Preparation and evaluation of formalized bivalent Newcastle and Salmonella poultry vaccine

This study was conducted to prepare and evaluate the potency of different inactivated vaccine formulations that protect chickens against Salmonella Enteritidis and Newcastle disease virus using Montanide as adjuvant. Protection and the humoral immune response of prepared vaccines against Salmonella Enteritidis and Newcastle disease virus was evaluated and compared to imported vaccine. In this study, different formulae of Salmonella Enteritidis and Newcastle disease vaccines were prepared and compared with the imported one by measuring the antibody titer against Newcastle disease virus by hemagglutination inhibition test and the antibody titer against Salmonella Enteritidis using Enzyme Linked Immunosorbent Assay. On the other hand, the protection percentages against Newcastle disease and Salmonella Enteritidis were recorded to determine the best effective formula. The highest hemagglutination inhibition antibody level against NDV at first week was recorded for the prepared combined Newcastle disease and Salmonella Enteritidis vaccine (4.2 log2) followed by the prepared monovalent Newcastle disease (3.4 log2); the lowest antibody level (3.1 log2) was obtained with the imported vaccine. A gradual increase was observed in all groups to 7.1 log2, 6.8 log2 and 6.4 log2 at fourth week post vaccination, respectively. The antibody titer against Salmonella Enteritidis was 552 for the prepared combined Salmonella Enteritidis and Newcastle disease, followed by the prepared monovalent Salmonella Enteritidis (477) at first week post vaccination; the antibody titer obtained for the imported vaccine was 477. There was a gradual increase to 1456, 1406 and 1130 at fourth week post vaccination, respectively. Prepared combined vaccines gave the highest protection percentage, followed by prepared monovalent types and finally imported vaccines. Vaccination by the prepared combined Salmonella Enteritidis and Newcastle disease vaccine may be a way to increase the resistance of birds to Salmonella and Newcastle and to decrease the shedding rate(AU)

Este estudio se llevó a cabo para preparar y evaluar la potencia de diferentes formulaciones de vacunas inactivadas que protegen a los pollos contra Salmonella Enteritidis y el virus de la enfermedad de Newcastle utilizando Montanide como adyuvante. Se evaluó la protección y la respuesta inmune humoral de las vacunas preparadas contra Salmonella Enteritidis y el virus de la enfermedad de Newcastle y se comparó con la vacuna importada. En este estudio se prepararon diferentes fórmulas de vacunas contra Salmonella Enteritidis y la enfermedad de Newcastle y se compararon con la importada midiendo el título de anticuerpos contra el virus de la enfermedad de Newcastle mediante la prueba de inhibición de la hemaglutinación y el título de anticuerpos contra Salmonella Enteritidis mediante ELISA. Por otra parte, se registraron los porcentajes de protección contra la enfermedad de Newcastle y Salmonella Enteritidis para determinar la fórmula más eficaz. El mayor nivel de anticuerpos inhibidores de la hemaglutinación contra el virus de la enfermedad de Newcastle, en la primera semana, se registró con la vacuna combinada preparada contra la enfermedad de Newcastle y Salmonella Enteritidis (4,2 log2), seguida de la vacuna monovalente preparada contra la enfermedad de Newcastle (3,4 log2); el menor nivel de anticuerpos (3,1 log2) se obtuvo con la vacuna importada. Se observó un aumento gradual en todos los grupos hasta alcanzar 7,1 log2, 6,8 log2 y 6,4 log2 en la cuarta semana tras la vacunación, respectivamente. El título de anticuerpos contra Salmonella Enteritidis fue de 552 para la vacuna combinada preparada contra la Salmonella Enteritidis y enfermedad de Newcastle, seguida por la vacuna monovalente preparada contra Salmonella Enteritidis (477) en la primera semana después de la vacunación; el título de anticuerpos obtenido con la vacuna importada fue de 477. Hubo un aumento gradual hasta 1456, 1406 y 1130 en la cuarta semana después de la vacunación, respectivamente. Las vacunas combinadas preparadas dieron el mayor porcentaje de protección, seguidas por los tipos monovalentes preparados y, por último, por las vacunas importadas. La vacunación con la vacuna combinada preparada contra la Salmonella Enteritidis y la enfermedad de Newcastle puede ser una forma de aumentar la resistencia de las aves a la Salmonella y Newcastle y de disminuir la tasa de excreción(AU)

Salmonella enteritidis abdominal aorta mycotic aneurysm presented with acute cholestatic jaundice: A case report and literature review

@#Mycotic aneurysm is one of the extra-intestinal manifestations of Salmonella Enteritidis infection. The diagnosis of this condition is challenging owed to its variation in clinical presentations. We presented a case of a 54-year-old man with underlying diabetes mellitus and chronic smokers presented with acute right flank pain and fever associated with mild jaundice. The initial laboratory investigations suggested features of obstructive jaundice and urinary tract infection. The contrast enhancing computed tomography of the abdomen revealed the presence of saccular mycotic aneurysm located at the infrarenal abdominal aorta. The blood culture grew Salmonella Enteritidis which was susceptible to ceftriaxone, trimethoprim-sulfamethoxazole, ciprofloxacin, ampicillin, and amoxicillin-clavulanic acid. Intravenous ceftriaxone was initiated, and he underwent open surgery and artery repair at day 8 of admission. He responded well to the treatment given and subsequently discharged home after completed three weeks of intravenous ceftriaxone.

Salmonella entérica serovar Enteritidis biofilm lifestyle induces lower pathogenicity and reduces inflammatory response in a murine model compared to planktonic bacteria / Salmonella entérica serovar Enteritidis en biopelícula induce una menor patogenicidad y respuesta inflamatoria en un modelo murino que su contraparte planctónica

Abstract Salmonella enterica serovar Enteritidis (S. Enteritidis) is the most frequent serovar involved in human salmonellosis. It has been demonstrated that about 80% of infections are related to biofilm formation. There is scant information about the pathogenicity of S. Enteritidis and its relationship to biofilm production. In this regard, this study aimed to investigate the differential host response induced by S. Enteritidis biofilm and planktonic lifestyle. To this purpose, biofilm and planktonic bacteria were inoculated to BALB/c mice and epithelial cell culture. Survival studies revealed that biofilm is less virulent than planktonic cells. Reduced signs of intestinal inflammation and lower bacterial translocation were observed in animals inoculated with Salmonella biofilm compared to the planktonic group. Results showed that Salmonella biofilm was impaired for invasion of non-phagocytic cells and induces a lower inflammatory response in vivo and in vitro compared to that of planktonic bacteria. Taken together, the outcome of Salmonella-host interaction varies depending on the bacterial lifestyle.

Resumen Salmonella entérica serovar Enteritidis (S. Enteritidis) es la serovariedad más frecuentemente aislada en la salmonelosis humana. Se ha demostrado que alrededor del 80% de las infecciones están relacionadas con la formación de biopelículas. Sin embargo, la información disponible acerca de la patogenicidad de S. Enteritidis y su relación con la producción de biopelículas es escasa. Este trabajo tuvo como objetivo investigar la respuesta diferencial del huésped frente a S. Enteritidis en sus 2 estilos de vida: biopelícula y planctónico. Para ello, se inocularon bacterias en estado de biopelícula o planctónico en ratones BALB/c y cultivo de células epiteliales. Los estudios de supervivencia revelaron que Salmonella en biopelícula fue menos virulenta que su contraparte planctónica. Los animales inoculados con biopelículas presentaron una mayor conservación estructural del intestino y una menor translocación bacteriana que el grupo planctónico. Asimismo, Salmonella en biopelícula mostró una capacidad deficiente para invadir células no fagocíticas e indujo una menor respuesta inflamatoria in vivo e in vitro que las bacterias planctónicas. Se concluye que el resultado de la interacción Salmonella-huésped depende del estilo de vida bacteriano.

Etiological characteristics of a foodborne diseases outbreak of Salmonella enteritidis in Suzhou / 上海预防医学

ObjectiveTo determine the pathogenic cause in a foodborne diseases outbreak of Salmonella enteritidis in a company in Suzhou City， and provide evidence for epidemiological investigation and guidance for clinical treatment. MethodsRelevant specimens were examined for Salmonella， Shigella， Staphylococcus aureus and Vibrio parahaemolyticus. Furthermore， for the isolated Salmonella enteritidis， a micro broth dilution method was used for antimicrobial susceptibility testing， and pulsed field gel electrophoresis （PFGE） was used for molecular typing. ResultsA total of 44 strains of Salmonella enteritidis were detected from 43 anal swabs of the patients in the outbreak， 7 anal swabs of canteen employees， 31 retained food specimens and 6 environmental specimens. A total of 15 antimicrobial susceptibility testings showed that the 44 strains had the same antimicrobial resistance spectrum， which was 100% resistant to cefazolin， ampicillin， ampicillin/sulbactam， polymyxin E and nalidixic acid， suggesting a multi-drug resistance to more than three antibiotics. PFGE cluster analysis showed that the 44 strains had a 100% of genetic similarity. ConclusionThe outbreak is caused by the consumption of food contaminated with Salmonella enteritidis. The isolated strains have multi-drug resistance， which could guide appropriate antimicrobial treatment based on the antimicrobial susceptibility testing.

Meningitis por Salmonella Enteritidis en un lactante: comunicación de un caso y revisión de la literatura / Salmonella Enteritidis meningitis in an infant: case report and literature review

Resumen La meningitis por Salmonella spp. es infrecuente en pediatría, pero debe sospecharse especialmente en menores de seis meses. Salmonella subespecie enterica es la más frecuente a nivel mundial, siendo el serovar Enteritidis el más importante. Tiene una alta tasa de complicaciones, predominando las colecciones y abscesos, por lo que se recomienda realizar una neuroimagen a todos los pacientes. A pesar que el manejo no está estandarizado, hay consenso en que el tratamiento de elección son las cefalosporinas de tercera generación. La resistencia antimicrobiana es infrecuente pero va en aumento, principalmente en países subdesarrollados. Presentamos el primer caso publicado en Chile de una meningitis por S. Enteritidis en un lactante de 11 meses, que se presentó con fiebre y una convulsión focalizada, con hemocultivos y cultivo de líquido cefalorraquídeo positivos. El paciente completó tratamiento con ceftriaxona y ciprofloxacina por cuatro semanas con buena respuesta.

Abstract Salmonella spp meningitis is rare in pediatrics. However, it should be especially suspected in children younger than 6 months. The enteric subspecies is the most frequent worldwide, with the Enteritidis serovar being the most important. The complication rate is high, with collections and abscesses predominating. For this reason, neuroimaging is currently recommended for all patients. Even though management is not standardized, there is consensus that first line treatment should be with third generation cephalosporins. Antibiotic resistance is infrequent but increasing, mainly in underdeveloped countries. We present the first case reported in Chile of meningitis by Salmonella Enteritidis in an 11-month-old infant presenting with fever and focused epileptic crisis. Blood culture and cerebrospinal fluid culture were positive. The patient completed four weeks of treatment with ceftriaxone and ciprofloxacin with a good response.

Antibacterial activity of chitosan biofilm for the conservation of fertile and table eggs / Atividade antibacteriana de biofilme de quitosana para conservação de ovos férteis e de mesa

The aim of this study was to develop a chitosan biofilm against Salmonella enteritidis, for the conservation of fertile and table eggs. Two experiments were performed. Experiment 1: 400 specific pathogen-free table eggs were divided in a completely randomized design into four treatments, five replicates and each replicate with 20 table eggs. Experimental groups were assigned to control and 1, 5 and 10% chitosan treatment. The eggs were immersed in the chitosan solution. They were then exposed to Salmonella enteritidis and stored for 1, 24, 96 and 168h at 4ºC. The eggs were then washed with 10mL of physiological saline solution. Experiment 2: 80 specific pathogen-free fertile eggs were tested, the assays were assigned to control and 1, 5 and 10% chitosan treatment. Each treatment had 20 fertile eggs. The eggs were immersed in the chitosan solution. They were individually weighed and incubated. Egg weight, humidity loss, and hatchability (weight and length of newly hatched chicks) characteristics were assessed. In Experiment 1, comparison between treatments showed differences (P< 0.05) in the total recovered of Salmonella enteritidis on eggshell, with the lower values in 5 y 10% chitosan treatment at 96 y 168h respectively. In Experiment 2, chitosan did not show any effect on the egg weight and chick weight, where the average was 57.44 and 38.23g respectively. The humidity loss and chick length showed differences (P< 0.05), with the lower values in 5 y 10% chitosan treatment. The antibacterial activity of chitosan biofilm provide a practical tool against Salmonella enteritidis in fertile and table eggs because the chitosan did not affect egg weight and chick weight, relevant parameters in the poultry industry.(AU)

O presente estudo teve como objetivo desenvolver um biofilme de quitosana contra Salmonella enteritidis, para conservação de ovos férteis e de mesa. Dois experimentos foram realizados. Experimento 1: 400 ovos de mesa livres de patógenos especificados foram divididos em delineamento inteiramente casualizado em quatro tratamentos, cinco repetições e cada réplica contendo 20 ovos de mesa. Grupos experimentais foram designados para controle e 1, 5 e 10% de tratamento com quitosana. Os ovos foram imersos em solução de quitosana. Em seguida foram expostos a Salmonella enteritidis, e armazenados por 1, 24, 96 e 168h a 4ºC. Após, os ovos foram lavados com 10mL de solução salina fisiológica. Experimento 2: 80 ovos férteis livres de patógenos especificados foram testados. Os ensaios foram atribuídos a controle e 1, 5 e 10% de tratamento com quitosana. Cada tratamento teve 20 ovos férteis. Os ovos foram imersos em solução de quitosana. Em seguida foram individualmente pesados e incubados. Peso dos ovos, perda de umidade e características de eclodibilidade (peso e comprimento dos pintinhos recém-nascidos) foram avaliados. No Experimento 1, a comparação entre tratamentos mostrou diferenças (P< 0,05) na quantidade total recuperada de Salmonella enteritidis na casca, com os menores valores em 5 e 10% de tratamento com quitosana a 96 e 168h respetivamente. No experimento 2, a quitosana não mostrou nenhum efeito no peso do ovo e no peso do pintinho, onde a média foi de 57,44 e 38,23g respetivamente. A perda de umidade e comprimento do pintinho apresentaram diferenças (P< 0,05), com os menores valores em 5 e 10% de tratamento com quitosana. A atividade antibacteriana do biofilme de quitosana, fornece uma ferramenta prática contra Salmonella enteritidis em ovos férteis e de mesa, pois a quitosana não afetou o peso do ovo e peso do pintinho, parâmetros relevantes na indústria avícola.(AU)

Salmonella enteritidis como causa de gangrena de Fournier / Fournier's gangrene caused by Salmonella enteritidis

La gangrena de Fournier es una fascitis necrotizante perineal grave atribuida a la acción sinérgica de diversos patógenos asociados a factores predisponentes del huésped, como la inmunosupresión. A pesar de que se han descripto manifestaciones extraintestinales de salmonelosis, es infrecuente su identificación como agente causal de infecciones de partes blandas; menos común aún es su implicación en la gangrena de Fournier. Nuestro objetivo es describir la presentación, manejo y desenlace de un caso de gangrena de Fournier con cultivos positivos para Salmonella enteritidis.

Fournier's Gangrene is a severe perineal necrotizing fasciitis attributed to the synergistic action of various pathogens associated with host predisposing factors, such as immunosuppression. Although extraintestinal manifestations of salmonellosis have been described, its identification as a causative agent of soft tissue infections is infrequent and its involvement in the Fournier's Gangrene is even less common. Our objective is to describe the presentation, management and outcome of a Fournier's Gangrene case with positive cultures for Salmonella enteritidis.

Salmonella Enteritidis forms biofilm under low temperatures on different food industry surfaces / Salmonella Enteritidis forma biofilme sob baixas temperaturas em diferentes superfícies da indústria de alimentos

ABSTRACT: We evaluated the influence of temperature on the ability of Salmonella Enteritidis (SE) to form biofilms on stainless steel, polyethylene, and polyurethane surfaces under different hygiene procedures. These materials were placed on SE culture and incubated at 42±1 ºC, 36±1 ºC, 25±1 ºC, 9±1 ºC, and 3±1 ºC for 4, 8, 12, and 24 h. Hot water at 45 ºC and 85 ºC, 0.5% peracetic acid solution, and 1% quaternary ammonia were used for hygienization. Biofilm formation occurred at all temperatures evaluated, highlighting at 3 ºC which has not been reported as an ideal temperature for the adhesion of SE to these materials. The SE adhered more often to polyethylene surfaces than to polyurethane and stainless steel surfaces (P<0.05). Peracetic acid and water at 85 ºC had similar hygienization efficiency (P<0.05) followed by quaternary ammonia whereas water at 45 ºC was not effective. SE adhered to these materials under low temperatures which to date have been deemed safe for food preservation.

RESUMO: Avaliou-se o efeito da temperatura na capacidade de Salmonella Enteritidis (SE) formar biofilme em superfícies de aço inoxidável, polietileno e poliuretano e diferentes processos de higienização. Corpos de prova destes materiais foram postos frente a culturas de SE e incubados a 42±1 ºC, 36±1 ºC, 25±1 ºC, 9±1 ºC e 3±1 ºC por 4, 8, 12 e 24 horas. Para a higienização foram testados água aquecida a 45ºC e 85 ºC e soluções de ácido peracético 0,5% e amônia quaternária 1%. Verificou-se a formação de biofilmes em todas as temperaturas avaliadas, ressaltando-se a 3 ºC, ainda não citada como propícia para adesão de SE. Houve maior adesão ao polietileno do que ao poliuretano e ao aço inoxidável (P<0.05). Para higienização, o ácido peracético e a água a 85 ºC tiveram ação semelhante (P<0.05), seguidos por amônia quaternária, enquanto que a água a 45 ºC não foi eficaz. Todos os materiais avaliados propiciaram a aderência de SE, mesmo sob temperaturas baixas, consideradas até então seguras para a conservação dos alimentos.

Comparison of three diagnostic methods for Salmonella enterica serovars detection in chicken rinse / Comparação de três métodos diagnósticos para detecção de Salmonella enterica em lavados de carcaça de frango

Salmonella detection is a key point in food safety testing, because of the frequent association of this pathogen with food poisoning in humans. The standard bacteriological tests currently used for Salmonella-detection are time-consuming; therefore, there is a need to develop alternative methods to accelerate the detection. In order to accelerate Salmonella diagnosis, we used the immunomagnetic separation assay associated with bacteriophage P22 for the rapid detection of the following Salmonella serovars in chicken rinses of drumsticks, artificially contaminated with 5, 10, and 100 CFU/25mL of bacteria: Salmonella enterica subsp. enterica serovar Heidelberg (S. Heidelberg), Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis) and Salmonella enterica subsp. enterica serovar Typhimurium (S. Typhimurium). The efficiency of the technique, represented by the time required for detection of positive and negative samples, was compared with that of the standard diagnostic tests used for this pathogen, the bacteriological assay and the polymerase chain reaction (PCR)-based test. This study confirmed the ability of the bacteriophage-associated immunomagnetic separation assay to identify 99.6% of Salmonella-positive samples of the three serovars tested. In contrast, the bacteriological assay and PCR-based test detected 95.1% and 98.5% of the Salmonella-positive samples respectively.(AU)

A detecção de Salmonella é um ponto crucial para a segurança alimentar, devido a frequente associação deste patógeno com infecções alimentares em humanos. O método padrão para detecção de Salmonella é o bacteriológico, mas o tempo requerido para o processamento das amostras e o diagnóstico final é longo, por isso existe a necessidade de desenvolvimento de métodos alternativos que visem acelerar esta etapa. Para isto utilizamos a separação imunomagnética associada ao bacteriófago P22 como técnica de detecção rápida para os seguintes sorovares de Salmonella: Salmonella enterica subsp. enterica sorovar Heidelberg (S. Heidelberg), Salmonella enterica subsp. enterica sorovar Enteritidis (S. Enteritidis) e Salmonella enterica subsp. enterica sorovar Typhimurium (S. Typhimurium), os quais foram inoculados artificialmente em lavados de sobre-coxas de frango nas seguintes concentrações: 5, 10 e 100 UFC/25mL. A eficiência da técnica, representada pelo tempo requerido para detecção de amostras positivas ou negativas, foi comparado com os testes rotineiramente utilizados para detecção de Salmonella, o exame bacteriológico e a reação em cadeia da polimerase (PCR). Este estudo confirmou a capacidade do teste de separação imunomagnética associado a bacteriófago, o qual identificou 99,6% das amostras positivas para Salmonella, dos três sorovares testados. Já o bacteriológico e PCR identificaram respectivamente 95,1% e 98,5% das amostras positivas.(AU)

Infección por Salmonella enteritidis en pacientes con lupus eritematoso sistémico / Salmonella enteritidis infection in patients with systemic lupus erythematosus

RESUMEN La infección por Salmonella enteritidis (ISE) es rara en pacientes con lupus eritematoso sistémico (LES), y comúnmente se confunde con exacerbaciones del LES. ISE en pacientes con LES se ha relacionado con malos desenlaces. Este artículo describe los hallazgos clínicos y desenlaces de 5 pacientes con LES y diagnóstico de ISE, en una cohorte de 325 pacientes (1,53%). Todos los pacientes fueron mujeres, con una edad promedio de 28,2 (14 a 37 años). La duración promedio del LES antes de la ISE clínica fue de 3,2 años (1 a 7 años). No hubo evidencia de infección recurrente. Los pacientes fueron tratados con ciprofloxacino o ceftriaxona, con resolución de las manifestaciones clínicas en 4 pacientes. Un paciente murió debido a falla orgánica multisistémica. Nuestro reporte remarca las manifestaciones clínicas inespecíficas de la ISE que asemeja actividad lúpica. Un adecuado diagnóstico y una terapia antibiótica temprana son esenciales para mejorar los desenlaces de estos pacientes.

ABSTRACT Salmonella enteritidis infection (SEI) is rare in systemic lupus erythematosus (SLE) patients, and it is often mistaken with SLE flares. SEI in SLE patients has been associated with a poor outcome. A description is presented of the clinical features and outcomes of five patients with an SEI from a cohort of 325 patients with SLE (1.53%). All patients were women, with a mean age of 28.2 years (14 to 37 years). Mean duration of SLE before clinical SEI was 3.2 years (1 to 7 years). There was no evidence of recurrent infection. The patients were treated with ciprofloxacin and/or ceftriaxone, with resolution of clinical manifestations in four of them. One patient died due to multisystem organ failure. Our report highlights the unspecific clinical manifestations of SEI that resembles lupus flare-up. An accurate diagnosis and early antibiotic treatment are essential to improve their outcomes.

Defined and undefined commercial probiotics cultures in the prevention of Salmonella Enteritidis in broilers / Probióticos comerciais de culturas definidas e indefinidas na prevenção de Salmonella Enteritidis em frangos de corte

This study aimed to evaluate the efficacy of probiotics from different formations, defined and undefined cultures, applied in the control of Salmonella Enteritidis in broilers, identifying the compositions and states for which the probiotics are more effective. For that, 390 broilers were inoculated orally with 1.00 ml of Salmonella Enteritidis at a concentration of 1.2x109 CFU (Colony Forming Units). The experimental design used was randomized blocks with 5 treatments and 6 replications, totaling 30 boxes with 13 birds/box (13 birds/m2). The treatments were provided via drinking water 1 hour after inoculation, keeping a daily treatment of 12 hours with probiotics, for 3 consecutive days (birds at 1, 2 and 3 days of age). In general, the five treatments conducted were: T1 - Control without probiotic, T2 - Probiotic A (defined culture - lyophilized form, strain 7), T3 - Probiotic B (defined culture - lyophilized form, strain 11), T4 - Probiotic C (undefined culture liquid form), T5 - Probiotic D (undefined culture - liquid form). After treatments, performance was evaluated through average body weight, feed conversion and mortality counting. Microbiological analysis and Salmonella isolation were performed using MPN (Most Probable Number) and selective enrichment technique methods, respectively. Samples of ileum and liver pool, cecal tonsils, cecum, heart and spleen pool were collected at 5 and 31 days of age. No differences were observed on growth performance and isolation of Salmonella Enteritidis (p≥0.05). All probiotics applied were effective on reducing Salmonella Enteritidis colonization in the ileum, cecal tonsils, and cecum at 5 days of life. Probiotics T2 and T5 has shown effectiveness in reducing colonization at 31 days, being considered the most efficient on Salmonella Enteritidis control, for the intestines segments evaluated. It was not possible to affirm which probiotics formation, defined or undefined, is more efficient for Salmonella Enteritidis control.(AU)

O objetivo deste trabalho foi avaliar a eficácia dos probióticos de diferentes constituições: de culturas definidas e de culturas indefinidas no controle de Salmonella Enteritidis em frangos de corte, identificando qual a constituição e qual ou quais probióticos testados é mais eficaz. Foram inoculados 390 frangos de corte com 1ml de Salmonella Enteritidis, via oral, na concentração de 1,2 x 109 UFC (Unidades Formadoras de Colônia). O delineamento experimental utilizado foi o de blocos casualizados com 5 tratamentos e 6 repetições cada, totalizando 30 boxes com 13 aves/boxe (13 aves/m2). Os tratamentos foram fornecidos via água de bebida 1 hora após a inoculação, com 12 horas de tratamento com probióticos por dia, durante 3 dias consecutivos (1º, 2º e 3º dia de idade das aves). Os cinco tratamentos foram: T1 - Controle sem probiótico, T2 - Probiótico A (cultura definida - forma liofilizada, 7 cepas), T3 - Probiótico B (cultura definida - forma liofilizada, 11 cepas), T4 - Probiótico C (cultura indefinida - forma líquida), T5 - Probiótico D (cultura indefinida - forma liofilizada). O desempenho zootécnico foi avaliado usando o peso médio, a conversão alimentar e a mortalidade. Análises microbiológicas foram realizadas utilizando o método NMP (NMP/g)e isolamento de Salmonella através técnica de enriquecimento seletivo. Amostras de pool de íleo, tonsilas cecais e cecos e pool de fígado, coração e baço foram coletadas aos 5 dias e aos 31 dias de idade. Para desempenho zootécnico e isolamento de Salmonella Enteritidis não foram observadas diferenças (p≥0,05). Todos os probióticos utilizados foram eficazes na redução da colonização de Salmonella Enteritidis no íleo, tonsilas cecais e cecos aos 5 dias de idade e somente os probióticos do T2 (cultura definida) e T5 (cultura indefinida) reduziram a colonização aos 31 dias sendo considerados os mais eficazes no controle de Salmonella Enteritidis nestes segmentos intestinais avaliados. Não se pode afirmar quais das constituições de probióticos, culturas definidas ou indefinidas, são mais eficazes no controle de Salmonella Enteritidis.(AU)

Characteristics of drug resistance and molecular typing of Salmonella enteritidis isolated in Shanxi Province / 中华微生物学和免疫学杂志

Objective@#To investigate the antimicrobial resistance status and pulsed field gel electrophoresis (PFGE) patterns of Salmonella enteritidis (S.enteritidis) in Shanxi Province in order to provide references for the treatment of Salmonella infection and for tracing the source of outbreaks of foodborne diseases.@*Methods@#Sixty-four S. enteritidis strains were collected by monitoring sites for foodborne diseases from April 2015 to March 2018. Biochemical identification system and serotyping analysis were used for bacterial identification. Drug susceptibility patterns were analyzed with micro-broth dilution method. PFGE was used for molecular typing.@*Results@#The antimicrobial resistance rate of 64 S. enteritidis strains to nalidixic acid (95.31%) was the highest, followed by that to ampicillin (90.63%) and to ampicillin/sulbactam (81.25%). They had lower resistance rates to cefazolin, cefotaxime, tetracycline, ceftazidime, trimethoprim/sulfamethoxzole and ciprofloxacin (3.13%-23.44%) and were all sensitive to chloramphenicol, cefotaxime, azithromycin, imipenem and gentamicin. No statistically significant difference in drug resistance rates was found between the sporadic strains and the outbreak strains (P>0.05). All 64 S. enteritidis strains digested with XbaⅠwere divided into 33 molecular patterns by PFGE. The numbers of bacteria contained in each pattern ranged from 1 to 10 strains. The similarity among patterns was between 54.6% and 100%.@*Conclusion@#More attention should be paid to the drug resistance status of S. enteritidis in Shanxi Province. It is necessary to strengthen the standardized administration of antibiotics. The PFGE patterns of S. enteritidis reveal the presence of significant genetic polymorphism. PFGE is of great significance in analyzing the genetic relationship among S. enteritidis strains and in identifying and tracing the source of outbreaks of foodborne diseases.

Production of recombinant flagellin to develop ELISA-based detection of Salmonella Enteritidis

ABSTRACT Food-borne diseases, caused by the pathogenic bacteria, are highly prevalent in the world. Salmonella is one of the most important bacterial genera responsible for this. Salmonella Enteritidis (SE) is one of the non-typhoid Salmonellae that can be transmitted to human from poultry products, water, and contaminated food. In recent years, new and rapid detection methods such as enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) have been developed. In this study, recombinant FliC (rFliC) was produced to be used as an antigen. The immunization was conducted in mice with the purified recombinant FliC (rFliC). The mice were subcutaneously immunized with rFliC and elicited significant rFliC specific serum IgG antibodies. An indirect ELISA system was established for the detection of Salmonella Enteritidis. Our results confirmed that the recombinant flagellin can be one of the excellent indicators for the detection of Salmonella Enteritidis.

Phenotypic and genotypic characterization of Salmonella spp. isolated from foods and clinical samples in Brazil

ABSTRACT Increasing antimicrobial resistance in Salmonella species has been a serious problem for public health worldwide. This study examines Salmonella spp. recovered from foods and clinical samples on serotype, antimicrobial resistance and PFGE genotypes. It identified 91 salmonellae, belonging to 31 different serotypes, from 36 isolates from food and 55 clinical samples. Salmonella Infantis (16.5%) and Salmonella Enteritidis (13.7%) are the most common among food isolates, whereas Salmonella Enteritidis (29.0%) and Salmonella Typhimurium (16.0%) mainly causes human salmonellosis. Antimicrobial susceptibility data showed that 63.0% of the isolates were fully susceptible to 12 antibiotics tested. Nalidixic acid showed high resistance rates, 32.7% and 25.0% of the clinical isolates and food, respectively. Three main PFGE types: A (Salmonella Enteritidis), B (Salmonella Infantis) and C (Salmonella Schwarzengrund) comprised isolates recovered from foods and clinical samples. Our results indicate that the clonal groups were both causing diseases and food contamination, emphasizing the need to maintain a system of surveillance for foodborne disease.

Surto de doença trasmitida por alimentos nos municipios de Mauá e Ribeirão Pires - SP / Outbreak of foodborne diseases in the municipalities of Maua and Ribeirao Pires - SP

Doenças de transmissão hídrica e alimentar (DTHA) acarretam importantes problemas econômicos e de saúde pública no mundo atual. Este estudo relata um surto de Doença Transmitida por Alimento - DTA que envolveu 12 pessoas de duas residências localizadas na Região do ABC paulista em dezembro de 2012. Quatro pessoas de uma residência tiveram sintomas de diarreia, cólica abdominal, náusea, vômito, febre e prostração, sendo que apenas duas consumiram o bolo preparado em Ribeirão Pires, SP - Brasil. Outras oito pessoas consumiram o mesmo alimento no município de Mauá e, além dos sintomas citados, houve também registro de insuficiência renal e parada cardiorrespiratória. Dentre os envolvidos, uma menina de oito anos veio a óbito após convulsão e bronco-aspiração. O período variou entre 2 e 22 horas após o consumo do alimento. A amostra de bolo foi analisada segundo a metodologia preconizada pelo BAM-FDA e teve como resultados: Coliformes termotolerantes (NMP = 4,6x104/g); Bacillus cereus (1,5x105 U.F.C./g) e presença de Salmonella Enteritidis em 25 gramas. Clostridium perfringens, Staphylococcus aureus e Listeria monocytogenes não foram isolados. Foram realizadas duas coproculturas que apresentaram resultados positivos para Salmonella Enteritidis. As cepas de Salmonella spp isoladas, tanto no alimento como nas fezes dos pacientes, apresentaram similaridade genética e mesmo perfil de suscetibilidade aos antimicrobianos. Assim, foi constatado o envolvimento do bolo como veiculador de patógenos e ressaltada a importância do trabalho em conjunto das vigilâncias sanitárias e epidemiológicas de ambos os municípios e o laboratório de referência em saúde pública, fundamental na elucidação deste surto.

Enteropatógenos predominantes en diarreas agudas y variables asociadas en niños atendidos en el Hospital Regional Lambayeque, Perú / Predominant enteropathogens in acute diarrhea and associated variables in children at the Lambayeque Regional Hospital, Peru

Objetivo: Determinar el tipo y frecuencia de enteropatógenos predominantes en diarreas agudas y sus características asociadas en niños atendidos en el Hospital Regional Lambayeque (HRL) - Perú. Materiales y métodos: Se realizó un estudio analítico transversal entre marzo y mayo del 2015 en 70 muestras fecales. Las muestras se estudiaron mediante coprocultivo e inmunocromatografía para la detección de bacterias y virus enteropatógenos respectivamente. Mientras que los enteroparásitos se buscaron mediante examen microscópico directo, tinción de Kinyoun y ELISA para coproantígenos (Entamoeba histolytica, Giardia lamblia y Cryptosporidium spp.). Asimismo se realizó conteo de leucocitos y pruebas químicas (Benedict, Thevenon y Sudan III) para el estudio funcional de la enfermedad diarreica. Resultados: En el 48,6% de muestras se detectó la etiología infecciosa de la diarrea, siendo predominante la causa parasitaria (25,8%), seguida de la bacteriana (17,1%) y viral (5,8%). Los enteropatógenos más frecuentes fueron G. lamblia (18,6%) y Salmonella Enteritidis (10,0%). Se observó asociación entre la cantidad de leucocitos mayor a 100 con la etiología bacteriana (p=0,027), mientras que un número menor de 10 por campo (p=0,002) y el Sudan III positivo (p=0,003) con la etiología parasitaria. Conclusiones: En más de la mitad de muestras (51,4%) no se demostró etiología infecciosa de la diarrea, mientras que Giardia lamblia fue la más frecuente causa de diarrea en la población estudiada. No obstante, es necesaria la implementación de técnicas más sensibles y específicas para la detección de un rango mayor de enteropatógenos con el que se mejore el diagnóstico y tratamiento de la enfermedad

Objective: To determine the type and frequency of predominant enteropathogens in acute diarrhea and their associated characteristics in children treated at Hospital Regional Lambayeque (HRL) - Peru. Materials and methods: A cross-sectional analytical study was carried out in 70 fecal samples between March and May 2015. These samples were studied by coproculture and immunochromatography for the detection of enteropathogenic bacteria and viruses, respectively, while enteroparasites were sought by direct microscopic examination, Kinyoun staining method and ELISA for the detection of coproantigens (Entamoeba histolytica, Giardia lamblia and Cryptosporidium spp). Leukocyte count and chemical tests (Benedict, Thevenon and Sudan III) were also performed for the functional study of the diarrheal disease. Results: In 48.6% of the samples, the infectious etiology of diarrhea was detected, prevailing the parasitic cause (25.8%), followed by the bacterial (17.1%) and viral (5.8%) ones. The most common enteropathogens were G. lamblia (18.6%) and Salmonella enteritidis (10.0%). An association between greater than 100 fecal leukocytes per field and the bacterial etiology (p=0.027) was observed, while less than 10 fecal leukocytes per field (p=0.002) and a positive Sudam III test (p=0.003) were associated with the parasitic etiology. Conclusions: In more than half of the samples (51.4%) the infectious etiology of diarrhea could not be proven, whereas Giardia lamblia was the most frequent cause of diarrhea in the studied population. However, it is necessary to implement more sensitive and specific techniques for the detection of a greater range of enteropathogens with which to improve the diagnosis and treatment of the disease

Characteristics of drug resistance and molecular typing for Salmonella enteritidis isolated in He'nan Province from 2013 to 2015 / 中华传染病杂志

Objective To investigate the etiological,antimicrobial resistance status and pulsed field gel electrophoresis (PFGE) patterns of S.enteritidis strains in Henan Province.Methods Totally 82 strains of S.enteritidis strains were isolated from seven sentiel hospitals which were able to detect multiple pathogens from April,2013 to December,2015.According to Salmonella K-B drug susceptibility testing and molecular typing method published by the USA clinical and Laboratory Standards Institute (2015),the drug sensitivity to 7 kinds of antimicrobial agents and PFGE molecule characteristics of 82 S.enteritidis strains were tested.The PFGE patterns were analyzed by BioNumerics 6.0 software based on international PulseNet bacterial infectious disease monitoring network.Results Forty-seven strains of S.enteritidis were isolated from male patients,35 strains isolated from female patients.A total of 56 S.enteritidis strains were isolated from young children aged from 0 to 5 years old,including 29 strains isolated from 6 months to 2 years old.The strains were mainly isolated between May and November of the year,and 15 (18.3％) strains were isolated between March and April,27(32.9％) strains between May and July,34 (41.5％) strains between August and October,6 (7.3％) strains were isolated in the rest time of the year,with a typical seasonal characteristics of summer and autumn.Sxity-four (78.0％)strains of S.enteritidis were resistant to ampicillin;23 (28.0％) strains were resistant to ceftazidime;39 (47.6％) strains were resistant to cefotaxime:13 (15.9％) strains were resistant to cefepime;55 (67.1％) strains were resistant to nalidixic acid;24 (29.3％) strains were resistant to ciprofloxacin;32 (39.0 ％) strains were resistant to gentamicin;14 (17.1 ％) strains were resistant to chloramphenicol;47 (57.3％) strains were resistant to methicillin benzyl ammonium;13 (15.9％) strains were resistant to sulfamethoxazole and trimethoprim;21 (25.6 ％) strains were resistant to tetracycline.All strains weremuhi-drug resistant,and 21 (25.6％) isolates were resistant to 3-4 kinds of antibiotics,32 (39.0％)isolates were resistant to 5-6 kinds of antibiotics, 29 (35.4 ％) isolates were resistant to 7-8 kinds ofantibiotics.These 82 strains of S.enteritidis were divided into 25 molecular patterns by digestion withXba Ⅰ enzyme.Each pattern contained 1 to 26 strains with similarity ranged from 59.33％-100.00％.EN1 and EN2 were the main PFGE belt types and included 26 and 16 strains,respectively.Conclusions The drug resistance of clinical isolates of S.enteritidis in Henan province is serious.PFGE patterns show polymorphism and the dominant patterns,part of which are associated with drug-resistance spectrum and show aggregation.

Analysis of the characteristics of Salmonella enteritidis virulence genes from different origins / 中国食品卫生杂志

Objective To investigate the differences and characteristics of virulence genes carried by Salmonella enteritidis from different sources in Shijiazhuang City.Methods One hundred and twenty-four strains of Salmonella enteritidis isolated from morning markets of raw and poultry stalls,slaughterhouses and food poisoning specimens in Shijiazhuang area were collected.Eight virulence genes (invA,sopE,agfA,spvR,hilA,stn,pefA,shdA) were detected by polymerase chain reaction (PCR).Results Salmonella enteritidis might have different virulence gene profiles.The above eight virulence genes were detected in different strains.The carrying rate of virulence genes invA,sopE,stn,hilA,spvR and pefA in the food poisoning strains was higher than 94％.There was no difference in the carrying rate of 8 virulence gene between the morning raw poultry stalls isolates and the patient strains,but was different with the slaughterhouse strains.Conclusion There were more risks of food poisoning caused by Salmonella enteritidis from morning markets,and the hygiene supervision should be strengthened to prevent and control foodborne disease.

Assessment of probiotic potential of Lactobacillus reuteri MD5-2 isolated from ceca of Muscovy ducks

Lactic acid bacteria (LAB) are commonly used as probiotics in poultry. The present study employed in vitro and in vivo methods to select and test LAB isolated from Muscovy duck ceca as potential probiotics. In the in vitro study, 50 LAB were isolated from Muscovy duck ceca and tested for growth inhibition against Salmonella (S.) Enteritidis. Eleven isolates strongly inhibited S. Enteritidis and only 1 isolate (MD5-2) showing the strongest inhibition was selected for identification. This isolate was called as Lactobacillus (L.) reuteri MD5-2. For the in vivo investigation, 90 1-day-old Muscovy ducks were randomly assigned into three groups of 30 animals each (group 1, control; group 2, treated with 108 colony-forming unit (CFU) of L. reuteri MD5-2 orally once on day 1; and group 3, treated with 108 CFU of L. reuteri MD5-2 orally once daily from days 1 to 5). The ducks were housed in three large cages and raised for 50 days, after which body weight, duodenal villus height and crypt depth were measured. Both villus height and villus height to crypt depth ratio were significantly greater in group 3 than in groups 1 and 2. In conclusion, further investigation of L. reuteri MD5-2 as a potential probiotic strain is warranted.

Avaliação dos efeitos de 5-hidroxitriptofano em-hidroxibenzilhidrazine associados a Lactobacillus spp. na morfometria intestinal e imunomarcação de serotonina em frangos de corte desafiados com Salmonella Enteridis / Assessment of the effects of 5-hydroxytryptophan and m-hydroxybenzyl hydrazine associated with Lactobacillus spp. in intestinal morphology and immunostaining of serotonin in broilers challenged with Salmonella Enteritidis.

As células enterocromafins são um dos componentes da mucosa intestinal que liberam serotonina para o lúmen, promovendo atividades secretórias e crescimento celular de vários tecidos, incluindo vilosidades intestinais. O presente estudo avaliou as influências do 5-hidroxitriptofano (5HTP) e do m-hidroxibenzilhidrazine (NSD1015), associados a Lactobacillus spp., sobre o peso corporal e o desenvolvimento das vilosidades intestinais na porção proximal do duodeno de frangos de corte desafiados com Salmonella Enteritidis. Verificou-se também se a presença de Lactobacillus spp. e Salmonella Enteritidis influenciaram a imunomarcação de serotonina no duodeno e, para isso, o estudo foi dividido em dois experimentos, com e sem desafio por S. Enteritidis. No Experimento 1, em aves sem desafio, os pesos corporais não diferiram significantemente (p>0,05) e, no Experimento 2, aves com desafio, os tratamentos com o precursor isolado e associado a Lactobacillus spp. determinaram maior peso corporal das aves. Nos dois experimentos, as aves tratadas com 5HTP apresentaram aumento na densidade e altura das vilosidades no duodeno, sugerindo a atuação de 5HTP como um agente trófico. A administração de Lactobacillus spp. também determinou altura maior de vilosidades duodenais. Quanto a imunomarcação de serotonina, as aves tratadas com Lactobacillus spp. no Experimento 1 e as aves tratadas com Lactobacillus spp. e desafiadas com S. Enteritidis no Experimento 2, apresentaram valores superiores aos demais tratamentos, sugerindo que a presença destas bactérias promove maior liberação de serotonina para o duodeno, porém o mecanismo exato de como este processo ocorre necessita ser mais elucidado...

Enterochromaffin cells are components of the intestinal mucosa to release serotonin lumen, promoting cell growth and secretory activity of various tissues, including intestinal villi. This study evaluated the influence of 5-hydroxytryptophan (5HTP) and m-hidroxibenzilhidrazine (NSD1015) associated with Lactobacillus spp. on body weight and development of intestinal villi in the proximal duodenum of broilers challenged with Salmonella Enteritidis. It was found that the presence of Lactobacillus spp. and Salmonella Enteritidis influenced immunostaining of serotonin in the duodenum. The study was divided into two experiments with and without challenge by S. Enteritidis. In Experiment 1, birds without challenge, body weights did not differ significantly (p>0.05), and in Experiment 2, the treatments with precursor and precursor associated with Lactobacillus spp. determined higher body weight of the birds. In both experiments the birds treated with 5HTP showed increased density and villus height in the duodenum, suggesting the presence of 5HTP as a trophic agent. The use of Lactobacillus spp. also determined greater duodenal villus height. The immunostaining of serotonin, birds treated with Lactobacillus spp. in Experiment 1, and the birds treated with Lactobacillus spp. and challenged with S. Enteritidis in Eperiment 2 showed higher values, suggesting that the presence of these bacteria promotes greater release of serotonin into the duodenum. The exact mechanism of how this process occurs needs to be further elucidated...