En production avicole, S. Senftenberg connue pour être souvent associée au couvoir, est devenue très fréquente dans les élevages de volaille. L’objectif de la thèse est donc de comprendre les causes de cette persistance. Les résultats présentés suggèrent que l’existence de souches persistantes au sein du sérovar Senftenberg peut être à l’origine de l’augmentation de sa prévalence dans les élevages. Nous avons en effet identifié des souches présentant des phénotypes distincts en termes de persistance dans les caeca de poulets. Les souches persistantes ont la capacité d’induire un portage intestinal similaire à S. Enteritidis.Toutefois, les résultats in vivo ont montré qu’à la différence de S. Enteritidis, les souches de S.Senftenberg sont incapables d’induire une forte infection systémique chez le poulet et la souris,probablement due à leur faible capacité à résister aux cellules immunitaires. La comparaison entre les souches persistantes et non-persistantes n’a montré aucune différence de survie dans le contenu et le mucus caecal. Cependant, les souches persistantes ont une meilleure capacité à coloniser et persister dans les tissus d’animaux ce qui pourrait être une explication possible à l’augmentation de leur persistance. Ce caractère pourrait donc présenter un risque pour la santé humaine étant donné que ces bactéries peuvent être présentes chez les animaux avant l'abattage et la transformation des aliments. / In poultry production, S. Senftenberg was associated to the hatchery, but has recently become morefrequent in poultry farms. The aim of our study is to explain the increased persistence of this serovar. Our findings suggest that the existence of persistent strains within the serovar Senftenberg could explain its recent emergence in poultry flocks. We identified strains showing different persistence phenotypes inchicken caeca. The persistent strains are able to induce an intestinal asymptomatic carrier state in chicks similar to S. Enteritidis. However, the in vivo analysis showed that S. Senftenberg strains, contrary to S.Enteritidis are unable to induce a strong systemic infection in infected mice and chickens which could be in part related to their low capacity to resist to immune cells. The comparison between persistent and nonpersistents trains showed no difference in their ability to grow in the caecal content and mucus. However,persistent strains are more able to colonize and persist in chickens and mice tissues, which could be a possible explanation for their increased persistence. This trait might thus present a human health risk as these bacteria could be present in animals before slaughter and during food processing.

Made available in DSpace on 2015-03-26T13:51:56Z (GMT). No. of bitstreams: 1 texto completo.pdf: 462563 bytes, checksum: 8c4d15a05d7b212d40f89724706d1c9a (MD5) Previous issue date: 2006-09-19 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / Considering that the acid tolerance is an important virulence factor related to survival at low pH of gastric barrier of foodborne pathogens and the expressive increasing of prevalence of Salmonella serovar Enteritidis in recent years as etiologic agent of human salmonelosis, this work aimed to evaluate the effect of acid and storage under low temperatures on Salmonella serovar Enteritidis. The growth of S. Enteritidis CCS3 isolated of swine carcass and S. Enteritidis ATCC 13076 at Trypticasein and Soya Broth (TSB) with pH values between 3.5 and 7.2 and the survival in the same media with pH values between 1.5 and 3.5 were investigated. The tolerance to simulated gastric fluid (SGF) of these cells previously sublethal acid treatment pH 4.0, 4.5 and 5.0 inoculated in pork maintained storage by 7 days at 4 ± 1°C and by 84 days at freezing temperature (-18°C ± 1) was also evaluated. S. Enteritidis CCS3 presented higher values of specific growth rate in acidified TSB and this culture had longer time survival in pH < 3,5 than S. Enteritidis ATCC 13076. The storage in pork by seven days at 4 ± 1°C did not effect the populations of S. Enteritidis studied. After 84 days of cold storage, the media of population reductions of S. Enteritidis CCS3 and ATCC 13076 were 0.8 and 1.5 log cycle, respectively. The previous acid treatment did not enhance the survival of both cultures at low temperatures. After low temperature storage, the cells of S. Enteritidis CCS3 were tolerant until three hours SFG exposure. Instead, cells of S. Enteritidis ATCC 13076 lost culturability after 10 minutes of SFG challenge. S. Enteritidis CCS3 cells submitted to acid treatment at pH 4.0 showed are more tolerant to 180 minutes SFG exposure (15% of population reduction) than cells submitted to other acid treatments (pH 4.5 and 5.0) and cells of control (without acid treatment) with 30% of population reduction. / Considerando que a tolerância ao ácido é um importante fator de virulência relacionado à sobrevivência ao baixo pH da secreção gástrica em patógenos veiculados por alimentos e o expressivo aumento da prevalência de Salmonella sorovar Enteritidis nos últimos anos, como agente etiológico de salmonelose humana, este trabalho teve como objetivo avaliar o efeito de ácido e da estocagem sob temperaturas baixas em Salmonella sorovar Enteritidis. O crescimento de S. Enteritidis CCS3, isolada de carcaça suína, e S. Enteritidis ATCC 13076 foi determinado em Caldo Tripticaseína e Soja (TSB) com valores de pH que variaram de 3,5 a 7,2. A sobrevivência foi avaliada no mesmo meio com pH entre 1,5 e 3,5. A tolerância ao Fluido Gástrico Simulado (FGS) dessas bactérias previamente submetidas a tratamento ácido subletal em pH 4,0; 4,5 e 5,0 inoculadas em carne suína estocada por sete dias a 4 ± 1°C e por 84 dias sob congelamento a -18 ± 1°C foi também avaliada. S. Enteritidis CCS3 apresentou valores maiores de velocidade específica de crescimento em TSB acidificado e sobreviveu por mais tempo em pH inferior a 3,5 do que S. Enteritidis ATCC 13076. A estocagem em carne suína por sete dias a 4 ± 1°C não afetou as populações de S. Enteritidis investigadas. Após 84 dias de congelamento, as reduções médias das populações de S. Enteritidis CCS3 e ATCC 13076 foram de 0,8 e 1,5 ciclos logarítmicos, respectivamente. O tratamento ácido aplicado previamente não teve efeito sobre a sobrevivência das duas culturas sob baixas temperaturas. Após a estocagem sob temperaturas baixas, as células de S. Enteritidis CCS3 apresentaram tolerância à exposição ao FGS por até três horas. Contrariamente, células de S. Enteritidis ATCC 13076 perderam a culturabilidade após 10 minutos de desafio ao FGS. Células de S. Enteritidis CCS3 submetidas a tratamento ácido prévio em pH 4,0 mostraram-se mais tolerantes à exposição por 180 minutos ao FGS (redução de 15%) que células submetidas aos tratamentos ácidos em pH 4,5 e 5,0 e células do tratamento controle (sem tratamento ácido) com redução de 30% da população.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The objective of this study work was to evaluate the effect of two organic acids (formic acid and propionic acid) and of one mannanoligosaccharide added to the diet to control the intestinal infection caused by Salmonella Enteritidis in broilers. In these 39 days of study it was used 150 birds, 1-day-old, of the Cobb lineage, both sexes and free of Salmonella Enteritidis. They were divided in 6 different treatments (T) with 25 birds each, where: T1 (diet and no infection), T2 (diet + organic acids + no infection), T3 (diet + organic acids + mannanoligosaccharide and no infection), T4 (diet + organic acids and infection with Salmonella Enteritidis) T5 (diet + organic acids + mannanoligosaccharide + and infection with Salmonella Enteritidis) T6 (diet and infection with Salmonella Enteritidis). After housing, the chicken litter was instilled on the 4th day with Salmonella Enteritidis and every seven days, five birds from each group were killed through cervical dislocation. The necropsy was performed and also the bacteriological exams to detect Salmonella Enteritidis using the feces collected over the chicken litter of the groups bacteriological analysis of the cecal tonsils was done as well. On the 18th day only 60% of birds were infected in treatments T4 and T5; on the 25th day, 40% of birds in T4 and 20% in the T5 were infected; on the 32nd day, 100% of tested samples were negative in both treatments. The T6 group was 100% positive until the 32nd day, but on the 39th day, it got reduced in 20% of the number of infected animals. In the experimental conditions of this study, the organic acids and the mannanoligosaccharide added to the diet possibly contributted to control the infection caused by Salmonella Enteritidis on tested birds. / O objetivo deste estudo foi avaliar a ação de dois ácidos orgânicos (ácido fórmico e ácido propiônico) e de um mananoligossacarídeo (MOS) adicionados à dieta no controle da infecção intestinal por Salmonella Enteritidis (SE) em frangos de corte. Neste estudo de 39 dias foram utilizadas 150 aves, de um dia de idade, da linhagem Cobb, lote misto, livre de SE, divididos em seis tratamentos (T) com 25 animais cada, onde: T1 (dieta e ausência de infecção), T2 (dieta + ácidos orgânicos e ausência de infecção), T3 (dieta + ácidos orgânicos + MOS e ausência de infecção), T4 (dieta + ácidos orgânicos e infecção com SE), T5 (dieta + ácidos orgânicos + MOS e infecção com SE) e T6 (dieta e infecção com SE). No 4º dia após o alojamento, a cama foi instilada, com SE e, a cada sete dias, cinco aves por grupo foram submetidas à eutanásia por deslocamento cervical, necropsiadas e realizados os exames bacteriológicos para SE, utilizando-se fezes coletadas sobre a cama de maravalha dos grupos, e das tonsilas cecais dos animais necropsiados. No 18º dia, somente 60% das aves estavam infectadas nos tratamentos T4 e T5; no 25º dia, 40% das aves no T4 e 20% no T5 estavam infectadas; no 32º dia 100% das amostras testadas foram negativos em ambos os tratamentos (T4 e T5). Constatou-se que o T6 foi 100% positivo até o 32º dia, e no 39º dia reduziu em 20% o número de animais infectados. Dentro dos parâmetros de avaliação deste experimento, os ácidos orgânicos e o mananoligossacarídeo adicionados à dieta, possivelmente contribuíram no controle da infecção por SE nas aves testadas.

Made available in DSpace on 2014-07-29T15:07:29Z (GMT). No. of bitstreams: 1 Dissertacao_Dunya_Moraes.pdf: 432826 bytes, checksum: 63ffc1c5c83c7f760eb90cca2350b35f (MD5) Previous issue date: 2010-03-26 / The objective of this research to investigate the presence of Salmonella sp. in raw materials of animal origin used in the manufacture of feed for broilers, in diets collected directly from bird feeders, and in organs ceca contents and liners carry case for newly hatched chicks, in environmental samples, in samples from swabs of the hands of officials of the farm and slaughterhouse samples and classify and determine the resistance of strains of Salmonella sp. found, before the action of chemotherapeutic nine. For data analysis was descriptive frequency results. 1200 samples were collected from flour and Salmonella sp. was found in 10.5% of samples with a predominance of serovar Enteritidis. The frequency of bacteria in meat meal was 12%, 6.8% in blood, the feathers of 4.3% and 14.6% in the viscera. Were also isolated Salmonella Cerro, Salmonella Montevideo, Salmonella anatum, Salmonella Tennessee, and Salmonella typhimurium among others. Regarding the resistance of strains found in the various categories of flour was observed resistance to sulfonamides, neomycin, tetracycline, sulphamethoxazole-to trimetopim and florfenicol. Of the three strains of bacteria isolated from two diets were of a Salmonella enteritidis and S. Anatum, showing resistance to sulfonamides and neomycin. Of the 32 batches of newly hatched chicks 9.4% were positive for Salmonella and 32 batches of liners carrying case 9.4%. Environmental samples, before bed accommodation, swabs of feeder and drinker and the drinking water of birds tested negative for Salmonella sp .. Drag swabs of poultry manure, and samples Alphitobius diaperinus swabs from the hands of officials of the farms had a frequency of 12.5%, 12.5% and 6.5% respectively. In samples from swabs of drag serovar Enteritidis was the most frequent and catfishes of the samples and swabs of hands was the only serovar isolated. In samples from slaughterhouse 26.7% of the lots from crop and 33.3% of batches of ceca were positive for Salmonella sp .. Regarding the resistance of strains, there was resistance sulfonamides, the amoxicillin and enrofloxacin in samples of newly hatched chicks and amoxicillin in the samples of liners carrying case. In drag swabs and samples of catfishes, the bacteria were resistant to sulfonamides. In samples from crop, were resistant to sulfonamide and enrofloxacin and the caeca to sulfonamides, the trimetopim-sulphamethoxazole, tetracycline, amoxicillin and ampicillin to. / Objetivou-se com esta pesquisa investigar a presença de Salmonella sp. em matérias primas de origem animal utilizadas na fabricação de rações de frangos de corte, em rações coletadas diretamente dos comedouros das aves, em órgãos e conteúdos de cecos e forros de caixa de transporte de pintos de um dia, em amostras ambientais, em amostras de suabes de mãos de funcionários de granjas e em amostras de abatedouro bem como tipificar e determinar o perfil de resistência das cepas de Salmonella sp. encontradas, frente à ação de nove quimioterápicos. Para análise dos dados foi feita freqüência descritiva dos resultados encontrados. Coletou-se 1200 amostras de farinhas e Salmonella sp. foi encontrada em 10,5% das amostras com predominância do sorovar Enteritidis. A freqüência da bactéria em farinhas de carne foi de 12%, nas de sangue 6,8%, nas de penas 4,3% e nas de vísceras 14,6%. Foram isoladas também Salmonella Cerro, Salmonella Montevideo, Salmonella Anatum, Salmonella Tennessee e Salmonella Typhimurium entre outras. Em relação ao perfil de resistência das cepas encontradas nas diversas categorias de farinhas foi observado resistência à sulfonamidas, à neomicina, à tetraciclina, ao trimetopim-sulfametoxasol e ao florfenicol. Das três cepas da bactéria isoladas de rações duas eram de Salmonella Enteritidis e uma de S. Anatum, apresentando resistência à sulfonamidas e à neomicina. Dos 32 lotes de pintos de um dia 9,4% apresentaram positividade para Salmonella e dos 32 lotes de forros de caixa de transporte 9,4%. As amostras ambientais, cama antes do alojamento, suabes de comedouro e de bebedouro e água de bebida das aves apresentaram resultados negativos para Salmonella sp.. Suabes de arrasto de cama de aviários, amostras de Alphitobius diaperinus e suabes de mãos de funcionários das granjas apresentaram freqüência de 12,5%, 12,5% e 6,5% respectivamente. Nas amostras de suabes de arrasto o sorovar Enteritidis foi o mais freqüente e nas amostras de cascudinhos e suabes de mãos foi o único sorovar isolado. Nas amostras de abatedouro 26,7% dos lotes de inglúvio e 33,3% dos lotes de cecos foram positivos para Salmonella sp.. Com relação ao perfil de resistência das cepas isoladas, observou-se resistência à sulfonamidas, à amoxacilina e à enrofloxacina nas amostras de pintos de um dia e à amoxacilina nas amostras de forros de caixa de transporte. Nos suabes de arrasto e amostras de cascudinhos, as bactérias foram resistentes à sulfonamidas. Nas amostras de inglúvios, foi observada resistência à sulfonamidas e à enrofloxacina e nos cecos à sulfonamidas, ao trimetopim-sulfametoxasol, à tetraciclina, à amoxacilina e à ampicilina.

Orientadores: Marcelo Brocchi, Gustavo Bueno Gregoracci / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-24T09:27:26Z (GMT). No. of bitstreams: 1 SartiSprogis_AdrianeCristina_M.pdf: 1852459 bytes, checksum: 397af9ee41d085346189ca76eee83f96 (MD5) Previous issue date: 2014 / Resumo: A salmonelose é uma zoonose que representa um sério problema de saúde pública mundial, devido: à sua alta prevalência, à dificuldade de seu controle, ao seu caráter endêmico, à morbidade e à mortalidade. O conhecimento da ocorrência das diferentes sorovariedades de S. enterica em diferentes regiões e países pode ajudar no rastreamento e reconhecimento de patógenos emergentes, e assim implementar políticas de tratamento e prevenção. A grande maioria das sorovariedades expressa dois tipos diferentes de antígenos flagelares codificados pelos genes fliC (fase 1) e fljB (fase 2), sendo assim denominadas bifásicas. Contudo, algumas sorovariedades expressam apenas uma das fases, e são denominadas monofásicas. É possível que a variação de fase flagelar em S. enterica esteja associada a uma função de escape do sistema imunológico, por aumentar o repertório de antígenos expressos pela célula bacteriana, evitando temporariamente a resposta imune celular. Assim sendo, S. enterica bifásicas possuiriam uma vantagem seletiva sobre as monofásicas, porém isso não é totalmente verificado nos estudos epidemiológicos, pois no Estado de São Paulo, S. enterica I,4,[5],12:i:-, (monofásica) é uma das mais comumente associadas aos casos de diarreia e/ou infecções sistêmicas em pacientes humanos. De fato, a partir da década de 1990 houve um aumento significativo da S. enterica I,4,[5],12:i:- em muitos países. Sequências de profagos são muito comuns em S. enterica, sendo de conhecimento que esses fagos codificam vários fatores que contribuem para patogenicidade, diversidade genética e/ou características que aumentam o fitness. Coculturas experimentais de linhagens de S. enterica podem induzir espontaneamente profagos, que matam bactérias sensíveis, e assim a indução espontânea de fagos em uma população lisogênica acentua a competitividade entre populações. Neste estudo foram analisadas culturas puras de S. enterica Enteritidis (bifásica) adicionadas de fagos líticos induzidos de S. enterica I,4,[5],12:i:-, bem como coculturas entre as duas sorovariedades citadas, nas quais foram observadas induções espontâneas de fagos associados à alta densidade populacional e alterações das taxas de crescimento em ambos os estudos, corroborando a hipótese de que S. enterica monofásica pode alterar a dinâmica populacional a seu favor, pela liberação de fagos líticos à outra sorovariedade, interferindo no crescimento populacional de S. enterica Enteritidis, e que o sucesso evolutivo de S. enterica I,4,[5],12.:i:- pode estar associado a fagos líticos atuando como um regulador na ecologia bacteriana. Esses dados podem mudar nosso conhecimento sobre a interação bactéria-fago de uma simples relação parasita-hospedeiro para uma coevolução de duas vias entre seus genomas / Abstract: Salmonellosis is a zoonosis that is a serious public health problem worldwide, due to its high prevalence, difficulty controlling, their endemicity, morbidity and mortality. The knowledge of the occurrence of different serovars of S. enterica in different regions and countries can help in tracking and recognition of emerging pathogens and thus implement policies for treatment and prevention. The majority of serovars express two different types of flagellar antigens encoded by genes: fliC (phase 1) and fljB (phase 2), so called biphasic. However, some serovars express only one of the phases and are termed monophasic. It is possible that flagellar phase variation of S. enterica is associated with an escape function of the immune system to increase the repertoire of antigens expressed by the bacterial cell temporarily preventing cellular immune response. Thus, S. enterica biphasic would have a selective advantage over monophasic, but this is not fully verified in epidemiologic studies, because in the State of São Paulo, S. enterica I,4,[5],12:i:-, (monophase) is the one most commonly associated with cases of diarrhea and/or systemic infections in human patients, in fact, from the 1990s there was a significant increase of S. enterica I,4,[5],12:i:- in many countries. Prophages sequences are very common in S. enterica, with the knowledge that these phages encode several factors that contribute to pathogenicity, genetic diversity and/or characteristics that increase fitness. Cocultures experimental strains of S. enterica prophages can induce spontaneous, killing susceptible bacteria, and thus the spontaneous induction in a population of lysogenic phage enhances the competitiveness between populations. This study analyzed pure cultures of S. enterica Enteritidis ( biphasic ) added lytic phage induced from S. enterica I,4,[5],12:i:-, as well as cocultures between the two serovars cited where inductions were observed spontaneous phage associated with high population density and changes in growth rates in both studies, supporting the hypothesis that S. enterica monophase can alter the population dynamics to their advantage by releasing lytic phage to another serovar, interfering with the population growth of S. enterica Enteritidis and the evolutionary success of S. enterica I,4,[5],12:i:- may be associated with lytic phages acting as a regulator in bacterial ecology. These data may change our understanding of bacteria- phage from a simple parasite-host coevolution for a two-way between their genomes / Mestrado / Clinica Medica / Mestra em Clínica Médica

In der vorliegenden Arbeit wurde einerseits der Verlauf der Schutzwirkung der zurzeit in Sachsens Legehennenbeständen überwiegend angewandten Impfschemata gegen Salmonella Enteritidis (SE) untersucht. Andererseits wurden die Impfschemata, die ausschließlich modifizierte Lebendimpfstoffe (MLV) umfassen, und Impfschemata, die aus einer Kombination von MLV und Inaktivatimpfstoffen (KV) bestehen, auf Unterschiede in der Wirksamkeit geprüft. Um die Wirksamkeit der Impfschemata im Verlauf der Legeperiode und im Vergleich miteinander untersuchen zu können, wurden zu drei verschiedenen Zeitpunkten der Legeperiode (39., 54. und 69. Lebenswoche (LW)) Infektionsversuche mit einem Nalidixinsäure-resistenten SE-Stamm durchgeführt. Es wurden insgesamt 180 Legehennen verwendet, die in fünf verschiedenen Herkunftsbetrieben etwa gleicher Größe aufgezogen und geimpft worden waren. In jedem der Herkunftsbetriebe wurde eines von fünf Impfschemata (A bis E) angewendet. Die Impfschemata A und C beinhalteten ausschließlich MLV, die Impfschemata B, D und E eine Kombination aus MLV und KV. Zu den genannten Zeitpunkten (39., 54. und 69. LW) wurden jeweils zwölf Tiere aus den Betrieben in den Infektionsstall des Instituts für Tierhygiene und Öffentliches Veterinärwesen verbracht und eingestallt. Nach der Adaptionsphase von einer Woche und der Prüfung der Tiere auf Salmonellenfreiheit wurde jedes Tier mit 1,0 x109 KbE SE oral infiziert. Zwei und sieben Tage post infectionem (p.inf). wurden jeweils sechs Hennen euthanasiert und seziert. Caeca, Leber-, Ovar- und Oviduktproben wurden entnommen und gemäß anerkannter quantitativer und qualitativer Untersuchungsmethoden auf den Infektionsstamm untersucht. Zur Kontrolle der Erregerausscheidung wurden ein, drei und fünf Tage p.inf. Kloakentupferproben von jedem Tier entnommen und entsprechend auf SE untersucht. Die Ergebnisse dieser Untersuchungen variierten in Abhängigkeit von Versuchs- und Sektions- bzw. Kloakentupferentnahmezeitpunkt, untersuchtem Organ sowie quantitativer und qualitativer Untersuchung. Die ausschließlich mit MLV geimpften Gruppen A und C wiesen im Vergleich zu den Gruppen D und E, die mit demselben MLV und zusätzlich mit einem KV geimpft worden waren, in den Kloakentupfer- und Caecumproben in der Regel qualitativ und quantitativ weniger Salmonellen auf. Der Salmonellennachweis in den Leberproben und den vereinzelt besiedelten Reproduktionsorganen variierte nur geringfügig zwischen den Impfgruppen. Da die Tiere der vier Impfgruppen aus jeweils anderen Herkunftsbetrieben stammten, sind haltungsbedingte Unterschiede anzunehmen. Das äußere Erscheinungsbild (Befiederung, Bemuskelung, makroskopische pathologische Veränderungen) und das Sozialverhalten der Tiere variierten, was mit Unterschieden in der Immunität einhergehen kann. Bei den Tieren der Gruppe A bzw. C waren in der 69. LW und in 54. LW, respektive, ein ausgeprägtes kannibalistisches Verhalten und dessen Konsequenzen (reduzierte Wasser- und Futteraufnahme der gepickten Tiere, Hackverletzungen) zu beobachten. Tendenziell waren die Tiere der Gruppen B bis E in der 54. LW stärker mit SE belastet als in der 39. und 69. LW. Ein Einfluss der unter Umständen erhöhten Umgebungstemperaturen in den Betrieben sowie des hohen Leistungsstresses während der mittleren Phase der Legeperiode auf die Immunität der 54 LW alten Tiere, die im August 2010 infiziert wurden, ist nicht auszuschließen. Auch eine sich ausbildende Altersresistenz könnte die bessere Salmonellenabwehr der 69 LW alten Tiere erklären. Tiere der Gruppe A waren jedoch in der 69. LW am stärksten mit SE belastet, was auf ein Nachlassen der durch die Impfung induzierten Immunität und möglicherweise auf den im Vergleich zu den jüngeren Tieren allgemein schwächeren Zustand der Tiere zurückzuführen ist. Die Ergebnisse einer Kontrollgruppe zur Beurteilung der von der Impfung unabhängigen Faktoren fehlen. Da in den sächsischen Legehennenhaltungen aufgrund der hohen Tierzahlen entsprechend der Hühner-Salmonellen Verordnung (Impfpflicht für Betriebe mit mehr als 350 Tieren) gegen SE geimpft wird, hätten zur Bildung einer ungeimpften Kontrollgruppe Tiere aus kleineren Betrieben oder Zuchttierhaltungen mit völlig anderen Haltungsstrukturen verwendet werden müssen. Damit wären die wissenschaftlichen Ansprüche an eine Kontrollgruppe jedoch nicht erfüllt worden. Entsprechend der Ergebnisse und Umstände dieser Studie scheint eine Zusatzimpfung mit einem KV im Vergleich zu einer Impfung mit ausschließlich MLV keinen Vorteil im Schutz vor SE zu bieten. Es konnte gezeigt werden, dass die Immunität der gemäß der Impfschemata B bis E geimpften Legehennen gegen SE am Ende der Legeperiode nicht nachlässt. Die Impfung allein kann den Erreger nicht eliminieren und muss daher stets in ein verantwortungsvolles und vielseitiges Bekämpfungsprogramm integriert werden.

Published Article / The development of tolerance to low pH levels and the existence of cross-resistance may promote survival of bacteria in acidic foodstuff and in acidic environments such as the human stomach, in so doing escalating the probability of food poisoning. Similar to antimicrobial resistance developing, there is growing concern that effectiveness of organic acids may decrease as a result of the emergence of acid-tolerant food-borne pathogens. The objectives of this study were to determine the development of acid tolerance in selected food-borne pathogenic bacteria and to explore the activity of organic acids against acid tolerant pathogens. Bacterial strains were screened for acid-tolerance and susceptible strains were induced through exposure to increasing concentrations of an inorganic acid, as well as acidic foodstuffs. Susceptibility to six organic acids at various pH levels was assessed in order to evaluate the possible relationship between altered antimicrobial activity and acid tolerance. Salmonella enterica sv. Enteritidis ATCC 13076 and Escherichia coli ATCC 25922 were found to rapidly develop acid tolerance, while intrinsic acid tolerance was noted in Salmonella enterica sv. Typhimurium ATCC 14028. Pseudomonas aeruginosa ATCC 27853 demonstrated intermediate intrinsic acid tolerance. As expected, pH played a significant role in inhibitory activity of the organic acids as these compounds exhibit optimum antimicrobial activity at a lower pH (pH ≤5). It is, however, necessary to further elucidate the two-way role of pH in foodstuff concomitant to the addition of an organic acid.

Salmonella enteritidis (S. enteritidis) is a bacterial pathogen of chickens, and is currently one of the leading causes of human food poisoning in the world. It is believed that contaminated poultry products, especially eggs and egg products, have been responsible for the dramatic increase in the incidence of this Salmonella serotype. Detection of S. entertidis has conventionally involved bacteriological examination of samples, yet these procedures are time-consuming which could lead to the rapid spread of S. enteritidis through commercial flocks and potentially cause a human health risk. A number of alternative detection techniques, mostly based on serological methods, have been reported as effective diagnostic assays. However, some of these reports have not been supported by representations of SDS-PAGE gels or Western blots. The objective of this study was the evaluation of these serological techniques as well as a PCR amplification technique, which has been reported to show promising results as a diagnostic method. The techniques discussed in these reports were evaluated with regards to how rapid they were, their specificity and their potential for use in local diagnostic laboratories. Antigens from the outer surface of S. enteritidis were purified by several methods and their antigenicity was tested by separating the antigens by means of SDS-PAGE, followed by Western blotting using sera of chickens infected with S. enteritidis. A high degree of cross reactivity was observed with many of the antigens tested, especially the lipopolysaccharides (LPSs) and outer membrane proteins (OMPs) which had previously been reported as containing antigens which could be used for specific detection of S. enteritidis. This cross-reactivity could be explained by the conserved nature of many of the LPS and OMP antigens among the Salmonella serotypes tested. A fimbrial antigen, SEF14, which has been reported as a novel antigen, was seen as a prominent band at 14.3 kDa and was found to react with antibodies against S. enteritidis, yet not to the specificity levels described in previous reports. PCR amplification of the sefA gene sequence, which encodes for the SEF14 fimbrial antigen, was found to give a predicted product of 310 bp when using a previously described oligonucleotide primer pair. This amplified product was found to be specific for S. enteritidis and other serogroup D Salmonella serotypes that are not poultry pathogens The cross-reactivity observed with many of the serological techniques used in this study, meant that detection of S. enteritidis infection in chickens was considerably hindered. However, the identification of further novel antigens by serological means, could result in the development of new vaccines. The specificity and speed afforded by PCR amplification indicated that this technique showed excellent potential for use in local diagnostic laboratories. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 2003.

Memoria para optar al Título Profesional de Médico Veterinario / Salmonella enterica es considerada una de las principales causas de enfermedades transmitidas por los alimentos alrededor del mundo, teniendo un amplio rango de hospederos incluyendo los animales silvestres, lo cual ha generado un esfuerzo constante en la vigilancia epidemiológica de la enfermedad. Sin embargo, las aves silvestres presentan un riesgo mayor, ya que pueden recorrer largas distancias, incluso entre países. En este estudio se aisló S. enterica en un 4,95% (46/928) mediante tórula de arrastre, desde aves silvestres acuáticas, principalmente la Gaviota Dominicana (Larus dominicanus), en ocho sitios de muestreo a lo largo del país. Los serovares identificados fueron: S. Enteritidis con un 69,56% (32/46), S. Heildeberg 8,69% (4/46), S. Seftenberg 4,34% (2/46) y por último, otros serovares en menores proporciones como S. Anatum, Havana, Agona, Infantis, Dublin, y una cepa Grupo B (I 4, 5, 12:b:), en donde cada uno presentó una frecuencia de 2,17% (1/46). Además, fueron detectados doce combinaciones de genes asociados a virulencia (virulotipos). Estos resultados sugieren que en Chile, la infección por Salmonella enterica en aves silvestres acuáticas podría tener impacto en la salud pública y animal / Financiamiento: Proyecto Fondecyt 11110398

Memoria para optar al Título Profesional de Médico Veterinario / Las enfermedades transmitidas por alimentos, provocadas por el género Salmonella spp, son un importante problema de salud pública a nivel mundial, siendo el serotipo Salmonella Enteritidis (SE) uno de los principales causantes de gastroenteritis asociadas al consumo de productos de origen aviar. La industria avícola ha implementado diversas medidas para enfrentar esto, tales como probióticos, vacunas y antimicrobianos, pero dado que la enfermedad persiste, las investigaciones actualmente se dirigen, complementariamente, al control del patógeno en el alimento. En este contexto, se está analizando el uso de bacteriófagos líticos en diferentes alimentos como herramienta de biocontrol altamente específica y efectiva. Estos son virus que infectan bacterias, se replican en ellas y las lisan. Son inocuos para las células eucariotas y no alteran la calidad organoléptica de los alimentos. En base a lo anterior, este estudio pretende evaluar la efectividad de una mezcla de cinco bacteriófagos líticos nativos en la reducción del crecimiento de SE en carne fresca de pollo. Para ello, se contaminaron muestras con 5,7x103UFC/mL de SE y se trataron con 107UFP de cada fago/mL, o con 5,7x105UFC/mL y 109UFP/mL, e incubaron durante 10 días a temperatura ambiente controlada (18°C) y de refrigeración (2-8°C), respectivamente. Los resultados demostraron que la mezcla de fagos logró reducciones significativas (p<0,05) de 0,88log10UFC/g a temperatura ambiente y 1,66log10UFC/g a temperatura de refrigeración. Esto evidencia que los bacteriófagos líticos, aplicados con una MOI de 104, pueden controlar eficazmente la presencia de SE en carne fresca de pollo mantenida a distintas temperaturas / Financiamiento: Proyecto Fondecyt 1110038