Étude de l’efficacité de la vaccination à Salmonella Enteritidis chez la poule pondeuse et de la protection contre l’infection

Les infections à Salmonella Enteritidis chez les humains sont associées à la consommation d’œufs ou d’ovoproduits contaminés. La vaccination est un outil utilisé pour diminuer les risques d’infection à SE chez la volaille, mais avec des résultats variables. Au Canada deux bactérines, MBL SE4C et Layermune, sont couramment utilisées pour lutter contre SE. Cependant, leur efficacité n’a pas été complètement déterminée chez les poules pondeuses plus âgées. Par ailleurs, la capacité de ces vaccins à prévenir la transmission verticale et horizontale n’a pas encore été étudiée. L’objectif principal de cette étude était d’évaluer l’effet des deux bactérines sur la réponse immunitaire chez les poules pondeuses, de vérifier la protection conférée par ces vaccins contre l’infection expérimentale à SE, et d’identifier des protéines immunogènes afin de développer un vaccin sous-unitaire. Les oiseaux ont été vaccinés avec deux protocoles d’immunisation en cours d’élevage (soit à 12 et 18, ou à 16 semaines d’âge). Le groupe contrôle a été injecté avec la solution saline. Les oiseaux ont été inoculés per os avec 2 x 109 CFU de la souche SE lysotype 4 à 55 ou à 65 semaines d’âge. Les anticorps (IgG et IgA) ont été mesurés à différents temps avec un ELISA maison en utilisant l’antigène entier de SE. La phagocytose, flambée oxydative, les populations des splénocytes B et T ont été analysées en utilisant la cytométrie en flux. Les signes cliniques, l’excrétion fécale, la contamination des jaunes d’œufs et l’invasion des salmonelles dans les organes ont été étudiés pour évaluer l’efficacité de protection. La transmission horizontale a aussi été étudiée en évaluant l’infection à SE chez les oiseaux mis en contact avec les oiseaux inoculés. Les protéines immunogènes ont été identifiées par SDS-PAGE et Western blot à l’aide d’antisérums prélevés suite à la vaccination et/ou à l’infection expérimentale/naturelle, puis caractérisées par la spectrométrie de masse. Le protocole de vaccination avec deux immunisations a généré un niveau élevé de séroconversion à partir de 3 jusqu’à 32-34 semaines post-vaccination par rapport à celui avec une seule immunisation (p < 0.02), mais il n’y avait plus de différence entre les groupes à 54 et 64 semaines d’âge. Il n’y a pas eu de corrélation entre les niveaux d’IgG et les taux d’isolement des salmonelles dans les organes et des jaunes d’œuf. La production des IgA n’a été observée que chez les oiseaux vaccinés avec 2 injections de MBL SE4C (p ≤ 0.04). Après l’infection expérimentale, la production des IgA a été significativement plus élevée aux jours 1 et 7 p.i dans l’oviducte des oiseaux vaccinés (sauf pour le groupe vacciné avec 2 injections de Layermune) par comparaison avec le groupe contrôle (p ≤ 0.03). Seule la bactérine MBL SE4C a eu un effet protecteur contre la contamination des jaunes d’œuf chez les oiseaux infectés. Ce vaccin réduit partiellement en utilisant deux immunisations, le taux d’excrétion fécale des salmonelles chez les oiseaux inoculés et les oiseaux horizontalement infectés (p ≤ 0.02). Cinq des protéines identifiées par la spectrométrie de masse sont considérées comme des protéines potentiellement candidates pour une étude plus approfondie de leur immonogénicité: Lipoamide dehydrogenase, Enolase (2-phosphoglycerate dehydratase) (2-phospho-D-glycerate hydro-lyase), Elongation factor Tu (EF-Tu), Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) et DNA protection during starvation protein. En général, les bactérines ont induit une immunité humorale (IgG et IgA) chez les poules pondeuses. Cette réponse immunitaire a protégé partiellement les oiseaux quant à l’élimination des salmonelles, la contamination des jaunes d’œuf, ainsi que la transmission horizontale. Dans cette étude, la bactérine MBL SE4C (avec deux immunisations) s’est montrée plus efficace pour protéger les oiseaux que la bactérine Layermune. Nos résultats apportent des informations objectives et complémentaires sur le potentiel de deux bactérines pour lutter contre SE chez les poules pondeuses. Étant donné la protection partielle obtenue en utilisant ces vaccins, l’identification des antigènes immunogènes a permis de sélectionner des protéines spécifiques pour l’élaboration éventuelle d’un vaccin plus efficace contre SE chez les volailles.

Zoning of the commercial poultry industry in Ontario as a method of avian influenza mitigation

L’Organisation mondiale de la santé animale (OIE) est l’institution internationale responsable de la mise en place des mesures sanitaires associées aux échanges commerciaux d’animaux vivants. Le zonage est une méthode de contrôle recommandée par l’OIE pour certaines maladies infectieuses, dont l’influenza aviaire. Les éclosions d’influenza aviaire été extrêmement coûteuses pour l’industrie avicole partout dans le monde. Afin d’évaluer la possibilité d’user de cette approche en Ontario, les données sur les sites de production avicole ont été fournies par les fédérations d’éleveurs de volailles ce cette province. L’information portant sur les industries associées à la production avicole, soit les meuneries, les abattoirs, les couvoirs, et les usines de classification d’œufs, a été obtenue par l’entremise de plusieurs sources, dont des représentants de l’industrie avicole. Des diagrammes de flux a été crée afin de comprendre les interactions entre les sites de production et les industries associées à ceux-ci. Ces industries constituaient les éléments de bas nécessaires au zonage. Cette analyse a permis de créer une base de données portant sur intrants et extrants de production pour chaque site d’élevage avicole, ainsi que pour les sites de production des industries associées à l’aviculture. À l’aide du logiciel ArcGIS, cette information a été fusionnée à des données géospatiales de Statistique Canada de l’Ontario et du Québec. La base de données résultante a permis de réaliser les essais de zonage. Soixante-douze essais ont été réalisés. Quatre ont été retenus car celles minimisaient de façon similaire les pertes de production de l’industrie. Ces essais montrent que la méthode utilisée pour l’étude du zonage peut démontrer les déficits et les surplus de production de l’industrie avicole commerciale en Ontario. Ceux-ci pourront servir de point de départ lors des discussions des intervenants de l’industrie avicole, étant donné que la coopération et la communication sont essentielles au succès du zonage.

Utilisation of Giant African snail (Achatina fulica) meal as protein source by laying hens

A 12-week experiment was carried out to investigate the effects of substituting Giant African snail meal for fish meal in laying hens diet. Four diets were formulated to contain snail meal as replacement for fish meal at 0 (control), 33, 67 and 100%. A total of 120 Shaver Brown pullets aged 18 weeks were allocated to the dietary treatments in a randomised design. Each treatment consisted of three replicates and ten birds per replicate. Feed intake increased only for the 33% treatment as compared to the 67% replacement diet but did not differ from the other treatments. There were no significant treatment effects on egg performance parameters observed (egg production, egg weight, total egg mass, feed conversion ratio and percent shell). The overall feed cost of egg production reduced on the snail meal-based diets. The organoleptic evaluation of boiled eggs revealed no difference between the treatments. Based on these results it was concluded that total replacement of fish meal with cooked snail meat meal does not compromise laying performance or egg quality. The substitution is beneficial in terms of production cost reduction and the reduction of snails will have a beneficial impact especially where these snails are a serious agricultural pest. The manual collection and processing of snails can also become a source of rural income.

What limits insect fecundity? Body size- and temperature-dependent egg maturation and oviposition in a butterfly

1. Large female insects usually have high potential fecundity. Therefore selection should favour an increase in body size given that these females get opportunities to realize their potential advantage by maturing and laying more eggs. However, ectotherm physiology is strongly temperature-dependent, and activities are carried out sufficiently only within certain temperature ranges. Thus it remains unclear if the fecundity advantage of a large size is fully realized in natural environments, where thermal conditions are limiting. 2. Insect fecundity might be limited by temperature at two levels; first eggs need to mature, and then the female needs time for strategic ovipositing of the egg. Since a female cannot foresee the number of oviposition opportunities that she will encounter on a given day, the optimal rate of egg maturation will be governed by trade-offs associated with egg- and time-limited oviposition. As females of different sizes will have different amounts of body reserves, size-dependent allocation trade-offs between the mother’s condition and her egg production might be expected. 3. In the temperate butterfly Pararge aegeria , the time and temperature dependence of oviposition and egg maturation, and the interrelatedness of these two processes were investigated in a series of laboratory experiments, allowing a decoupling of the time budgets for the respective processes. 4. The results show that realized fecundity of this species can be limited by both the temperature dependence of egg maturation and oviposition under certain thermal regimes. Furthermore, rates of oviposition and egg maturation seemed to have regulatory effects upon each other. Early reproductive output was correlated with short life span, indicating a cost of reproduction. Finally, large females matured more eggs than small females when deprived of oviposition opportunities. Thus, the optimal allocation of resources to egg production seems dependent on female size. 5. This study highlights the complexity of processes underlying rates of egg maturation and oviposition in ectotherms under natural conditions. We further discuss the importance of temperature variation for egg- vs. time-limited fecundity and the consequences for the evolution of female body size in insects.

The efficacy of Salenvac, a Salmonella enterica subsp. Enterica serotype Enteritidis iron-restricted bacterin vaccine, in laying chickens

The protective effect of two vaccination regimes using Salenvac, a commercially available iron-restricted Salmonella enterica subsp. Enterica serotype Enteritidis PT4 bacterin vaccine, was verified in laying birds. Immunization was intramuscular at 1 day old and again at 4 weeks of age (V2), or at 1 day and 4 weeks with a third dose at 18 weeks of age (V3). Challenge S. Enteritidis (5 to 7.5) x 10(7) colony forming units) was given intravenously at 8, 17, 23, 30 and 59 weeks of age. For all age groups, both vaccination regimes reduced significantly the number of tissues and faecal samples that were culture positive for the challenge strain. For laying birds, fewer eggs (P < 0.001) were culture positive for S. Enteritidis after challenge from vaccinated laying birds ( 56/439 batches of eggs) than unvaccinated birds (99/252 batches). The data give compelling evidence that the vaccine is efficacious and may contribute to the reduction of layer infection and egg contamination.

Evidence for systemic spread of the potentially zoonotic intestinal spirochaete Brachyspira pilosicoli in experimentally challenged laying chickens

Brachyspira pilosicoli is a potentially zoonotic anaerobic intestinal spirochaete that is one of several species causing avian intestinal spirochaetosis. The aim of this study was to develop a reproducible model of infection in point-of-lay chickens and compare the virulence of two strains of B. pilosicoli in a model using experimentally challenged laying chickens. Seventeen-week-old commercial laying chickens were experimentally challenged by oral gavage with either B. pilosicoli strain B2904 or CPSp1, following an oral dose of 10 % sodium bicarbonate to neutralize acidity in the crop. Approximately 80 % of the chickens became colonized and exhibited increased faecal moisture content, reduced weight gain and delayed onset of lay. Tissues sampled at post-mortem examination were analysed to produce a quantitative output on the number of spirochaetes present and hence, the extent of colonization. The liver and spleen were colonized, and novel histopathology was observed in these tissues. The infection model we report here has potential use in studies to improve our understanding of the mechanisms by which Brachyspira elicit disease in poultry and in testing novel intervention strategies.

Oral treatment of chickens with Lactobacillus reuteri LM1 reduces Brachyspira pilosicoli-induced pathology

Avian intestinal spirochaetosis (AIS) results from the colonization of the caeca and colon of poultry by pathogenic Brachyspira, notably Brachyspira pilosicoli. Following the ban on the use of antibiotic growth promoters in the European Union in 2006, the number of cases of AIS has increased, which, alongside emerging antimicrobial resistance in Brachyspira, has driven renewed interest in alternative intervention strategies. Lactobacillus-based probiotics have been shown to protect against infection with common enteric pathogens in livestock. Our previous studies have shown that Lactobacillus reuteri LM1 antagonizes aspects of the pathobiology of Brachyspira in vitro. Here, we showed that L. reuteri LM1 mitigates the clinical symptoms of AIS in chickens experimentally challenged with B. pilosicoli. Two groups of 15 commercial laying hens were challenged experimentally by oral gavage with B. pilosicoli B2904 at 18 weeks of age; one group received unsupplemented drinking water and the other received L. reuteri LM1 in drinking water from 1 week prior to challenge with Brachyspira and thereafter for the duration of the study. This treatment regime was protective. Specifically, B. pilosicoli was detected by culture in fewer birds, bird weights were higher, faecal moisture contents were significantly lower (P<0.05) and egg production as assessed by egg weight and faecal staining score was improved (P<0.05). Also, at post-mortem examination, significantly fewer B. pilosicoli were recovered from treated birds (P<0.05), with only mild–moderate histopathological changes observed. These data suggest that L. reuteri LM1 may be a useful tool in the control of AIS.

Desempenho comparativo entre duas linhagens de roedeiras comerciais debicadas em diferentes idades na fase de recria

O experimento foi realizado com o objetivo de comparar o desempenho produtivo de duas linhagens comerciais de poedeiras debicadas em diferentes idades na fase de recria. Foram utilizadas 250 aves da linhagem Lohmann (LSL) e 250 aves da linhagem Isa Babcock (IB), debicadas em diferentes idades. A primeira debicagem foi realizada no 8º dia de idade e a segunda na 8ª, 10ª, 12ª ou 14ª semana de idade. Um grupo de 32 aves permaneceu com o bico intacto e outro não foi submetido à segunda debicagem. O delineamento utilizado foi em blocos ao acaso, em esquema fatorial 2x6 (linhagens x idade de debicagem). Não foi verificado efeito significativo da idade de debicagem sobre os parâmetros produtivos analisados, exceto a melhoria significativa na produção de ovos. As aves da linhagem LSL apresentaram melhores resultados para maturidade sexual, peso médio dos ovos, produção de ovos/ave dia, porcentagem de ovos quebrados e conversão alimentar, quando comparadas às aves da linhagem IB. Embora a prática da debicagem tenha melhorado a porcentagem de postura, os resultados evidenciaram que os parâmetros produtivos analisados foram mais influenciados pela linhagem do que pela idade de debicagem.

Influence of egg pre-storage heating period and storage length on incubation results

This experiment aimed at evaluating the influence of different heating times of settable eggs of Cobb 500® broiler breeders before submitting them to different storage periods on egg weight loss, embryo mortality, and hatchability. A total number of 1,980 eggs were distributed in a completely randomized experimental design with a 3 x 3 factorial arrangement, comprising nine treatments with 22 replicates of 10 eggs each. The following factors were analyzed: pre-storage heating periods (0, 6, 12 hours at 36.92°C) and storage periods (4, 9, 14 days at 12.06°C). After storage, eggs were incubated under usual conditions, and were transferred to the hatcher at 442 hours of incubation. Eggs were weighed before heating, incubation, and transference to determine weight loss. Partial hatchability was determined at 480 hours, and total hatchability at 498 hours of incubation. Embryo mortality was determined in non-hatched eggs. It was concluded that heating eggs for six hour before storage improves incubation results as it decreases incubation length and late embryo mortality, therefore its use can be indicated in commercial operations. Storing eggs for 14 days and pre-heating for 14 days and pre-heating for 12 hours severely impair incubation results, and therefore are not recommended.

Experimental infection of laying hens with Salmonella enterica serovar Gallinarum

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Experimental Salmonella Gallinarum infection in light laying hen lines

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Control of Salmonella enterica serovar Enteritidis in laying hens by inactivated Salmonella Enteritidis vaccines

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

INFLUENCE of ASCORBIC ACID on EGG QUALITY

A qualidade dos ovos pode ser influenciada por alguns nutrientes adicionados à dieta das aves e entre esses nutrientes se inclui o ácido ascórbico, que segundo alguns pesquisadores, além de melhorar a qualidade, pode aumentar a produção e o peso dos ovos e diminuir a quantidade de ovos trincados e o consumo de ração. Com a finalidade de tentar contribuir com mais algumas informações, desenvolveu esse experimento com o objetivo de avaliar a influência do ácido ascórbico sobre a qualidade de ovos brancos e marrons mantidos sob condições de ambiente (26,7 ± 2,2°C e 60 ± 5%UR), por um período de 28 dias. Foram utilizados 1440 ovos, sendo 720 de casca branca e 720 de casca marrom. Foram adicionadas à ração de aves, durante 60 dias, concentrações de 100, 150 e 200mg/kg de ácido ascórbico. A qualidade dos ovos foi determinada de 7 em 7 dias, avaliando-se a unidade Haugh e o índice gema. Os dados obtidos permitiram concluir que a suplementação com o ácido ascórbico não foi suficiente para diminuir a perda de qualidade dos ovos. Independentemente dos níveis de vitaminas utilizados, os ovos marrons mostraram-se qualitativamente superiores aos brancos.

Influência da interação fósforo disponível × fitase da dieta sobre o desempenho, os níveis plasmáticos de fósforo e os parâmetros ósseos de poedeiras comerciais

Dois experimentos foram realizados para avaliar a interação fósforo disponível (Pd) × fitase (UF) da dieta sobre o desempenho, os níveis plasmáticos de fósforo e os parâmetros ósseos de poedeiras. No experimento 1, utilizaram-se 240 aves com 40 semanas de idade distribuídas em esquema fatorial 5 × 2, composto de cinco níveis de fósforo disponível (0,094; 0,194; 0,294; 0,394 e 0,494%) e dois de fitase (0 e 300 unidades de fitase - UF). No experimento 2, utilizaram-se 288 aves com 44 semanas de idade, distribuídas também em esquema fatorial 3 × 4, composto de três níveis de fósforo disponível (0,094; 0,294 e 0,494%) e quatro de fitase (0; 300; 600 e 1.200 UF). No experimento 1, não houve interação fósforo disponível × fitase, mas o nível de fósforo disponível influenciou, de forma quadrática, a produção (PR) e a massa (MO) de ovos, os níveis plasmáticos de fósforo e a conversão por massa de ovos (CMO) e, de forma linear, o teor de cinzas na tíbia (CT) e a resistência óssea (RO). A adição de 300 UF elevou a disponibilidade do fósforo (de 4,034 para 4,784 mg/dL), o teor de CT (de 41,55 para 42,90%) e a resistência óssea (de 9,678 para 11,135 kgf/mm). No experimento 2, a produção aumentou de forma linear com os níveis de fósforo disponível da ração, que aumentaram linearmente com o aumento do nível de fitase. Os teores de cinzas na tíbia aumentaram até o nível de 567 UF. O aumento do fósforo disponível no nível de 600 UF melhorou de forma linear a massa de ovos, a conversão por massa de ovos e a conversão por dúzia de ovos. A suplementação da ração com menor teor de fósforo disponível (0,094%) com 300 a 1.200 UF melhorou a resistência óssea das aves. Poedeiras exigem em média 0,31 a 0,34% Pd. A adição de 300 UF ou o aumento do fósforo disponível em rações com 600 UF melhora a conversão por massa de ovos. A suplementação de rações deficientes em fósforo disponível com pelo menos 300 UF aumenta os níveis plasmáticos de fósforo, os teores de cinzas na tíbia e a resistência óssea de poedeiras comerciais.

Passionflower supplementation in diets of Japanese qualis at rearing and laying periods

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)