RESUMO
The aim of this study was to determine whether Southern Blot/Hybridization (SB) associated to Polymerase Chain Reaction (PCR) improves the sensitivity in the detection of hemoplasma DNA in domestic cats (Felis catus). Whole blood was collected in tubes containing the anticoagulant ethylenediamine tetra-acetic acid and DNA extracted from 149 animals. PCR was performed using species specific primers to amplify the 16S ribosomal RNA subunit of Mycoplasma haemofelis and 'Candidatus M. haemominutum' from these samples. Hybridization was performed using a 16S rDNA probes chemically labeled and the results were visualized using a chemiluminescent substrate addition followed by autoradiography. Eighteen (12.1%) of the 149 tested samples had a positive PCR result for hemoplasma species DNA. SB/hybridization technique showed that 24/149 (16.1%) samples were positive for hemoplasmas, confirming the 18 PCR-positive results and reveling six additional positive animals (p<0.001). SB/hybridization method with specific probes was more sensitive than PCR performed alone, being complimentary to this technique to diagnose infections caused by feline hemoplasmas.
Assuntos
Animais Domésticos , Southern Blotting , Doenças do Gato/diagnóstico , Infecções por Mycoplasma/veterinária , Reação em Cadeia da Polimerase , Animais , Gatos , Infecções por Mycoplasma/diagnóstico , Sensibilidade e EspecificidadeRESUMO
In Brazil ostriches farming began at the end of 20th Century, to supply the international market of meat, feathers and leather. Feathers are economically important to industry and artisanal sectors. Lice can cause intense pruritus that depending on the intensity of infestation can lead to depreciation of feathers. In Rio de Janeiro there are unknown species of lice infesting ostriches. Thus the objective of this study was to identify the lice that happen in birds from commercial farms in the Municipality of Três Rios. From 2003 to 2006 ostriches were monthly examined for the presence of ectoparasites. When infestations were detected some feathers were removed, placed into plastic bags and sent to the laboratory for further analysis. After removed from feathers, ectoparasites were examined using a stereoscope, followed by clarification in 10% potassium hydroxide and dehydrated in ethanol. Permanent slides were mounted in natural Canada balsam for observation in optical microscope. Based on observed characteristics, it was possible to determine that collected specimens belong to the species Struthiolipeurus rheae.
Assuntos
Infestações por Piolhos/veterinária , Ftirápteros , Struthioniformes/parasitologia , Animais , BrasilRESUMO
The aim of this study was to determine whether Southern Blot/Hybridization (SB) associated to Polymerase Chain Reaction (PCR) improves the sensitivity in the detection of hemoplasma DNA in domestic cats (Felis catus). Whole blood was collected in tubes containing the anticoagulant ethylenediamine tetra-acetic acid and DNA extracted from 149 animals. PCR was performed using species specific primers to amplify the 16S ribosomal RNA subunit of Mycoplasma haemofelis and 'Candidatus M. haemominutum' from these samples. Hybridization was performed using a 16S rDNA probes chemically labeled and the results were visualized using a chemiluminescent substrate addition followed by autoradiography. Eighteen (12.1%) of the 149 tested samples had a positive PCR result for hemoplasma species DNA. SB/hybridization technique showed that 24/149 (16.1%) samples were positive for hemoplasmas, confirming the 18 PCR-positive results and reveling six additional positive animals (p 0.001). SB/hybridization method with specific probes was more sensitive than PCR performed alone, being complimentary to this technique to diagnose infections caused by feline hemoplasmas.
O objetivo deste trabalho foi verificar se a técnica de Southern Blot/Hibridização (SB) em associação à reação de polimerização em cadeia (PCR) aumenta a sensibilidade na detecção de DNA de hemoplasmas em gatos domésticos (Felis catus). O sangue total foi coletado em tubos contendo o anticoagulante ácido etilenodiamino tetra-acético, o DNA extraído a partir de 149 animais e a PCR realizada com o uso de sequências iniciadoras espécie-específicas, para amplificar subunidade 16S do RNA ribossomal de Mycoplasma haemofelis e 'Candidatus M. haemominutum' dessas amostras. Para a hibridização, foram utilizadas sondas específicas quimicamente marcadas, e os resultados visualizados por meio da adição de substrato quimiluminescente seguida de autoradiografia. Dezoito (12,1%) das 149 amostras testadas apresentaram resultado PCR-positivo para o DNA de hemoplasmas. A técnica de SB mostrou que 24/149 (16,1%) amostras apresentaram resultado positivo para hemoplasmas, confirmando os 18 resultados PCR-positivos, além de revelar seis outros adicionais (p 0,001). O método de SB com sondas específicas mostrou-se mais sensível do que a PCR realizada isoladamente, sendo complementar para o diagnóstico das infecções causadas pelos hemoplasmas felinos.
RESUMO
In Brazil ostriches farming began at the end of 20th Century, to supply the international market of meat, feathers and leather. Feathers are economically important to industry and artisanal sectors. Lice can cause intense pruritus that depending on the intensity of infestation can lead to depreciation of feathers. In Rio de Janeiro there are unknown species of lice infesting ostriches. Thus the objective of this study was to identify the lice that happen in birds from commercial farms in the Municipality of Três Rios. From 2003 to 2006 ostriches were monthly examined for the presence of ectoparasites. When infestations were detected some feathers were removed, placed into plastic bags and sent to the laboratory for further analysis. After removed from feathers, ectoparasites were examined using a stereoscope, followed by clarification in 10% potassium hydroxide and dehydrated in ethanol. Permanent slides were mounted in natural Canada balsam for observation in optical microscope. Based on observed characteristics, it was possible to determine that collected specimens belong to the species Struthiolipeurus rheae.
No Brasil, a criação de avestruzes foi iniciada no final do Século XX, para suprir o mercado internacional de carne, plumas e couro. As plumas têm importância econômica nos setores industrial e artesanal. Os piolhos podem causar prurido intenso, que, dependendo da intensidade de infestação, pode levar à depreciação das plumas. No Estado do Rio de Janeiro, são desconhecidas as espécies de piolhos que infestam avestruzes. Assim, o objetivo deste trabalho foi identificar os piolhos que ocorrem nas aves pertencentes a um criatório comercial no Município de Três Rios. Nos anos de 2003 a 2006, mensalmente, as avestruzes foram examinadas, para verificar a presença de ectoparasitos. Quando constatada a infestação, algumas plumas foram retiradas, colocadas em sacos plásticos e enviadas ao laboratório para exame. Após serem coletados das plumas, os ectoparasitos foram examinados ao microscópio estereoscópio, clarificados em hidróxido de potássio a 10%, desidratados em álcool etílico e montados em lâmina com balsamo do Canadá natural, para observação em microscópio óptico. Baseando-se na observação das características, foi possível determinar os espécimes coletados como pertencentes à espécie Struthiolipeurus rheae.
