Use of the FLOTAC technique as a new coproparasitological diagnostic method in aquatic mammals and comparison with traditional methods.

The inadequate choice of a diagnostic method or the option for techniques that have low sensitivity and specificity may limit the diagnosis of parasitic agents that affect aquatic mammals. The aim of this study was to evaluate the performance of the FLOTAC technique and compare it with three traditional methods (Willis, sedimentation and centrifugation- flotation) used in the diagnosis of gastrointestinal parasites in aquatic mammals. For this, 129 fecal samples from 12 species were collected. Each sample was submitted to laboratory processing using the Willis, Hoffman techniques, Faust method and FLOTAC. Sensitivity, specificity, real prevalence, estimated prevalence, positive predictive value, negative predictive value, correct classification (accuracy) and incorrect classification were evaluated to compare the different diagnostic methods. The highest frequency of positive samples occurred using FLOTAC (46.51%), compared to Hoffman (23.25%), Faust (10.07%) and Willis techniques (6.97%). In the samples analyzed, the occurrence of Strongylidae eggs and Eimeriidae oocysts was frequently observed. The FLOTAC technique proved to be the most appropriate technique and due to its efficacy, is strongly recommended for coproparasitological evaluations in aquatic mammals.

Molecular and serological investigation of infectious diseases in captive and free-range crab-eating fox (Cerdocyon thous - Linnaeus, 1776) from northeastern Brazil.

The aim of this study was to detect DNA and antibodies anti-Leishmania spp., Neospora caninum and Toxoplasma gondii in captive and free-range crab-eating fox (Cerdocyon thous) from northeastern Brazil. Twenty-five crab-eating foxes from different states of northeastern Brazil were sampled by this study. Blood samples were collected by cephalic or jugular vein punctures. The whole blood was submitted to PCR, and the sera samples to the serological analysis as follows: MAT for T. gondii, NAT for N. caninum, and ELISA for L. chagasi. The frequence of antibodies anti-T. gondii was 50% and 29.41% for free-range and captive wild canids, respectively. The frequence of antibodies anti-N. caninum observed by this study was 62.50% and 23.52% for free-range and captive wild canids, respectively. The frequence of antibodies anti-L. chagasi was 4.0% for captive wild canids. Co-infections cases were identified as follows: one captive wild canid seropositive for T. gondii and L. chagasi and two free-range animals seropositive for T. gondii and N. caninum. All PCR assays performed were negative for the pathogens analyzed. This study describes the presence of antibodies anti-T. gondii, N. caninum e L. chagasi in wild canids from northeastern Brazil and highlights the necessity of further studies on infectious diseases in free-range and captive wild canids.

Soroprevalência de Toxoplasma gondii em equídeos do Nordeste do Brasil / Seroprevalence of Toxoplasma gondii in equidae from Northeast Brazil

Este estudo teve como objetivo determinar a soroprevalência da toxoplasmose em equídeos mantidos em diferentes formas de manejo no estado de Pernambuco. Para tanto, um total de 400 amostras de soro sanguíneo de equídeos clinicamente saudáveis foram analisados através do teste de aglutinação modificado (MAT) considerando-se cut-off de 1:25. Dados referentes às características dos animais e dos rebanhos, sistema de criação, presença de outros animais, idade, sexo, raça, aptidão, condição física foram coletados por meio de questionários investigativos. Anticorpos IgG anti-Toxoplasma gondii foram detectados em 12,5% (50/400) dos animais analisados. Dos 12 municípios estudados, houve positividade em 91,67% (11/12) com variação entre 4,4% e 33,3%. Quando avaliados os fatores de risco, apenas o fator mesorregião (p=0,029) apresentou associação com a infecção, particularmente Zona da Mata (OR=3), seguida de Região Metropolitana do Recife (OR=2,2), Agreste (OR=1,7) e Sertão (OR=1). Os resultados revelam a presença do parasito na área estudada, o que pode representar um elo na cadeia de transmissão da toxoplasmose a qual tem repercussão em saúde pública tendo em vista que o Brasil é o oitavo maior exportador de carne equina do mundo.(AU)

This paper reports seroprevalence of toxoplasmosis in horses kept in different forms of breeding system in the state of Pernambuco. For that, 400 blood serum samples from clinically healthy horses were analyzed through the test of modified agglutination (MAT) considering cut-off of 1:25. Data related to the characteristics of the animals and herds, breeding system, presence of other animals, age, gender, breed, aptitude, and physical conditions were collected throughout investigative surveys. IgG anti-Toxoplasma gondii antibodies were detected in 12.5% (50/400) of the analyzed animals. In the 12 studied towns, there was a positivity in 91. 67% (11/12) with a variation between 4% and 33.3%. When the risk factors were evaluated, only the mesoregion factor (p=0.029) had an association with the infection, particularly the Zona da Mata region (OR=3), followed by the Recife Metropolitan Area (OR=2.2), Agreste region (OR=1.7) and Sertão region (OR=1). The results shows the presence of the parasites on the studied area, which may represent a link with the transmission chain of toxoplasmosis which has influence on the public health system, considering that Brazil is the eighth greatest exporter of equine meat in the world.(AU)

SEROPREVALENCE OF TOXOPLASMA GONDII IN CAPTIVE ANTILLEAN MANATEE (TRICHECHUS MANATUS MANATUS) IN BRAZIL.

Antillean manatees ( Trichechus manatus manatus) are aquatic mammals that inhabit marine waters from Central America to the northeastern region of Brazil, and they are an endangered species. Infection with Toxoplasma gondii through intake of water or food contaminated with oocysts has been reported among marine mammals. The present study aimed to evaluate the prevalence of antibodies to T. gondii in West Indian manatees living in captivity in northeastern Brazil. Serum samples from 55 West Indian manatees from three different captive groups were tested for T. gondii antibodies by means of the modified agglutination test using a cutoff of 1:25. The samples were screened at dilutions of 1:25, 1:50, and 1:500, and positive samples were end-titrated using twofold serial dilutions; antibodies were found in six Antillean manatees (10.9%) with titers of 1:50 in three, 1:500 in one, 1:3,200 in one, and 1:51,200 in one manatee. This study is the first report of T. gondii antibodies in captive Antillean manatees in Brazil.

Novel markers of inflammatory response and hepatic dysfunction in canine leishmaniasis.

Dogs are the main host of Leishmania infantum, and the clinical presentation may range from asymptomatic to systemic manifestations. The immune mechanisms in infected, but clinically healthy dogs, prevails Th1 response mediated by cytokines. In this sense, adenosine deaminase (ADA) and butyrylcholinesterase (BChE) are considered as key enzymes in several physiological processes, including the modulation of inflammatory process. Considering the variable immune response against Leishmania and the known participation of ADA and BChE, the aim of this study was to assess the relation between these two enzymes with the inflammatory response as well as hepatic function in dogs naturally infected with L. infantum. For this purpose, the activity of ADA and BChE was assessed in sera of 24 dogs naturally infected with L. infantum, plus 17 healthy dogs. The naturally infected dogs had clinical signs compatible with leishmaniasis and sera activities of ADA (P<0.01) and BChE (P<0.05) decreased, when compared to the healthy group. The reduction of ADA activity probably represented an effect on inflammatory response, especially due to the decreased hydrolysis of extracellular adenosine, might in order to protect against tissue damage and, also, setting a down-regulation on pro-inflammatory cytokines. BChE enzyme had no effect on modulating the immune response in leishmaniasis, but it decreased, a fact may related to deficiency of synthesis in the liver. Therefore, ADA and BChE activities reduced probably in order to protect against extra tissue damage and due failure in synthesis, respectively.

In vitro efficacy of oil from the seed of Carapa guianensis (andiroba) in the control of Felicola subrostratus

This study was perfomed to assess the in vitro effect of oil from the seed of andiroba (Carapa guianensis Aubl.) on Felicola subrostratus (Burmeister, 1838) (Mallophaga: Trichodectidae). Six hundred specimens of F. subrostratus from neighborhood of Jordão, Recife-PE, Brazil, were collected by hand directly from the fur of cats infested naturally. The lice were transported in plastic recipients to the Laboratory of Parasitic Diseases of Domesticated Animals of the Department of Veterinary Medicine of the Universidade Federal Rural de Pernambuco (Brazil) for the immersion test. Four dilutions of andiroba oil (100, 50, 25 and 10%) in distilled water were tested, using Tween 80 as dispersant; two control groups (one with distilled water and the other with monosulfiram) were also formed; four replicates were performed, with 25 specimens for each dilution, totaling 100 lice per treatment. After the test, the insects were kept at room temperature and observed for mortality rates for 72 h. The biological activity of the product achieved 100% mortality of the insects in the first hour at concentrations of 100 and 50% and in the third hour at concentrations of 25 and 10%. The results demonstrate the possibility of controlling F. subrostratus throught the use of oil extracted from andiroba seeds.

Situação epidemiológica da infecção por Toxoplasma gondii em equídeos na microrregião do Brejo Paraibano / Epidemiological situation of Toxoplasma gondii infection in equids from Brejo Paraibano microregion, Brazil

Objetivou-se com o estudo caracterizar a situação epidemiológica da infecção por Toxoplasma gondii em equídeos na microrregião do Brejo Paraibano, região Nordeste do Brasil. Anticorpos contra T. gondii foram pesquisados em 257 amostras de equídeos (204 equinos, 46 muares e sete asininos) em 26 propriedades. Para o diagnóstico sorológico utilizou-se a Reação de Imunofluorescência Indireta (RIFI) e um ponto de corte de 1:64. O número de focos encontrado foi de 46,1%. Nas amostras analisadas, a prevalência geral foi de 7,8% (I.C. 4,8-8,8). A prevalência foi de 8,3% (I.C. 4,9-13,0) para os equinos, 2,2% (I.C. 0,1-11,5) para os muares e 28,6% (I.C. 3,7-71,0) entre os asininos. Na regressão logística das variáveis observou-se que a fonte de água foi um fator de risco, pois naquelas propriedades que forneciam água corrente para os animais o risco de infecção foi 4,4 vezes maior do que naquelas propriedades que forneciam água parada (OR 4,4; I.C. 1,0-19,0). Este é o primeiro relato da presença de anticorpos contra T. gondii em equídeos nessa microrregião do estado da Paraíba. Para diminuir os riscos de infecção nestas espécies, deve-se fornecer aos animais uma água de boa qualidade, bem como evitar acesso de gatos a fontes de água e instalações onde os animais são mantidos.

The objective of the study was to characterize the epidemiological situation of Toxoplasma gondii infection in equids from Brejo Paraibano microregion, Northeastern Brazil. Antibodies against T. gondii were investigated in samples of 257 equids (204 horses, 46 mules and seven donkeys) in 26 properties. For serological diagnosis was used the Indirect Fluorescent Antibody Test (IFAT) and a cut-off of 1:64. The number of foci was found to be 46.1%. In the samples analyzed, the overall prevalence was 7.8% (C.I. 4.8-8.8). The prevalence was 8.3% (C.I. 4.9-13.0) for horses, 2.2% (C.I. 0.1-11.5) for mules and 28.6% (C.I. 3.771.0) among donkeys. Logistic regression of the variables showed that the water source was a risk factor, because in those properties that supplied running water to the animals the risk of infection was 4.4 times higher than in those properties which provided standing water (OR 4.4; C.I. 1.0-19.0). This is the first report of the presence of antibodies against T. gondii in equids in this micro-region of Paraiba State. To reduce the risk of infection in these species, good quality water should be given to the animals, as well as access of cats to water sources and facilities where animals are kept must be avoided.

Genetic conservation of potentially immunogenic proteins among Brazilian isolates of Babesia bovis.

Bovine babesiosis caused by Babesia bovis remains an important constraint for the development of cattle industries worldwide. Effective control can be achieved by vaccination with live attenuated phenotypes of the parasite. However, these vaccines have a number of drawbacks, which justifies the search for better, safer vaccines. In recent years, a number of parasite proteins with immunogenic potential have been discovered. However, there is little information on the genetic conservation of these proteins among different parasite isolates, which hinders their assessment as immunogens. The aim of the present study was to evaluate the conservation of the genes ama-1, acs-1, rap-1, trap, p0 and msa2c among five Brazilian isolates of B. bovis. Through polymerase chain reaction, genetic sequencing and bioinformatics analysis of the genes, a high degree of conservation (98-100%) was found among Brazilian isolates of B. bovis and the T2Bo isolate. Thus, these genes are worth considering as viable candidates to be included in a recombinant cocktail vaccine for cattle babesiosis caused by B. bovis.

Structural and ultrastructural changes in the lungs of cats Felis catus (Linnaeus, 1758) experimentally infected with D. immitis (Leidy, 1856).

Clinical signs are seldom observed in feline heartworm disease, and the pathophysiological changes in the lungs of infected animals remain undefined. The goal of this study was to evaluate the structural and ultrastructural changes in the lungs of cats experimentally infected with Dirofilaria immitis. Six healthy cats were each infected with two adult heartworms by intravenous transplantation (Receptor Group, RG). The control group consisted of two uninfected animals kept under the same conditions as the RG. At 42 days after transplantation, all cats were euthanized and necropsied for worm recovery and collection of lung samples for examination by light microscopy (LM) and transmission electron microscopy. By LM, lung sections from the six infected cats exhibited bronchial and bronchiolar lesions. Alterations in all tissues of the pulmonary arteries were observed in the infected animals. In conclusion, cats infected experimentally with D. immitis developed lesions in their lungs as a consequence of arterial disease and intense interstitial pneumonia.

Real-time polymerase chain reaction based on msa2c gene for detection of Babesia bovis.

This paper reports a quantitative real-time polymerase chain reaction (q-PCR) based on the msa2c gene and standardized with Platinum SYBR Green/ROX for the detection of Babesia bovis in cattle. The msa2c q-PCR amplified a DNA fragment with average dissociation temperature of 77.41°C (± 0.25°C). No amplification was detected when DNA from B. bigemina, A. marginale or Bos taurus was used as the template. The detection limit of the msa2c q-PCR was 1000 copies per ml of blood sample, with a linear correlation between the number of msa2c copies and threshold cycle. The comparison between msa2c q-PCR and conventional PCR for cytochrome b revealed 88.8% agreement, with a Kappa index of 0.75. In the comparison between msa2c q-PCR and an enzyme-linked immunosorbent assay (ELISA) with semi-purified B. bovis antigen, agreement was 96.3% and the Kappa index was 0.91. The agreement between three tests was 85.8%. The msa2c q-PCR detected a higher number of positive cattle than conventional PCR in an enzootically stable area, but did not differ significantly from ELISA. No significant differences were detected between the three diagnostic tests with cattle from an enzootically unstable area. All animals raised on a tick-free facility were negative for B. bovis in the three tests. These results suggest that msa2c q-PCR is a useful test for the detection of B. bovis infection.

Prevalência de anticorpos anti-neospora caninum (Apicomplexa: Sarcocystidae) em bovinos leiteiros de propriedades rurais em três microrregiões no estado do Maranhão / Prevalence of anti-neospora caninum (Apicomplexa: Sarcocystidae) antibodies in dairy cattle in rural properties of three microrregions of Maranhão, Brazil

Objetivou-se, no presente estudo, pesquisar a prevalência de anticorpos anti-Neospora caninum em 812 amostras de soros sangüíneos de bovinos leiteiros procedentes de propriedades rurais de sete municípios das microrregiões de Itapecuru-Mirim, Médio Mearim e Presidente Dutra, estado do Maranhão, Brasil. Para o cálculo do tamanho da amostra, considerou-se um soroprevalência de 34,7 por cento para N. caninum, com erro máximo de 9,5 por cento e intervalo de confiança de 95 por cento. Para a detecção da presença de anticorpos da classe IgG, utilizou-se a técnica de Imunofluorescência Indireta (IFI), com ponto de corte 1:200, usando como antígeno, taquizoítos da cepa NC-1, mantida em cultura celular no Laboratório de Diagnóstico das Parasitoses dos Animais da Escola de Medicina Veterinária da UFBA. Do total de amostras analisadas, encontrou-se uma prevalência de 50,74 por cento. Os títulos variaram de 1:200 a 1:6400, assim distribuídos: 108 (26,21 por cento) amostras de soro apresentaram título de 1:200; 132 (32,04 por cento) 1:400; 94 (22,81 por cento) 1:800; 46 (11,16 por cento) 1:1600; 23 (5,58 por cento) 1:3200 e nove (2,18 por cento) com títulos de 1:6400. Dentre as microrregiões, a Itapecuru-Mirim apresentou o menor percentual de animais soropositivos (20,69 por cento) e Presidente Dutra o maior (47,66 por cento). Com relação à variável sexo, observou-se maior prevalência de sororreagentes nas fêmeas (46,80 por cento) do que nos machos (52,46 por cento). Não se verificou diferença significativa (P>0,05) para as variáveis microrregiões, sexo e idade. Conclui-se que os bovinos leiteiros das regiões estudadas estão expostos à infecção por N. caninum.

The objective in the present study was to research the prevalence of anti-Neospora caninum in 812 samples of blood serum of dairy cattle from farms of seven municipalities of microrregions of Itapecuru-Mirim, Middle Mearim and President Dutra, state of Maranhão, Brazil. For the calculation of sample size, it was considered a seroprevalence of 34.7 percent for N. caninum, with a maximum error of 9.5 percent and a confidence interval of 95 percent. To detect antibodies, it was used the technique of Indirect Immunofluorescence (IFI), with the cut-off of 1:200, using as antigen, tachyzoites strain NC-1, maintained in cell culture in the Laboratory of Diagnosis of Parasitism of the Animals, School of Veterinary Medicine of the Federal University of Bahia, Brazil. Of the total samples, it was obtained a prevalence of 50.74 percent. The titles ranged from 1:200 to 1:6400, distributed as follows: 108 (26.21 percent) serum samples showed title of 1:200; 132 (32.04 percent) 1:400; 94 (22.81 percent) 1:800; 46 (11.16 percent) of 1:1600; 23 (5.58 percent) of 1:3200 and nine (2.18 percent) with titers of 1:6400. Among the microrregiões the Itapecuru-Mirim showed the lowest percentage of animals seropositive (20.69 percent) and President Dutra the largest (47.66 percent). It was observed higher prevalence of seropositives in females (46.80 percent) than in males (52.46 percent). There was no significant difference (P> 0.05) for the microrregions variables, sex and age. Concluded that the dairy cattle of the regions studied are exposed to infection by N. caninum.

Detection of Leishmania infantum in Rhipicephalus sanguineus ticks from Brazil and Italy.

Canine leishmaniosis is a widespread disease caused by Leishmania parasites, which are transmitted by phlebotomine sand flies. However, in some areas where canine leishmaniosis is endemic, but the primary vectors have not been found, ticks have been suspected to play a role in transmitting the infection. Herewith, we report the detection of Leishmania infantum kinetoplast minicircle DNA (kDNA) in ticks collected from naturally infected dogs living in rural areas of Southern Italy (site A) and Northeastern Brazil (site B). Between March and October 2007, ticks were collected from 26 dogs positive to anti-Leishmania antibodies (one from site A and 25 from site B) and either placed directly into vials containing 70% ethanol or maintained alive for identification and subsequent dissection. All the 95 ticks collected were morphologically identified as Rhipicephalus sanguineus. After identification, their genomic DNA was extracted (either individually or in pools) and processed by polymerase chain reaction (PCR) for the detection of L. infantum kDNA. Two pools of salivary glands from ticks (one from five females and other from five males) found on a dog from site A and tested by a conventional PCR were positive. Amplicon sequencing confirmed the identity of the parasite. In addition, nine (12.3%) out of the 73 ticks found on dogs from site B and tested by a real-time PCR were positive, with a low parasite load (less than 1 parasite/ml). The retrieval of L. infantum kDNA in salivary glands of R. sanguineus ticks has been here reported for the first time. Therefore, further studies are needed to assess the competence of ticks as vectors of Leishmania parasites from dog to dog.

Comparação entre diversos antígenos para o diagnóstico de Anaplasma marginale por ELISA / Comparison between several antigens for diagnosis of Anaplasma marginale by ELISA

Anaplasmose bovina é uma doença com grande importância nas regiões tropicais e subtropicais do mundo por determinar perdas econômicas devido à mortalidade e redução da produtividade. É causada por Anaplasma marginale, uma riquétsia intraeritrocítica obrigatória cujo controle requer, além de uma vacina eficiente, uma acurada identificação de bovinos cronicamente infectados. Apesar de existirem atualmente diversos métodos de diagnóstico dessa riquétsia, os métodos sorológicos, em particular o ensaio de imunoadsorção enzimática-ELISAs, são os mais utilizados devido à sua versatilidade e praticidade. No entanto, devido ao grande número de antígenos disponíveis, atualmente torna-se necessária uma avaliação para definir quais antígenos apresentam um melhor desempenho no diagnóstico da anaplasmose. Soros de bovinos positivos e negativos para A. marginale por PCR, e soros de animais provenientes do Brasil e Costa Rica, foram testados em ELISAs baseados em MSP1a, MSP2 e MSP5 recombinantes, um pool das três proteínas recombinantes, e antígeno de lisado de corpúsculos iniciais da riquétsia (CI). Utilizando soro de bovinos positivos para A. marginale por PCR, uma maior sensibilidade foi observada no ELISA CI. No entanto, uma maior especificidade, com soro de bovinos negativos a PCR, foi observada com os ELISAs recombinantes. O porcentual de bovinos positivos do Brasil e Costa Rica foi maior com ELISA CI. Razões para essas diferenças são discutidas.

Bovine anaplasmosis is a major disease in tropical and subtropical regions of the world by determine economical loss due mortality and productive reduction. The disease is caused by Anaplasma marginale, an intraerythrocytic rickettsia whose control requires, besides an efficient vaccine, the accurate identification of chronically infected cattle. Although the existence of diverse methods of diagnosis of this rickettsia, the serological methods, in particular the enzyme immunosorbent assays (ELISAs), are the most used due to its versatility and practice. However, due to the high number of antigens currently available, an evaluation becomes necessary to define which antigens present the better performance in the diagnosis of anaplasmosis. Sera from cattle positive or negative to A. marginale by PCR, and sera from cattle proceeding from Brazil and Costa Rica, were tested by ELISAs based in recombinant MSP1a, MSP2, and MSP5, a pool of the three recombinant proteins, and initial body lisate antigen (CI). Using sera from A. marginale positive cattle by PCR, the highest sensitivity was shown by CI ELISA. Nevertheless, the highest specificity, with sera from negative cattle by PCR, was shown by recombinants ELISAs. The percentiles of positive cattle from Brazil and Costa Rica were higher with CI ELISA. Reasons for such differences were discussed.

Molecular and antigenic characterisation of ribosomal phosphoprotein P0 from Babesia bovis.

Babesia bovis is a tick-borne pathogen that remains an important constraint for the development of cattle industries in tropical and subtropical regions of the world. Effective control can be achieved by vaccination with live attenuated phenotypes of the parasite. However, these phenotypes have a number of drawbacks, which justifies the search for new, more efficient immunogens based mainly on recombinant protein technology. In the present paper, ribosomal phosphoprotein P0 from a Brazilian isolate of B. bovis was produced and evaluated with regard to conservation and antigenicity. The protein sequence displayed high conservation between different Brazilian isolates of B. bovis and several Apicomplexa parasites such as Theileria, Neospora and Toxoplasma. IgG from cattle experimentally and naturally infected with B. bovisas well as IgG1 and IgG2 from naturally infected cattle reacted with the recombinant protein. IgG from cattle experimentally infected with Babesia bigemina cross-reacted with B. bovis recombinant P0. These characteristics suggest that P0 is a potential antigen for recombinant vaccine preparations against bovine babesiosis.

[Comparison of nested-PCR with blood smear examination in detection of Ehrlichia canis and Anaplasma platys in dogs]. / Comparação de nested-PCR com o diagnóstico direto na detecçáo de Ehrlichia canis e Anaplasma platys em cães.

The clinical signs of Ehrlichia canis and Anaplasma platys infection are similar, and the diagnosis of these pathogens made by stained blood smears is poor due sensibility and specificity. On the other hand, the molecular diagnosis is highly sensitive and specific and nested-PCR have been optimized for accurate diagnosis these pathogens in dogs. At the veterinary teaching hospital, whole-blood samples with EDTA were obtained from 100 dogs and smears were made from blood samples for evaluation for intracellular parasites. For each sample, DNA was extracted and submitted to nPCR analysis for detection of E. canis and A. platys. The results of stained blood smears showed 9% of the animals were positive for E. canis and 21% for A. platys. Regarding of nPCR analysis, 57 and 55% of dogs were positive for E. canis and A. platys respectively. As compared to a nested PCR, the stained blood smears revealed false-negative results for both E. canis and A. platys. The results indicate that the nPCR is highly sensitive and specific for detection of both pathogens and the molecular diagnosis could be more useful at veterinary hospital.

Eficácia do extrato concentrado contendo Saccharum officinarum L: Poaceae, Azadirachta indica A. Juss. Meliaceae, e Eucaliptus spp Myrtaceae, sobre Pediculus capitis De Geer, (Anoplura: Pediculidae) / Effectiveness of concentrate extract containing Saccharum Officinarum L: Poaceae, Azadirachta Indica A. Juss. Meliaceae, And Eucaliptus Spp Myrtaceae Against Pediculus Capitis De Geer, (Anoplura: Pediculidae)

O objetivo deste trabalho foi avaliar a eficácia do extrato concentrado contendo Saccharum officinarum L. Poaceae, Azadirachta indica A. Juss. Meliaceae, e Eucaliptus spp Myrtaceae sobre Pediculus capitis De Geer Pediculidae. Foram coletados mil e trinta e cinco piolhos da cabeça de crianças infestadas por Pediculus capitis de creches da Região Metropolitana do Recife, Estado de Pernambuco, Brasil. Para realização do teste utilizou-se três grupos com trezentos e quarenta e cinco piolhos cada referentes aos produtos, extrato concentrado (EC), inseticida piretróide (PI) e controle (C). Os piolhos foram imersos em solução dos produtos por três minutos e em seguida secos. A mortalidade foi monitorada em diferentes momentos, por um período de vinte e quatro horas. O extrato concentrado matou todos os piolhos antes de doze horas e uma média de 60,28 por cento (208/345) de mortalidade do EC ocorreu entre três e seis horas, a maior parte da mortalidade do PI foi observada 24 h após o tratamento. Os resultados mostraram que o extrato contendo Saccharum officinarum, Azadirachta indica, e Eucaliptus spp constitui-se como uma potente ferramenta no controle do Pediculus capitis.

The aim of this work was to evaluate the effectiveness of concentrate extract containing Saccharum officinarum L. Poaceae, Azadirachta indica A. Juss. Meliaceae, and Eucaliptus spp Myrtaceae against Pediculus capitis De Geer Pediculidae. A thousand and thirty five head lice were collected from children with Pediculus capitis infestation from some day care centers at Metropolitan Region of Recife, Pernambuco State, Brazil. The tests were performed in three groups with three hundred forty five lice each one according to product, concentrate extract (CE), pyrethroid insecticide (PI) and control (C). The immersing head lice in the diluted products for three minutes, washing off products and dry the insects were used. The mortality of lice was monitored at different points in time, for a period of twenty four hours. Concentrate extract killed all head lice after twelve hours and an average of 60.28 percent (208/345) of lice mortality of the CE occurred between three and six hours, while the mortality of PI was observed 24 h after treatment. The results showed the extract containing Saccharum officinarum, Azadirachta indica, and Eucaliptus spp could be a potent tool in the control of Pediculus capitis.

Comparação de nested-PCR com o diagnóstico direto na detecção de Ehrlichia canis e Anaplasma platys em cães / Comparison of nested-PCR with blood smear examination in detection of Ehrlichia canis and Anaplasma platys in dogs

Os sinais clínicos das infecções por Ehrlichia canis e Anaplasma platys são similares, e o diagnóstico desses patógenos feito por esfregaços sanguíneos corados é difícil devido à sensibilidade e especificidade. Por outro lado, os diagnósticos moleculares são altamente sensíveis e específicos, e nested-PCRs têm sido otimizadas para o diagnóstico preciso desses patógenos em cães. Em um Hospital Veterinário Escola, amostras de sangue total com EDTA foram obtidas de 100 cães, e esfregaços foram feitos das amostras de sangue para busca dos parasitos intracelulares. Para cada amostra, DNA foi extraído e submetido à nPCR para detecção de E. canis e A. platys. Os resultados dos esfregaços sanguíneos mostraram que 9% dos animais foram positivos para E. canis e 21% para A. platys. Com relação à nPCR, 57 e 55% dos cães foram positivos para E. canis e A. platys, respectivamente. Quando comparados com a nPCR, os esfregaços sanguíneos corados revelaram resultados falso-negativos para E. canis e A. platys. Os resultados indicam que a nPCR é altamente sensível e específica para detecção de ambos os patógenos, e os diagnósticos moleculares podem ser mais úteis nos Hospitais Veterinários.

The clinical signs of Ehrlichia canis and Anaplasma platys infection are similar, and the diagnosis of these pathogens made by stained blood smears is poor due sensibility and specificity. On the other hand, the molecular diagnosis is highly sensitive and specific and nested-PCR have been optimized for accurate diagnosis these pathogens in dogs. At the veterinary teaching hospital, whole-blood samples with EDTA were obtained from 100 dogs and smears were made from blood samples for evaluation for intracellular parasites. For each sample, DNA was extracted and submitted to nPCR analysis for detection of E. canis and A. platys. The results of stained blood smears showed 9% of the animals were positive for E. canis and 21% for A. platys. Regarding of nPCR analysis, 57 and 55% of dogs were positive for E. canis and A. platys respectively. As compared to a nested PCR, the stained blood smears revealed false-negative results for both E. canis and A. platys. The results indicate that the nPCR is highly sensitive and specific for detection of both pathogens and the molecular diagnosis could be more useful at veterinary hospital.

Molecular and antigenic characterisation of ribosomal phosphoprotein P0 from Babesia bovis

Babesia bovis is a tick-borne pathogen that remains an important constraint for the development of cattle industries in tropical and subtropical regions of the world. Effective control can be achieved by vaccination with live attenuated phenotypes of the parasite. However, these phenotypes have a number of drawbacks, which justifies the search for new, more efficient immunogens based mainly on recombinant protein technology. In the present paper, ribosomal phosphoprotein P0 from a Brazilian isolate of B. bovis was produced and evaluated with regard to conservation and antigenicity. The protein sequence displayed high conservation between different Brazilian isolates of B. bovis and several Apicomplexa parasites such as Theileria, Neospora and Toxoplasma. IgG from cattle experimentally and naturally infected with B. bovisas well as IgG1 and IgG2 from naturally infected cattle reacted with the recombinant protein. IgG from cattle experimentally infected with Babesia bigemina cross-reacted with B. bovis recombinant P0. These characteristics suggest that P0 is a potential antigen for recombinant vaccine preparations against bovine babesiosis.

Comparação de nested-PCR com o diagnóstico direto na detecção de Ehrlichia canis e Anaplasma platys em cães / Comparison of nested-PCR with blood smear examination in detection of Ehrlichia canis and Anaplasma platys in dogs

Os sinais clínicos das infecções por Ehrlichia canis e Anaplasma platys são similares, e o diagnóstico desses patógenos feito por esfregaços sanguíneos corados é difícil devido à sensibilidade e especificidade. Por outro lado, os diagnósticos moleculares são altamente sensíveis e específicos, e nested-PCRs têm sido otimizadas para o diagnóstico preciso desses patógenos em cães. Em um Hospital Veterinário Escola, amostras de sangue total com EDTA foram obtidas de 100 cães, e esfregaços foram feitos das amostras de sangue para busca dos parasitos intracelulares. Para cada amostra, DNA foi extraído e submetido à nPCR para detecção de E. canis e A. platys. Os resultados dos esfregaços sanguíneos mostraram que 9% dos animais foram positivos para E. canis e 21% para A. platys. Com relação à nPCR, 57 e 55% dos cães foram positivos para E. canis e A. platys, respectivamente. Quando comparados com a nPCR, os esfregaços sanguíneos corados revelaram resultados falso-negativos para E. canis e A. platys. Os resultados indicam que a nPCR é altamente sensível e específica para detecção de ambos os patógenos, e os diagnósticos moleculares podem ser mais úteis nos Hospitais Veterinários.

The clinical signs of Ehrlichia canis and Anaplasma platys infection are similar, and the diagnosis of these pathogens made by stained blood smears is poor due sensibility and specificity. On the other hand, the molecular diagnosis is highly sensitive and specific and nested-PCR have been optimized for accurate diagnosis these pathogens in dogs. At the veterinary teaching hospital, whole-blood samples with EDTA were obtained from 100 dogs and smears were made from blood samples for evaluation for intracellular parasites. For each sample, DNA was extracted and submitted to nPCR analysis for detection of E. canis and A. platys. The results of stained blood smears showed 9% of the animals were positive for E. canis and 21% for A. platys. Regarding of nPCR analysis, 57 and 55% of dogs were positive for E. canis and A. platys respectively. As compared to a nested PCR, the stained blood smears revealed false-negative results for both E. canis and A. platys. The results indicate that the nPCR is highly sensitive and specific for detectionof both pathogens and the molecular diagnosis could be more useful at veterinary hospital.

[Occurrence of weevils (Insecta: Coleoptera) in pet food traded in the Metropolitan Region of Recife, Pernambuco State, Brazil]. / Ocorrência de coleópteros em alimentos industrializados para cães, comercializados na região metropolitana de Recife, PE.

With the purpose of verifying the occurrence of insect pests in dog food commercialized in the Metropolitan Region of Recife, samples from 15 different pet stores were submitted to the extraction of insects in a Berlese-Tullgren apparatus. Tribolium castaneum (Herbst) (Tenebrionidae) (55.2%) was the most frequent specie followed by Oryzaephilus surinamensis (L.) (Cucujidae) (31.3%), Rhyzopertha dominica (Fabricius) (Bostrichidae) (8.9%) and Lasioderma serricorne (Fabricius) (Anobiidae) (4.7%), all from Coleoptera. Recife showed the highest rate of infestation (53.6%), followed by Olinda (34.4%) and Jaboatão dos Guararapes (12.0%). The infestation by coleopters in the region occurs with high frequency and may represent a threat mainly in commercialized products in bulk.