Optimization of a molecular method for the diagnosis of canine babesiosis.

Babesiosis is a hemolytic disease caused by protozoans of the genus Babesia (Apicomplexa). This disease occurs worldwide and is transmitted by ticks to a variety of mammals, including humans. The objective of the present study was to optimize a molecular approach for the detection of a fragment of 18S rDNA of Babesia canis, Babesia vogeli, Babesia rossi or Babesia gibsoni based on a single semi-nested Polymerase Chain Reaction (PCR), and compare the efficiency of this approach with that of a simple PCR protocol. To this end, 100 blood samples collected from dogs with suspected hemoparasite infections were analyzed. A comparison of the results of simple PCR and semi-nested PCR indicated a highly significant difference (p value = 0.0000). While only five (5%) of the samples tested positive using the simple protocol, 22 (22%) were positive using the snPCR technique. The results of this study reinforce the findings of previous studies, which have demonstrated the greater sensitivity of tests based on nested or semi-nested PCR. Therefore, to avoid false-negative results due to low levels of parasitemia, we suggest the preferential use of this protocol in epidemiological studies of canine babesiosis, particularly those that require reliable estimates of the prevalence of infection.

Optimization of a molecular method for the diagnosis of canine babesiosis / Otimização de método molecular para diagnóstico da babesiose canina

Babesiosis is a hemolytic disease caused by protozoans of the genus Babesia (Apicomplexa). This disease occurs worldwide and is transmitted by ticks to a variety of mammals, including humans. The objective of the present study was to optimize a molecular approach for the detection of a fragment of 18S rDNA of Babesia canis, Babesia vogeli, Babesia rossi or Babesia gibsoni based on a single semi-nested Polymerase Chain Reaction (PCR), and compare the efficiency of this approach with that of a simple PCR protocol. To this end, 100 blood samples collected from dogs with suspected hemoparasite infections were analyzed. A comparison of the results of simple PCR and semi-nested PCR indicated a highly significant difference (p value = 0.0000). While only five (5%) of the samples tested positive using the simple protocol, 22 (22%) were positive using the snPCR technique. The results of this study reinforce the findings of previous studies, which have demonstrated the greater sensitivity of tests based on nested or semi-nested PCR. Therefore, to avoid false-negative results due to low levels of parasitemia, we suggest the preferential use of this protocol in epidemiological studies of canine babesiosis, particularly those that require reliable estimates of the prevalence of infection.

A babesiose é uma doença hemolítica de ocorrência mundial, causada por protozoários do gênero Babesia (Apicomplexa), que são transmitidos por carrapatos a diversos mamíferos, incluindo o homem. O objetivo deste estudo foi otimizar um método molecular para a detecção de fragmento do 18S rDNA de Babesia canis, Babesia vogeli, Babesia rossi ou Babesia gibsoni com base em uma única semi-nested (snPCR), comparando sua eficiência com um protocolo de PCR simples. Para isso, 100 amostras de sangue de cães com suspeita de hemoparasitoses foram analisadas e, enquanto o protocolo de PCR simples indicou somente 5% (5/100) de amostras positivas, o protocolo de snPCR, com 22% (22/100) de amostras positivas, apresentou maior sensibilidade (p valor = 0,0000). Este resultado está de acordo com outros estudos que mostram a maior sensibilidade de detecção dos testes baseado em nested ou snPCR. Assim, como uma forma de prevenir resultados falso-negativos devido à baixa parasitemia, sugere-se que este protocolo seja preferencialmente usado nos estudos epidemiológicos de babesiose canina, em especial naqueles que tratam da sua prevalência.

Optimization of a molecular method for the diagnosis of canine babesiosis / Otimização de método molecular para diagnóstico da babesiose canina

Babesiosis is a hemolytic disease caused by protozoans of the genus Babesia (Apicomplexa). This disease occurs worldwide and is transmitted by ticks to a variety of mammals, including humans. The objective of the present study was to optimize a molecular approach for the detection of a fragment of 18S rDNA of Babesia canis, Babesia vogeli, Babesia rossi or Babesia gibsoni based on a single semi-nested Polymerase Chain Reaction (PCR), and compare the efficiency of this approach with that of a simple PCR protocol. To this end, 100 blood samples collected from dogs with suspected hemoparasite infections were analyzed. A comparison of the results of simple PCR and semi-nested PCR indicated a highly significant difference (p value = 0.0000). While only five (5%) of the samples tested positive using the simple protocol, 22 (22%) were positive using the snPCR technique. The results of this study reinforce the findings of previous studies, which have demonstrated the greater sensitivity of tests based on nested or semi-nested PCR. Therefore, to avoid false-negative results due to low levels of parasitemia, we suggest the preferential use of this protocol in epidemiological studies of canine babesiosis, particularly those that require reliable estimates of the prevalence of infection.

A babesiose é uma doença hemolítica de ocorrência mundial, causada por protozoários do gênero Babesia (Apicomplexa), que são transmitidos por carrapatos a diversos mamíferos, incluindo o homem. O objetivo deste estudo foi otimizar um método molecular para a detecção de fragmento do 18S rDNA de Babesia canis, Babesia vogeli, Babesia rossi ou Babesia gibsoni com base em uma única semi-nested (snPCR), comparando sua eficiência com um protocolo de PCR simples. Para isso, 100 amostras de sangue de cães com suspeita de hemoparasitoses foram analisadas e, enquanto o protocolo de PCR simples indicou somente 5% (5/100) de amostras positivas, o protocolo de snPCR, com 22% (22/100) de amostras positivas, apresentou maior sensibilidade (p valor = 0,0000). Este resultado está de acordo com outros estudos que mostram a maior sensibilidade de detecção dos testes baseado em nested ou snPCR. Assim, como uma forma de prevenir resultados falso-negativos devido à baixa parasitemia, sugere-se que este protocolo seja preferencialmente usado nos estudos epidemiológicos de babesiose canina, em especial naqueles que tratam da sua prevalência.

Detection of Ehrlichia canis and Anaplasma platys DNA using multiplex PCR.

We hereby propose a novel sensitive, specific, and cost-efficient method to detect Ehrlichia canis and Anaplasma platys DNA from canine whole blood samples by multiplex PCR. Blood samples from hemoparasited dogs attending the Veterinary Hospital at the Universidade Federal Rural da Amazônia-UFRA, Belém, Brazil, were collected in tubes containing EDTA. Amplification of E. canis and A. platys 16S rDNA by nested (n) PCR was successfully achieved by using primers specific to the Anaplasmataceae in the first round of PCR, followed by a second round of PCR using E. canis-specific primers in conjunction with A. platys-specific primers. The amplicons obtained were cloned and sequenced, yielding sequences of 478 and 473 bp (including primers) pertaining to regions of the 16S rDNA of E. canis and A. platys, respectively. The protocol we here propose may help to measure the prevalence of canine monocytic ehrlichiosis (CME) and canine cyclic thrompocytopenia, not only in northern Brazil, where there is no data available, but also elsewhere.

Bloqueo epidural para laparotomía en síndrome carcinoide / Epidural blockade for a laparotomy in a patient with carcinoid syndrome

El síndrome carcinoide es una entidad rara caracterizada por la liberación de péptidos vasoactivos por un tumor carcinoide. Cuando éste se diagnostica generalmente ya ha dado metástasis hepáticas. Estos tumores carcinoides son más frecuentes en el tracto gastrointestinal, existiendo también en otros órganos siendo el de ovario uno de los más raros con una frecuencia de 0.3 por ciento, la asociación de enfermedad cardiaca carcinoide es aún más rara y sólo existen 14 casos reportados en la literatura. Los pacientes con síndrome carcinoide son difíciles de manejar durante la anestesia y la enfermedad es de tal rareza que es inusual que un individuo obtenga experiencia en el manejo, con la excepción de una serie, la experiencia reciente muestra estar basada en casos únicos. Describimos el manejo anestésico con bloqueo epidural de una paciente con síndrome carcinoide para laparotomía por un carcinoide de ovario asociado con enfermedad cardiaca carcinoide. Previo a la cirugía la paciente fue manejada con digoxina, maleato de enalapril y furosemide. Medicación pre-anestésica, ranitidina e hidrocortisona, el monitoreo incluyó oximetría del pulso, presión venosa central, tensión arterial, EKG. Después de la infusión de lactato de Ringer, se aplicó bloqueo epidural con lidocaína al 2 por ciento c/ep., sin complicaciones; se lograron condiciones óptimas para la cirugía. Se sugiere el bloqueo epidural como una alternativa viable en el manejo anestésico en este tipo de pacientes

Incidence of memory of transoperative events during total intravenous general anesthesia with propofol

The incidence of memories of events during the total intravenous general anesthesia is studied in a 400 adult patient group in order to establish the influence of the included doses of drugs in this scheme, upon the mentioned incidence. The anesthetic method included propofol, fentanyl and vecuronium. Two groups of patients were constituted according to the propofol infusion rate (PIR) required for the anesthetic maintenance: group I required a PIR<6.0 mg.kg-1.h-1, and group II required a PIR>6.0 mg.kg-1.h-1. Tranasnesthetic monitoring was done with cardiac frequency, systolic and diastolic arterial pressures, ECG, pulse oximeter oxygen saturation and tidal CO2. The memory incidence of transoperative events was obtained through postoperative interviews about intraoperative events. Patients in group I had a significantly greater remembrance incidence both in real events and in dreams than the patients in gropu II. We clonclude that a PIR of propofol<6.0 mg. kg-1.h1 can be associated with a high incidence of remembrances of transoperative events