Genetic variability of Mycoplasma synoviae detected in commercial layers in southeastern Brazil / Variabilidade genética de Mycoplasma synoviae detectado em poedeiras comerciais do sudeste do Brasil

Este estudo teve como objetivo caracterizar genótipos de M. synoviae circulantes em poedeiras comerciais, na Região Sudeste do Brasil. Para estabelecer a relação evolutiva entre as cepas de MS, foi analisada a sequência genética do gene vlhA de 11 cepas de MS identificadas no Rio de Janeiro e em São Paulo, de 10 outras cepas de MS identificadas no Brasil e cujas sequências foram depositadas no banco de dados GenBank, e da cepa vacinal MSH. O método da máxima verossimilhança e o modelo de Kimura com dois parâmetros foram utilizados para comparar as cepas. As sequências obtidas foram depositadas no Genbank, sob os números de acesso OP279775 a OP279785. Foi possível verificar a presença de diferentes cepas circulantes no Brasil, com alta similaridade entre as cepas do Rio de Janeiro pela análise do gene vlhA. As duas cepas paulistas detectadas no presente estudo possuem o baixo percentual (68%) de similaridade, demonstrando a variabilidade das cepas dessa localidade.

Detection of Mycoplasma Synoviae and Other Pathogens in Laying Hens with Respiratory Signs in the Rearing and Production Phases

The present study aimed to investigate the occurrence of Mycoplasma synoviae (MS), M. gallisepticum (MG), Ornitobacterium rhinotracheale (OR), Avibacterium paragallinarum (AP), Pasteurella multocida (PM) and Infectious Bronchitis Virus (IBV) in laying hens with respiratory clinical signs in two phases of production. 140 tracheal swabs and 140 blood samples were collected from laying hens in the rearing and production phases, the chickens belonged to six farms (A-F) located in the state of São Paulo, Brazil. The samples were analyzed by PCR for MG, MS, OR, AP, PM and IBV and by ELISA for MG and MS. The highest frequencies observed by PCR were for MS at farms B and C with 95 and 100% positivity, followed by MG at farms D and E with 35% and 65%, IBV with 35% at farm F and ORT with 15% at farm A. All flocks were positive for MG and MS in serology. Although MG and IBV have been detected, this can be explained by the vaccination protocols, since live attenuated vaccines are widely used for immunization against these pathogens. It was also possible to detect OR and AP thorugh PCR in some flocks. The occurrence of several etiological agents that cause respiratory diseases in laying hens was confirmed by PCR and serology, with MS being the most prevalent and being present in all farms studied.(AU)

Detection of Mycoplasma Synoviae and Other Pathogens in Laying Hens with Respiratory Signs in the Rearing and Production Phases

The present study aimed to investigate the occurrence of Mycoplasma synoviae (MS), M. gallisepticum (MG), Ornitobacterium rhinotracheale (OR), Avibacterium paragallinarum (AP), Pasteurella multocida (PM) and Infectious Bronchitis Virus (IBV) in laying hens with respiratory clinical signs in two phases of production. 140 tracheal swabs and 140 blood samples were collected from laying hens in the rearing and production phases, the chickens belonged to six farms (A-F) located in the state of São Paulo, Brazil. The samples were analyzed by PCR for MG, MS, OR, AP, PM and IBV and by ELISA for MG and MS. The highest frequencies observed by PCR were for MS at farms B and C with 95 and 100% positivity, followed by MG at farms D and E with 35% and 65%, IBV with 35% at farm F and ORT with 15% at farm A. All flocks were positive for MG and MS in serology. Although MG and IBV have been detected, this can be explained by the vaccination protocols, since live attenuated vaccines are widely used for immunization against these pathogens. It was also possible to detect OR and AP thorugh PCR in some flocks. The occurrence of several etiological agents that cause respiratory diseases in laying hens was confirmed by PCR and serology, with MS being the most prevalent and being present in all farms studied.

Proposal of an iELISA for Mycoplasma bovis diagnosis in dairy cattle and associated risk factors / [Proposta de um iELISA para diagnóstico de Mycoplasma bovis em bovinos leiteiros e fatores de risco associados]

Mycoplasma bovis is a highly contagious agent associated with several pathologies in cattle. The detection of reactive antibodies to M. bovis by Indirect Enzyme-Linked Immunosorbent Assay (iELISA) identifies if there was an exposure to the microorganism. The current study aimed to optimize an iELISA from M. bovis total cell antigen, applying it to bovine serum samples, and to evaluate risk factors. Serum samples were obtained from 400 cows from 17 herds from Southeast Brazil. In the optimization of iELISA, the following was established: 2 µg/mL of antigen, sera dilution 1:300, and conjugate dilution 1:15000. The frequency was 62.3% (249/400) of reactive animals and 100% (17/17) of reactive herds. Risk factors were: herds with more than 100 animals (OR= 3.1; CI= 95%); Holstein breed (OR= 72.5; CI= 95%); cows (OR= 29.7; CI= 95%); intensive breeding system (OR= 3.3; CI= 95%); associated small ruminant production (OR= 4.4; CI= 95%); milk production above 500L (OR= 2.9; CI= 95%); no quarantine (OR= 1.5; CI= 95%); mechanical milking (OR= 5.5; CI= 95%) and cases of mastitis (OR= 5.5; CI= 95%). The proposed iELISA was able to detect antibodies reactive to M. bovis in bovine serum. Knowledge of these risk factors can assist in the implementation of prophylactic measures.(AU)

Mycoplasma bovis é um agente altamente contagioso relacionado a várias patologias em bovinos. A detecção de anticorpos reativos a M. bovis por Ensaio de Imunoadsorção Enzimática Indireto (iELISA) identifica se houve exposição ao microrganismo. O presente estudo teve como objetivo otimizar um iELISA de antígeno celular total de M. bovis, aplicando-o a amostras de soro bovino, bem como avaliar fatores de risco. Amostras de soro foram obtidas de 400 vacas de 17 rebanhos da Região Sudeste do Brasil. Na otimização do iELISA foram obtidos: 2µg/mL de antígeno, diluição dos soros 1:300 e do conjugado 1:15000. A frequência de animais reativos foi de 62,3% (249/400) e de 100% (17/17) para os rebanhos. Os fatores de risco foram: rebanhos com mais de 100 animais (OR= 3,1; IC= 95%); raça Holandesa (OR= 72,5;IC= 95%); vacas (OR= 29,7;IC= 95%); sistema intensivo (OR= 3,3; C= 95%); produção de pequenos ruminantes (OR= 4,4;IC=95%); produção de leite acima de 500L (OR= 2,9;IC= 95%); sem quarentena (OR= 1,5;IC= 95%); ordenha mecânica (OR= 5,5;IC= 95%) e casos de mastite (OR= 5,5;IC= 95%). O iELISA proposto foi capaz de detectar anticorpos reativos a M. bovis no soro bovino. O conhecimento desses fatores de risco pode auxiliar na implementação de medidas profiláticas.(AU)

Qualitative and Quantitative Analysis of Salmonella spp. in Broilers Technological Processing and Determination of a Performance Objective (PO) for Frozen Chicken Breast

The present study investigated the frequency, level of contamination and serotyping of Salmonella strains isolated from broiler flocks in different processing sites and the fulfillment of a Performance Objective (PO) in frozen chicken breasts, as a risk assessment to measure the efficacy of prevention and control programs applied to reduce the risk of Salmonella spp. in raw poultry meat that contribute to reach food safety and public health goals. From 1,800 samples of cloacal swabs, carcasses before and after immersion chilling and frozen breasts derived from 20 broiler flocks slaughtered at two processing plants located in the mid-west and southern regions of Brazil, 278 samples were positive for Salmonella spp. by polymerase chain reaction (PCR) automated BAX System (DUPONT QUALICOM, USA), and 118 were enumerated by miniaturized most probable number technique. 122 Salmonella spp. strains were serotyped at the National Reference Laboratory of Cholera and Enteric Diseases of Oswaldo Cruz Institute Foundation (FIOCRUZ), showing a dominance of Salmonella Minnesota in every processing steps of the slaughterhouse located in the Brazilian mid-west region. Only 1 lot failed to reach the expected result for the Performance Objective (PO), using a maximum of 10% positivity acceptance for Salmonella spp. in frozen chicken breasts. Qualitative and quantitative results combined may be considered an effective tool to evaluate the effect of prevention and control programs for Salmonella spp. on the safety of the final product.(AU)

Qualitative and Quantitative Analysis of Salmonella spp. in Broilers Technological Processing and Determination of a Performance Objective (PO) for Frozen Chicken Breast

The present study investigated the frequency, level of contamination and serotyping of Salmonella strains isolated from broiler flocks in different processing sites and the fulfillment of a Performance Objective (PO) in frozen chicken breasts, as a risk assessment to measure the efficacy of prevention and control programs applied to reduce the risk of Salmonella spp. in raw poultry meat that contribute to reach food safety and public health goals. From 1,800 samples of cloacal swabs, carcasses before and after immersion chilling and frozen breasts derived from 20 broiler flocks slaughtered at two processing plants located in the mid-west and southern regions of Brazil, 278 samples were positive for Salmonella spp. by polymerase chain reaction (PCR) automated BAX System (DUPONT QUALICOM, USA), and 118 were enumerated by miniaturized most probable number technique. 122 Salmonella spp. strains were serotyped at the National Reference Laboratory of Cholera and Enteric Diseases of Oswaldo Cruz Institute Foundation (FIOCRUZ), showing a dominance of Salmonella Minnesota in every processing steps of the slaughterhouse located in the Brazilian mid-west region. Only 1 lot failed to reach the expected result for the Performance Objective (PO), using a maximum of 10% positivity acceptance for Salmonella spp. in frozen chicken breasts. Qualitative and quantitative results combined may be considered an effective tool to evaluate the effect of prevention and control programs for Salmonella spp. on the safety of the final product.

Prevalência e estudo genético de Mycoplasma gallisepticum e M. synoviae em poedeiras comerciais, na região centro-oeste do estado de São Paulo, Brasil / [Prevalence and genetic study of Mycoplasma gallisepticum and M. synoviae strains from commercial laying hens in the Midwest region of Sao Paulo state, Brazil]

O objetivo deste trabalho foi estudar a prevalência de MG e MS e a filogenia das cepas circulantes, comparando-as com outras já descritas em poedeiras comerciais no Brasil. Foram coletados 140 suabes traqueais de poedeiras comerciais com sinais respiratórios em seis granjas da região centro-oeste de São Paulo. As amostras foram avaliadas por PCR, com posterior sequenciamento e análise filogenética das cepas identificadas. Das 140 amostras, 16,4% foram positivas para MG e 68,6% para MS. Houve diferença significativa nas frequências de MG e MS por granja, segundo o teste G de independência (P<0,05). Todas as cepas identificadas de MG e MS de granjas distintas apresentaram similaridade tanto pela lipoproteína para MG quanto pela região 16s rRNA para MS. Neste estudo, foi possível observar altas prevalências dos agentes estudados, sendo a de MS maior que a de MG. Foi detectada infecção mista por MG e MS em 11,4% das amostras e sabe-se que esses micoplasmas podem agir de forma sinérgica, agravando o quadro respiratório. As cepas circulantes identificadas, pela análise das regiões gênicas da lipoproteína para MG e 16S rRNA para MS, são similares em todas as granjas estudadas.(AU)

The aim of this study was to evaluate the prevalence of MG and MS and the phylogeny of the circulating strains, comparing them with others already described in commercial laying hens from Brazil. A total of 140 tracheal swabs were collected from commercial laying hens with respiratory signs in six farms from the western region of São Paulo state. The samples were analyzed by PCR with subsequent sequencing and phylogenetic analysis of the identified strains. From the 140 samples, 68.6% were positive for MS and 16.4% for MG. There was a significant difference in the frequencies of MG and MS per farm according to G Test of independence (P<0.05). All strains identified as MG and MS from distinct farms presented similarity both by lipoprotein to MG and by 16s rRNA region to MS. In this study, it was possible to observe a high prevalence of MS compared to MG. Mixed MG and MS infection was detected in 11.4% of the samples. These mycoplasmas may act synergistically, worsening the respiratory signs. The circulating strains identified by analysis of the lipoprotein for MG and 16S rRNA for MS are similar on all poultry farms studied.(AU)

Occurrence of Avian Mycoplasmas in Free-Living Muscovy-Ducks (Cairina Moschata)

The present study aimed to investigate, by culture and PCR, the occurrence of Mollicutes, Mycoplasma gallisepticum and Mycoplasma synoviae in free-living Muscovy-ducks (Cairina moschata) from the Rio Zoo, RJ, Brazil. Tracheal swabs were obtained from 82 asymptomatic ducks and the samples were submitted to culture of mycoplasmas and PCR for identification of Mollicutes Class, Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS). Samples were also analyzed directly by PCR, without prior culture, for Mollicutes, MG and MS. Eighteen (18/82) Muscovy-ducks were positive for Mollicutes by culture, all isolates were confirmed as Mollicutes and seven were identified as MG. Of the samples analyzed directly by PCR, without prior culture, 17,1% (14/82) was positive for Mollicutes, being 35,7% (5/14) identified as MG and 21,4% (3/14) as MS. The occurrence of Mollicutes class bacteria was detected in Muscovy-ducks. MG and MS were identified in these animals suggesting the circulation of these agents in the Rio de Janeiro Zoo and may present a risk for the health status of the other birds.(AU)

Prevalência e estudo genético de Mycoplasma gallisepticum e M. synoviae em poedeiras comerciais, na região centro-oeste do estado de São Paulo, Brasil / [Prevalence and genetic study of Mycoplasma gallisepticum and M. synoviae strains from commercial laying hens in the Midwest region of Sao Paulo state, Brazil]

O objetivo deste trabalho foi estudar a prevalência de MG e MS e a filogenia das cepas circulantes, comparando-as com outras já descritas em poedeiras comerciais no Brasil. Foram coletados 140 suabes traqueais de poedeiras comerciais com sinais respiratórios em seis granjas da região centro-oeste de São Paulo. As amostras foram avaliadas por PCR, com posterior sequenciamento e análise filogenética das cepas identificadas. Das 140 amostras, 16,4% foram positivas para MG e 68,6% para MS. Houve diferença significativa nas frequências de MG e MS por granja, segundo o teste G de independência (P<0,05). Todas as cepas identificadas de MG e MS de granjas distintas apresentaram similaridade tanto pela lipoproteína para MG quanto pela região 16s rRNA para MS. Neste estudo, foi possível observar altas prevalências dos agentes estudados, sendo a de MS maior que a de MG. Foi detectada infecção mista por MG e MS em 11,4% das amostras e sabe-se que esses micoplasmas podem agir de forma sinérgica, agravando o quadro respiratório. As cepas circulantes identificadas, pela análise das regiões gênicas da lipoproteína para MG e 16S rRNA para MS, são similares em todas as granjas estudadas.(AU)

The aim of this study was to evaluate the prevalence of MG and MS and the phylogeny of the circulating strains, comparing them with others already described in commercial laying hens from Brazil. A total of 140 tracheal swabs were collected from commercial laying hens with respiratory signs in six farms from the western region of São Paulo state. The samples were analyzed by PCR with subsequent sequencing and phylogenetic analysis of the identified strains. From the 140 samples, 68.6% were positive for MS and 16.4% for MG. There was a significant difference in the frequencies of MG and MS per farm according to G Test of independence (P<0.05). All strains identified as MG and MS from distinct farms presented similarity both by lipoprotein to MG and by 16s rRNA region to MS. In this study, it was possible to observe a high prevalence of MS compared to MG. Mixed MG and MS infection was detected in 11.4% of the samples. These mycoplasmas may act synergistically, worsening the respiratory signs. The circulating strains identified by analysis of the lipoprotein for MG and 16S rRNA for MS are similar on all poultry farms studied.(AU)

High Prevalence of Multidrug-Resistant Nontyphoidal Salmonella Recovered from Broiler Chickens and Chicken Carcasses in Brazil

The extensive use of antimicrobial agents has contributed to the emergence of antimicrobial resistance and multidrug resistance (MDR) in Salmonella, an important zoonotic pathogen that causes outbreaks and sporadic cases of gastroenteritis in humans. The study aimed to investigate the antimicrobial resistance profile of Salmonella strains isolated from poultry in Brazil. A total of 230 Salmonella strains, isolated from cloacal swabs (n=56) and broiler carcasses swabs (n=174) before and after chilling from slaughterhouses under Federal Inspection Service within the period 2012-2017, were analyzed. Serotyping and antibiotic susceptibility testing were performed on all the isolates. Serotyping results showed that 41% of the strains were Salmonella Heidelberg, 29% S. Minnesota, 12% S. Saintpaul, 6.5% S. Enteritidis, 3.9% S. Anatum, 2.2% S. Cerro, 2.2% S. Senftenberg, 1.7% S. Newport, 0.4% S. Ealing, 0.4% S. O:4,5 and 0.4% S. O:9,12. MDR rates of the isolates were 67.4%. S. Heidelberg 89.5%, S. Minnesota 51.5%, S. Saintpaul 82.1%, S. Anatum 66.7%, S. Cerro 60%, S. Senftenberg 40%. Out of the 230 strains, 41.3% presented resistance to Penicillins + beta-lactamase inhibitor, Penicillin, 1st and 2nd Generation Cephalosporin, 3rd and 4th Generation Cephalosporin, Tetracycline and Sulfonamide. Salmonella Heidelberg, S. Saintpaul, S. Anatum, S. Cerro, S. Senftenberg and S. Minnesota were isolated after chilling tank highlighting a food safety concern for the industry of poultry and poultry products indicating a risk to collective health. The high prevalence of MDR nontyphoidal Salmonella obtained in this study limit the options available to treat infectious disease in humans and animals.(AU)

High Prevalence of Multidrug-Resistant Nontyphoidal Salmonella Recovered from Broiler Chickens and Chicken Carcasses in Brazil

The extensive use of antimicrobial agents has contributed to the emergence of antimicrobial resistance and multidrug resistance (MDR) in Salmonella, an important zoonotic pathogen that causes outbreaks and sporadic cases of gastroenteritis in humans. The study aimed to investigate the antimicrobial resistance profile of Salmonella strains isolated from poultry in Brazil. A total of 230 Salmonella strains, isolated from cloacal swabs (n=56) and broiler carcasses swabs (n=174) before and after chilling from slaughterhouses under Federal Inspection Service within the period 2012-2017, were analyzed. Serotyping and antibiotic susceptibility testing were performed on all the isolates. Serotyping results showed that 41% of the strains were Salmonella Heidelberg, 29% S. Minnesota, 12% S. Saintpaul, 6.5% S. Enteritidis, 3.9% S. Anatum, 2.2% S. Cerro, 2.2% S. Senftenberg, 1.7% S. Newport, 0.4% S. Ealing, 0.4% S. O:4,5 and 0.4% S. O:9,12. MDR rates of the isolates were 67.4%. S. Heidelberg 89.5%, S. Minnesota 51.5%, S. Saintpaul 82.1%, S. Anatum 66.7%, S. Cerro 60%, S. Senftenberg 40%. Out of the 230 strains, 41.3% presented resistance to Penicillins + beta-lactamase inhibitor, Penicillin, 1st and 2nd Generation Cephalosporin, 3rd and 4th Generation Cephalosporin, Tetracycline and Sulfonamide. Salmonella Heidelberg, S. Saintpaul, S. Anatum, S. Cerro, S. Senftenberg and S. Minnesota were isolated after chilling tank highlighting a food safety concern for the industry of poultry and poultry products indicating a risk to collective health. The high prevalence of MDR nontyphoidal Salmonella obtained in this study limit the options available to treat infectious disease in humans and animals.

Occurrence of Avian Mycoplasmas in Free-Living Muscovy-Ducks (Cairina Moschata)

The present study aimed to investigate, by culture and PCR, the occurrence of Mollicutes, Mycoplasma gallisepticum and Mycoplasma synoviae in free-living Muscovy-ducks (Cairina moschata) from the Rio Zoo, RJ, Brazil. Tracheal swabs were obtained from 82 asymptomatic ducks and the samples were submitted to culture of mycoplasmas and PCR for identification of Mollicutes Class, Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS). Samples were also analyzed directly by PCR, without prior culture, for Mollicutes, MG and MS. Eighteen (18/82) Muscovy-ducks were positive for Mollicutes by culture, all isolates were confirmed as Mollicutes and seven were identified as MG. Of the samples analyzed directly by PCR, without prior culture, 17,1% (14/82) was positive for Mollicutes, being 35,7% (5/14) identified as MG and 21,4% (3/14) as MS. The occurrence of Mollicutes class bacteria was detected in Muscovy-ducks. MG and MS were identified in these animals suggesting the circulation of these agents in the Rio de Janeiro Zoo and may present a risk for the health status of the other birds.

Caracterização macroscópica e microscópica da pododermatite em relação à classificação industrial de pés de frango para consumo humano / Pododermatitis macroscopic and microscopic characterization related to industrial classification of chicken feet for human consumption

O presente trabalho teve como objetivo identificar e classificar a pododermatite em frangos de corte, comparando as lesões microscópicas com os aspectos macroscópicos utilizados pela inspeção sanitária. Foi realizada coleta de pés de frangos de corte, em matadouro de aves sob inspeção sanitária, após classificação utilizada nos padrões para exportação. Foram coletados 30 pés tipo A, 30 pés tipo B e 33 pés tipo C, escolhidos aleatoriamente dentro de cada grupo. Para análise histopatológica, foram desenvolvidos escores de acordo com a gravidade das lesões, variando de 0 a 2. Foi observada associação (qui-quadrado, P<0,05) entre a classificação macroscópica (A, B e C) e as alterações histológicas (0, 1 e 2). A classificação A diferiu significativamente (ANOVA, Tukey-Kramer, P<0,05) das classificações B e C quanto aos escores histopatológicos observados. Não houve diferença no comprometimento dos pés pelas lesões que justificassem a separação entre os pés classificados em B e C, uma vez que ambos apresentaram delimitação linear das lesões, sugerindo superficialidade e restrição ao epitélio queratinizado. Portanto, sugere-se o aproveitamento dos pés para consumo humano após remoção mecânica do "calo de pé", uma vez que este produto não oferece riscos ao consumidor.(AU)

The aim of this study was to identify and classify pododermatitis in broilers, comparing the microscopic lesions with the macroscopic aspects used by the Sanitary Inspection. Broiler chicken feet were collected at a Poultry slaughterhouse under Sanitary Inspection, after classification according to the exportation standards. The chicken feet were randomly selected in each group, 30 feet type A, 30 feet type B and 33 feet type C. For the histopathological analysis, scores were developed according to the severity of the lesions, varying from 0 to 2. There was association (Chi-square, P< 0.05) between the macroscopic classification (A, B and C) and histological changes (0, 1 and 2). The A classification differed significantly (ANOVA, Tukey-Kramer, P< 0.05) from the B and C classifications regarding the histopathological scores observed. There was no difference in feet lesions that justified the separation between the feet classified in B and C, since both presented a linear delimitation of the lesions, suggesting superficiality and restriction to the keratinized epithelium. Therefore, the use of the feet for human consumption after mechanical removal of the footpad lesions is suggested since the product does not pose risks to the consumer.(AU)

Caracterização macroscópica e microscópica da pododermatite em relação à classificação industrial de pés de frango para consumo humano / Pododermatitis macroscopic and microscopic characterization related to industrial classification of chicken feet for human consumption

O presente trabalho teve como objetivo identificar e classificar a pododermatite em frangos de corte, comparando as lesões microscópicas com os aspectos macroscópicos utilizados pela inspeção sanitária. Foi realizada coleta de pés de frangos de corte, em matadouro de aves sob inspeção sanitária, após classificação utilizada nos padrões para exportação. Foram coletados 30 pés tipo A, 30 pés tipo B e 33 pés tipo C, escolhidos aleatoriamente dentro de cada grupo. Para análise histopatológica, foram desenvolvidos escores de acordo com a gravidade das lesões, variando de 0 a 2. Foi observada associação (qui-quadrado, P<0,05) entre a classificação macroscópica (A, B e C) e as alterações histológicas (0, 1 e 2). A classificação A diferiu significativamente (ANOVA, Tukey-Kramer, P<0,05) das classificações B e C quanto aos escores histopatológicos observados. Não houve diferença no comprometimento dos pés pelas lesões que justificassem a separação entre os pés classificados em B e C, uma vez que ambos apresentaram delimitação linear das lesões, sugerindo superficialidade e restrição ao epitélio queratinizado. Portanto, sugere-se o aproveitamento dos pés para consumo humano após remoção mecânica do "calo de pé", uma vez que este produto não oferece riscos ao consumidor.(AU)

The aim of this study was to identify and classify pododermatitis in broilers, comparing the microscopic lesions with the macroscopic aspects used by the Sanitary Inspection. Broiler chicken feet were collected at a Poultry slaughterhouse under Sanitary Inspection, after classification according to the exportation standards. The chicken feet were randomly selected in each group, 30 feet type A, 30 feet type B and 33 feet type C. For the histopathological analysis, scores were developed according to the severity of the lesions, varying from 0 to 2. There was association (Chi-square, P< 0.05) between the macroscopic classification (A, B and C) and histological changes (0, 1 and 2). The A classification differed significantly (ANOVA, Tukey-Kramer, P< 0.05) from the B and C classifications regarding the histopathological scores observed. There was no difference in feet lesions that justified the separation between the feet classified in B and C, since both presented a linear delimitation of the lesions, suggesting superficiality and restriction to the keratinized epithelium. Therefore, the use of the feet for human consumption after mechanical removal of the footpad lesions is suggested since the product does not pose risks to the consumer.(AU)

Serotyping and genotyping of salmonella strains isolated from broilers, chicken carcasses before and after chilling, and frozen chicken breasts produced in the states of Mato Grosso do Sul and Santa Catarina, Brazil

The present study investigated the effectiveness of a single Salmonella prevention and control program applied in two different processing plants, located in the states of Mato Grosso do Sul (plant A) and Santa Catarina (Plant B), belonging to the same company, and identified Salmonella strain subtypes isolated from broilers, carcasses before and after chilling, and frozen chicken breasts. The Salmonella prevention and control program was 90% effective in plant A and 100% in plant B, considering a level of 10% positive samples per frozen chicken breast batch acceptable. A total of 128 strains were serotyped, being 10 from drag swabs, 31 from cloacal swabs, 83 from carcasses, and 4 from frozen chicken breasts. After serotyping analysis, 30 strains isolated at different processing steps and drag swabs, and three Salmonella Minnesota strains isolated in 2012 in plant A, were genotyped by PFGE. In plant A, the most frequently strain isolated was Salmonella Minnesota (90.35%), followed by Salmonella Newport (8.77%), and in Plant B, Salmonella Senftenberg (80%). Salmonella Minnesota strains were differentiated by PFGE into 19 pulsotypes distributed in three clusters. The phenotypic identification by serotyping of four strains diverged from their PFGE genotypic results. Most Salmonella Minnesota strains genotyped in plant A and the strains isolated from environmental samples in 2012 in the same broiler processing plant belong to a single cluster, confirming the dominance and persistence of this clone over time.(AU)

Serotyping and genotyping of salmonella strains isolated from broilers, chicken carcasses before and after chilling, and frozen chicken breasts produced in the states of Mato Grosso do Sul and Santa Catarina, Brazil

The present study investigated the effectiveness of a single Salmonella prevention and control program applied in two different processing plants, located in the states of Mato Grosso do Sul (plant A) and Santa Catarina (Plant B), belonging to the same company, and identified Salmonella strain subtypes isolated from broilers, carcasses before and after chilling, and frozen chicken breasts. The Salmonella prevention and control program was 90% effective in plant A and 100% in plant B, considering a level of 10% positive samples per frozen chicken breast batch acceptable. A total of 128 strains were serotyped, being 10 from drag swabs, 31 from cloacal swabs, 83 from carcasses, and 4 from frozen chicken breasts. After serotyping analysis, 30 strains isolated at different processing steps and drag swabs, and three Salmonella Minnesota strains isolated in 2012 in plant A, were genotyped by PFGE. In plant A, the most frequently strain isolated was Salmonella Minnesota (90.35%), followed by Salmonella Newport (8.77%), and in Plant B, Salmonella Senftenberg (80%). Salmonella Minnesota strains were differentiated by PFGE into 19 pulsotypes distributed in three clusters. The phenotypic identification by serotyping of four strains diverged from their PFGE genotypic results. Most Salmonella Minnesota strains genotyped in plant A and the strains isolated from environmental samples in 2012 in the same broiler processing plant belong to a single cluster, confirming the dominance and persistence of this clone over time.

Pathological Aspects of a Subclinical Marek's Disease Case in Free-Range Chickens

The present report aims at describing the occurrence of Marek's Disease (MD) in a free range poultry farm based on macroscopic and microscopic lesions. For this purpose, seven free-range chickens were evaluated by gross and microscopic examination. Tumor lesions were observed in the liver and ovary, in addition of enlarged kidney and spleen, skin thickening and whitish lesions in streak form in the breast muscle. Fragments of the affected organs were collected and placed in 10% formalin for processing by the usual histopathology techniques, and stained with hematoxylin-eosin. Under microscopic evaluation, we observed pleomorphic populations of neoplastic lymphoreticular cells with pyknotic nuclei in the same organs, all compatible with MD. The macroscopic and microscopic lesions presented in this study were consistent with MD, meaning that the MD virus is present in the rearing environment of the studied chicken farm.

Pathological Aspects of a Subclinical Marek's Disease Case in Free-Range Chickens

The present report aims at describing the occurrence of Marek's Disease (MD) in a free range poultry farm based on macroscopic and microscopic lesions. For this purpose, seven free-range chickens were evaluated by gross and microscopic examination. Tumor lesions were observed in the liver and ovary, in addition of enlarged kidney and spleen, skin thickening and whitish lesions in streak form in the breast muscle. Fragments of the affected organs were collected and placed in 10% formalin for processing by the usual histopathology techniques, and stained with hematoxylin-eosin. Under microscopic evaluation, we observed pleomorphic populations of neoplastic lymphoreticular cells with pyknotic nuclei in the same organs, all compatible with MD. The macroscopic and microscopic lesions presented in this study were consistent with MD, meaning that the MD virus is present in the rearing environment of the studied chicken farm.(AU)

Bacterial microbiota present in the gallbladder of cattle and antimicrobial resistance of Staphylococcus isolates / Microbiota bacteriana presente na vesícula biliar de bovinos e resistência antimicrobiana de isolados de Staphyloccocus

Pathogenic microorganisms can reside transiently or permanently in the gallbladder of cattle. Thus, during slaughter, more attention should be given to the gastrointestinal tract, especially to the accessory organ, the gallbladder. The main aim of this study was to characterize the bacterial microbiota present in bile and gallbladder epithelium of cattle slaughtered in a slaughtering plant under sanitary conditions and to evaluate the antimicrobial resistance in strains of the genus Staphylococcus. Thirty intact gallbladders were collected and the in bile and epithelium were researched for the presence of Aerobic Mesophilic Heterotrophic Bacteria (AMHB), Staphylococcus spp., total Enterobacteriaceae, Enterococcus spp. and Salmonella spp.The frequency of isolation of the microorganism mentioned above were, respectively: 23.02 percent, 14.39 percent, 13.67 percent, 24.46 percent, 0 percent and 24.46 percent. Concerning both gallbladder environments, the frequency of isolation of the microorganisms in the epithelium was 64.03 percent, and in the bile 35.97 percent, with no statistical difference, but with significant difference between the population averages. In antimicrobial susceptibility testing, strains of Staphylococcus from both bile and gallbladder epithelium showed sensitivity to the antimicrobials: penicillin G, ceftriaxone, chloramphenicol and gentamicin. The observation that the gallbladder supports a high frequency of microorganisms brings us to the possible fact that cattle might be a persistent carrier of pathogens of great importance to public health.(AU)

Microrganismos patogênicos podem residir temporariamente ou permanentemente na vesícula biliar de bovinos. Assim, durante o abate, maior atenção deve ser dada ao trato gastrointestinal, especialmente para o órgão acessório, a vesícula biliar. O principal objetivo deste estudo foi caracterizar a microbiota bacteriana presente na bile e no epitélio de vesículas biliares de bovinos abatidos em matadouro frigorífico sob inspeção sanitária e avaliar a resistência antimicrobiana de estirpes do gênero Staphyloccocus. Foram coletadas 30 vesículas biliares íntegras e foi pesquisada na bile e no epitélio do órgão a presença de bactérias heterotróficas aeróbias mesófilas (BHAM), Staphylococcus spp. e Enterobacteriaceae totais, Escherichia coli, Enterococcus spp. e Salmonella spp. A frequência de isolamento dos microrganismos citados acima foi, respectivamente: 23,02 por cento, 14,39 por cento, 13,67 por cento, 24,46 por cento, 0por cento e 24,46 por cento. Em relação aos dois ambientes da vesícula, a frequência de isolamento dos microrganismos no epitélio foi de 64,03 por cento, e na bile 35,97 por cento, não sendo diferente estatisticamente, mas com diferença significativa entre as médias populacionais.No teste de susceptibilidade aos antimicrobianos, as estirpes de Staphylococcus isoladas a partir da bile e do epitélio da vesícula biliar apresentaram sensibilidade a: penicilina G, ceftriaxona, cloranfenicol e gentamicina. A observação de que a vesícula biliar comporta microrganismos em elevadas frequências atenta para o fato de que o bovino possa ser um portador persistente de patógenos de grande importância em saúde pública.(AU)

Eggshell apex abnormalities in a free-range hen farm with mycoplasma synoviae and infectious bronchitis virus in Rio de Janeiro state, Brazil

A farm with 3,000 free-range hens between 24 and 65 weeks of age was investigated. These hens were separated in small flocks of 400 to 700 birds, presenting 10 to 23% egg production reduction. Twenty serum samples were collected during the period of drop in egg production and three weeks later for the investigation of Mycoplasma synoviae (MS), M. gallisepticum (MG) and Infectious Bronchitis Virus (IBV) antibodies using ELISA. At the time of the second collection, egg production had resumed to normal levels; however, with 10.23% of the eggs showed eggshell abnormalities limited to the apex. Eggshell strength was significantly different between normal and those with eggshell apex abnormalities, but not other egg-quality parameters. ELISA tests showed that MS and IBV titers increased during the evaluated period. MS infection was confirmed by culture and by PCR of tracheal swabs. All samples were negative for MG by ELISA and PCR. Further studies with larger samples to ensure the occurrence of this disease in industrial layer flocks in Brazil are under way.