Avian biodiversity assessment studies in a Neotropical wetland - the combination of sampling methods makes the difference

In studies of avian diversity, many different methods have been applied. Since methodological approaches may affect research results, the choice of a given methodology must be consistent with the scientific objectives. The aim of this study was to investigate how different methodologies with their intrinsic limitations help detect and monitor birds to evaluate how they complement each other in the survey of species. Three different assessment methods, mist nets, point counts, and autonomous acoustic recordings were used to serve this purpose in a study of different Pantanal habitats, such as savannas and forests. The point counts detected more species (126 species) than the two other methods autonomous acoustic recordings (113 species) and mist nets (79 species). We observed significant differences in the number of species detected by mist nets and the other two methods. Each survey method identified exclusive species. When comparing habitats, all three methods showed significant differences in bird species composition. Savannas were richer in bird species than forests, and replacement was the main driver responsible for the differences in beta diversity between the habitats. The three methodologies, when applied together, proved to be complementary in avian species detection.

Prevalence and association of mycoplasma infection in the development of coronary artery disease

Abstract Coronary heart disease (CHD) has been associated with significant morbidity and mortality worldwide. Although remain controversial, several studies have demonstrated the association of M. pneumoniae infections with atherosclerosis. We evaluated the possible association of mycoplasma infections in patients diagnosed with atherosclerosis by ELISA and PCR methods. Atherosclerotic tissue samples and blood samples were collected for the detection of mycoplasma antibodies (IgA) by ELISA from the 97 patients with coronary artery disease (CAD). M. pneumoniae specific IgA, IgG and IgM were measured by using the Anti-M. pneumoniae IgA/IgG/IgM ELISA. Detection of M. pneumoniae targeting the P1 adhesion gene was performed by PCR Acute infection of M. pneumoniae was diagnosed in 43.3% (42) of patients by PCR. The M. pneumoniae specific antibodies were detected in 36.1% (35) of patients. Twenty-five (25.8%) cases had IgG antibodies, 15 (15.5%) cases had IgM antibodies, 3 (3.1%) cases had IgA antibodies, 10 (10.3%) cases had both IgM + IgG antibodies and 1 (1%) case of each had IgM + IgA and IgG + IgA antibodies. None of the cases was positive for all three antibodies. A Pearson correlation coefficient analysis revealed an excellent correlation between the PCR and the serological results (r=0.921, p 0.001). A majority (17, 40.5%) of the M. pneumoniae positive patients are within the 41-50 years of age group, followed by 10 (23.8%) patients in the age group of 61-70 years and 2 (4.8%) patients were >70 years of age. Our study reported an unusually higher prevalence of M. pneumoniae by serological tests (36.1%) and PCR (43.3%). Although the hypothesis of the association of M. pneumoniae and CAD is yet to be proven, the unusually high prevalence of M. pneumoniae in CAD patients indicates an association, if not, in the development of atherosclerosis.

Resumo A doença coronariana (DCC) tem sido associada a significativa morbidade e mortalidade em todo o mundo. Embora ainda sejam controversos, vários estudos têm demonstrado a associação de infecções por M. pneumoniae com aterosclerose. Avaliamos a possível associação de infecções por micoplasma em pacientes com diagnóstico de aterosclerose pelos métodos ELISA e PCR. Amostras de tecido aterosclerótico e amostras de sangue foram coletadas para a detecção de anticorpos contra micoplasma (IgA) por ELISA de 97 pacientes com doença arterial coronariana (DAC). IgA, IgG e IgM específicos para M. pneumoniae foram medidos usando o Anti-M. pneumoniae IgA / IgG / IgM ELISA. A detecção de M. pneumoniae visando o gene de adesão P1 foi realizada por PCR. A infecção aguda por M. pneumoniae foi diagnosticada em 43,3% (42) dos pacientes pela PCR. Os anticorpos específicos para M. pneumoniae foram detectados em 36,1% (35) dos pacientes. Vinte e cinco (25,8%) casos tinham anticorpos IgG, 15 (15,5%) casos tinham anticorpos IgM, 3 (3,1%) casos tinham anticorpos IgA, 10 (10,3%) casos tinham anticorpos IgM + IgG e 1 (1%) caso de cada um tinha anticorpos IgM + IgA e IgG + IgA. Nenhum dos casos foi positivo para os três anticorpos. A análise do coeficiente de correlação de Pearson revelou uma excelente correlação entre o PCR e os resultados sorológicos (r = 0,921, p 0,001). A maioria (17, 40,5%) dos pacientes positivos para M. pneumoniae está na faixa etária de 41-50 anos, seguida por 10 (23,8%) pacientes na faixa etária de 61-70 anos e 2 (4,8%) pacientes tinham > 70 anos de idade. Nosso estudo relatou uma prevalência incomumente maior de M. pneumoniae por testes sorológicos (36,1%) e PCR (43,3%). Embora a hipótese da associação de M. pneumoniae e DAC ainda não tenha sido comprovada, a prevalência incomumente alta de M. pneumoniae em pacientes com DAC indica uma associação, se não, no desenvolvimento de aterosclerose.

Neorickettsia sp. in coatis (Nasua nasua) in Brazil / Neorickettsia sp. em quatis (Nasua nasua) no Brasil

The genus Neorickettsia comprises trematode-associated bacteria that can cause diseases in animals and humans. Despite detection of Neorickettsia antigens in the intestine of coatis kept in captivity in southern Brazil through immunohistochemistry, the molecular identity of the bacteria in South American procyonids remains elusive. The aim of the present study was to investigate the occurrence of Neorickettsia sp. in blood samples from coatis in central-western Brazil. Between March 2018 and January 2019, animals were captured and recaptured in two areas of the Cerrado (Parque Estadual do Prosa, PEP; and Vila da Base Aérea, VBA) located in the city of Campo Grande, state of Mato Grosso do Sul, central-western Brazil. All captures were performed according to convenience. DNA from 97 blood samples was subjected to nested PCR (nPCR) targeting a fragment of the 16S rRNA gene of Neorickettsia sp. Six samples (3.6%; five from VBA and one from PEP) from different coatis were positive in nPCR based on the 16S rRNA. The sequences obtained (~500 bp) showed ˃ 99% similarity to N. risticii. Phylogenetic analysis clustered the sequences detected in the present study in a clade with N. risticii. This is the first molecular detection of Neorickettsia sp. in coatis in Brazil.(AU)

O gênero Neorickettsia compreende bactérias associadas a trematódeos que podem causar doenças em animais e humanos. Apesar da detecção de antígenos de Neorickettsia por imuno-histoquímica no intestino de quatis mantidos em cativeiro no sul do Brasil, a identidade molecular da bactéria em procionídeos da América do Sul permanece indefinida. O presente trabalho teve como objetivo investigar a ocorrência de Neorickettsia sp. em amostras de sangue de quatis do Centro-Oeste do Brasil. Entre março de 2018 e janeiro de 2019, os animais foram capturados em duas áreas de Cerrado (Parque Estadual do Prosa PEP e Vila da Base Aérea VBA) localizadas na cidade de Campo Grande, Mato Grosso do Sul, Centro-Oeste do Brasil. Todas as capturas e recapturas foram realizadas por conveniência. O DNA de 97 amostras de sangue foi submetido a "nested" (nPCR), baseada em um fragmento do gene 16S rRNA de Neorickettsia sp. Seis (3,6% - 5 de VBA e 1 de PEP) amostras de quatis diferentes foram positivas na nPCR, baseada no rRNA 16S. As sequências obtidas (~500 pb) apresentaram ˃99% de identidade com N. risticii. A inferência filogenética agrupou as sequencias detectadas no presente estudo em um clado com N. risticii. Esta é a primeira detecção molecular de Neorickettsia sp. em quatis do Brasil.(AU)

Overview of the use of genomic data in animal breeding / Panorama do uso de dados genômicos no melhoramento genético animal

ABSTRACT: The use of molecular information in breeding programs contributed to important advances in the improvement of traits of economic interest in livestock production. The advent of single nucleotide polymorphism (SNP) panels applied to genome-wide selection (GWS) and genome-wide association studies (GWAS), along with computational advances (e.g., use of powerful software and robust analyses) allowed a better understanding of the genetic architecture of farm animals and increased the selection efficiency. In this context, the statistic method single-step GBLUP has been frequently used to perform GWS, and more recently GWAS analyses, providing accurate predictions and QTL detection, respectively. Nevertheless, in developing countries, species such as sheep and goats, whose genomic data are more difficult to be obtained, the use of data simulation has been efficient in the study of the major factors involved in the selection process, such as size of training population, density of SNP chips, and genotyping strategies. The effects of these factors are directly associated with the prediction accuracy of genomic breeding values. In this review we showed important aspects of the use of genomics in the genetic improvement of production traits of animals, the main methods currently used for prediction and estimation of molecular marker effects, the importance of data simulation for validation of those methods, as well as the advantages, challenges and limitations of the use of GWS and GWAS in the current scenario of livestock production.

RESUMO: Em programas de melhoramento genético, o uso de informações moleculares garantiu importantes avanços para a melhoria de características de interesse econômico, no âmbito da produção animal. O advento da tecnologia de painéis de SNPs aplicados à seleção genômica ampla (GWS) e associação genômica ampla (GWAS), aliado ao avanço computacional, com o uso de softwares e análises robustas, permitiram melhor compreensão sobre a arquitetura genética dos animais de produção e, consequentemente, maior eficiência na seleção. Nesse contexto, o método estatístico single-step GBLUP tem sido utilizado, frequentemente, na execução da GWS e, mais recentemente, em GWAS, possibilitando predições acuradas e detecção de QTLs, respectivamente. No entanto, em países em desenvolvimento e, em espécies como os ovinos e caprinos, que existe maior dificuldade para a aquisição de dados genômicos, o uso da simulação de dados tem se mostrado eficiente para estudar os principais fatores envolvidos no processo de seleção, como o tamanho da população de treinamento, densidade de chipde SNPs e estratégias de genotipagem, cujos efeitos estão diretamente associados à acurácia da predição de valores genéticos genômicos. Nesta revisão, serão abordados pontos importantes sobre o uso da genômica no melhoramento genético de características produtivas em animais, principais métodos de predição e estimação de efeitos de marcadores moleculares na atualidade, a importância da simulação de dados para a validação desses métodos, bem como as vantagens, os desafios e as limitações no cenário atual da produção animal com o uso da seleção e associação genômica ampla.

Molecular detection of Sarcocystis spp. in the breast muscle of captive birds from the southern region of Brazil / Detecção molecular de Sarcocystis spp. no músculo peitoral de aves de cativeiro da região sul do Brasil

Studies on diseases of wild birds are essential in the context of public health, as these animals act as sentinels, allowing information regarding a determined geographic area. In addition, birds are food protein sources for animals, and therefore play an important role in the life cycle of the protozoan Sarcocystis spp. This study aimed to identify the Sarcocystis spp. in breast muscle samples of naturally infected captive birds. The breast muscle of 89 birds were sampled, and the DNA amplified by PCR targeting the 18S ribosomal RNA gene to detect Sarcocystis spp. PCR products were sequenced and 5.61% (5/89) samples showed 100% similarity with Sarcocystis spp. (one Cyanoliseus patagonus, one Psittacula krameri, two Pyrrhura frontalis, and one Ramphastos dicolorus). The large number of naturally infected species analyzed by molecular methods allowed the detection of Sarcocystis spp. in different bird species, corroborating the epidemiology of Sarcocystis spp.

Estudos sobre doenças de aves silvestres são essenciais no contexto da saúde pública, pois esses animais atuam como sentinelas, permitindo obter informações sobre uma determinada área geográfica. Além disso, as aves são fontes de proteína alimentar para os animais e, portanto, desempenham um papel importante no ciclo de vida do Sarcocystis. Este estudo teve como objetivo identificar Sarcocystis spp. nos músculos do peito de aves de cativeiro naturalmente infectadas. Os músculos do peito de 89 aves foram coletados, e o DNA amplificado pela PCR do gene RNA ribossômico 18S para detecção de Sarcocystis spp. Os produtos da PCR foram sequenciados e 5,61% (5/89) amostras apresentaram 100% de similaridade com o Sarcocystis spp. (um Cyanoliseus patagonus, um Psittacula krameri, dois Pyrrhura frontalis e um Ramphastos dicolorus). O grande número de espécies naturalmente infectadas analisadas por métodos moleculares permitiu a detecção de Sarcocystis spp. em diferentes espécies de aves, corroborando a epidemiologia de Sarcocystis spp.

Molecular survey of hemotropic mycoplasmas in crab-eating raccoons (Procyon cancrivorus) in southern Brazil / Levantamento molecular de micoplasma hemotrópico em mão-pelada (Procyon cancrivorus) do sul do Brasil

Hemoplasmas are non-cultivable bacterial parasites of erythrocytes that infect domestic and wild animals, as well as humans. Their means of transmission and pathogenesis remain contentious issues and difficult to evaluate in wild animals. Procyon cancrivorus is a South American carnivore and occurs in all Brazilian biomes. In this study, we aimed to investigate occurrences of hemoplasmas infecting P. cancrivorus and to identify their 16S rRNA gene, in southern Brazil. DNA was extracted from spleen and blood samples of P. cancrivorus (n = 9) from different locations. Hemoplasma DNA was detected in six samples, based on 16S rRNA gene amplification and phylogenetic analysis. Four of the six sequences belonged to the "Mycoplasma haemofelis group", which is closely related to genotypes detected in Procyon lotor from the USA; one was within the "Mycoplasma suis group", closely related to "Candidatus Mycoplasma haemominutum"; and one was within the intermediate group between these clusters. Thus, these sequences showed that the molecular identity of hemoplasmas in the population studied was very variable. In five positive animals, Amblyomma aureolatum ticks and a flea (Ctenocephalides felis felis) were collected. The present study describes the first molecular detection of mycoplasmas in P. cancrivorus.(AU)

Os micoplasmas hemotrópicos (hemoplasmas) são parasitas bacterianos não-cultiváveis de eritrócitos que infectam tanto animais domésticos e selvagens, como seres humanos. A transmissão e a patogênese são discutíveis e difíceis de avaliar em animais selvagens. O mão pelada (Procyon cancrivorus) é um carnívoro Sul-americano, que ocorre em todos os biomas brasileiros. O objetivo do presente estudo é o de investigar a ocorrência de hemoplasmas infectando P. cancrivorus e identificar seu gene 16S rRNA no Sul do Brasil. O DNA foi extraído do baço e amostras de sangue de P. cancrivorus (n= 9). O DNA de hemoplasma foi detectado em seis amostras, com base na amplificação do gene 16S rRNA e na análise filogenética. Quatro das seis sequências pertencem ao "Grupo Mycoplasma haemofelis", que estão intimamente relacionadas aos genótipos detectados no Procyon lotor dos EUA; uma dentro do "Grupo Mycoplasma suis", que está intimamente relacionado ao "Candidatus Mycoplasma haemominutum", e uma dentro do grupo intermediário entre esses clusters, mostrando assim que há uma diversidade genética de hemoplasmas na população estudada. Em cinco animais positivos, foram coletados carrapatos Amblyomma aureolatum e uma pulga Ctenocephalides felis. O presente estudo traz a primeira detecção molecular de micoplasmas em P. cancrivorus.(AU)

How does reproduction account for dairy farm sustainability?

Sustainability - the new hype of the 21st century has brought discomfort for the government and society. Sustainable agriculture is essential to face our most concerning challenges: climate change, food security, and the environmental footprint, all of which add to consumers' opinions and choices. Improvements in reproductive indexes can enhance animal production and efficiency, guaranteeing profit and sustainability. Estrus detection, artificial insemination (AI), embryo transfer (ET), estrus synchronization (ES), and multiple ovulations are some strategies used to improve animal reproduction. This review highlights how reproductive strategies and genetic selection can contribute to sustainable ruminant production. Improved reproductive indices can reduce the number of nonproductive cows in the herd, reducing methane emissions and land use for production while preserving natural resources.(AU)

Molecular diagnostic for a screening investigation method of tick-borne pathogens in Didelphis albiventris road-killed in north of Paraná, Brazil

The synanthropization of wild animals puts public health at risk by promoting the circulation of zoonotic agents, found naturally in the wild, in the anthropic environment. The objective of this work was to carry out screening by molecular detection of pathogens of the Anaplasmatacea family in Didelphis albiventris, a specie characterized as having a synanthropic habit. Opossums that were dead (n = 25) after being road-killed were collected in the North of Paraná state, southern Brazil during the 2016 and 2018 years, through active search. A questionnaire was filled out with information about the animal and collected place. Biological samples of spleen and liver were collected. The genetic material extracted from the spleen and liver was submitted to molecular diagnosis through PCR for amplification of dsb of Ehrlichia and 16S genes for the other agents of the Anaplasmataceae family. One animal was positive for the genus Ehrlichia in semi-nested PCR for amplification of the 349 bp fragment of the dsb gene in extracted from the liver samples. In PCR for the 16S target no animal was positive. These are preliminary results that reinforce the circulation of Ehrlichia in opossums. To improve the knowledge of these agents in opossums more studies are necessary.(AU)

A sinantropização de animais silvestres coloca em risco a saúde pública por propiciar a circulação de agentes zoonóticos, encontrados naturalmente em ambiente silvestre, no ambiente antrópico. O trabalho teve como objetivo realizar a triagem por detecção molecular de patógenos da família Anaplasmataceae em Didelphis albiventris, espécie caracterizada como de hábito sinantrópico. Gambás mortos (n=25) por atropelamento durante os anos de 2016 e 2018 foram coletados na região norte do Paraná, sul do Brasil, por meio de busca ativa. Realizou-se o preenchimento de formulário com informações sobre a espécie do animal e o local do atropelamento. Foi realizada a necrópsia e coleta de amostras biológicas, de baço e fígado. O material genético extraído de baço e fígado foi submetido a diagnóstico molecular, por meio de PCR, para amplificação dos genes dsb de Ehrlichia sp. e 16S para os demais agentes da família Anaplasmataceae. Um animal foi positivo para o gênero Ehrlichia em semi-nested PCR para a amplificação do fragmento de 349 pb do gene dsb, extraído de fígado. Na PCR para detecção do gene 16S nenhum dos animais foi positivo. Esses resultados preliminares reforçam a circulação de Ehrlichias em gambás. Para melhorar o conhecimento desses agentes em gambás mais estudos são necessários.(AU)

Molecular detection of Toxoplasma gondii, Neospora caninum and Sarcocystis spp in tissues of Sus scrofa slaughtered in southern Brazil / Detecção molecular de Toxoplasma gondii, Neospora caninum e Sarcocystis spp. em tecidos de Sus scrofa abatidos no Sul do Brasil

The aim of this study was to determine the presence of deoxyribonucleic acid (DNA) from Toxoplasma gondii, Sarcocystis spp. and Neospora caninum, in tissues of wild boars slaughtered in southern Brazil. A total of 156 samples were collected from different organs of 25 wild boars, and DNA from at least one of the protozoa investigated was detected in 79 samples. To differentiate between infectious agents, restriction fragment length polymorphism was performed using the restriction enzymes DdeI and HpaII. For N. caninum, conventional PCR was performed with specific primers. The DNA of at least one of the studied pathogens was detected in each animal: 26.58% for T. gondii, 68.36% for Sarcocystis spp. and 5.06% for N. caninum. Coinfection between T. gondii and Sarcocystis spp. occurred in 14 animals, between T. gondii and N. caninum in only one male animal, between Sarcocystis spp. and N. caninum in a female, while co-infection with the three agents was equally observed in only one male animal. Considering the high frequency of detection and its zoonotic risk, especially T. gondii, it appears that wild boars can be potential sources of transmission of infectious agents and the adoption of monitoring measures in these populations should be prioritized.(AU)

O objetivo deste estudo foi determinar a presença de ácido desoxirribonucléico (DNA) de Toxoplasma gondii, Sarcocystis spp. e Neospora caninum, em tecidos de javalis abatidos no sul do Brasil. Foram coletadas 156 amostras de diferentes órgãos de 25 javalis, sendo detectado o DNA de pelo menos um dos protozoários pesquisados em 79 amostras. Para diferenciar entre os agentes infecciosos, o polimorfismo do comprimento do fragmento de restrição, foi realizado usando-se as enzimas de restrição DdeI e HpaII. Para N. caninum, a PCR convencional foi realizada com "primers" específicos. O DNA de pelo menos um dos patógenos estudados foi detectado em cada animal: 26,58% para T. gondii, 68,36% para Sarcocystis spp. e 5,06% para N. caninum. Coinfecção entre T. gondii e Sarcocystis spp. ocorreu em 14 animais; entre T. gondii e N. caninum em apenas um animal macho; entre Sarcocystis spp. e N. caninum em uma fêmea, enquanto a coinfecção com os três agentes foi observada igualmente em apenas um animal macho. Considerando-se a alta frequência de detecção e seu risco zoonótico, especialmente T. gondii, constata-se que os javalis podem ser potenciais fontes de transmissão de agentes infecciosos, e a adoção de medidas de monitoramento nessas populações devem ser priorizadas.(AU)

Overview of the use of genomic data in animal breeding / Panorama do uso de dados genômicos no melhoramento genético animal

The use of molecular information in breeding programs contributed to important advances in the improvement of traits of economic interest in livestock production. The advent of single nucleotide polymorphism (SNP) panels applied to genome-wide selection (GWS) and genome-wide association studies (GWAS), along with computational advances (e.g., use of powerful software and robust analyses) allowed a better understanding of the genetic architecture of farm animals and increased the selection efficiency. In this context, the statistic method single-step GBLUP has been frequently used to perform GWS, and more recently GWAS analyses, providing accurate predictions and QTL detection, respectively. Nevertheless, in developing countries, species such as sheep and goats, whose genomic data are more difficult to be obtained, the use of data simulation has been efficient in the study of the major factors involved in the selection process, such as size of training population, density of SNP chips, and genotyping strategies. The effects of these factors are directly associated with the prediction accuracy of genomic breeding values. In this review we showed important aspects of the use of genomics in the genetic improvement of production traits of animals, the main methods currently used for prediction and estimation of molecular marker effects, the importance of data simulation for validation of those methods, as well as the advantages, challenges and limitations of the use of GWS and GWAS in the current scenario of livestock production.

Em programas de melhoramento genético, o uso de informações moleculares garantiu importantes avanços para a melhoria de características de interesse econômico, no âmbito da produção animal. O advento da tecnologia de painéis de SNPs aplicados à seleção genômica ampla (GWS) e associação genômica ampla (GWAS), aliado ao avanço computacional, com o uso de softwares e análises robustas, permitiram melhor compreensão sobre a arquitetura genética dos animais de produção e, consequentemente, maior eficiência na seleção. Nesse contexto, o método estatístico single-step GBLUP tem sido utilizado, frequentemente, na execução da GWS e, mais recentemente, em GWAS, possibilitando predições acuradas e detecção de QTLs, respectivamente. No entanto, em países em desenvolvimento e, em espécies como os ovinos e caprinos, que existe maior dificuldade para a aquisição de dados genômicos, o uso da simulação de dados tem se mostrado eficiente para estudar os principais fatores envolvidos no processo de seleção, como o tamanho da população de treinamento, densidade de chipde SNPs e estratégias de genotipagem, cujos efeitos estão diretamente associados à acurácia da predição de valores genéticos genômicos. Nesta revisão, serão abordados pontos importantes sobre o uso da genômica no melhoramento genético de características produtivas em animais, principais métodos de predição e estimação de efeitos de marcadores moleculares na atualidade, a importância da simulação de dados para a validação desses métodos, bem como as vantagens, os desafios e as limitações no cenário atual da produção animal com o uso da seleção e associação genômica ampla.

Prevalence and association of mycoplasma infection in the development of coronary artery disease / Prevalência e associação de infecção por micoplasma no desenvolvimento de doença arterial coronariana

Coronary heart disease (CHD) has been associated with significant morbidity and mortality worldwide. Although remain controversial, several studies have demonstrated the association of M. pneumoniae infections with atherosclerosis. We evaluated the possible association of mycoplasma infections in patients diagnosed with atherosclerosis by ELISA and PCR methods. Atherosclerotic tissue samples and blood samples were collected for the detection of mycoplasma antibodies (IgA) by ELISA from the 97 patients with coronary artery disease (CAD). M. pneumoniae specific IgA, IgG and IgM were measured by using the Anti-M. pneumoniae IgA/IgG/IgM ELISA. Detection of M. pneumoniae targeting the P1 adhesion gene was performed by PCR Acute infection of M. pneumoniae was diagnosed in 43.3% (42) of patients by PCR. The M. pneumoniae specific antibodies were detected in 36.1% (35) of patients. Twenty-five (25.8%) cases had IgG antibodies, 15 (15.5%) cases had IgM antibodies, 3 (3.1%) cases had IgA antibodies, 10 (10.3%) cases had both IgM + IgG antibodies and 1 (1%) case of each had IgM + IgA and IgG + IgA antibodies. None of the cases was positive for all three antibodies. A Pearson correlation coefficient analysis revealed an excellent correlation between the PCR and the serological results (r=0.921, p70 years of age. Our study reported an unusually higher prevalence of M. pneumoniae by serological tests (36.1%) and PCR (43.3%). Although the hypothesis of the association of M. pneumoniae and CAD is yet to be proven, the unusually high prevalence of M. pneumoniae in CAD patients indicates an association, if not, in the development of atherosclerosis.

A doença coronariana (DCC) tem sido associada a significativa morbidade e mortalidade em todo o mundo. Embora ainda sejam controversos, vários estudos têm demonstrado a associação de infecções por M. pneumoniae com aterosclerose. Avaliamos a possível associação de infecções por micoplasma em pacientes com diagnóstico de aterosclerose pelos métodos ELISA e PCR. Amostras de tecido aterosclerótico e amostras de sangue foram coletadas para a detecção de anticorpos contra micoplasma (IgA) por ELISA de 97 pacientes com doença arterial coronariana (DAC). IgA, IgG e IgM específicos para M. pneumoniae foram medidos usando o Anti-M. pneumoniae IgA / IgG / IgM ELISA. A detecção de M. pneumoniae visando o gene de adesão P1 foi realizada por PCR. A infecção aguda por M. pneumoniae foi diagnosticada em 43,3% (42) dos pacientes pela PCR. Os anticorpos específicos para M. pneumoniae foram detectados em 36,1% (35) dos pacientes. Vinte e cinco (25,8%) casos tinham anticorpos IgG, 15 (15,5%) casos tinham anticorpos IgM, 3 (3,1%) casos tinham anticorpos IgA, 10 (10,3%) casos tinham anticorpos IgM + IgG e 1 (1%) caso de cada um tinha anticorpos IgM + IgA e IgG + IgA. Nenhum dos casos foi positivo para os três anticorpos. A análise do coeficiente de correlação de Pearson revelou uma excelente correlação entre o PCR e os resultados sorológicos (r = 0,921, p 70 anos de idade. Nosso estudo relatou uma prevalência incomumente maior de M. pneumoniae por testes sorológicos (36,1%) e PCR (43,3%). Embora a hipótese da associação de M. pneumoniae e DAC ainda não tenha sido comprovada, a prevalência incomumente alta de M. pneumoniae em pacientes com DAC indica uma associação, se não, no desenvolvimento de aterosclerose.

Diagnóstico ultrassonográfico intrauterino de cardiomegalia fetal em felino / Intrauterine ultrasound diagnosis of fetal cardiomegaly in a feline

A ultrassonografia é um dos principais métodos de diagnóstico gestacional, no qual se realiza: biometria, monitoramento de batimentos e organogênese, detecção de anormalidades e avaliação da circulação sanguínea materno-fetal. Nesse quesito, o diagnóstico ultrassonográfico intrauterino de anormalidades fetais vem adquirindo espaço com o desenvolvimento de equipamentos mais avançados, tendo potencial para se tornar uma ferramenta de triagem para tal. Baseado na escassez observada nesse aspecto, visa-se relatar o diagnóstico ultrassonográfico intrauterino de uma alteração fetal em uma felina gestante. Foi atendida uma felina da raça persa, 3 anos, com histórico de monta natural há 40 dias. Na ultrassonografia visibilizou-se quatro fetos vivos com aproximadamente 38 dias. Uma segunda avaliação ultrassonográfica ocorreu após 12 dias, notando-se um feto com cardiomegalia, oscilação da frequência cardíaca e sofrimento fetal, enquanto os demais fetos apresentavam-se dentro da normalidade. O terceiro exame foi feito após quatro dias, visibilizando ausência de batimento cardíaco e presença de líquido em espaço pleural no feto em questão, confirmando o óbito. O parto natural ocorreu após uma semana, com nascimento de três filhotes vivos e um natimorto. Ao exame necroscópico do natimorto, confirmou-se cardiomegalia generalizada. Conclui-se que a ultrassonografia é um método padrão-ouro para diagnóstico de anormalidades fetais, permitindo planejar o parto e interceder de maneira precoce conforme a situação. Assim, esse trabalho enriquece a literatura com maiores informações relacionadas a malformações fetais observadas antes do parto, contribuindo assim com as condutas obstétricas em pequenos animais.(AU)

Ultrasonography is one of the main methods of gestational diagnosis, in which it performs: biometry, monitoring of beats and organogenesis, detection of abnormalities and evaluation of maternal-fetal blood circulation. In this regard, intrauterine sonographic diagnosis of fetal abnormalities has been gaining space with the development of more advanced equipment, and has the potential to become a screening tool for this purpose. Based on the scarcity observed in this aspect, this study aimsto report the intrauterine ultrasound diagnosis of a fetal abnormality in a pregnant feline. The patient was a 3-year-old Persian female with a history of natural mounting for 40 days. At ultrasonography, four live fetuses were visualized at approximately38 days of age. Asecond ultrasonographic evaluation was performed after 12 days, and one fetus with cardiomegaly, heart rate oscillation and fetal distress was observed, while the other fetuses were within normal limits. The third scan was performed after four days, showing absence of heartbeat and presence of fluid in the pleural space in the fetus in question, confirming the death. Natural delivery occurred after one week, with the birth of three live pups and one stillborn. At necroscopic examination of the stillborn, generalized cardiomegaly was confirmed. We conclude that ultrasonography is a gold standard method for diagnosing fetal abnormalities, allowing birth planning and early intervention according to the situation. Thus, this study enriches theliterature with more information related to fetal malformations observed before delivery, thus contributing to obstetric management in small animals.(AU)

Sarcocystis spp. detection in cattle using different diagnostic methods / Detecção de Sarcocystis spp. em bovinos usando diferentes métodos de diagnóstico

Cattle are considered intermediate hosts of Sarcocystis, which can cause clinical signs and lower performance in the acute phase of infection. Sarcocystis spp. are usually not visible to the naked eye during the post mortem inspection. Moreover, fresh microscopic examination and transmission electron microscopy techniques are difficult to apply to large samples. Therefore, extensive studies on Sarcocystis infection in cattle using molecular and serological methods are required. Here, we investigated Sarcocystis spp. infection in cattle using fresh microscopic examination and polymerase chain reaction of myocardium samples and compared the results with the presence of antibodies against Sarcocystis spp. in corresponding serum samples detected using indirect fluorescent antibody test. Microscopic Sarcocystis were observed in 100% of the myocardial samples, and Sarcocystis DNA was present in 86% (43/50) of these samples. Antibodies against Sarcocystis spp. were detected in 96% (48/50) and 80% (40/50) of the serum samples at 1:25 and 1:200 dilutions, respectively. The three associated methods (fresh microscopic examination, PCR and serology) showed good sensitivity and detection for Sarcocystis spp. compared with fresh microscopic examination (only), and they may facilitate diagnosis in live animals on a large scale as well as monitoring of the herd status.

Os bovinos são considerados hospedeiros intermediários de Sarcocystis, podendo causar sinais clínicos e menor desempenho na fase aguda da infecção. Sarcocystis spp. geralmente não são visíveis a olho nu durante a inspeção post mortem. Além disso, o exame microscópico a fresco e as técnicas de microscopia eletrônica de transmissão são difíceis de aplicar a uma amostras de grande tamanho. Portanto, são necessários extensos estudos sobre a infecção por Sarcocystis em bovinos usando métodos moleculares e sorológicos. Aqui, investigamos a infecção de Sarcocystis spp. em bovinos por meio de exame microscópico a fresco e reação em cadeia da polimerase de amostras de miocárdio e comparado os resultados com a presença de anticorpos contra Sarcocystis spp. em amostras de soro correspondentes detectadas usando o teste de anticorpos fluorescentes indiretos. Sarcocistos microscópicos foram observados em 100% das amostras de miocárdio, e o DNA de Sarcocystis estava presente em 86% (43/50) dessas amostras. Anticorpos contra Sarcocystis spp. foram detectados em 96% (48/50) e 80% (40/50) das amostras de soro nas diluições 1:25 e 1:200, respectivamente. Os três métodos associados (exame microscópico a fresco, PCR e sorologia) mostraram boa sensibilidade e detecção para Sarcocystis spp. em comparação com o exame microscópico fresco (apenas) e podem facilitar o diagnóstico em animais vivos em larga escala, bem como o monitoramento do status do rebanho.

Bovine viral diarrhea virus infection in cattle - antioxidant status and some biochemical parameters

Background: Bovine viral diarrhea virus (BVDV) infections in cattle result in significant economic losses due to reproductive performance deficiencies caused by gastrointestinal, respiratory system infections, and transplacental infections. BVDV is one of the most important and widespread pathogens in cattle worldwide, including Turkey. Methods such as virus neutralization, enzyme-linked immunosorbent assay (ELISA), reverse transcriptase and polymerase chain reaction (RT-PCR) are used for the detection of the disease. The diagnosis of the disease in its subclinical form is challenging due to the lengthy and costly procedures involved. Investigating oxidative stress parameters in ruminants with various diseases contributes significantly to diagnosis and prognosis. This study aimed to investigate some oxidative stress and biochemical parameters in cattle infected with BVDV. Materials, Methods & Results: In the study, blood samples were collected from 80 Simmental breed cows aged between approximately 4 and 8 years to determine the presence of BVDV antibodies using the ELISA method. Based on the results obtained, study groups were organized. The study included a group of 10 animals with positive antibody levels as the infected group, and a group of 10 animals with negative antibody levels as the healthy group. Blood samples were taken from the animals, and serum separation was ensured. In the obtained serum samples, levels of vitamin E, vitamin A, ß-Carotene, catalase, GSH-Px, and MDA were determined using spectrophotometric methods. In addition, serum total protein, albumin, alkaline phosphatase (ALP), aspartate aminotransferase (AST), glucose, low-density lipoprotein (LDL), high-density lipoprotein (HDL), calcium (Ca), and phosphorus (P) were measured using commercial test kits and an autoanalyzer. In the study, it was observed that the differences in serum MDA, vitamin E, vitamin A, ß-carotene, and catalase levels were statistically significant between the healthy and BVDV-infected groups (P < 0.001). The activity of GSH-Px was also found to be statistically different between the groups (P < 0.01). Among the biochemical parameters, HDL, LDL, and AST levels were found to be statistically significant between the healthy and BVDV-infected groups (P < 0.001). Additionally, ALP and glucose levels were found to be statistically significant (P < 0.01). However, although there were differences in the levels of total protein, albumin, Ca, and P between the groups, these results were not statistically significant. Discussion: Although the diagnosis of the disease was partially made based on clinical observations in BVDV infections, the ELISA method was used for accurate diagnosis. Furthermore, it was found that there was a significant difference in MDA concentration between the healthy and infected groups, indicating oxidative damage caused by the virus. Similarly, significant differences in vitamin E, vitamin A, ß-carotene, GSH-Px, and catalase levels were observed between the groups, indicating a decrease in antioxidant values due to the infection. In addition, differences in ALP, AST, glucose, LDL, and HDL levels were found between the groups. This difference is thought to be related to the effects of the disease agent on the liver and systemically. This study demonstrates that, in addition to the viral pathogen, antioxidant and biochemical values are important criteria in the detection of the disease.

Effect of GnRH administration on pregnancy-associated glycoproteins in dairy sheep with different reproductive status

Background: Measurement of pregnancy-associated glycoproteins (PAGs) by radioimmunoassay or enzyme-linked immunosorbent assay has been commonly used for early pregnancy diagnosis in ruminants. However, an accurately pregnancy detection depends on test antibody, breed and number of embryos. Only few studies have conducted to detect or predict animals at risk of late embrionic mortality (LEM) and to use hormonal interventions for embryo losses reducing, but this area is still open in sheep. The current study aimed to evaluate the effect of gonadotropin releasing hormone (GnRH) administration on Day 4 after artificial insemination on the pregnancy-associated glycoproteins in dairy sheep with different reproductive status in relation to pregnancy and late embrionic mortality detection. Materials, Methods & Results: Sixty-five East Friesian sheep were divided in 2 groups - I (Control group, n = 35) and II (GnRH group, n = 30) and sublected to estrus synchronization and artificial insemination (AI). Group I was not treated and Group II received 50 µg GnRH on Day 4 after AI. PAGs in blood serum were measured by Alertys Ruminant Pregnancy test on Days 4, 12, 20, 25 and 35 after AI and ultrasound pregnancy test was conducted on Days 20, 25, 35 and 60. Reproductive status (non-pregnant, pregnant and animals with LEM) was determined by ultrasound and the results between different groups were compared. The PAGs mean values according to reproductive status and Day after AI were analysed. Accuracy, sensitivity and specificity of the PAGs test for pregnancy diagnosis on Day 35 were calculated. On Day 20 after AI the pregnat sheep (83.3%) in group I tended to be higher than those (77.1%) in group II with decrease of 25.7% and 20% for the same groups on Day 25. On day 25 LEM was recorded in 33.3% and 24% in group I and II, respectively. The ultrasound exams on Day 60 confirmed the results from Day 25 after AI. A total value of non-pegnant, pregnant animals and LEM was 20%, 56.9% and 28.8%, respectively. The mean values of PAGs in animals with the same reproductive status in group I and II no differed statistically between Days 4 and 35 after AI. On Day 25 the PAGs values in LEM groups (0.126 ± 0.072 and 0.179 ± 0.029) were higher than those (0.062 ± 0.038 and 0.083 ± 0.023) in the non-pregnant groups, but no significan difference was deternined. On Day 35 after AI the mean values of PAGs (0.414 ± 0.125 and 0.421 ± 0.121) for the pregnant groups were significantly (P < 0.05) higher than those (0.078 ± 0.053 and 0.093 ± 0.034) for the non-pregnant groups. The values of PAGs in LEM groups on Day 25 (0.099 ± 0.062 and 0.113 ± 0.058) were decreased and close to the mean values in non-pregnant sheep. A significant effect of the day after AI on the PAGs values was evidenced in pregnant animals only (control r = 0.97 and GnRH r = 0.98; P < 0.05). The PAGs started to rise rapidly between Days 25 and 35 after AI. On Day 35 the accuracy (98%), the sensitivity (97.3%) and the specificity (100%) of the PAG test for pregnancy diagnosis were similar to the values (100%) for ultrasound method. Discussion: The gonadotropin treatment on Day 4 after AI tended to improve the reproductive performance in the sheep, but no significant effect of GnRH on the PAGs values in the animals with different reproductive status was found. The PAGs profiles in non-pregnant and LEM sheep were close and distinguishing between non-pregnant and LEM ewes on Day 35 was impossible. The PAGs values in pregnant sheep showed significantly (P < 0.05) increase between Days 25 and 35 after AI with higher mean values in pregnant than non-pregnant groups on Day 35 (P < 0.05). Alertys Ruminant Pregnancy test was a reliable for pregnancy detection in sheep on Day 35 after artificial insemination.

Prevalence and association of mycoplasma infection in the development of coronary artery disease / Prevalência e associação de infecção por micoplasma no desenvolvimento de doença arterial coronariana

Abstract Coronary heart disease (CHD) has been associated with significant morbidity and mortality worldwide. Although remain controversial, several studies have demonstrated the association of M. pneumoniae infections with atherosclerosis. We evaluated the possible association of mycoplasma infections in patients diagnosed with atherosclerosis by ELISA and PCR methods. Atherosclerotic tissue samples and blood samples were collected for the detection of mycoplasma antibodies (IgA) by ELISA from the 97 patients with coronary artery disease (CAD). M. pneumoniae specific IgA, IgG and IgM were measured by using the Anti-M. pneumoniae IgA/IgG/IgM ELISA. Detection of M. pneumoniae targeting the P1 adhesion gene was performed by PCR Acute infection of M. pneumoniae was diagnosed in 43.3% (42) of patients by PCR. The M. pneumoniae specific antibodies were detected in 36.1% (35) of patients. Twenty-five (25.8%) cases had IgG antibodies, 15 (15.5%) cases had IgM antibodies, 3 (3.1%) cases had IgA antibodies, 10 (10.3%) cases had both IgM + IgG antibodies and 1 (1%) case of each had IgM + IgA and IgG + IgA antibodies. None of the cases was positive for all three antibodies. A Pearson correlation coefficient analysis revealed an excellent correlation between the PCR and the serological results (r=0.921, p<0.001). A majority (17, 40.5%) of the M. pneumoniae positive patients are within the 41-50 years of age group, followed by 10 (23.8%) patients in the age group of 61-70 years and 2 (4.8%) patients were >70 years of age. Our study reported an unusually higher prevalence of M. pneumoniae by serological tests (36.1%) and PCR (43.3%). Although the hypothesis of the association of M. pneumoniae and CAD is yet to be proven, the unusually high prevalence of M. pneumoniae in CAD patients indicates an association, if not, in the development of atherosclerosis.

Resumo A doença coronariana (DCC) tem sido associada a significativa morbidade e mortalidade em todo o mundo. Embora ainda sejam controversos, vários estudos têm demonstrado a associação de infecções por M. pneumoniae com aterosclerose. Avaliamos a possível associação de infecções por micoplasma em pacientes com diagnóstico de aterosclerose pelos métodos ELISA e PCR. Amostras de tecido aterosclerótico e amostras de sangue foram coletadas para a detecção de anticorpos contra micoplasma (IgA) por ELISA de 97 pacientes com doença arterial coronariana (DAC). IgA, IgG e IgM específicos para M. pneumoniae foram medidos usando o Anti-M. pneumoniae IgA / IgG / IgM ELISA. A detecção de M. pneumoniae visando o gene de adesão P1 foi realizada por PCR. A infecção aguda por M. pneumoniae foi diagnosticada em 43,3% (42) dos pacientes pela PCR. Os anticorpos específicos para M. pneumoniae foram detectados em 36,1% (35) dos pacientes. Vinte e cinco (25,8%) casos tinham anticorpos IgG, 15 (15,5%) casos tinham anticorpos IgM, 3 (3,1%) casos tinham anticorpos IgA, 10 (10,3%) casos tinham anticorpos IgM + IgG e 1 (1%) caso de cada um tinha anticorpos IgM + IgA e IgG + IgA. Nenhum dos casos foi positivo para os três anticorpos. A análise do coeficiente de correlação de Pearson revelou uma excelente correlação entre o PCR e os resultados sorológicos (r = 0,921, p < 0,001). A maioria (17, 40,5%) dos pacientes positivos para M. pneumoniae está na faixa etária de 41-50 anos, seguida por 10 (23,8%) pacientes na faixa etária de 61-70 anos e 2 (4,8%) pacientes tinham > 70 anos de idade. Nosso estudo relatou uma prevalência incomumente maior de M. pneumoniae por testes sorológicos (36,1%) e PCR (43,3%). Embora a hipótese da associação de M. pneumoniae e DAC ainda não tenha sido comprovada, a prevalência incomumente alta de M. pneumoniae em pacientes com DAC indica uma associação, se não, no desenvolvimento de aterosclerose.

Radiological measures of patellofemoral alignment in healthy small-breed dogs: reference values and sex variability

Background: The evaluation of vertical kneecap position within the trochlear groove is essential for understanding the anatomical changes that may lead to canine patellar luxation. Unlike other important diagnostic imaging measurements (e.g. measures of pelvic limb alignment), no data on patellofemoral congruence parameters of canine stifles (congruence angle, linear axial patellar displacement, lateral patellofemoral angle, patellar tilt angle) are reported. The aim of this study was to present metric values of these parameters in healthy dogs from small breeds from both sexes in order to use them for identification of animals with increased risk for medial patellar luxation. Materials, Methods & Results: The study was performed on 44 stifle joints from 22 healthy dogs of both sexes from 4 small breeds (Pinscher, Pomeranian, Chihuahua and Yorkshire terrier). Inclusion criteria were negative patellar displacement and dancing patella tests after orthopaedic examination, lack of femoral and gluteal muscles asymmetry after palpation, lack of signs of long-term proprioceptive dysfunction, normal patellar and withdrawal reflexes after neurological examination. The translation of the patella in the axial plane was evaluated by means of congruence angle and axial linear patellar displacement. The rotation of the patella in the axial plane was evaluated through measurement of lateral patellofemoral angle and patellar tilt angle. Reference ranges of parameters were analysed in agreement with the guidelines of the American College of Veterinary Pathologists for the determination of reference intervals in veterinary species for sample sizes of 40-120 with non-Gaussian distribution - robust method, 95% confidence interval (CI) with 90 % CI for upper and lower reference limits. All measured patellofemoral congruence angles were negative. Their reference range was from -8.06o to -0.58o (median value -3o ). The axial patellar displacement was negligible, with values ranging from 0.1 to 0.8 mm. Lateral patellofemoral angles in all dogs were open to the lateral side (positive); the computed reference interval was from 21.61o to 41.80o (median 31.5o ). Patellar tilt angles ranged from 2o to 8o (median 5o ). A statistically significant difference between sexes was demonstrated for angles describing the rotation of the patella in the axial plane: the lateral patellofemoral angle (P = 0.0248) and the patellar tilt angle (P = 0.0004) were greater in stifles of male dogs. Discussion: This is the first study presenting reference values of parameters describing patellofemoral alignment in healthy dogs from small breeds on tangential radiographs. The values of one of most commonly measured parameters on tangential radiographs (the lateral patellofemoral angle) in stifles of small breed dogs were higher than values in humans, supposedly due to the specific joint anatomy. The patellar tilt angle is considered as more objective and not influenced by limb rotation. For the detection of patellar misalignment, the tilt angle was reported to be almost as specific as the congruence angle but more sensitive and more accurate. It is the most specific and sensitive parameter determining the direction of patellar luxation. In male dogs, the median lateral patellofemoral angle was greater than in females (P = 0.0248). Similar tendency was shown for patellar tilt angle that was also greater in male stifles (P = 0.0004). The reported statistically significant between-sex differences in lateral patellofemoral angle and patellar tilt angle with higher values of both angles in stifles of male dogs could be attributed to the better developed femoral muscles, in particular m. vastus medialis and m. sartorius, and to differences in trochlear groove and kneecap morphology. The presented reference intervals may be useful for early detection of subjects with increased risk to medial patellar luxation.

Prevalence and association of mycoplasma infection in the development of coronary artery disease / Prevalência e associação de infecção por micoplasma no desenvolvimento de doença arterial coronariana

Coronary heart disease (CHD) has been associated with significant morbidity and mortality worldwide. Although remain controversial, several studies have demonstrated the association of M. pneumoniae infections with atherosclerosis. We evaluated the possible association of mycoplasma infections in patients diagnosed with atherosclerosis by ELISA and PCR methods. Atherosclerotic tissue samples and blood samples were collected for the detection of mycoplasma antibodies (IgA) by ELISA from the 97 patients with coronary artery disease (CAD). M. pneumoniae specific IgA, IgG and IgM were measured by using the Anti-M. pneumoniae IgA/IgG/IgM ELISA. Detection of M. pneumoniae targeting the P1 adhesion gene was performed by PCR Acute infection of M. pneumoniae was diagnosed in 43.3% (42) of patients by PCR. The M. pneumoniae specific antibodies were detected in 36.1% (35) of patients. Twenty-five (25.8%) cases had IgG antibodies, 15 (15.5%) cases had IgM antibodies, 3 (3.1%) cases had IgA antibodies, 10 (10.3%) cases had both IgM + IgG antibodies and 1 (1%) case of each had IgM + IgA and IgG + IgA antibodies. None of the cases was positive for all three antibodies. A Pearson correlation coefficient analysis revealed an excellent correlation between the PCR and the serological results (r=0.921, p<0.001). A majority (17, 40.5%) of the M. pneumoniae positive patients are within the 41-50 years of age group, followed by 10 (23.8%) patients in the age group of 61-70 years and 2 (4.8%) patients were >70 years of age. Our study reported an unusually higher prevalence of M. pneumoniae by serological tests (36.1%) and PCR (43.3%). Although the hypothesis of the association of M. pneumoniae and CAD is yet to be proven, the unusually high prevalence of M. pneumoniae in CAD patients indicates an association, if not, in the development of atherosclerosis.(AU)

A doença coronariana (DCC) tem sido associada a significativa morbidade e mortalidade em todo o mundo. Embora ainda sejam controversos, vários estudos têm demonstrado a associação de infecções por M. pneumoniae com aterosclerose. Avaliamos a possível associação de infecções por micoplasma em pacientes com diagnóstico de aterosclerose pelos métodos ELISA e PCR. Amostras de tecido aterosclerótico e amostras de sangue foram coletadas para a detecção de anticorpos contra micoplasma (IgA) por ELISA de 97 pacientes com doença arterial coronariana (DAC). IgA, IgG e IgM específicos para M. pneumoniae foram medidos usando o Anti-M. pneumoniae IgA / IgG / IgM ELISA. A detecção de M. pneumoniae visando o gene de adesão P1 foi realizada por PCR. A infecção aguda por M. pneumoniae foi diagnosticada em 43,3% (42) dos pacientes pela PCR. Os anticorpos específicos para M. pneumoniae foram detectados em 36,1% (35) dos pacientes. Vinte e cinco (25,8%) casos tinham anticorpos IgG, 15 (15,5%) casos tinham anticorpos IgM, 3 (3,1%) casos tinham anticorpos IgA, 10 (10,3%) casos tinham anticorpos IgM + IgG e 1 (1%) caso de cada um tinha anticorpos IgM + IgA e IgG + IgA. Nenhum dos casos foi positivo para os três anticorpos. A análise do coeficiente de correlação de Pearson revelou uma excelente correlação entre o PCR e os resultados sorológicos (r = 0,921, p < 0,001). A maioria (17, 40,5%) dos pacientes positivos para M. pneumoniae está na faixa etária de 41-50 anos, seguida por 10 (23,8%) pacientes na faixa etária de 61-70 anos e 2 (4,8%) pacientes tinham > 70 anos de idade. Nosso estudo relatou uma prevalência incomumente maior de M. pneumoniae por testes sorológicos (36,1%) e PCR (43,3%). Embora a hipótese da associação de M. pneumoniae e DAC ainda não tenha sido comprovada, a prevalência incomumente alta de M. pneumoniae em pacientes com DAC indica uma associação, se não, no desenvolvimento de aterosclerose.(AU)

The influence of primer choice on archaeal phylogenetic analyses based on 16S rRNA gene PCR / A influência da escolha de iniciadores na filogenia de Archaea em ensaios de PCR para o gene rRNA 16S

Polymerase chain reaction (PCR) assays targeting 16S rRNA genes followed by DNA sequencing are still important tools to characterize microbial communities present in environmental samples. However, despite the crescent number of deposited archaeal DNA sequences in databases, until now we do not have a clear picture of the effectiveness and specificity of the universal primers widely used to describe archaeal communities from different natural habitats. Therefore, in this study, we compared the phylogenetic profile obtained when Cerrado lake sediment DNA samples were submitted to 16S rDNA PCR employing three Archaea-specific primer sets commonly used. Our findings reveal that specificity of primers differed depending on the source of the analyzed DNA. Furthermore, archaeal communities revealed by each primer pair varied greatly, indicating that 16S rRNA gene primer choice affects the community profile obtained, with differences in both taxon detection and operational taxonomic unit (OTU) estimates.

A amplificação de genes que codificam o rRNA 16S por reação em cadeia da polimerase (PCR) e o seu sub sequentesequenciamento consistem em uma ferramenta importante na caracterização de comunidades microbianas presentes em amostras ambientais. No entanto, apesar do crescente número de sequências de DNA de Archaea depositadas em bancos de dados, a especificidade e efetividade dos iniciadores de PCR descritos como universais e amplamente utilizados na descrição desse grupo ainda não está clara. Neste estudo foram comparados os perfis filogenéticos de comunidades de arqueias obtidos a partir amostras de DNA de sedimentos lacustres do Cerrado submetidas a ensaios de PCR empregando três pares de iniciadores específicos para Archaea, comumente utilizados neste tipo de estudo. Nossos resultados indicam que as comunidades de arqueias detectadas com cada par de iniciadores apresentaram grande variação filogenética, sugerindo que a escolha de iniciadores dirigidos ao gene de rRNA 16S tem efeito significativo no perfil da comunidade descrita, com diferenças tanto em relação aos táxons detectados, como nas estimativas de unidades taxonômicas operacionais (OTU).

Cross-genera amplification and identification of Colpodella sp. with Cryptosporidium primers in fecal samples of zoo felids from northeast China / Amplificação cruzada de gêneros e identificação de Colpodella sp. com iniciadores de Cryptosporidium em amostras fecais de zoofilia do nordeste da China

Abstract The protozoans include many intracellular human pathogens. Accurate detection of these pathogens is necessary to treat the diseases. In clinical epidemiology, molecular identification of protozoan is considered a more reliable and rapid method for identification than microscopy. Among these protozoans, Cryptosporidium considered being one of the important water-borne zoonotic pathogens and a major cause of a diarrheal disease named cryptosporidiosis in humans, domestic animals, and wild animals. This study was aimed to identify Cryptosporidium in zoo felids (N= 56) belonging to different zoo of China, but accidentlly Colpodella was encountered in the zoo felids sample and phylogenetic data confirmed this unexpected amplification from fecal samples using two-step nested-PCR. Phylogenetic analysis revealed the fact about the specific primers used previously by many researchers and cross-genera amplification. We came to know that genetically sequenced amplicon gives more accurate identification of species. This study suggests more investigation on Colpodella which has been neglected previously but gains the attention of researchers after identified from humans and animals and has been known to correlate with neurological symptoms in patients.

Resumo Os protozoários incluem muitos patógenos humanos intracelulares. A detecção acurada desses patógenos é necessária para tratar as doenças. Na epidemiologia clínica, a identificação molecular de protozoários é considerada o método de identificação mais confiável e rápido do que a microscopia. Entre esses protozoários, o Cryptosporidium é considerado um dos importantes patógenos zoonóticos transmitidos pela água e uma das principais causas de uma doença diarreica denominada criptosporidiose em humanos, animais domésticos e selvagens. Este estudo teve como objetivo identificar Cryptosporidium em zoofelídeos (N = 56) pertencentes a diferentes zoológicos da China, mas acidentalmente Colpodella foi encontrada na amostra de zoofelídeos e os dados filogenéticos confirmaram essa amplificação inesperada de amostras fecais usando nested-PCR em duas etapas. A análise filogenética revelou o fato sobre os primers específicos usados ​​anteriormente por muitos pesquisadores e a amplificação entre gêneros. Ficamos sabendo que o amplicon sequenciado geneticamente fornece uma identificação mais acurada das espécies. Este estudo sugere mais investigação sobre Colpodella, que foi negligenciada anteriormente, mas ganha a atenção dos pesquisadores depois de identificada em humanos e animais e é conhecida por se correlacionar com sintomas neurológicos em pacientes.