic losses for rainbow trout (Oncorhynchus mykiss) farms worldwide. This bacterial disease is likely the most common among trout in Peru; however, no commercial vaccine is available nationally, which is, in part, due to a lack of information on the bacterium. The aim of the current study was to characterize 29 Y. ruckeri isolates sampled from seven cage-reared farms in the Puno Region, the focal point for aquaculture activities in Peru. For this, samples were taken from fish with clinical signs (i.e. haemorrhages, uni- or bilateral exophthalmia, hyphaemia and/or melanosis). Notable among our findings was the existence of both Y. ruckeri biotype 1 (9 isolates) and biotype 2 (20 isolates; negative for sorbitol and Tween 80). The isolates further differed in API profiles 5307100 (21 isolates), 1307100 (4 isolates), 1305100 (2 isolates), 1307120 (1 isolate) and 5305100 (1 isolate), with the main differences being in the tests for lysine decarboxylase, gelatine hydrolysis and D-saccharose fermentation. Despite these differences, all isolates shared identical ERIC-PCR and REP-PCR profiles and belonged to the O1a serotype. Fingerprints were identical to the reference strain CECT 955 (serotype O1a). The information obtained will be used for epidemiological purposes by health authorities and for the development of a vaccine against Y. ruckeri, a prominent request made by fish farmers in Peru.

Occurrence and Abundance of Pathogenic Vibrio Species in Raw Oysters at Retail Seafood Markets in Northwestern Mexico.

Seafood has frequently been associated with foodborne illness because pathogens are easily introduced during seafood cultivation, handling, and processing. Vibrio parahaemolyticus and Vibrio cholerae are human pathogens that cause gastroenteritis and cholera, respectively, and Vibrio vulnificus can cause fatal wound infections and septicemia. However, information about the occurrence of these pathogens in oysters from the Pacific coast of Mexico is limited to V. parahaemolyticus. In the present study, we evaluated the presence and abundance of these three Vibrio species in 68 raw oysters (Crassostrea corteziensis) obtained from retail seafood markets in Sinaloa, Mexico. The most probable number (MPN)-PCR assay was used for amplification of the tlh (thermolabile hemolysin), ompW (outer membrane protein), and vvhA (hemolytic cytolysin) genes that are specific to V. parahaemolyticus, V. cholerae, and V. vulnificus, respectively. All oyster samples were positive for at least one Vibrio species. V. parahaemolyticus, V. cholerae, and V. vulnificus prevalences were 77.9, 8.8, and 32.3% overall, respectively, and most species were present in all sample periods with increased prevalence in period 3. The tdh (thermostable direct hemolysin) gene was detected in 30.1%, trh (TDH-related hemolysin) was detected in 3.7%, and tdh/trh was detected in 7.5% of the total tlh-positive samples (53 of 68), whereas the pandemic serotype O3:K6 (orf8 positive) was detected in only 1 sample (1.8%). The total prevalence of tdh and/or trh was 41.5%. In none of the samples positive for V. cholerae were the cholera toxin (ctxA) and cholix (chxA) toxigenic genes or the rfb gene encoding the O1 and O139 antigens amplified, suggesting the presence of non-O1 non-O139 V. cholerae strains. Our results clearly indicated a high prevalence of pathogenic Vibrio species in raw oysters from retail seafood markets in Mexico. Consumption of these raw oysters carries the potential risk of foodborne illness, which can be limited by cooking.

Infecciones por Escherichia coli productora de toxina Shiga O121: H19 en pacientes atendidos en Mar del Plata / Infections caused by Escherichia coli producing Shiga O121: H19 toxin in patients treated in Mar del Plata

Escherichia coli productora de toxina Shiga (STEC) O157:H7 es el serotipo más frecuentemente identificado como agente causal de colitis hemorrágica y síndrome urémico hemolítico (SUH), aunque se han descripto más de 100 serotipos con potencial patogénico similar. El objetivo del trabajo fue describir casos de enfermedad humana asociados a la infección por STEC O121:H19, atendidos en la ciudad de Mar del Plata y establecer la relación genética de los aislamientos mediante técnicas de epidemiología molecular. Se observó un amplio espectro en la severidad clínica de los ocho casos estudiados: dos fueron asintomáticos (contactos de SUH), un paciente tuvo diarrea sanguinolenta, y cinco presentaron SUH. Uno de los pacientes con SUH falleció. Las cepas O121:H19 portadoras del genotipo stx2a/eae/ehxA fueron sensibles a los antibióticos ensayados y presentaron por electroforesis en gel de campo pulsado (Xbal-PFGE) distintos patrones de macrorrestricción, con similitud del 84,25%. El patrón AREXKX01.0072, detectado en un SUH y en su contacto, es nuevo en la Base de Datos Nacional de STEC no-O157 de la Argentina. La utilización de métodos estandarizados de detección y tipificación de STEC permite a los laboratorios de referencia monitorear la frecuencia temporal y la distribución geográfica de las cepas circulantes para la prevención y control de estos patógenos asociados a enfermedad humana.

Shiga toxin-producing Escherichia coli (STEC) O157:H7 is the most frequent serotype identified as causative agent of sporadic cases and outbreaks of diarrhea with or without blood, hemorrhagic colitis and hemolytic uremic syndrome (HUS), although more than 100 serotypes have been described of similar pathogenic potential. The aim of the study was to describe cases of human disease associated with STEC O121:H19 infections, assisted in Mar del Plata City, and to establish the genetic relationship of the isolates by molecular epidemiology techniques. A wide spectrum was observed in the clinical severity of the eight cases studied: two were asymptomatic (contacts of HUS), one patient had bloody diarrhea, and five cases presented HUS. One HUS case died. All STEC O121:H19 strains carried the stx2a/eae/ehxA genotype, were sensitive to all antibiotics tested and showed different macrorestriction patterns by pulsed-field gel electrophoresis (Xbal-PFGE), with 84.25% similarity. The pattern AREXKX01.0072, detected in a HUS case and in his contact, is new in the Argentine National Database of non-O157 STEC. The use of standardized methods for detection and typing of STEC allows reference laboratories to monitor the temporal frequency and geographical distribution of circulating strains for the prevention and control of these pathogens associated with human diseases.

Escherichia coli produtora de toxina Shiga (STEC) O157:H7 é o sorotipo mais frequentemente identificado como o agente causador de colite hemorrágica e síndrome hemolítica urêmica (SHU), embora tenham sido descritas mais de 100 sorotipos com potencial patogênico semelhantes. O objectivo foi o de descrever os casos de doença humana associadas com a infecção por STEC O121:H19, assistido, na cidade de Mar del Plata e estabelecer relação genética de isolados utilizando epidemiologia molecular. Um amplo espectro foi observado na severidade clínica dos oito casos estudados, dois eram assintomáticos (contacto SHU), uma paciente teve diarreia com sangue, e cinco tiveram SHU. Um caso de SHU faleceu. As cepas O121:H19 portaram o genótipo stx2a/eae/ehxA, foram sensíveis aos antibióticos testados e apresentaram, por eletroforese em gel de campo pulsado (Xbal-PFGE), diferentes padrões de macrorestrição, com similaridade de 84,25%. O padrão AREXKX01.0072 detectado em SHU e em seu contato, é novo para a Base de Dados Nacional de STEC não-O157 na Argentina. O uso de métodos padrão de detecção e tipagem de STEC permite os laboratórios de referência monitorar frequência temporal e distribuição geográfica de estirpes circulantes para a prevenção e controlo destes agentes patogénicos associados com a doença humana.

Fatores de virulência em linhagens de Escherichia coli isoladas de mastite bovina / Virulence factors in Escherichia coli strains isolated from bovine mastitis

Avaliou-se a ocorrência de fatores de virulência e do sorotipo O157:H7 em 120 linhagens de Escherichia coli, isoladas de 80 casos de mastite clínica bovina e 40 de mastite subclínica. Verificou-se alfa-hemolisina em oito (6,7%) linhagens, isoladas de cinco casos de mastite clínica e três de mastite subclínica e em nenhuma das estirpes detectou-se enteroemolisina. A presença de sideróforos foi encontrada em 11 (9,2%) linhagens, sete de mastite clínica e quatro de subclínica. Em duas (1,7%) estirpes isoladas de mastite subclínica, identificou-se enterotoxina STa. Observou-se efeito citopático em células vero compatível com a produção de verotoxina-VT em cinco (4,2%) linhagens, duas de mastite clínica e três subclínicas. Em uma (0,8%) linhagem isolada de mastite clínica, detectou-se efeito citopático compatível com o fator necrosante citotóxico. Nenhuma estirpe apresentou-se sorbitol-negativa no MacConkey-sorbitol, tampouco aglutinou com o sorotipo O157:H7. Os antimicrobianos mais efetivos foram polimixina B (97,5%) e norfloxacina (95,8%). Observou-se multi-resistência a dois ou mais antimicrobianos em 24 (20%) estirpes, principalmente com o uso de ampicilina e ceftiofur.(AU)

The occurrence of different virulence factors and O157:H7 serotype investigation in 120 Escherichia coli strains isolated from clinical (80 cases) and subclinical (40 cases) bovine mastitis was evaluated. Alpha-haemolysin was detected in 8 (6.7%) strains (5 clinical and 3 subclinical cases). None strain showed enterohaemolysin production. E. coli growth under iron restriction conditions (siderophores production) was observed in 11 (9.2%) strains (7 clinical and 4 subclinical cases). STa enterotoxin was detected in 2 (1.7%) strains from subclinical cases. Cytotoxic effect in vero cells compatible with verotoxin-VT production was observed in 5 (4.2%) strains (2 clinical and 3 subclinical cases). One strain (0.8%) isolated from clinical mastitis showed cytophatic effect in vero cells compatible with cytotoxic-necrotic-factor production. None strain showed non-sorbitol fermenting colonies using sorbitol MacConkey agar and agglutination with specific anti-sera for E. coli O157:H7 serotype investigation. Polymixin B (97.5%) and norfloxacin (95.8%) were the most effective drugs. Multiple-drug resistance for 2 or more antimicrobials was observed in 24 (20.0%) strains, mainly with use of ampicillin and ceftiofur.(AU)