EFFECTS OF MYCOPLASMA GALLISEPTICUM INFECTION ON PREENING BEHAVIORS AND FEATHER QUALITY IN HOUSE FINCHES (HAEMORHOUS MEXICANUS).

Infections can have far-reaching sublethal effects on wildlife, including reduced maintenance of external structures. For many wildlife taxa, daily maintenance of external structures (termed preening in birds) is critical to fitness, but few studies have examined how infections alter such maintenance. Mycoplasma gallisepticum is a common pathogen in free-living House Finches (Haemorhous mexicanus), where it causes mycoplasmal conjunctivitis. Despite documented behavioral changes associated with M. gallisepticum infections in finches, no studies have examined how preening behavior may change with infection and how potential differences in preening may affect feather quality. To test this, we experimentally inoculated captive House Finches with M. gallisepticum or a control treatment, and we collected behavioral and feather quality data to detect potential changes in feather maintenance due to infection. We found that finches infected with M. gallisepticum preened significantly less often, and within the infected treatment, birds with the highest conjunctivitis severity preened the least often. However, there was no difference in the quality scores for secondary flight feathers collected from control versus infected birds. We also assayed feather water retention and found that the degree of water retention correlated with our feather quality scores, such that feathers with poor scores retained more water. However, as with quality scores, feather water retention did not differ with infection; this may be due to the controlled environment that the birds experienced while in captivity. Our data suggest that, in addition to sickness behaviors previously observed in finches, M. gallisepticum infection decreases other behaviors critical to survival, such as preening. While the consequences of reduced preening on feather maintenance were not apparent in captive conditions, further work is needed to determine whether House Finches in the wild that are infected with M. gallisepticum experience a fitness cost, such as increases in ectoparasite loads, due to this reduced feather maintenance.

Molecular detection of Mycoplasma gallisepticum in different poultry breeds of Abbottabad and Rawalpindi, Pakistan / Detecção molecular de Mycoplasma gallisepticum em diferentes raças de aves de Abbottabad e Rawalpindi, Paquistão

Abstract The poultry sector in Pakistan is contributing mainly in bridging gap between demand and supply for protein. Mycoplasma gallisepticum is an emerging bacterium causing serious problems in poultry industry of Pakistan. A cross-sectional study was conducted to evaluate the M. gallisepticum load in poultry populated regions of Pakistan. Total 600 serum and 600 swab samples were collected, 200 from each broiler, layers and breeders poultry in Rawalpindi and Abbottabad districts. Serum samples were analyzed through ELISA for seroprevalence. Swabs were cultured on Frey's medium followed by PCR and partial mgc2 gene sequencing. Results of seroprevalence of M. gallisepticum showed that layers (75%, n=150) are more positive as compared to breeders (70%, n=140) and broilers (50%, n=100). Typical colonies of the M. gallisepticum were observed in breeder (26.5%), followed by layer (21%) and broilers (9%). A total of 37.1% (n=42) samples were identified positive through PCR out of total 113 cultured based positive samples. A total of six M. gallisepticum isolates of current study showed 98-99 percent similarity with previously reported isolates on the basis of mgc2 gene partial sequencing. The M. gallisepticum was found highly prevalent in different poultry breads. Results of this study would add into basic data and provide a direction for livestock sector to strengthen a control strategy for mycoplasmosis in poultry farms.

Resumo O setor avícola do Paquistão está contribuindo principalmente para preencher a lacuna entre a demanda e a oferta de proteína. Mycoplasma gallisepticum é uma bactéria emergente que causa sérios problemas na indústria avícola do Paquistão. Um estudo transversal foi conduzido para avaliar a carga de M. gallisepticum em regiões de avicultura do Paquistão. Um total de 600 amostras de soro e 600 amostras de esfregaço foi coletado, 200 de cada frango de corte, poedeiras e aves reprodutoras nos distritos de Rawalpindi e Abbottabad. Amostras de soro foram analisadas por ELISA para soroprevalência. As zaragatoas foram cultivadas em meio Frey, seguido de PCR e sequenciação parcial do gene mgc2. Os resultados da soroprevalência de M. gallisepticum mostraram que as poedeiras (75%, n = 150) são mais positivas em comparação com matrizes (70%, n = 140) e frangos de corte (50%, n = 100). Colônias típicas de M. gallisepticum foram observadas em reprodutoras (26,5%), seguidas de poedeiras (21%) e frangos de corte (9%). Um total de 37,1% (n = 42) das amostras foi identificado como positivas por PCR de um total de 113 amostras positivas baseadas em cultura. Um total de seis isolados de M. gallisepticum do estudo atual mostrou 98-99% de similaridade com isolados relatados anteriormente com base no sequenciamento parcial do gene mgc2. O M. gallisepticum foi encontrado com alta prevalência em diferentes pães de aves. Os resultados deste estudo acrescentariam dados básicos e forneceriam orientação para o setor pecuário fortalecer uma estratégia de controle da micoplasmose em granjas avícolas.

Molecular detection of Mycoplasma gallisepticum in different poultry breeds of Abbottabad and Rawalpindi, Pakistan.

The poultry sector in Pakistan is contributing mainly in bridging gap between demand and supply for protein. Mycoplasma gallisepticum is an emerging bacterium causing serious problems in poultry industry of Pakistan. A cross-sectional study was conducted to evaluate the M. gallisepticum load in poultry populated regions of Pakistan. Total 600 serum and 600 swab samples were collected, 200 from each broiler, layers and breeders poultry in Rawalpindi and Abbottabad districts. Serum samples were analyzed through ELISA for seroprevalence. Swabs were cultured on Frey's medium followed by PCR and partial mgc2 gene sequencing. Results of seroprevalence of M. gallisepticum showed that layers (75%, n=150) are more positive as compared to breeders (70%, n=140) and broilers (50%, n=100). Typical colonies of the M. gallisepticum were observed in breeder (26.5%), followed by layer (21%) and broilers (9%). A total of 37.1% (n=42) samples were identified positive through PCR out of total 113 cultured based positive samples. A total of six M. gallisepticum isolates of current study showed 98-99 percent similarity with previously reported isolates on the basis of mgc2 gene partial sequencing. The M. gallisepticum was found highly prevalent in different poultry breads. Results of this study would add into basic data and provide a direction for livestock sector to strengthen a control strategy for mycoplasmosis in poultry farms.

Flow-Cytometric Method for Viability Analysis of Mycoplasma gallisepticum and Other Cell-Culture-Contaminant Mollicutes.

Mycoplasma is the smallest self-replicating bacteria, figuring as common contaminant of eukaryotic cell cultures. Production inputs and operator's manipulation seem to be the main sources of such contamination. Many analytical approaches have been applied for mycoplasma detection in cell cultures and also in biological products. However, unless they were validated, only indicator cell culture and bacteriological culture are considered as compendial methods for quality control of biological products. Nano-flow cytometry has been pointed out as an alternative technique for addressing prokaryotic and eukaryotic cell viability being a substantial tool for reference material production. In this study, a viability-flow-cytometry assay was standardized for M. gallisepticum and then applied to other cell-culture-contaminant mycoplasmas. For this, M. galliseticum's growth rate was observed and different treatments were evaluated to establish low viability cultures (cell death-induced control). Distinct viability markers and their ideal concentrations (titration) were appraised. Ethanol treatment showed to be the best death-inducing control. CFDA and TOPRO markers revealed to be the best choice for detecting live and dead mycoplasma frequencies, respectively. The standardized methodology was applied to Mycoplasma arginini, M. hyorhinis, M. orale, Spiroplasma citri and Acholeplasma laidlawii. Significant statistical difference was observed in the percentage of viable cells in comparison to ethanol treatment for A. laidlawii in CFDA and in both markers for M. gallisepticum, M. hyorhinis and S. citri. In summary, we standardized a flow cytometry assay for assessing M. gallisepticum - and potentially other species - viability and ultimately applied for reference material production improving the quality control of biological products.

Incidence of Mycoplasma gallisepticum and Mycoplasma synoviae in broiler flocks at the Federal District of Brazil and its surrounding areas / Incidência de lotes de frangos afetados por Mycoplasma gallisepticum e Mycoplasma synoviae no Distrito Federal e Entorno, Brasil

Mycoplasma is an important avian pathogen that can cause both respiratory disease and synovitis in birds, resulting in considerable economic losses to the poultry industry worldwide. This study aimed to determine the incidence of Mycoplasma gallisepticum and M. synoviae in broiler flocks at the Federal District and its surrounding regions using polymerase chain reaction (PCR). All slaughtered lots (57 flocks) were analyzed from July to November in one of the two slaughterhouses at the Federal District with Federal inspection services. Approximately 10 samples of broiler tracheae per slaughtered batch were collected from the evisceration line. The results obtained from the accumulated incidence over the study period were 7.02% and 35.09% for M. gallisepticum and M. synoviae, respectively. A greater concentration of flocks affected by M. synoviae was observed during October. The sample design as well as the PCR technique assisted in detecting both agents in the broiler batches in the first epidemiological study of these two agents in the region.(AU)

Os micoplasmas são importantes patógenos aviários, que podem causar doenças respiratórias e sinovites em aves, resultando em consideráveis perdas econômicas para a indústria avícola em todo o mundo. O objetivo deste estudo foi determinar a incidência de Mycoplasma gallisepticum e Mycoplasma synoviae em lotes de frangos na região do Distrito Federal e Entorno, por meio da reação em cadeia de polimerase (PCR). Todos os lotes abatidos (57 lotes) foram analisados durante os meses de julho a novembro, em um dos dois abatedouros frigoríficos do Distrito Federal com Serviço de Inspeção Federal. Na linha de evisceração foram coletadas cerca de 10 amostras de traqueia de frangos de corte por lote abatido. Os resultados obtidos da incidência acumulada no período de estudo foram de 7.02% para M. gallisepticum e 35,09% M. synoviae. Uma maior concentração do número de lotes afetados por M. synoviae foi observada durante o mês de outubro. O desenho amostral, assim como a técnica de PCR, permitiu a detecção de ambos os agentes nos lotes de frangos de corte analisados, sendo este o primeiro estudo epidemiológico desses dois agentes na região de estudo.(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)

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)

Investigation of Mycoplasma spp. in birds of the Rio de Janeiro Zoo by isolation and PCR / Investigação de Mycoplasma spp. em aves do Zoológico do Rio de Janeiro por isolamento e PCR

Brazil is one of the countries with the most abundant avifauna in the world. The confinement of birds associated with close contact with other animals and humans favor the spread of agents of respiratory diseases. Among them, mycoplasmas can cause asymptomatic or apparent disease that manifests in birds by coughing, sneezing, rales, conjunctivitis, ocular and nasal discharge. Several described mycoplasmas cause disease in birds, especially Mycoplasma gallisepticum (MG) and Mycoplasma synoviae(MS). The diagnosis of Mycoplasma spp. can be done by clinical observation and laboratory analysis. Molecular diagnosis by PCR was boosted by its speed, sensitivity, and low cost of agent isolation techniques that take up to 21 days to complete. This study aimed to verify the occurrence of Mycoplasma spp. in birds of the Rio de Janeiro Zoo (Rio Zoo), by isolation and PCR. Of the total 635 birds from the Rio Zoo, 81 were studied for detection of Mycoplasma spp., when taken for routine health assessment exams. These birds belonged to the following orders: Psittaciformes (45), Accipitriformes (18), Galliformes (7), Piciformes (5), Strigiformes (4), Falconiformes (1) and Cariamiformes (1), all individuals already identified by microchip or leg-ring. There was no isolation of mycoplasmas in any of the samples tested, whereas, in the PCR, 62.96% (51/81) were positive, with 1.96% (1/51) identified as MG and 19.61% (10/51) as MS, representing 1.23% (1/81) and 12.34% (10/81) of the total population studied. PCR was shown to be a more effective technique than isolation in the detection of Mycoplasma spp. in birds. It was possible to detect mycoplasmas in birds from Riozoo with no clinical respiratory signs, with higher MS prevalence than MG. The positivities for Mycoplasma spp., MS, and MG were different among the orders studied, being the highest occurrence in birds of prey, followed by Galliformes and Piciformes...(AU)

O Brasil é um dos países com maior avifauna do mundo. O confinamento de aves associado ao contato próximo a outros animais e seres humanos favorece a disseminação de agentes etiológicos causadores de doenças respiratórias. Dentre eles, os micoplasmas podem causar doença assintomática ou aparente que se manifesta em aves por espirros, estertores, conjuntivite, corrimentos oculares e nasais. São diversos os micoplasmas descritos causadores de doença em aves, com destaque para Mycoplasma gallisepticum (MG) e Mycoplasma synoviae (MS). O diagnóstico de Mycoplasma spp. pode ser feito pela observação clínica e análises laboratoriais. O diagnóstico molecular pela Reação em Cadeia da Polimerase (PCR) ganhou impulso por sua rapidez, sensibilidade e baixo custo em relação às técnicas de isolamento do agente que levam até 21 dias para conclusão do gênero Mycoplasma. Objetivou-se verificar a ocorrência da infecção por Mycoplasma spp. em aves no Zoológico do Rio de Janeiro (Rio Zoo), por isolamento e PCR. Do plantel de 635 aves do Rio Zoo, foram estudadas 81 para detecção de Mycoplasma spp., quando contidas para exames rotineiros de avaliação da condição de saúde. Essas aves eram pertencentes às ordens Psittaciformes (45), Accipitriformes (18), Galliformes (7), Piciformes (5), Strigiformes (4), Falconiformes (1) e Cariamiformes (1), todas já identificadas por microchip ou por anilha. Não houve isolamento de micoplasmas em nenhuma das amostras testadas, enquanto na PCR, 62,96% (51/81) foram positivas, sendo 1,96% (1/51) identificadas como MG e 19,61% (10/51) como MS, representando 1,23% (1/81) e 12,34% (10/81) da população total estudada. A PCR demonstrou ser uma técnica mais efetiva que o isolamento na detecção de Mycoplasma spp. em aves. Foi possível detectar micoplasmas nas aves do Riozoo sem sinal clínico respiratório, tendo MS maior prevalência do que MG...(AU)

Investigation of Mycoplasma spp. in birds of the Rio de Janeiro Zoo by isolation and PCR / Investigação de Mycoplasma spp. em aves do Zoológico do Rio de Janeiro por isolamento e PCR

Brazil is one of the countries with the most abundant avifauna in the world. The confinement of birds associated with close contact with other animals and humans favor the spread of agents of respiratory diseases. Among them, mycoplasmas can cause asymptomatic or apparent disease that manifests in birds by coughing, sneezing, rales, conjunctivitis, ocular and nasal discharge. Several described mycoplasmas cause disease in birds, especially Mycoplasma gallisepticum(MG) andMycoplasma synoviae(MS). The diagnosis ofMycoplasmaspp. can be done by clinical observation and laboratory analysis. Molecular diagnosis by PCR was boosted by its speed, sensitivity, and low cost of agent isolation techniques that take up to 21 days to complete. This study aimed to verify the occurrence ofMycoplasmaspp. in birds of the Rio de Janeiro Zoo (Rio Zoo), by isolation and PCR. Of the total 635 birds from the Rio Zoo, 81 were studied for detection ofMycoplasmaspp., when taken for routine health assessment exams. These birds belonged to the following orders: Psittaciformes (45), Accipitriformes (18), Galliformes (7), Piciformes (5), Strigiformes (4), Falconiformes (1) and Cariamiformes (1), all individuals already identified by microchip or leg-ring. There was no isolation of mycoplasmas in any of the samples tested, whereas, in the PCR, 62.96% (51/81) were positive, with 1.96% (1/51) identified as MG and 19.61% (10/51) as MS, representing 1.23% (1/81) and 12.34% (10/81) of the total population studied. PCR was shown to be a more effective technique than isolation in the detection ofMycoplasmaspp. in birds. It was possible to detect mycoplasmas in birds from Riozoo with no clinical respiratory signs, with higher MS prevalence than MG. The positivities forMycoplasmaspp., MS, and MG were different among the orders studied, being the highest occurrence in birds of prey, followed by Galliformes and Piciformes. The presence of MG and MS in birds of Rio de Janeiro Zoo confirms the circulation of these agents and the need for further studies on the dissemination of mycoplasmas in zoos for the epidemiological analysis of these bacteria in these places.(AU)

O Brasil é um dos países com maior avifauna do mundo. O confinamento de aves associado ao contato próximo a outros animais e seres humanos favorece a disseminação de agentes etiológicos causadores de doenças respiratórias. Dentre eles, os micoplasmas podem causar doença assintomática ou aparente que se manifesta em aves por espirros, estertores, conjuntivite, corrimentos oculares e nasais. São diversos os micoplasmas descritos causadores de doença em aves, com destaque para Mycoplasma gallisepticum (MG) e Mycoplasma synoviae (MS). O diagnóstico de Mycoplasma spp. pode ser feito pela observação clínica e análises laboratoriais. O diagnóstico molecular pela Reação em Cadeia da Polimerase (PCR) ganhou impulso por sua rapidez, sensibilidade e baixo custo em relação às técnicas de isolamento do agente que levam até 21 dias para conclusão do gênero Mycoplasma. Objetivou-se verificar a ocorrência da infecção por Mycoplasma spp. em aves no Zoológico do Rio de Janeiro (Rio Zoo), por isolamento e PCR. Do plantel de 635 aves do Rio Zoo, foram estudadas 81 para detecção de Mycoplasma spp., quando contidas para exames rotineiros de avaliação da condição de saúde. Essas aves eram pertencentes às ordens Psittaciformes (45), Accipitriformes (18), Galliformes (7), Piciformes (5), Strigiformes (4), Falconiformes (1) e Cariamiformes (1), todas já identificadas por microchip ou por anilha. Não houve isolamento de micoplasmas em nenhuma das amostras testadas, enquanto na PCR, 62,96% (51/81) foram positivas, sendo 1,96% (1/51) identificadas como MG e 19,61% (10/51) como MS, representando 1,23% (1/81) e 12,34% (10/81) da população total estudada. A PCR demonstrou ser uma técnica mais efetiva que o isolamento na detecção de Mycoplasma spp. em aves. Foi possível detectar micoplasmas nas aves do Riozoo sem sinal clínico respiratório, tendo MS maior prevalência do que MG. As positividades para Mycoplasma spp., MG e MS foram diferentes entre as ordens de aves estudadas, sendo a maior ocorrência nas aves de rapina, seguida dos Galliformes e dos Piciformes. A presença de MG e MS nas aves do Rio de Janeiro Zoo confirma a circulação destes agentes e a necessidade de mais estudos sobre a disseminação de micoplasmas em zoológicos para análise epidemiológica dessas bactérias nesse local.(AU)

Comparison of Mycoplasma gallisepticum Infection in Different Samples and Ages of Chicken Breeder Flocks

This study aimed to compare method-based and newly developed sample-based methods for Mycoplasma gallisepticum (MG) detection in different samples of breeder flocks suffering from respiratory disease problems by using culture, real-time PCR (rPCR) and ELISA from chicks and embryonated eggs. Overall, 450 samples of 19-day-old chicken embryos trachea, 450 samples of 8-day-old chicken tracheal swabs and 900 blood samples of 20-, 27-, 34-, 40- and 46-week-old breeder chickens from 5 flocks were sampled for 26 weeks, and were all tested for MG by culture, MG-rPCR and MG-ELISA. Culturing assays and rPCR were applied to 450 mixture samples from 19-day-old chicken embryos trachea and 450 tracheal swab samples (each pooled into groups of 3) from 8-day-old chicks from the same flocks. Also, 900 blood samples from the same 5 breeder flocks suffering from respiratory disease problems were tested by MG-ELISA. In individual sample-based analyses, 55 (18.3%) of the 300 pooled swab samples were positive for MG using culture methods, and 106 (35.3%) of the same samples were found positive by rPCR (sensitivity, specificity). The ELISAs indicated that 252 (28%) of the 900 breeding blood samples were MG seropositive. Using age-based analyses, the most positive period was 46 weeks, followed by 40 weeks, 34 weeks, 27 weeks and at least 20 weeks, in order of decreasing seropositivity. When comparing the culture and rPCR results of the two different sampling methods, chicken embryos trachea yielded more positive results than did tracheal swabs from the same flocks. In conclusion, rPCR is a highly specific, sensitive and reliable method for MG identification.

Comparison of Mycoplasma gallisepticum Infection in Different Samples and Ages of Chicken Breeder Flocks

This study aimed to compare method-based and newly developed sample-based methods for Mycoplasma gallisepticum (MG) detection in different samples of breeder flocks suffering from respiratory disease problems by using culture, real-time PCR (rPCR) and ELISA from chicks and embryonated eggs. Overall, 450 samples of 19-day-old chicken embryos trachea, 450 samples of 8-day-old chicken tracheal swabs and 900 blood samples of 20-, 27-, 34-, 40- and 46-week-old breeder chickens from 5 flocks were sampled for 26 weeks, and were all tested for MG by culture, MG-rPCR and MG-ELISA. Culturing assays and rPCR were applied to 450 mixture samples from 19-day-old chicken embryos trachea and 450 tracheal swab samples (each pooled into groups of 3) from 8-day-old chicks from the same flocks. Also, 900 blood samples from the same 5 breeder flocks suffering from respiratory disease problems were tested by MG-ELISA. In individual sample-based analyses, 55 (18.3%) of the 300 pooled swab samples were positive for MG using culture methods, and 106 (35.3%) of the same samples were found positive by rPCR (sensitivity, specificity). The ELISAs indicated that 252 (28%) of the 900 breeding blood samples were MG seropositive. Using age-based analyses, the most positive period was 46 weeks, followed by 40 weeks, 34 weeks, 27 weeks and at least 20 weeks, in order of decreasing seropositivity. When comparing the culture and rPCR results of the two different sampling methods, chicken embryos trachea yielded more positive results than did tracheal swabs from the same flocks. In conclusion, rPCR is a highly specific, sensitive and reliable method for MG identification.(AU)

Investigations of pathological immunohistochemical and immunocytochemical findings in natural infection with Mycoplasma gallisepticum in laying hens

Background: Mycoplasmosis is an infectious disease caused by Mycoplasma gallisepticum (MG), usually seen in therespiratory system of chickens, chick and turkeys, that causing great economic loss. The disease is characterized by respiratory system lesions such as sinusitis, tracheitis, airsacculitis, pneumonia and other symptoms such as loss of yield, arthritis,tenosynovitis. In this study, it was aimed to investigate diagnose of the disease by pathologic and molecular techniquesin hens that naturally infected with MG as well as the usability of immunocytochemical (ICC) method in diagnose of thedisease.Materials, Methods & Results: For this purpose, 98 hens were collected from 10 different coops that serologically positive.After necropsy, routine pathological procedures were performed to samples taken from nose, sinus, larynx, trachea, lungand air sacs. Scraping samples taken from lungs and tracheas were evaluated by ICC. Immunohistochemical (IHC) stainingwas performed to samples taken from nose, sinus, larynx, trachea, lung and air sacs. Indirect immunoperoxidase methodwas applied in the both IHC and ICC staining. Rabbit polyclonal anti MG antibody was used as primer antibody in theIHC and ICC staining. Additionally, culture and PCR techniques were applied to tracheas of all hens for MG. The GPO3and MGSO genes were made for PCR analysis. In the tracheal examinations, 23 cases were positive for PCR, 17 casesICC positive, 16 cases IHC positive and 10 culture samples found positive. All of culture positive cases were also positivefor other three methods. When findings in all organs were evaluated, in 37 cases were detected positive by IHC (38%) and23 cases were positive by ICC (23.5%). In the IHC positive cases, the first order was trachea in 16 cases followed by in 11cases in sinus, in 8 cases in lung, in 6 cases air sac and 4 cases in nose, respectively. In 8 cases, IHC positivity was foundin at least two organs...(AU)

Investigations of pathological immunohistochemical and immunocytochemical findings in natural infection with Mycoplasma gallisepticum in laying hens

Background: Mycoplasmosis is an infectious disease caused by Mycoplasma gallisepticum (MG), usually seen in therespiratory system of chickens, chick and turkeys, that causing great economic loss. The disease is characterized by respiratory system lesions such as sinusitis, tracheitis, airsacculitis, pneumonia and other symptoms such as loss of yield, arthritis,tenosynovitis. In this study, it was aimed to investigate diagnose of the disease by pathologic and molecular techniquesin hens that naturally infected with MG as well as the usability of immunocytochemical (ICC) method in diagnose of thedisease.Materials, Methods & Results: For this purpose, 98 hens were collected from 10 different coops that serologically positive.After necropsy, routine pathological procedures were performed to samples taken from nose, sinus, larynx, trachea, lungand air sacs. Scraping samples taken from lungs and tracheas were evaluated by ICC. Immunohistochemical (IHC) stainingwas performed to samples taken from nose, sinus, larynx, trachea, lung and air sacs. Indirect immunoperoxidase methodwas applied in the both IHC and ICC staining. Rabbit polyclonal anti MG antibody was used as primer antibody in theIHC and ICC staining. Additionally, culture and PCR techniques were applied to tracheas of all hens for MG. The GPO3and MGSO genes were made for PCR analysis. In the tracheal examinations, 23 cases were positive for PCR, 17 casesICC positive, 16 cases IHC positive and 10 culture samples found positive. All of culture positive cases were also positivefor other three methods. When findings in all organs were evaluated, in 37 cases were detected positive by IHC (38%) and23 cases were positive by ICC (23.5%). In the IHC positive cases, the first order was trachea in 16 cases followed by in 11cases in sinus, in 8 cases in lung, in 6 cases air sac and 4 cases in nose, respectively. In 8 cases, IHC positivity was foundin at least two organs...

Characterization and differentiation of chicken mycoplasma isolates using 16S-23S intergenic spacer region sequencing

The objective of this study was to identify the species and characterize the genetic relationships among mycoplasma isolates from commercial layer hen flocks using 16S-23S rDNA intergenic spacer region (IGSR) sequencing. Twenty-one isolates were obtained from samples collected from commercial layer flocks in four Brazilian states: São Paulo, Minas Gerais, Rio de Janeiro and Espírito Santo. The isolates were recovered from the São Paulo, Rio de Janeiro and Espírito Santo states. Eleven isolates were originated from tracheal swabs, five from shell gland swabs and five from ovary fragment collection. The 16S-23S rDNA IGSR of isolates were amplified by PCR, and the obtained products were subsequently sequenced. The consensus of each isolate was compared to the available sequences using Nucleotide BLAST® to determine the mycoplasma species. A phylogenetic analysis of the Mycoplasma gallisepticum (MG) sequences was performed. Pairwise analyses showed homologies of 99% to 100% with the previously characterized sequences listed in GenBank®. Four Mycoplasma gallinaceum were isolated from three flocks and seven M. pullorum isolates were obtained from a single flock. The other 10 isolates were all identified as MG and were obtained from four flocks. The 16S-23S rDNA IGSR sequencing was a good method to identify Mycoplasma species isolated from field samples, providing fast and reliable results at relatively low costs. The results were also satisfactory for the single-locus sequence typing of MG isolates.

Characterization and differentiation of chicken mycoplasma isolates using 16S-23S intergenic spacer region sequencing

The objective of this study was to identify the species and characterize the genetic relationships among mycoplasma isolates from commercial layer hen flocks using 16S-23S rDNA intergenic spacer region (IGSR) sequencing. Twenty-one isolates were obtained from samples collected from commercial layer flocks in four Brazilian states: São Paulo, Minas Gerais, Rio de Janeiro and Espírito Santo. The isolates were recovered from the São Paulo, Rio de Janeiro and Espírito Santo states. Eleven isolates were originated from tracheal swabs, five from shell gland swabs and five from ovary fragment collection. The 16S-23S rDNA IGSR of isolates were amplified by PCR, and the obtained products were subsequently sequenced. The consensus of each isolate was compared to the available sequences using Nucleotide BLAST® to determine the mycoplasma species. A phylogenetic analysis of the Mycoplasma gallisepticum (MG) sequences was performed. Pairwise analyses showed homologies of 99% to 100% with the previously characterized sequences listed in GenBank®. Four Mycoplasma gallinaceum were isolated from three flocks and seven M. pullorum isolates were obtained from a single flock. The other 10 isolates were all identified as MG and were obtained from four flocks. The 16S-23S rDNA IGSR sequencing was a good method to identify Mycoplasma species isolated from field samples, providing fast and reliable results at relatively low costs. The results were also satisfactory for the single-locus sequence typing of MG isolates.(AU)

Burden of exposure to infectious bursal disease virus, infectious bronchitis virus, Newcastle disease virus, Mycoplasma gallisepticum, and intestinal parasites in introduced broiler chickens on the Galapagos.

Diseases in introduced broilers can possibly spill over to wild birds on the Galapagos. Knowledge about the current burden of exposure to pathogens in broilers on the Galapagos is very limited. The objective of the study reported here was to measure the burden of exposure to infectious bursal disease virus (IBDV), infectious bronchitis virus (IBV), Newcastle disease virus (NDV), Mycoplasma gallisepticum (MG), and intestinal parasites in a sample of broiler chickens on 13 farms on Santa Cruz Island and San Cristobal Island in July 2017. Blood serum samples were tested for detection of antibodies to IBDV, IBV, NDV, and MG by using an IDEXX Enzyme-linked Immunosorbent Assay. In addition, fecal samples and pen bedding environmental samples were processed and analyzed for diagnosis of intestinal parasite eggs under a compound light microscope. The frequency of seropositive broilers to IBDV was 74/130 or 56% (95% CI = 48, 65%), to IBV was 27/130 or 20% (14, 28%), and to NDV was 1/130 or 0.7% (0.1, 4%). All broilers tested negative to MG antibodies. Eimeria spp. infection was common in study broilers. Finally, we observed interaction between broiler chickens and wild birds (finches) inside broiler pens, as well as the presence of backyard chickens inside property limits of study farms. This study produced evidence that exposure to IBDV, IBV, and intestinal parasites in broilers on Santa Cruz Island and San Cristobal Island is important. Study results are relevant because (i) they provide new baseline data on the burden of exposure to avian pathogens in broiler farms, (ii) justify the need to verify standard operating procedures in hatcheries that supply (non-vaccinated) day-old chicks to the Galapagos and (iii) to implement enhanced biosecurity standards on broiler chicken farms to mitigate risk of disease transmission between broilers, backyard poultry, and wild birds on the Galapagos.

Detection of Mycoplasma gallisepticum in House Finches ( Haemorhous mexicanus) from Arizona.

In 1994, an endemic poultry pathogen, Mycoplasma gallisepticum (MG), was identified as the causative agent of a novel disease in house finches ( Haemorhous mexicanus). After an initial outbreak in Maryland, MG spread rapidly throughout eastern North American populations of house finches. Subsequently, MG spread slowly through the northern interior of North America and then into the Pacific Northwest, finally reaching California in 2006. Until 2009, there were no reports of MG in the southwestern United States east of California. In August 2011, after reports of house finches displaying conjunctivitis characteristic of MG infection in Arizona, we trapped house finches at bird feeders in central Arizona (Tempe) and southern Arizona (Tucson and Green Valley) to assay for MG infection. Upon capture, we noted whether birds exhibited conjunctivitis, and we collected choanal swabs to test for the presence of MG DNA using PCR. We detected MG in finches captured from Green Valley (in ∼12% of birds captured), but not in finches from Tucson or Tempe. Based on resampling of house finches at these sites in July 2014, we suggest that central Arizona finches likely remain unexposed to MG. We also suggest that low urban connectivity between arid habitats of southern and central Arizona or a reduction in the prevalence of MG after its initial arrival in Arizona may be limiting the spread of MG from south to north in Arizona. In addition, the observed conjunctivitis-like signs in house finches that were negative for MG by PCR may be caused primarily by avian pox virus.

A incidência de aerossaculite na produção de frango de corte / The incidence of airsacculitis in the production of broiler

Esta revisão de literatura foi conduzida com o objetivo de contribuir para o conhecimento a respeito de uma das principais infecções respiratórias que podem acometer os frangos de corte, denominada aerossaculite. Essa patologia é de extrema importância para as empresas avícolas pois, uma vez que causa o acometimento dos sacos aéreos, desencadeia grandes perdas econômicas para o produtor em decorrência da condenação de sua carcaça. Dessa forma, foi discutido a incidência que a aerossaculite tem sobre os sistemas de criação, as principais causas que podem levar a contaminação e a disseminação dessa patogenia e como o produtor pode prevenir a exposição dos animais a essa infecção, visando assim a melhor qualidade e aceitação desse produto no mercado interno e externo.

This literature review was conducted with the objective of contributing to the knowledge of one of the main respiratory infections that can be accompanied by broilers, called airsacculitis. This pathology is of extreme importance for poultry companies, since it causes the attack of the air sacs, it triggers great economic losses for the producer due to the condemnation of its carcass. Thus, it was discussed the incidence of airsacculitis on breeding systems, as the main causes that can lead to a contamination and a dissemination of the pathogenesis and how the producer can prevent an exposure of the animals to this infection, aiming at the best quality and acceptance of the product in the internal and external market.

A incidência de aerossaculite na produção de frango de corte / The incidence of airsacculitis in the production of broiler

Esta revisão de literatura foi conduzida com o objetivo de contribuir para o conhecimento a respeito de uma das principais infecções respiratórias que podem acometer os frangos de corte, denominada aerossaculite. Essa patologia é de extrema importância para as empresas avícolas pois, uma vez que causa o acometimento dos sacos aéreos, desencadeia grandes perdas econômicas para o produtor em decorrência da condenação de sua carcaça. Dessa forma, foi discutido a incidência que a aerossaculite tem sobre os sistemas de criação, as principais causas que podem levar a contaminação e a disseminação dessa patogenia e como o produtor pode prevenir a exposição dos animais a essa infecção, visando assim a melhor qualidade e aceitação desse produto no mercado interno e externo.(AU)

This literature review was conducted with the objective of contributing to the knowledge of one of the main respiratory infections that can be accompanied by broilers, called airsacculitis. This pathology is of extreme importance for poultry companies, since it causes the attack of the air sacs, it triggers great economic losses for the producer due to the condemnation of its carcass. Thus, it was discussed the incidence of airsacculitis on breeding systems, as the main causes that can lead to a contamination and a dissemination of the pathogenesis and how the producer can prevent an exposure of the animals to this infection, aiming at the best quality and acceptance of the product in the internal and external market.(AU)

HOUSE FINCH ( HAEMORHOUS MEXICANUS)-ASSOCIATED MYCOPLASMA GALLISEPTICUM IDENTIFIED IN LESSER GOLDFINCH ( SPINUS PSALTRIA) AND WESTERN SCRUB JAY ( APHELOCOMA CALIFORNICA) USING STRAIN-SPECIFIC QUANTITATIVE PCR.

: In 1994 Mycoplasma gallisepticum was found to be the etiologic agent of House Finch ( Haemorhous mexicanus) conjunctivitis, a rapidly expanding epidemic caused by a genetically discrete, House Finch-associated strain of M. gallisepticum (HFMG). While most prominent in House Finches, HFMG has been reported in other members of the family Fringillidae, including American Goldfinches ( Spinus tristis), Purple Finches ( Haemorhous purpureus), Pine Grosbeaks ( Pinicola enucleator), and Evening Grosbeaks ( Coccothraustes vespertinus). Herein we report two new potential host species of HFMG strain, the Lesser Goldfinch ( Spinus psaltria), belonging to the Fringillidae family, and the Western (California) Scrub Jay ( Aphelocoma californica), belonging to the Corvidae family. The latter is one of only two reports of HFMG being found outside the Fringillidae family, and of these is the only one reported outside of captivity. Furthermore, non-HFMG M. gallisepticum was identified in an American Crow ( Corvus brachyrhynchos), indicating presence of additional strains in wild birds. Strain typing of M. gallisepticum isolates was done via HFMG-specific quantitative PCR analysis and validated using random amplified polymorphic DNA analysis. Our results suggested an expanded host range of HFMG strain, and further suggested that the host range of HFMG was not limited to members of the family Fringillidae.