Fowl Aviadenovirus E associated with hepatitis-hydropericardium syndrome in broiler breeders / Síndrome da hepatite-hidropericárdio por Aviadenovirus e em reprodutores de corte

ABSTRACT: Fowl aviadenovirus (FAdV) is an important pathogen in the global poultry industry and the etiology of inclusion body hepatitis-hydropericardium syndrome (HHS) in chickens. Since the 1990s, several outbreaks of HHS have occurred in poultry producing areas, including South America. The coinfection of FAdV and chicken anemia virus (CAV) may markedly impact the incidence of HHS. This study describes an outbreak of HHS in coinfection with CAV in industrial broiler breeders and characterizes the FAdV isolate. The three-week-old male broiler breeders had pale bone marrow, enlarged and yellowish liver, splenomegaly, and atrophied thymus; one chicken was also found with hydropericardium. Virus isolation was performed in SPF chicken embryos of liver and thymus. Tissues of the naturally infected chickens and the inoculated embryos were evaluated by PCR and histopathology. All affected chickens and inoculated embryos were positive for FAdV and CAV. The inoculated embryos had enlarged, greenish and hemorrhagic livers, and 30% died within 7 days of inoculation. Phylogenetic analysis of the FAdV isolate hexon gene partial sequence enabled grouping with E species. The E species has recently become a relevant species in several countries. The association of FAdV with CAV in breeders is of further concern due to both being capable of vertical transmission. Within the last decade, a worldwide upsurge of HHS in broiler breeders owing to failed biosecurity has occurred. In this episode, the failure on biosecurity may have enabled challenge with both FAdV and CAV, with pathological synergism. The CAV-impaired adaptive immunity may have benefited the FAdV infection.

RESUMO: Adenovírus aviário (FAdV) é um importante patógeno na indústria avícola global e a etiologia da síndrome da hepatite por corpúsculo de inclusão-hidropericárdio (SHH) em galinhas. Desde a década de 1990, vários surtos de SHH foram descritos em todas as áreas de produção de aves, incluindo na América do Sul, e a coinfecção entre FAdV e vírus da anemia das galinhas (CAV) pode ser agravante para todos os aspectos da SHH. Objetiva-se descrever um surto de SHH em matrizes de frangos corte, caracterizar a estirpe de FAdV envolvida e destacar a coinfecção com CAV. Foram avaliados machos reprodutores de corte com três semanas de idade, com medula óssea pálida, fígado aumentado e amarelado e esplenomegalia, timo atrofiado e um com hidropericárdio. Fígado e timo foram coletados para isolamento do vírus em embriões de galinhas SPF, PCR e histopatologia. Todas as aves acometidas e embriões inoculados foram positivos para FAdV e CAV. Os embriões inoculados tiveram óbito de 30% em até sete dias após a inoculação e alterações hepáticas por fígados esverdeados e aumentados. A análise filogenética de FAdV com base em parte da sequência do gene que codifica a proteína hexon revelou identidade com a espécie E, que se tornou disseminada em vários países. A coinfecção de FAdV e CAV resulta em maior intensidade de lesões, maior morbidade e mortalidade e em reprodutores tem alta relevância epidemiológica, em razão da transmissão vertical de ambos e da ampla distribuição geográfica das progênies infectadas. Na última década, ocorreu um aumento mundial na ocorrência de SHH em frangos de corte relacionado a falhas em biosseguridade, especialmente em reprodutores, condição que pode ter ocorrido neste episódio. A presença de FAdV e CAV em reprodutores é motivo para preocupação por reflexos negativos à imunidade e viabilidade das progênies.

Fowl Aviadenovirus E associated with hepatitis-hydropericardium syndrome in broiler breeders / Síndrome da hepatite-hidropericárdio por Aviadenovirus e em reprodutores de corte

Fowl aviadenovirus (FAdV) is an important pathogen in the global poultry industry and the etiology of inclusion body hepatitis-hydropericardium syndrome (HHS) in chickens. Since the 1990s, several outbreaks of HHS have occurred in poultry producing areas, including South America. The coinfection of FAdV and chicken anemia virus (CAV) may markedly impact the incidence of HHS. This study describes an outbreak of HHS in coinfection with CAV in industrial broiler breeders and characterizes the FAdV isolate. The three-week-old male broiler breeders had pale bone marrow, enlarged and yellowish liver, splenomegaly, and atrophied thymus; one chicken was also found with hydropericardium. Virus isolation was performed in SPF chicken embryos of liver and thymus. Tissues of the naturally infected chickens and the inoculated embryos were evaluated by PCR and histopathology. All affected chickens and inoculated embryos were positive for FAdV and CAV. The inoculated embryos had enlarged, greenish and hemorrhagic livers, and 30% died within 7 days of inoculation. Phylogenetic analysis of the FAdV isolate hexon gene partial sequence enabled grouping with E species. The E species has recently become a relevant species in several countries. The association of FAdV with CAV in breeders is of further concern due to both being capable of vertical transmission. Within the last decade, a worldwide upsurge of HHS in broiler breeders owing to failed biosecurity has occurred. In this episode, the failure on biosecurity may have enabled challenge with both FAdV and CAV, with pathological synergism. The CAV-impaired adaptive immunity may have benefited the FAdV infection.

Adenovírus aviário (FAdV) é um importante patógeno na indústria avícola global e a etiologia da síndrome da hepatite por corpúsculo de inclusão-hidropericárdio (SHH) em galinhas. Desde a década de 1990, vários surtos de SHH foram descritos em todas as áreas de produção de aves, incluindo na América do Sul, e a coinfecção entre FAdV e vírus da anemia das galinhas (CAV) pode ser agravante para todos os aspectos da SHH. Objetiva-se descrever um surto de SHH em matrizes de frangos corte, caracterizar a estirpe de FAdV envolvida e destacar a coinfecção com CAV. Foram avaliados machos reprodutores de corte com três semanas de idade, com medula óssea pálida, fígado aumentado e amarelado e esplenomegalia, timo atrofiado e um com hidropericárdio. Fígado e timo foram coletados para isolamento do vírus em embriões de galinhas SPF, PCR e histopatologia. Todas as aves acometidas e embriões inoculados foram positivos para FAdV e CAV. Os embriões inoculados tiveram óbito de 30% em até sete dias após a inoculação e alterações hepáticas por fígados esverdeados e aumentados. A análise filogenética de FAdV com base em parte da sequência do gene que codifica a proteína hexon revelou identidade com a espécie E, que se tornou disseminada em vários países. A coinfecção de FAdV e CAV resulta em maior intensidade de lesões, maior morbidade e mortalidade e em reprodutores tem alta relevância epidemiológica, em razão da transmissão vertical de ambos e da ampla distribuição geográfica das progênies infectadas. Na última década, ocorreu um aumento mundial na ocorrência de SHH em frangos de corte relacionado a falhas em biosseguridade, especialmente em reprodutores, condição que pode ter ocorrido neste episódio. A presença de FAdV e CAV em reprodutores é motivo para preocupação por reflexos negativos à imunidade e viabilidade das progênies.

Hepatic changes in Gallus gallus domesticus in Brazil / Alterações hepáticas em Gallus gallus domesticus no Brasil

The occurrence and the frequency of hepatic changes in chickens, including broiler (BC), layer (LC) and backyard chickens (BYC) were investigated. The retrospective and prospective study (2006-2021) was conducted with a total of 300 cases of liver disorders. Industrial poultry (BC and LC) were frequently affected (88%) and noninfectious changes were the most diagnosed (69%). Considering etiology or conditions, the hepatic changes were classified as follows: degenerative changes (42%), bacterial (28%), metabolic (15%), toxic (8%), viral (3%), neoplastic (2%), protozoal diseases (1.5%) and circulatory disorders (0.5%). Regarding the type of bird, degenerative, toxic changes and viral hepatitis were more frequent in BC. Circulatory and metabolic disorders, as well as bacterial hepatitis, were more frequently diagnosed in LC. Neoplastic and protozoal hepatitis occurred more frequently in BYC. The macroscopic examination in association with histopathology enabled the diagnosis of the hepatic changes in 59% of the cases. Considering bacterial hepatitis in commercial poultry, the etiological diagnosis is highly important, in view of the risk for public health, despite the obvious importance due to the productivity losses and condemnation at processing.

A ocorrência e a frequência de alterações hepáticas em aves, incluindo frangos de corte (FC), galinhas poedeiras (GP) e aves de subsistência (AS) foram investigadas. O estudo retrospectivo e prospectivo (2006-2021) foi realizado com um total de 300 casos de alterações hepáticas. Aves industriais (FC e GP) foram frequentemente acometidas (88%) e as alterações não infecciosas foram as mais comumente diagnosticadas (69%). Quanto à etiologia ou condição, as alterações hepáticas foram classificadas da seguinte forma: alterações degenerativas (42%), bacterianas (28%), metabólicas (15%), tóxicas (8%), virais (3%), neoplásicas (2%), bem como doenças por protozoários (1,5%) e distúrbios circulatórios (0,5%). Em relação ao tipo de ave, alterações degenerativas, tóxicas e hepatites virais foram mais frequentes nos FC. Distúrbios circulatórios e metabólicos, assim como hepatites bacterianas, foram diagnosticados com maior frequência nas GP. Neoplasias e hepatite por protozoário ocorreram com maior frequência em AS. O exame macroscópico associado à histopatologia possibilitou o diagnóstico da alteração hepática em 59% dos casos. Considerando a hepatite bacteriana em aves comerciais, o diagnóstico etiológico é de grande importância, em vista do risco para a saúde pública, apesar da óbvia importância devido às perdas de produtividade e condenação no processamento.

Gastrointestinal Parasites in Free-Range Chicken Raised under Extensive System from the Northeast of Brazil

Brazilian poultry production is growing, mainly due to the cost and benefit that chicken meat provides. The importance of free-range chickens and the susceptibility, to which they are exposed, makes it necessary to know about gastrointestinal parasites and the consequences that large infections can cause for them as such as weight loss. The present study aimed to determine the occurrence of gastrointestinal parasites in free-range hens raised in an extensive regime in the municipality of Santa Rita, state of Maranhão, Brazil. The studied population was composed of adult chickens (Gallus gallus domesticus) acquired from slaughterhouses in the region. The gastrointestinal organs of each chicken were separated and subsequently conditioned in flasks containing Railliet and Henrys solution and sent to the laboratory, to be analyzed and processed. Of the 100 chickens examined, 227 specimens of helminthes were identified, with a frequency of 32.6% for nematodes and 67.4% for cestodes. Among the nematode the following parasites were identified: Ascaridia galli (27.03%); Heterakis gallinarum (48.65%) and Subulura spp. (24, 32%). Raillietina echinobothrida (100%) was the only cestoda identified. The average infection rate by species of parasite was 1.18 for A. galli, 2.22 for H. gallinarum, 1.06 for Subulura spp. and 9.00 for R. echinobothrida. It is concluded that free-range chickens are parasitized by nematodes and cestoda.(AU)

Gastrointestinal Parasites in Free-Range Chicken Raised under Extensive System from the Northeast of Brazil

Brazilian poultry production is growing, mainly due to the cost and benefit that chicken meat provides. The importance of free-range chickens and the susceptibility, to which they are exposed, makes it necessary to know about gastrointestinal parasites and the consequences that large infections can cause for them as such as weight loss. The present study aimed to determine the occurrence of gastrointestinal parasites in free-range hens raised in an extensive regime in the municipality of Santa Rita, state of Maranhão, Brazil. The studied population was composed of adult chickens (Gallus gallus domesticus) acquired from slaughterhouses in the region. The gastrointestinal organs of each chicken were separated and subsequently conditioned in flasks containing Railliet and Henrys solution and sent to the laboratory, to be analyzed and processed. Of the 100 chickens examined, 227 specimens of helminthes were identified, with a frequency of 32.6% for nematodes and 67.4% for cestodes. Among the nematode the following parasites were identified: Ascaridia galli (27.03%); Heterakis gallinarum (48.65%) and Subulura spp. (24, 32%). Raillietina echinobothrida (100%) was the only cestoda identified. The average infection rate by species of parasite was 1.18 for A. galli, 2.22 for H. gallinarum, 1.06 for Subulura spp. and 9.00 for R. echinobothrida. It is concluded that free-range chickens are parasitized by nematodes and cestoda.

Performance, nutrient digestibility, and intestinal histomorphometry of broilers fed diet supplemented with guava extract standardized in phenolic compounds

The objective of this study was to evaluate the effects of guava extract standardized in phenolic compounds (SGE) on broiler performance, nutrient digestibility, and intestinal histomorphometry. A total of 300 one-day-old male Cobb-500® broiler chicks were distributed in a completely randomized design with five treatments (basal diet supplemented with 120 mg vitamin E/kg and basal diet supplemented with 0, 600, 800, or 1,000 mg SGE/kg) and six replicates of ten birds each. Performance was evaluated at seven and 21 days of age; digestibility of nutrients was determined by total excreta collection from 18 to 21 days of age; and histomorphometry of the small intestine was assessed at 21 days of age. Broilers fed diets supplemented with SGE or vitamin E had higher body weight and weight gain and better feed conversion than those fed unsupplemented diet at seven days of age. At the same age, there was a quadratic effect of SGE levels on body weight and weight gain, with better weights for 715 and 716 mg SGE/kg, respectively; and a decreasing linear effect for feed conversion. At 21 days of age, body weight and weight gain increased linearly with the inclusion of SGE in diet. Digestibility of feed nutrients was not influenced by treatments. Broilers fed diet supplemented with 800 or 1,000 mg SGE/kg had greater villus height and villus:crypt ratio of the duodenum than those fed unsupplemented diet. Villus height in the jejunum of broilers fed diet supplemented with 600 mg SGE/kg was lower than that of broilers that received vitamin E. Guava extract standardized in phenolic compounds can be used in diets for broilers in the starter phase, considering that the extract increases weight gain, reduces feed conversion, and helps in the development of the intestinal mucosa.

Thermal Response of Three Strains of Hens Housed in a Cage-Free Aviary at the Amazon Rainforest

This study aimed to evaluate the thermal response of three strains of hens housed in a cage-free system at the Amazon rainforest in order to evaluate how feather coverage influences thermal exchange with the environment. The experimental method was completely randomized and treatments comprised three strains of hens (Rhode Island Red (red feathers with feathers on the neck), alternative strain FCI (red feathers without feathers on the neck), and alternative strain FCIII (white feathers without feathers on the neck)), with 20 hens (replicates) analyzed per strain. Thermal images of each bird were captured in order to record the birds surface temperatures on five points in five targets. All data collected in this study were subjected to ANOVA and subsequently to the Tukey test at p≤0.01 and p≤0.05. The aviarys left wall presented a lower average temperature, indicating lower heat accumulation, while the floor presented higher heat accumulation. FCIII hens (white feathers) presented higher (p<0.05) heat accumulation on the head and legs, and lower (p<0.05) heat accumulation on the neck and back in relation to other analyzed hens, indicating increased heat exchange efficiency and high concentration of this process in specific body areas. FCI and FCIII hens (without feathers on the neck) presented lower (p<0.05) heat accumulation on the neck and higher (p<0.05) heat accumulation on the head and legs, indicating that the feather coverage directly influenced heat exchange mechanisms, and an increased area without feathers provided great heat exchange zones for birds in a tropical climate.(AU)

Thermal Response of Three Strains of Hens Housed in a Cage-Free Aviary at the Amazon Rainforest

This study aimed to evaluate the thermal response of three strains of hens housed in a cage-free system at the Amazon rainforest in order to evaluate how feather coverage influences thermal exchange with the environment. The experimental method was completely randomized and treatments comprised three strains of hens (Rhode Island Red (red feathers with feathers on the neck), alternative strain FCI (red feathers without feathers on the neck), and alternative strain FCIII (white feathers without feathers on the neck)), with 20 hens (replicates) analyzed per strain. Thermal images of each bird were captured in order to record the birds’ surface temperatures on five points in five targets. All data collected in this study were subjected to ANOVA and subsequently to the Tukey test at p≤0.01 and p≤0.05. The aviary’s left wall presented a lower average temperature, indicating lower heat accumulation, while the floor presented higher heat accumulation. FCIII hens (white feathers) presented higher (p<0.05) heat accumulation on the head and legs, and lower (p<0.05) heat accumulation on the neck and back in relation to other analyzed hens, indicating increased heat exchange efficiency and high concentration of this process in specific body areas. FCI and FCIII hens (without feathers on the neck) presented lower (p<0.05) heat accumulation on the neck and higher (p<0.05) heat accumulation on the head and legs, indicating that the feather coverage directly influenced heat exchange mechanisms, and an increased area without feathers provided great heat exchange zones for birds in a tropical climate.

An Overview on Mareks Disease Virus Evolution and Evidence for Increased Virulence in Brazil

Mareks disease virus (MDV) has been shown to be evolving to higher virulence. One of the genetic sites involved in virulence which enables such characterization is the 339-amino acid Meq protein encoding gene (meq). The reemergence of clinical Mareks disease (MD) in vaccinated flocks can be associated to changes in meq. Our studies have shown the presence of very virulent MDV strains in the Brazilian industrial and free-range poultry. We present an overview of MD increasing severity and indicate the necessity of using phylogenetic tools for best accompanying MDV evolution.(AU)

An Overview on Mareks Disease Virus Evolution and Evidence for Increased Virulence in Brazil

Mareks disease virus (MDV) has been shown to be evolving to higher virulence. One of the genetic sites involved in virulence which enables such characterization is the 339-amino acid Meq protein encoding gene (meq). The reemergence of clinical Mareks disease (MD) in vaccinated flocks can be associated to changes in meq. Our studies have shown the presence of very virulent MDV strains in the Brazilian industrial and free-range poultry. We present an overview of MD increasing severity and indicate the necessity of using phylogenetic tools for best accompanying MDV evolution.

Detection of efflux pump CmeABC in enrofloxacin resistant Campylobacter spp. strains isolated from broiler chickens (Gallus gallus domesticus) in the state of Rio de Janeiro, Brazil / Detecção da bomba de efluxo CmeABC em cepas de Campylobacter spp. resistentes à enrofloxacina isoladas de frangos de corte (Gallus gallus domesticus) no Estado do Rio de Janeiro, Brasil

Fowls are the main reservoirs of the highly important food-originating pathogen called Campylobacter spp. and broilers' meat and byproducts are the main vehicles of this microorganism. Increasing of Campylobacter spp. resistant strains to fluorquinolones, an antimicrobial class often employed in poultry farming and in human medicine has become a great concern to poultry breeders. In fact, several studies evaluated increasing bacterial resistance against these antimicrobial agents. The role of CmeABC efflux system has been underscored among the resistance mechanisms in Campylobacter spp. to fluorquinolones. This study investigated the occurrence of CmeABC efflux pump in 81 and 78 enrofloxacin resistant strains of Campylobacter jejuni and C. coli respectively, isolated from broilers collected from six abattoirs situated at São José do Vale do Rio Preto/RJ poultry center and from two commercial abattoirs situated at Metropolitan Region of Rio de Janeiro, from 2013 to 2016. The resistance to enrofloxacin was assessed by agar dilution to determine minimum inhibitory concentration (MIC). The CmeABC efflux system was investigated through the detection of genes genes cmeA, cmeB and cmeC by PCR. The activity of CmeABC efflux pump was investigated in 20 strains by using the efflux pump inhibitor Phenylalanine-Arginine ß-Naphthylamide (PAßN). The three genes cmeA, cmeB and cmeC were detected in 94.3% of the strains (C. jejuni = 80 and C. coli = 70), whereas the system was absent or incomplete in 5.7% of strains (C. jejuni = 1 and C. coli = 8). MIC varied between 0.5µg/ml and 64µg/ml, and 88.7% of strains were enrofloxacin resistant and 11.3% featuring intermediate resistance. The inhibition of the efflux pump by PAßN reduced the MIC to enrofloxacin up to eight times in fifteen strains (75%). These results indicate that this system is frequent and active in Campylobacter spp. Resistant strains in the presence of enrofloxacin.(AU)

As aves são os principais reservatórios de Campylobacter spp., importante patógeno de origem alimentar e a carne de frango e produtos derivados são os principais veículos desse microrganismo. O aumento de cepas de Campylobacter spp. resistentes às fluorquinolonas, uma classe antimicrobiana frequentemente empregada na avicultura e na medicina humana, tornou-se uma grande preocupação para os produtores de aves e vários estudos avaliaram o aumento da resistência bacteriana a esses antimicrobianos. O papel do sistema de efluxo CmeABC tem sido enfatizado entre os mecanismos de resistência em Campylobacter spp. à fluorquinolonas. O presente estudo investigou a ocorrência da bomba de efluxo CmeABC em 81 cepas de Campylobacter jejuni e 78 cepas de Campylobacter coli resistentes à enrofloxacina, isoladas de frangos de corte coletados em seis abatedouros situados no polo avícola de São José do Rio Preto/RJ e de dois abatedouros comerciais situados na Região Metropolitana do Rio de Janeiro, de 2013 a 2016. A resistência à enrofloxacina foi avaliada pelo método de diluição em ágar para determinar a concentração inibitória mínima (CIM). O sistema de efluxo CmeABC foi investigado através da detecção dos genes cmeA, cmeB e cmeC por PCR. A atividade da bomba de efluxo CmeABC foi investigada em 20 cepas utilizando o inibidor da bomba de efluxo Phenylalanine-Arginine ß-Naftilamida (PAßN). Os três genes cmeA, cmeB e cmeC foram detectados em 94,3% das cepas (C. jejuni = 80 e C. coli = 70), enquanto o sistema estava ausente ou incompleto em 5,7% das cepas (C. jejuni = 1 e C coli = 8). A CIM variou entre 0,5µg/ml e 64µg/ml e 88,7% das cepas foram resistentes à enrofloxacina, enquanto 11,3% apresentaram resistência intermediária. A inibição da bomba de efluxo pelo PAßN reduziu a CIM da enrofloxacina até oito vezes em quinze cepas (75%). Estes resultados indicam que este sistema é frequente e ativo em cepas resistentes de Campylobacter spp. na presença de enrofloxacina.(AU)

Detection of efflux pump CmeABC in enrofloxacin resistant Campylobacter spp. strains isolated from broiler chickens (Gallus gallus domesticus) in the state of Rio de Janeiro, Brazil / Detecção da bomba de efluxo CmeABC em cepas de Campylobacter spp. resistentes à enrofloxacina isoladas de frangos de corte (Gallus gallus domesticus) no Estado do Rio de Janeiro, Brasil

Fowls are the main reservoirs of the highly important food-originating pathogen called Campylobacter spp. and broilers' meat and byproducts are the main vehicles of this microorganism. Increasing of Campylobacter spp. resistant strains to fluorquinolones, an antimicrobial class often employed in poultry farming and in human medicine has become a great concern to poultry breeders. In fact, several studies evaluated increasing bacterial resistance against these antimicrobial agents. The role of CmeABC efflux system has been underscored among the resistance mechanisms in Campylobacter spp. to fluorquinolones. This study investigated the occurrence of CmeABC efflux pump in 81 and 78 enrofloxacin resistant strains of Campylobacter jejuni and C. coli respectively, isolated from broilers collected from six abattoirs situated at São José do Vale do Rio Preto/RJ poultry center and from two commercial abattoirs situated at Metropolitan Region of Rio de Janeiro, from 2013 to 2016. The resistance to enrofloxacin was assessed by agar dilution to determine minimum inhibitory concentration (MIC). The CmeABC efflux system was investigated through the detection of genes genes cmeA, cmeB and cmeC by PCR. The activity of CmeABC efflux pump was investigated in 20 strains by using the efflux pump inhibitor Phenylalanine-Arginine ß-Naphthylamide (PAßN). The three genes cmeA, cmeB and cmeC were detected in 94.3% of the strains (C. jejuni = 80 and C. coli = 70), whereas the system was absent or incomplete in 5.7% of strains (C. jejuni = 1 and C. coli = 8). MIC varied between 0.5µg/ml and 64µg/ml, and 88.7% of strains were enrofloxacin resistant and 11.3% featuring intermediate resistance. The inhibition of the efflux pump by PAßN reduced the MIC to enrofloxacin up to eight times in fifteen strains (75%). These results indicate that this system is frequent and active in Campylobacter spp. Resistant strains in the presence of enrofloxacin.(AU)

As aves são os principais reservatórios de Campylobacter spp., importante patógeno de origem alimentar e a carne de frango e produtos derivados são os principais veículos desse microrganismo. O aumento de cepas de Campylobacter spp. resistentes às fluorquinolonas, uma classe antimicrobiana frequentemente empregada na avicultura e na medicina humana, tornou-se uma grande preocupação para os produtores de aves e vários estudos avaliaram o aumento da resistência bacteriana a esses antimicrobianos. O papel do sistema de efluxo CmeABC tem sido enfatizado entre os mecanismos de resistência em Campylobacter spp. à fluorquinolonas. O presente estudo investigou a ocorrência da bomba de efluxo CmeABC em 81 cepas de Campylobacter jejuni e 78 cepas de Campylobacter coli resistentes à enrofloxacina, isoladas de frangos de corte coletados em seis abatedouros situados no polo avícola de São José do Rio Preto/RJ e de dois abatedouros comerciais situados na Região Metropolitana do Rio de Janeiro, de 2013 a 2016. A resistência à enrofloxacina foi avaliada pelo método de diluição em ágar para determinar a concentração inibitória mínima (CIM). O sistema de efluxo CmeABC foi investigado através da detecção dos genes cmeA, cmeB e cmeC por PCR. A atividade da bomba de efluxo CmeABC foi investigada em 20 cepas utilizando o inibidor da bomba de efluxo Phenylalanine-Arginine ß-Naftilamida (PAßN). Os três genes cmeA, cmeB e cmeC foram detectados em 94,3% das cepas (C. jejuni = 80 e C. coli = 70), enquanto o sistema estava ausente ou incompleto em 5,7% das cepas (C. jejuni = 1 e C coli = 8). A CIM variou entre 0,5µg/ml e 64µg/ml e 88,7% das cepas foram resistentes à enrofloxacina, enquanto 11,3% apresentaram resistência intermediária. A inibição da bomba de efluxo pelo PAßN reduziu a CIM da enrofloxacina até oito vezes em quinze cepas (75%). Estes resultados indicam que este sistema é frequente e ativo em cepas resistentes de Campylobacter spp. na presença de enrofloxacina.(AU)

Detection of efflux pump CmeABC in enrofloxacin resistant Campylobacter spp. strains isolated from broiler chickens (Gallus gallus domesticus) in the state of Rio de Janeiro, Brazil

ABSTRACT: Fowls are the main reservoirs of the highly important food-originating pathogen called Campylobacter spp. and broilers meat and byproducts are the main vehicles of this microorganism. Increasing of Campylobacter spp. resistant strains to fluorquinolones, an antimicrobial class often employed in poultry farming and in human medicine has become a great concern to poultry breeders. In fact, several studies evaluated increasing bacterial resistance against these antimicrobial agents. The role of CmeABC efflux system has been underscored among the resistance mechanisms in Campylobacter spp. to fluorquinolones. This study investigated the occurrence of CmeABC efflux pump in 81 and 78 enrofloxacin resistant strains of Campylobacter jejuni and C. coli respectively, isolated from broilers collected from six abattoirs situated at São José do Vale do Rio Preto/RJ poultry center and from two commercial abattoirs situated at Metropolitan Region of Rio de Janeiro, from 2013 to 2016. The resistance to enrofloxacin was assessed by agar dilution to determine minimum inhibitory concentration (MIC). The CmeABC efflux system was investigated through the detection of genes genes cmeA, cmeB and cmeC by PCR. The activity of CmeABC efflux pump was investigated in 20 strains by using the efflux pump inhibitor Phenylalanine-Arginine -Naphthylamide (PAN). The three genes cmeA, cmeB and cmeC were detected in 94.3% of the strains (C. jejuni = 80 and C. coli = 70), whereas the system was absent or incomplete in 5.7% of strains (C. jejuni = 1 and C. coli = 8). MIC varied between 0.5g/ml and 64g/ml, and 88.7% of strains were enrofloxacin resistant and 11.3% featuring intermediate resistance. The inhibition of the efflux pump by PAN reduced the MIC to enrofloxacin up to eight times in fifteen strains (75%). These results indicate that this system is frequent and active in Campylobacter spp. Resistant strains in the presence of enrofloxacin.

RESUMO: As aves são os principais reservatórios de Campylobacter spp., importante patógeno de origem alimentar e a carne de frango e produtos derivados são os principais veículos desse microrganismo. O aumento de cepas de Campylobacter spp. resistentes às fluorquinolonas, uma classe antimicrobiana frequentemente empregada na avicultura e na medicina humana, tornou-se uma grande preocupação para os produtores de aves e vários estudos avaliaram o aumento da resistência bacteriana a esses antimicrobianos. O papel do sistema de efluxo CmeABC tem sido enfatizado entre os mecanismos de resistência em Campylobacter spp. à fluorquinolonas. O presente estudo investigou a ocorrência da bomba de efluxo CmeABC em 81 cepas de Campylobacter jejuni e 78 cepas de Campylobacter coli resistentes à enrofloxacina, isoladas de frangos de corte coletados em seis abatedouros situados no polo avícola de São José do Rio Preto/RJ e de dois abatedouros comerciais situados na Região Metropolitana do Rio de Janeiro, de 2013 a 2016. A resistência à enrofloxacina foi avaliada pelo método de diluição em ágar para determinar a concentração inibitória mínima (CIM). O sistema de efluxo CmeABC foi investigado através da detecção dos genes cmeA, cmeB e cmeC por PCR. A atividade da bomba de efluxo CmeABC foi investigada em 20 cepas utilizando o inibidor da bomba de efluxo Phenylalanine-Arginine -Naftilamida (PAN). Os três genes cmeA, cmeB e cmeC foram detectados em 94,3% das cepas (C. jejuni = 80 e C. coli = 70), enquanto o sistema estava ausente ou incompleto em 5,7% das cepas (C. jejuni = 1 e C coli = 8). A CIM variou entre 0,5g/ml e 64g/ml e 88,7% das cepas foram resistentes à enrofloxacina, enquanto 11,3% apresentaram resistência intermediária. A inibição da bomba de efluxo pelo PAN reduziu a CIM da enrofloxacina até oito vezes em quinze cepas (75%). Estes resultados indicam que este sistema é frequente e ativo em cepas resistentes de Campylobacter spp. na presença de enrofloxacina.

Toxoplasma gondii molecular and immunological identification and risk factors associated with infection in chicken slaughtered at Triângulo Mineiro region, Minas Gerais, Brazil / Identificação molecular e imunológica de Toxoplasma gondii e fatores de risco associados com infecção em galinhas abatidas na região do Triângulo Mineiro, Minas Gerais, Brasil

This study aimed to determine the occurrence of anti-Toxoplasma gondii antibodies in the serum of slaughtered chickens in the region of Triângulo Mineiro, Minas Gerais, Brazil, to detect the parasite in tissues (heart and brain) of serologically positive chickens, based on molecular analysis, and to investigate risk variables associated with the infection. Sera from 417 chickens raised in extensive, semi-intensive, and intensive production systems were tested by an indirect immunofluorescent antibody test (IFAT) and indirect hemagglutination antibody test (IHA). Polymerase chain reaction (PCR) was performed to detect T. gondii DNA in brain and heart tissues. Antibody anti-T. gondii were found in 37.65% (157/417) of chickens by IFAT, and in 75.06% (313/417) by IHA. The Kappa index showed a weak concordance between the techniques (0.087). Association was observed between seropositivity and the variables, age (p 0.0001), type of feeding (p 0.0001) and collective raising with other animals species (p 0.0001). Association, based on IFAT, was not observed between seropositivity and the variables, sex (p = 0.0526), presence of cats (p > 0.9999), and presence of rats (p > 0.9999). Presence of parasite DNA was detected in brain samples from two chickens, which were raised in intensive and semi-intensive production systems. The results suggest(AU)

O objetivo do presente estudo foi determinar a frequência de anticorpos anti-Toxoplasma gondii em soro de galinhas abatidas na região do Triângulo Mineiro, Minas Gerais, detectar molecularmente o parasito em tecidos (coração e cérebro) de algumas das aves sorologicamente positivas e averiguar variáveis de risco associadas à infecção. Foram testados soros de 417 galinhas, criadas nos sistemas extensivo, semi-intensivo e intensivo. Para a pesquisa de anticorpos anti-T. gondii foi utilizada a Reação de Imunofluorescência Indireta (RIFI) e Hemaglutinação Indireta (HAI). A Reação em Cadeia da Polimerase (PCR) foi utilizada para detectar o DNA de T. gondii em fragmentos de cérebro e coração. Anticorpos foram detectados no soro de 37,65% (157/417) das aves pela RIFI e em 75,06% (313/417) pela HAI. O índice Kappa mostrou uma fraca concordância entre as técnicas (0,087). Baseado na RIFI, foi verificada associação estatisticamente significativa (p 0,0001) entre a soropositividade e as variáveis: idade, tipo de alimentação e criação em conjunto com outras espécies animais. Não foi observada associação estatística (p > 0,01) entre as variáveis: sexo, presença de gatos e presença de ratos. Pelo diagnóstico molecular DNA do parasito foi detectado em duas amostras de cérebro, de indivíduos diferentes criados em sistema intensivo e semi-intensivo.(AU)

Transportadores de cálcio e fósforo em aves de postura / Calcium and phosphorus transporters in laying hens

Todos os organismos vivos, tanto animais como vegetais, apresentam quantidades variáveis de minerais, que são necessários para manter seu metabolismo fisiológico, sendo esses nutrientes de grande importância para o desenvolvimento das espécies. Para poedeiras, o cálcio e o fósforo são minerais chaves para o crescimento e formação do ovo, e sua disponibilidade é mais crucial durante o período de postura, pois os dois estão envolvidos no processo de formação do ovo. Apesar da importância bioquímica, fisiológica e nutricional destes minerais pouco se conhece a respeito do seu processo de transporte intestinal. Portanto, considerando a importância do cálcio e fósforo na nutrição de poedeiras e a necessidade do desenvolvimento de estratégias que possibilitem seu melhor aproveitamento, objetivou-se com esta revisão descrever o funcionamento dos transportadores de cálcio e fósforo para aves de postura, bem como, os fatores nutricionais que interferem sobre os mesmos.

All living organisms, both animals and plants, have varying amounts of minerals, which are necessary to keep its physiological metabolism of these nutrients, are important for the development of species. For laying hens, calcium and phosphorus are key minerals for growth and egg formation, and its availability is more crucial during the laying period, because both are involved in egg formation process. Despite the biochemical, physiological and nutritional importance of these minerals little is known about the process of intestinal transport. Therefore, considering the importance of calcium and phosphorus in the nutrition of hens and the need to develop strategies that enable their better utilization, aimed with this review describe the operation of carriers calcium and phosphorus for laying hens, as well as the nutritional factors that interfere with about the same.

Toxoplasma gondii molecular and immunological identification and risk factors associated with infection in chicken slaughtered at Triângulo Mineiro region, Minas Gerais, Brazil / Identificação molecular e imunológica de Toxoplasma gondii e fatores de risco associados com infecção em galinhas abatidas na região do Triângulo Mineiro, Minas Gerais, Brasil

This study aimed to determine the occurrence of anti-Toxoplasma gondii antibodies in the serum of slaughtered chickens in the region of Triângulo Mineiro, Minas Gerais, Brazil, to detect the parasite in tissues (heart and brain) of serologically positive chickens, based on molecular analysis, and to investigate risk variables associated with the infection. Sera from 417 chickens raised in extensive, semi-intensive, and intensive production systems were tested by an indirect immunofluorescent antibody test (IFAT) and indirect hemagglutination antibody test (IHA). Polymerase chain reaction (PCR) was performed to detect T. gondii DNA in brain and heart tissues. Antibody anti-T. gondii were found in 37.65% (157/417) of chickens by IFAT, and in 75.06% (313/417) by IHA. The Kappa index showed a weak concordance between the techniques (0.087). Association was observed between seropositivity and the variables, age (p < 0.0001), type of feeding (p < 0.0001) and collective raising with other animal's species (p < 0.0001). Association, based on IFAT, was not observed between seropositivity and the variables, sex (p = 0.0526), presence of cats (p > 0.9999), and presence of rats (p > 0.9999). Presence of parasite DNA was detected in brain samples from two chickens, which were raised in intensive and semi-intensive production systems. The results suggest the meat of these slaughtered animals may serve as a transmission source of this protozoan to humans.(AU)

O objetivo do presente estudo foi determinar a frequência de anticorpos anti-Toxoplasma gondii em soro de galinhas abatidas na região do Triângulo Mineiro, Minas Gerais, detectar molecularmente o parasito em tecidos (coração e cérebro) de algumas das aves sorologicamente positivas e averiguar variáveis de risco associadas à infecção. Foram testados soros de 417 galinhas, criadas nos sistemas extensivo, semi-intensivo e intensivo. Para a pesquisa de anticorpos anti-T. gondii foi utilizada a Reação de Imunofluorescência Indireta (RIFI) e Hemaglutinação Indireta (HAI). A Reação em Cadeia da Polimerase (PCR) foi utilizada para detectar o DNA de T. gondii em fragmentos de cérebro e coração. Anticorpos foram detectados no soro de 37,65% (157/417) das aves pela RIFI e em 75,06% (313/417) pela HAI. O índice Kappa mostrou uma fraca concordância entre as técnicas (0,087). Baseado na RIFI, foi verificada associação estatisticamente significativa (p < 0,0001) entre a soropositividade e as variáveis: idade, tipo de alimentação e criação em conjunto com outras espécies animais. Não foi observada associação estatística (p > 0,01) entre as variáveis: sexo, presença de gatos e presença de ratos. Pelo diagnóstico molecular DNA do parasito foi detectado em duas amostras de cérebro, de indivíduos diferentes criados em sistema intensivo e semi-intensivo. Os resultados indicam a possibilidade de a carne dessas aves poderem atuar como fonte de infecção deste protozoário para o homem.(AU)

Transportadores de cálcio e fósforo em aves de postura / Calcium and phosphorus transporters in laying hens

Todos os organismos vivos, tanto animais como vegetais, apresentam quantidades variáveis de minerais, que são necessários para manter seu metabolismo fisiológico, sendo esses nutrientes de grande importância para o desenvolvimento das espécies. Para poedeiras, o cálcio e o fósforo são minerais chaves para o crescimento e formação do ovo, e sua disponibilidade é mais crucial durante o período de postura, pois os dois estão envolvidos no processo de formação do ovo. Apesar da importância bioquímica, fisiológica e nutricional destes minerais pouco se conhece a respeito do seu processo de transporte intestinal. Portanto, considerando a importância do cálcio e fósforo na nutrição de poedeiras e a necessidade do desenvolvimento de estratégias que possibilitem seu melhor aproveitamento, objetivou-se com esta revisão descrever o funcionamento dos transportadores de cálcio e fósforo para aves de postura, bem como, os fatores nutricionais que interferem sobre os mesmos.(AU)

All living organisms, both animals and plants, have varying amounts of minerals, which are necessary to keep its physiological metabolism of these nutrients, are important for the development of species. For laying hens, calcium and phosphorus are key minerals for growth and egg formation, and its availability is more crucial during the laying period, because both are involved in egg formation process. Despite the biochemical, physiological and nutritional importance of these minerals little is known about the process of intestinal transport. Therefore, considering the importance of calcium and phosphorus in the nutrition of hens and the need to develop strategies that enable their better utilization, aimed with this review describe the operation of carriers calcium and phosphorus for laying hens, as well as the nutritional factors that interfere with about the same.(AU)

Frequency of anti-Toxocara antibodies in broiler chickens in southern Brazil / Frequência de anticorpos anti-Toxocara em frangos de corte no Sul do Brasil

The aim of this study was to evaluate the presence of anti-Toxocara antibodies in naturally infected broiler chickens (n = 189) from the state of Paraná, southern Brazil. The chickens were reared in a semi-intensive system by small family farmers (n = 7). An enzyme-linked immunosorbent assay (ELISA) was performed to detect the presence of anti- Toxocara spp. IgY after serum adsorption with Ascaridia galli antigens. An overall seroprevalence of 67.7% (128/189; 95% CI = 61.1-74.4) was observed. The frequency of positive animals by farm ranged from 29.6% to 100%. The optical density and reactivity index values observed in ELISA test indicated the possible chronicity of infection of the evaluated chickens. Associations between the presence of antibodies and the area where the chickens were reared (p = 0.382) or the population density of dogs on the farm (p = 0.785) were not observed. This study shows a high prevalence of Toxocara spp. antibodies in broiler chickens reared in semi-intensive systems and provides evidence that chickens are a good indicator of environmental contamination by larva migrans agents. Further studies are necessary to assess the risk factors associated with poultry infection and the likelihood of toxocariasis transmission to humans via the ingestion of free-range chicken meat.(AU)

A finalidade do presente estudo foi avaliar a presença de anticorpos anti- Toxocara, em frangos de corte naturalmente infectados (n = 189), no Norte do Paraná, Sul do Brasil. Os frangos foram criados em sistema semi-intensivo, em pequenas propriedades rurais (n = 7). Os testes sorológicos foram realizados pela técnica de ELISA, para detecção de anticorpos IgY (IgG), com pré-adsorção do soro com antígenos de Ascaridia galli. Foi observada uma prevalência de 67,7% (128/189; IC 95% = 61,1-74,4). A frequência de animais soropositivos por propriedade variou de 29,6% a 100%. Os valores da Densidade Ótica e do Índice de Reatividade observados no teste de ELISA indicaram uma possível cronicidade de infecção dos frangos avaliados. Não foi observada correlação entre a positividade dos animais, quando comparada a área (p = 0,382) e a densidade populacional de cães por propriedade (p = 0,785). O presente estudo verificou uma alta prevalência de anticorpos anti-Toxocara em frangos de corte criados em sistema semi-intensivo e oferece dados que apontam esses animais como bons indicadores de contaminação ambiental por agentes de larva migrans . Estudos futuros são necessários para avaliar os fatores de risco associados e a possibilidade da transmissão de toxocaríase ao ser humano pela ingestão de carne de frango.(AU)

Serological diagnosis of influenza A subtype H1 on family poultry of Belo Horizonte, Minas Gerais and Santa Maria, Rio Grande Do Sul, in Brazil

Serum samples (n=687) from Gallus gallus domesticus were collected for the investigation of antibodies to avian influenza virus (AIV-A) in the family poultry of the surrounding counties of Santa Maria/RS and the metropolitan region of Belo Horizonte/MG, totaling twenty different counties. Additional samples of seventeen (n=17) free-flying ducks (C. moschata pure or hybrid with Anas platyrhynchos) were collected in Belo Horizonte. The chosen tests for the survey were performed as described by the World Organization for Animal Health (OIE), including agar gel immunodiffusion (AGID) for antibodies to AIV-A nucleoprotein (N) and haemagglutination- inhibition (HI) for antibodies to subtype H1. Out of the 704 serum tests performed by AGID, eight (8/704) were revealed positive for antibodies to AIV-A N protein, with six (6/704) retested positive for subtype H1. Two sera tested positive by AGID were shown to be non reactive to the H1 subtype, suggesting specificity to another subtype. A low occurrence of antibodies to influenza A (1.13%) was found, and mostly (75%) specific to subtype H1. This represents an approximately 0,85% overall occurrence for subtype H1 antibodies, with an unknown subtype specific antibodies detected in one free-flying anatid. The low occurrence of antibodies in the family poultry may suggest a low AIV-A activity during the period of study, information which remains to be confirmed by virus detection.(AU)

Serological diagnosis of influenza A subtype H1 on family poultry of Belo Horizonte, Minas Gerais and Santa Maria, Rio Grande Do Sul, in Brazil

Serum samples (n=687) from Gallus gallus domesticus were collected for the investigation of antibodies to avian influenza virus (AIV-A) in the family poultry of the surrounding counties of Santa Maria/RS and the metropolitan region of Belo Horizonte/MG, totaling twenty different counties. Additional samples of seventeen (n=17) free-flying ducks (C. moschata pure or hybrid with Anas platyrhynchos) were collected in Belo Horizonte. The chosen tests for the survey were performed as described by the World Organization for Animal Health (OIE), including agar gel immunodiffusion (AGID) for antibodies to AIV-A nucleoprotein (N) and haemagglutination- inhibition (HI) for antibodies to subtype H1. Out of the 704 serum tests performed by AGID, eight (8/704) were revealed positive for antibodies to AIV-A N protein, with six (6/704) retested positive for subtype H1. Two sera tested positive by AGID were shown to be non reactive to the H1 subtype, suggesting specificity to another subtype. A low occurrence of antibodies to influenza A (1.13%) was found, and mostly (75%) specific to subtype H1. This represents an approximately 0,85% overall occurrence for subtype H1 antibodies, with an unknown subtype specific antibodies detected in one free-flying anatid. The low occurrence of antibodies in the family poultry may suggest a low AIV-A activity during the period of study, information which remains to be confirmed by virus detection.