Genotype and antibiotic resistance analyses of Campylobacter isolates from ceca and carcasses of slaughtered broiler flocks

To obtain genetic information about Campylobacter jejuni and Campylobacter coli from broilers and carcasses at slaughterhouses, we analyzed and compared 340 isolates that were collected in 2008 from the cecum right after slaughter or from the neck skin after processing. We performed rpoB sequence-based identification, multilocus sequence typing (MLST), and flaB sequence-based typing; we additionally analyzed mutations within the 23S rRNA and gyrA genes that confer resistance to macrolide and quinolone antibiotics, respectively. The rpoB-based identification resulted in a distribution of 72.0% C. jejuni and 28.0% C. coli. The MLST analysis revealed that there were 59 known sequence types (STs) and 6 newly defined STs. Most of the STs were grouped into 4 clonal complexes (CC) that are typical for poultry (CC21, CC45, CC257, and CC828), and these represented 61.8% of all of the investigated isolates. The analysis of 95 isolates from the cecum and from the corresponding carcass neck skin covered 44 different STs, and 54.7% of the pairs had matching genotypes. The data indicate that cross-contamination from various sources during slaughter may occur, although the majority of Campylobacter contamination on carcasses appeared to originate from the slaughtered flock itself. Mutations in the 23S rRNA gene were found in 3.1% of C. coli isolates, although no mutations were found in C. jejuni isolates. Mutations in the gyrA gene were observed in 18.9% of C. jejuni and 26.8% of C. coli isolates, which included two C. coli strains that carried mutations conferring resistance to both classes of antibiotics. A relationship between specific genotypes and antibiotic resistance/susceptibility was observed.

Vertical transmission of a fluoroquinolone-resistant Escherichia coli within an integrated broiler operation

The epidemiology of an enrofloxacin-resistant Escherichia coli clone was investigated during two separate outbreaks of colibacillosis in the Danish broiler production. In total five flocks were reported affected by the outbreaks. Recorded first-week mortalities were in the range of 1.7-12.7%. The clone was first isolated from dead broilers and subsequently demonstrated in samples from associated hatchers and the parent flock with its embryonated eggs, suggesting a vertical transmission from the parents. The second outbreak involved two broiler flocks unrelated to the affected flocks from the first outbreak. However, the clone could not be demonstrated in the associated parent flock. Furthermore, samplings from grand-parent flocks were negative for the outbreak clone. The clonality was evaluated by plasmid profiling and pulsed-field gel electrophoresis. None of the recognized virulence factors were demonstrated in the outbreak clone by microarray and PCR assay. The molecular background for the fluoroquinolone-resistance was investigated and point mutations in gyrA and parC leading to amino-acid substitutions in quinolone-resistance determining regions of GyrA and ParC were demonstrated. Vertical transmission of enrofloxacin-resistant E. coli from healthy parents resulting in high first-week mortality in the offspring illustrates the potential of the emergence and spreading of fluoroquinolone-resistant bacteria in animal husbandry, even though the use of fluoroquinolones is restricted.

Campylobacter monitoring in German broiler flocks: an explorative time series analysis

Campylobacter, a major zoonotic pathogen, displays seasonality in poultry and in humans. In order to identify temporal patterns in the prevalence of thermophilic Campylobacter spp. in a voluntary monitoring programme in broiler flocks in Germany and in the reported human incidence, time series methods were used. The data originated between May 2004 and June 2007. By the use of seasonal decomposition, autocorrelation and cross-correlation functions, it could be shown that an annual seasonality is present. However, the peak month differs between sample submission, prevalence in broilers and human incidence. Strikingly, the peak in human campylobacterioses preceded the peak in broiler prevalence in Lower Saxony rather than occurring after it. Significant cross-correlations between monthly temperature and prevalence in broilers as well as between human incidence, monthly temperature, rainfall and wind-force were identified. The results highlight the necessity to quantify the transmission of Campylobacter from broiler to humans and to include climatic factors in order to gain further insight into the epidemiology of this zoonotic disease.

Harmonised monitoring of antimicrobial resistance in Salmonella and Campylobacter isolates from food animals in the European Union

Many Member States of the European Union (EU) currently monitor antimicrobial resistance in zoonotic agents, including Salmonella and Campylobacter. According to Directive 2003/99/EC, Member States shall ensure that the monitoring provides comparable data on the occurrence of antimicrobial resistance. The European Commission asked the European Food Safety Authority to prepare detailed specifications for harmonised schemes for monitoring antimicrobial resistance. The objective of these specifications is to lay down provisions for a monitoring and reporting scheme for Salmonella in fowl (Gallus gallus), turkeys and pigs, and for Campylobacter jejuni and Campylobacter coli in broiler chickens. The current specifications are considered to be a first step towards a gradual implementation of comprehensive antimicrobial resistance monitoring at the EU level. These specifications propose to test a common set of antimicrobial agents against available cut-off values and a specified concentration range to determine the susceptibility of Salmonella and Campylobacter. Using isolates collected through programmes in which the sampling frame covers all epidemiological units of the national production, the target number of Salmonella isolates to be included in the antimicrobial resistance monitoring per Member State per year is 170 for each study population (i.e., laying hens, broilers, turkeys and slaughter pigs). The target number of Campylobacter isolates to be included in the antimicrobial resistance monitoring per Member State per year is 170 for each study population (i.e., broilers). The results of the antimicrobial resistance monitoring are assessed and reported in the yearly national report on trends and sources of zoonoses, zoonotic agents and antimicrobial resistance.

Time-series analysis of Campylobacter incidence in Switzerland

SUMMARY Campylobacteriosis has been the most common food-associated notifiable infectious disease in Switzerland since 1995. Contact with and ingestion of raw or undercooked broilers are considered the dominant risk factors for infection. In this study, we investigated the temporal relationship between the disease incidence in humans and the prevalence of Campylobacter in broilers in Switzerland from 2008 to 2012. We use a time-series approach to describe the pattern of the disease by incorporating seasonal effects and autocorrelation. The analysis shows that prevalence of Campylobacter in broilers, with a 2-week lag, has a significant impact on disease incidence in humans. Therefore Campylobacter cases in humans can be partly explained by contagion through broiler meat. We also found a strong autoregressive effect in human illness, and a significant increase of illness during Christmas and New Year's holidays. In a final analysis, we corrected for the sampling error of prevalence in broilers and the results gave similar conclusions.

Apparent digestibility and metabolizable energy content of lipid sources in poultry

A trial was conducted to determine the apparent digestibility (ATTD) and AME content of different lipid sources in 21d-old broilers. There were a control diet based on corn and soybean meal without any supplemental fat and 6 additional diets forming a 3x2 factorial with 3 sources of fat (soy oil, SBO; reconstituted monoglyceride oil, RMG; and reconstituted triglyceride oil, RTG) included in the diet (3 or 6%) at expenses (wt:wt) of the basal diet.

Indicators of sustainability of agriculture and livestock in Spain - Indicadores de sostenibilidad de la agricultura y ganadería española

Sustainability is an adjective used to characterize agriculture according to the degree of fulfillment of goals. Those goals are related to agro-ecological, environmental and socio-economic dimensions. Sustainability is a dynamic and temporal character. In absolute terms there is not an ending value because it changes as its dimensions make it. Spain is one of the main agricultural countries of the European Union both in terms of crop land and value of productions. The object of this study is to present a methodology of sustainability account to be incorporated into national statistical and to assess their performance in the course of the years. For that reason the data sources used have been the statistics of the Department of Agriculture and from others database. We presented a set of indicators of sustainability and its evaluation in a time series of at least 30 years. The trend analysis offers the evolution of the numerical values of the indicators in terms of efficiency, physical units used for a unit of product or its value in euros. The analyzed crops have been: wheat, barley, maize, sunflower, sugar beet, wine grape, olive oil, citrus, melon and tomato. Physical indicators were: land, water, energy, erosion, soil organic matter, and carbon balance; socio-economic indicators were: agricultural final production, prices, income, employment and use of fertilizers. In general, all crops increased their productive efficiency, higher in irrigated than on dry land. Spanish agricultural carbon sequestration capacity has multiplied by five in the last seventy years, as a result of the increase in the productivity of crops, in terms of total biomass and the modification of the soil management techniques. Livestock sector presents data of pork, broilers and laying hen. Those showed an improvement in efficiency and economic indicators. Overall we can say that Spanish agriculture and livestock subsector have a tendency towards sustainability, being its main threats extreme meteorological factors and the instability of todays markets.

The effect of a mono component protease and different AMEn levels in broiler diets on growth performance from 1 to 18 days of age.

The objective of the present study was to evaluate the effect of the inclusion of a mono component serine protease (RONOZYME ProAct, DSM Nutritional Products) in diets with two different AMEn contents on apparent ileal digestibility (AID) of amino acids (AA) and growth performance in broilers from 1 to 18 days of age.

Influencia de la fuente y el nivel de glicerina cruda en el pienso sobre los paráemtros productivos en pollos de 1 a 21 días de edad.

Las cantidades de grasas y aceites destinadas a la producción de biodiesel han aumentado en los últimos años a nivel mundial (National Biodiesel Board, 2012). La glicerina cruda (Gly) es un subproducto de la industria del biodiesel que contiene cantidades variables de humedad, cenizas y glicerol. La Gly es una fuente atractiva de energía que se utiliza actualmente como sustituto parcial del grano de cereal en dietas para avicultura y otras especies domésticas (Swiatkiewicz y Koreleski, 2009). Se estima que el nivel máximo de Gly a utilizar en piensos para broilers varía entre 5 y 10% (Cerrate et al., 2006). A estos niveles de inclusión, la Gly no presenta efecto negativo alguno sobre la salud o los parámetros productivos de las aves. Sin embargo, la calidad de las Gly comerciales varía en función de su origen, composición y proceso de obtención. El objetivo de este estudio fue evaluar el efecto de la fuente y el nivel de glicerina cruda en el pienso sobre el rendimiento productivo en pollos de engorde de 1 a 21 días de edad.

Influencia de la presentación y del nivel de energía del pienso sobre los rendimientos productivos en pollitas de 1 a 35 días de edad.

Numerosos estudios (Brickett et al., 2007; Serrano et al., 2012) han demostrado que la granulación del pienso mejora el consumo, los IC y el PV en broilers. La granulación podría aumentar la digestibilidad de algunos de los componentes de la dieta (Abdollahi et al., 2011) y reducir las pérdidas de alimento en animales no rumiantes (Serrano et al., 2012; Berrocoso et al., 2013). Sin embargo, la información disponible sobre el efecto de la presentación del pienso sobre la productividad en pollitas es escasa. Debido a su menor consumo y menor velocidad de crecimiento, es de esperar que los beneficios de la granulación sean inferiores, en valores absolutos, en pollitas que en broilers. La densidad energética del pienso afecta al crecimiento y al IC en avicultura. Las aves comen para satisfacer sus necesidades energéticas (Nahashon et al, 2006) y por tanto, aumentan la ingesta voluntaria de pienso al disminuir el contenido de energía del mismo (Veldkamp et al, 2005). Sin embargo, las pollitas podrían no ser capaces de mantener constante la ingesta de energía con dietas muy diluidas (Pérez-Bonilla et al., 2012). Por otra parte, piensos muy energéticos son más palatables y tienden a aumentar el consumo voluntario, lo que podría resultar en mejoras de las GMD y de los IC (Frikha et al., 2009a; Perez-Bonilla et al., 2012). El objetivo del presente estudio fue comparar los efectos de la presentación del pienso y la concentración energética del mismo sobre los rendimientos productivos en pollitas de 1 a 35 d de edad.

Influencia del cereal y el nivel de inclusión de fibra en el pienso sobre la productividad en pollitas de 0 a 17 semanas de edad.

La fibra ha sido considerada como un diluyente del pienso en monogástricos con efectos negativos sobre consumo y la digestibilidad de los nutrientes (Jansen and Carré, 1985). Sin embargo, estudios recientes (Mateos et al., 2012) han mostrado que la inclusión de cantidades moderadas de fibra en el pienso podría mejorar tanto la digestibilidad de los nutrientes como el crecimiento de las aves. La inclusión de fibra en piensos para broilers podría incrementar el tiempo de retención de la digesta en la parte superior del tracto digestivo, estimulando el funcionamiento de la molleja (Hetland et al. 2005) y la producción de HCl en el proventrículo (Duke, 1986). Un descenso del pH de la molleja favorece la producción de pepsina e incrementa la solubilidad de las sales minerales (Jiménez-Moreno et al. 2009). Por lo tanto, la inclusión de cantidades adecuadas de fibra podría beneficiar el desarrollo del tracto gastrointestinal (Mateos et al., 2012). Sin embargo, la información disponible sobre los efectos beneficiosos de la fibra dietética en pollitas es escasa (Frikha et al., 2009). De hecho, los autores no han encontrado ningún estudio publicado sobre los efectos de la inclusión de fibra sobre el crecimiento de pollitas de 0 a 17 sem de edad. La influencia de fuentes de fibra tales como la cascarilla de avena, pulpa de remolacha (PRE) o cascarilla de soja sobre el desarrollo del tracto gastrointestinal ha sido bien estudiada en broilers (González-Alvarado el al., 2010). Sin embargo, la información disponible sobre el uso de paja de cereales (PCE) en dietas para avicultura es muy limitada; aun siendo una fuente de fibra abundante. La hipótesis del presente ensayo fue que la inclusión de cantidades moderadas de fibra en piensos de pollitas podría mejorar el rendimiento productivo con una mejora de la utilización de los nutrientes. El objetivo del presente trabajo fue estudiar el efecto de la inclusión de diversas fuentes de fibra a diferentes niveles en el pienso de pollitas sobre los rendimientos productivos de 0 a 17 sem de edad.

Influencia e la inclusión de fibra en el pienso sobre los rendimientos produtivos de pollitas rubias de 1 a 35 días de edad.

La fibra dietética representa la fracción indigestible de los ingredientes del pienso por lo que diluye el contenido en nutrientes del mismo (Rougière y Carré, 2010). Además, un aumento en el nivel de fibra del pienso puede reducir el consumo voluntario (Sklan et al., 2003), alterar el perfil de la flora gastrointestinal y aumentar la incidencia de procesos entéricos (Janssen and Carré, 1985). Sin embargo, estudios recientes muestran que la inclusión de ciertas fuentes de fibras a niveles adecuados podrían mejorar los rendimientos productivos y reducir la incidencia de problemas digestivos en aves (Mateos, 2012). En broilers, la inclusión de fibra adicional en el pienso mejoró la fisiología digestiva, aumentando el tamaño de la molleja, reduciendo el pH de la digesta de este órgano (Sacranie et al., 2012) y mejorando de los rendimientos productivos (González-Alvarado et al., 2010). Fuentes de fibra insoluble tales como la cascarilla de avena (CAV), la cascarilla de girasol (CGI) y la paja de cereales (PCE) podrían mejorar la fisiología del tracto gastrointestinal (TGI) y la digestibilidad de ciertos nutrientes cuando se incluyen en el pienso en cantidades moderadas (Hetland et al., 2003). Por otro lado, fuentes de fibra soluble, tal como la pulpa de remolacha (PRE), incrementan la viscosidad intestinal, lo que podría reducir el consumo de pienso (Jiménez-Moreno et al., 2010). Los autores no han encontrado estudio alguno sobre los efectos de la inclusión de fibra adicional en el pienso sobre los rendimientos productivos de pollitas comerciales. El objetivo del presente trabajo fue estudiar el efecto de incluir diversas fuentes de fibra a diferentes niveles en el pienso sobre los rendimientos productivos de pollitas de 1 a 35 d de edad.

Dietary xylanase increases hepatic vitamin E concentration of chickens fed wheat based diet

The study examined the effect of xylanase supplementation on apparent metabolizable energy (AME) and hepatic vitamin E and carotenoids in broiler chickens fed wheat based diets. A total of one hundred forty four male Ross 308 chickens were used in this study. Birds were randomly assigned to 3 dietary treatments (8 cages per treatment of 6 male broilers each) for 14 days from 7 to 21 day old. The control treatment was based on wheat-soyabean meal and was either unsupplemented or supplemented with either 1000 or 2000 xylanase units per kg diet. Orthogonal polynomial contrasts were used to test linear response to dietary xylanase activity. There was a positive linear relationship (P < 0.05) between dietary AME and doses of supplementary xylanase. A linear relationship (P < 0.05) was also observed between dosage of xylanase supplementation and hepatic vitamin E concentration and retention. In conclusion, xylanase supplementation improved dietary AME and increased hepatic vitamin E concentration which may have positive effects on the antioxidative status of the birds.

Painéis de partículas de saco de cimento e embalagem longa vida aplicados como forro em protótipos de aviários

No presente trabalho, considerou o estudo do potencial de utilização de sacos de cimento e embalagens de longa vida com o intuito de agregar valor a esses resíduos, mediante a fabricação de painéis de partículas para uso como forro em protótipos de galpões avícolas. Para fabricação desse material foi utilizado sacos de cimento descartados nas obras civis, embalagens longa vida residuais e resina poliuretana bicomponente à base de óleo de mamona. O estudo foi dividido em quatro etapas: 1) Caracterização da matéria-prima (sacos de cimento); 2) Efeito da densidade e teor de resina nas propriedades físicas, mecânicas e térmicas de painéis de partículas de saco de cimento; 3) Avaliação do desempenho de embalagens longa vida e verniz como revestimentos dos painéis selecionados na etapa anterior; 4) Determinação do desempenho térmico de protótipos de aviários executados em escala reduzida e distorcida com forro de painel de saco de cimento e embalagem longa vida. Os resultados obtidos indicaram: Em razão do painel com 0,6 g.cm-3 e 12% de resina ter apresentado melhor desempenho nas propriedades investigadas, essa combinação foi selecionada para avaliar o desempenho do material (físicas, mecânicas e térmicas) quando revestido com embalagens longa vida, adotando a testemunha e o verniz como um revestimento comparativo. Dentre os revestimentos avaliados, os painéis com embalagens de longa vida, foram superiores aos revestidos com verniz, quando comparados as suas propriedades físicas, mecânicas e térmicas. Dessa maneira, painéis com 0,6 g.cm-3 e 12% de resina revestidos embalagens longa vida, foram testados como forro quanto ao desempenho térmico (primavera, verão, outono e inverno) em protótipos em escala reduzida e distorcida, mediante a determinação de índices de conforto térmico (Entalpia e IAPfc) para aves de corte. A associação do forro sob o protótipo permitiu reduzir a temperatura interna do ar e dos índices de conforto térmico na primavera, verão e outono, enquanto que no inverno não foi constatado eficácia do material. Dentre as estações estudadas, o verão e a primavera, demonstraram serem as épocas mais críticas para criação de aves corte em instalações avícolas que apresentem características semelhantes aos protótipos experimentais.

Detecção e genotipagem de astrovírus em amostras fecais de aves de produção procedentes de diferentes criações comerciais brasileiras

Os astrovírus são agentes virais associados com enteropatias e infecções extra-intestinais em aves, ocasionando prejuízos no desenvolvimento produtivo e saúde animal. Porém, são escassos os estudos no Brasil que visem a detecção e caracterização deste vírus com maior amplitude. Nesse contexto, detectaram-se e caracterizaram-se cepas de astrovírus a partir de 60 amostras fecais de aves procedentes de diferentes criações comerciais brasileiras (postura, corte e matrizes), mediante o emprego de uma reação PanAstrovírus como triagem inicial, que consistiu numa reação de RT-hemi-nested-PCR, visando a amplificação e posterior sequenciamento de um segmento parcial do gene RdRp (RNA polimerase dependente de RNA) do gene ORF1b, uma região gênica conservada entre todos os astrovírus. Subsequentemente, nas amostras positivas na triagem, realizou-se a amplificação, clonagem e sequenciamento do gene ORF2 codificante do capsídeo viral, analisado mediante análise filogenética. Os dados obtidos demonstraram uma frequência de infecção por astrovírus em 48,3% (29/60) das amostras analisadas. As espécies virais detectadas foram as seguintes: Avastrovírus 2 (Avian Nephritis Virus, ANV) em 72,4% (21/29), Chicken Astrovírus (CAstV) em 6,8% (2/29) e Avastrovírus 1 (Turkey Astrovírus tipo 1, TAstV-1) em 20,8% (6/29) das amostras positivas. O sequenciamento total do gene ORF2 foi possível em oito amostras (8/21) positivas a ANV, em uma amostra (1/6) positiva a TAstV-1 e uma amostra (1/2) positiva para CAstV. A análise deste gene revelou, nas sequências ANV, a presença de três genótipos bem diferenciados, segundo critérios do Astroviridae Study Group, sendo que um deles, agrupou-se dentro do genótipo 5. Além disso, quando analisado com sequências procedentes de outras regiões, as sequências deste estudo formaram seis clusters, dois deles, relacionados com sequências procedentes de Austrália e Reino Unido. O análise do ORF2 em CAstV, revelou também um agrupamento com cepas procedentes do Reino Unido. Já o análise da ORF2 de TAstV-1, revelou divergência genética com cepas isoladas em Estados Unidos, que foram as únicas sequências disponíveis no GenBank. O presente estudo traz a luz vários dados epidemiológicos concernentes aos astrovírus aviário de origem brasileira o que permitirá a realização de outros estudos relacionados a epidemiologia deste vírus, seus possíveis riscos potenciais em saúde pública e a adoção de medidas de controle direcionadas a este agente