Detecção e quantificação do genoma de parvovirus de galinha (chpv) em frangos de corte saudáveis e com síndrome da má absorção

A síndrome da má absorção (SMA), caracterizada pelo mau desenvolvimento e desuniformidade do lote de aves, causa importantes prejuízos econômicos à avicultura comercial. No entanto, por se tratar de uma doença multifatorial e possivelmente polimicrobiana, o envolvimento do parvovírus de galinha (ChPV) na ocorrência da SMA ainda é pouco conhecido. Com o propósito de elucidar a possível associação entre a presença do ChPV e a ocorrência da SMA, foram desenvolvidas e aplicadas ferramentas moleculares para a detecção e quantificação do ChPV em amostras de frangos comerciais no Estado do Rio Grande do Sul. Uma PCR quantitativa foi desenvolvida para detectar e quantificar cópias do genoma (CG) do ChPV em amostras de suabes de cloaca de 59 frangos saudáveis e 68 frangos com sinais clínicos sugestivos de SMA. Os resultados revelaram que todas as amostras dos dois grupos investigados, continham o genoma do ChPV. No entanto, a carga viral em frangos com SMA foi significativamente (p≤0,0001) maior (1x105 CG/100 ng DNA) do que em frangos saudáveis (1,3x103 CG/100 ng DNA). Adicionalmente às amostras de cloaca, o ChPV também foi investigado em amostras de tecidos (fígado, timo, baço, bursa de Fabricius - BF e intestino) e soros provenientes de nove frangos saudáveis e 50 frangos com sinais indicativos da SMA. O ChPV foi encontrado tanto em aves saudáveis como nas aves com SMA, no entanto, observou-se uma diferença na distribuição deste agente nos tecidos analisados. O genoma do vírus foi mais frequentemente detectado na BF, baço e intestino das aves com SMA, sendo que o intestino foi o tecido que apresentou maior carga viral. Os resultados encontrados nestes estudos demonstraram que o genoma viral estava altamente disseminado nas aves investigadas. Além disso, observou-se uma maior carga viral em frangos de corte com SMA quando comparado com aves sadias. Com base nos resultados encontrados, sugere-se que a maior carga viral de ChPV existente em aves com SMA, em relação a aves saudáveis, seja um dos fatores que favoreça a ocorrência da síndrome.The malabsorption syndrome (MAS), characterized by the poor development and lack of uniformity of chicken flocks, causes significant economic losses to commercial poultry. However, because it is a multifactorial and possibly polymicrobial disease, the involvement of Chicken parvovirus (ChPV) in the MAS occurrence is still not clear. In order to elucidate the possible association between the presence of ChPV and the occurrence of MAS, molecular tools were developed and applied for the detection and quantification of ChPV DNA in commercial poultry samples in the state of Rio Grande do Sul. A quantitative PCR was developed to detect and quantify the ChPV genome copies (GC) in cloacal swab samples of 59 healthy broilers and 68 broilers with clinical signs suggestive of MAS. The results showed that all investigated samples of the two groups contained the genome ChPV. However, viral loads in MAS-affected animals were significantly (p≤0.0001) higher (1x105 GC/100 ng DNA) than in healthy broilers (1.3x103 GC/100 ng DNA). In addition to the cloacal samples, the presence of ChPV DNA was also investigated in tissue samples (liver, thymus, spleen, bursa of Fabricius - BF and intestine) and sera from nine healthy broilers and 50 broilers with signals indicative of MAS. The ChPV was found in healthy avian as well MAS-affected, however, there was a difference in the distribution of this agent in those tissues. The virus genome was more frequently detected in the BF, spleen and intestines of the MAS-affected broilers, and the intestines contained the highest viral loads, in comparison with other tissues. Our results demonstrated that the viral genome can be found in both healthy and MAS-affected broilers. In addition, higher viral loads were detected in broilers with signs suggestive of MAS compared to healthy birds. Based on these results, it is suggested that the greatest viral load of ChPV existing in MAS-affected broilers when compared to healthy birds, is one of the factors that favor the occurrence of the syndrome

Detecção e quantificação do genoma de parvovirus de galinha (chpv) em frangos de corte saudáveis e com síndrome da má absorção

A síndrome da má absorção (SMA), caracterizada pelo mau desenvolvimento e desuniformidade do lote de aves, causa importantes prejuízos econômicos à avicultura comercial. No entanto, por se tratar de uma doença multifatorial e possivelmente polimicrobiana, o envolvimento do parvovírus de galinha (ChPV) na ocorrência da SMA ainda é pouco conhecido. Com o propósito de elucidar a possível associação entre a presença do ChPV e a ocorrência da SMA, foram desenvolvidas e aplicadas ferramentas moleculares para a detecção e quantificação do ChPV em amostras de frangos comerciais no Estado do Rio Grande do Sul. Uma PCR quantitativa foi desenvolvida para detectar e quantificar cópias do genoma (CG) do ChPV em amostras de suabes de cloaca de 59 frangos saudáveis e 68 frangos com sinais clínicos sugestivos de SMA. Os resultados revelaram que todas as amostras dos dois grupos investigados, continham o genoma do ChPV. No entanto, a carga viral em frangos com SMA foi significativamente (p≤0,0001) maior (1x105 CG/100 ng DNA) do que em frangos saudáveis (1,3x103 CG/100 ng DNA). Adicionalmente às amostras de cloaca, o ChPV também foi investigado em amostras de tecidos (fígado, timo, baço, bursa de Fabricius - BF e intestino) e soros provenientes de nove frangos saudáveis e 50 frangos com sinais indicativos da SMA. O ChPV foi encontrado tanto em aves saudáveis como nas aves com SMA, no entanto, observou-se uma diferença na distribuição deste agente nos tecidos analisados. O genoma do vírus foi mais frequentemente detectado na BF, baço e intestino das aves com SMA, sendo que o intestino foi o tecido que apresentou maior carga viral. Os resultados encontrados nestes estudos demonstraram que o genoma viral estava altamente disseminado nas aves investigadas. Além disso, observou-se uma maior carga viral em frangos de corte com SMA quando comparado com aves sadias. Com base nos resultados encontrados, sugere-se que a maior carga viral de ChPV existente em aves com SMA, em relação a aves saudáveis, seja um dos fatores que favoreça a ocorrência da síndrome.The malabsorption syndrome (MAS), characterized by the poor development and lack of uniformity of chicken flocks, causes significant economic losses to commercial poultry. However, because it is a multifactorial and possibly polymicrobial disease, the involvement of Chicken parvovirus (ChPV) in the MAS occurrence is still not clear. In order to elucidate the possible association between the presence of ChPV and the occurrence of MAS, molecular tools were developed and applied for the detection and quantification of ChPV DNA in commercial poultry samples in the state of Rio Grande do Sul. A quantitative PCR was developed to detect and quantify the ChPV genome copies (GC) in cloacal swab samples of 59 healthy broilers and 68 broilers with clinical signs suggestive of MAS. The results showed that all investigated samples of the two groups contained the genome ChPV. However, viral loads in MAS-affected animals were significantly (p≤0.0001) higher (1x105 GC/100 ng DNA) than in healthy broilers (1.3x103 GC/100 ng DNA). In addition to the cloacal samples, the presence of ChPV DNA was also investigated in tissue samples (liver, thymus, spleen, bursa of Fabricius - BF and intestine) and sera from nine healthy broilers and 50 broilers with signals indicative of MAS. The ChPV was found in healthy avian as well MAS-affected, however, there was a difference in the distribution of this agent in those tissues. The virus genome was more frequently detected in the BF, spleen and intestines of the MAS-affected broilers, and the intestines contained the highest viral loads, in comparison with other tissues. Our results demonstrated that the viral genome can be found in both healthy and MAS-affected broilers. In addition, higher viral loads were detected in broilers with signs suggestive of MAS compared to healthy birds. Based on these results, it is suggested that the greatest viral load of ChPV existing in MAS-affected broilers when compared to healthy birds, is one of the factors that favor the occurrence of the syndrome

Hoja quincenal de precipitación: Quincena 1ª - Junio 1966

Although Pasteurella multocida is a member of the respiratory microbiota, under some circumstances, it is a primary agent of diseases , such as fowl cholera (FC), that cause significant economic losses. Experimental inoculations can be employed to evaluate the pathogenicity of strains, but the results are usually subjective and knowledge on the pathogenesis of this agent is still limited. The objective of this study was to establish a new methodology for classifying the pathogenicity of P. multocida by formulating a standard index. Strains isolated from FC cases and from swine with respiratory problems were selected. One hundred mL of a bacterial culture of each strain, containing 106 CFU, was inoculated in 10 one-day-old broilers. Mortality after inoculation, time of death (TD), and the presence of six macroscopic lesions were evaluated over a period of seven days post-inoculation (dpi). A Pathogenicity Index Per Bird (IPI), ranging 0 to 10, was calculated. Liver and heart fragments were collected to reisolate the bacteria. Blood was collected from the surviving birds, and an ELISA test was carried out to detect specific antibodies. The median of the pathogenicity indices, the number of lesions and the rate of bacteria reisolation were significantly different (p<0.05) among the origins of the isolates (p<0.05). The pathogenicity index developed in this study allows the classification of Pasteurella multocida pathogenicity and may be an alternative to the pathogenicity models currently used for screening

Antimicrobial susceptibility and pathogenicity of Escherichia coli strains of environmental origin

The study aimed to evaluate the antimicrobial susceptibility of 109 samples of Escherichia coli (E. coli) of environmental origin and to characterize these isolates according to the degree of pathogenicity in vivo, verifying a possible relationship between this variable and susceptibility to the active principles tested. The isolates were subjected to disc diffusion test to 14 antibiotics. From 16.5% to 90% of the samples were sensitive; 1 - 28.5% showed intermediate degree of susceptibility and between 9 to 78% of E. coli analyzed were resistant. The highest resistance percentages were seen in the class of quinolones and tetracyclines (&gt;75%), and for sensitivity in the class of amphenicols (68.8%). By inoculating 1- day - old chicks, the isolates were classified as highly pathogenic (2.7%), intermediate (10.1%), low (42.2%) and apathogenic (45%). It was observed a wide variation in the susceptibility profile of isolates in relation to antimicrobials. It was also found that most of the samples had pathogenic potential (55%), thus being considered as APEC (avian pathogenic E. coli). No relationship between pathogenicity and antimicrobial susceptibility (P≤0.05) was observed