Resumo A dirofilariose é um problema de saúde para cães e gatos, especialmente nas regiões costeiras tropicais e subtropicais do mundo. Alguns estudos compararam a eficácia das técnicas de diagnóstico usadas para detectar esta parasitose. Portanto, o objetivo deste estudo foi comparar a microscopia óptica (OM), técnicas sorológicas e moleculares para o diagnóstico de infecção por Dirofilaria immitis . Foram coletadas, entre julho de 2015 e abril de 2016, amostras de 103 cães em Cabo Frio, RJ, Brasil. O exame direto, distensão espessa, distensão delgada e o teste de Knott modificado foram usados para detectar microfilárias. O ELISA (Snap ™ 4Dx ® IDEXX) foi usado para detectar antígenos e a reação em cadeia da polimerase (PCR) foi usada para detectar DNA e o sequenciamento para diferenciação e confirmação de espécie. Das amostras, 19,4% foram positivas de acordo com a microscopia. Por PCR, 15,5% do total foram positivos. Utilizando o ELISA, a taxa de positividade foi de 29,1%. Dirofilariose oculta foi detectada em 11,6% das amostras. A sensibilidade ao ELISA mostrou-se superior à PCR ou microscopia (P = 0,001). O sequenciamento das amostras confirmou a presença de Dirofilaria immitis e Acanthocheilonema reconditum . O ELISA foi mais eficaz no diagnóstico sorológico de dirofilariose canina e deve ser usado em estudos clínicos e epidemiológicos.

Abstract Heartworm disease is a health problem for dogs and cats, especially in tropical and subtropical coastal regions of the world. Some studies have compared the efficacy of the diagnostic techniques used to detect this parasitosis. Therefore, the aim of this study was to compare parasitological optical microscopy (POM), serological and molecular techniques for diagnosing canine heartworm infection. Samples were collected between July 2015 and April 2016 from 103 dogs in Cabo Frio, RJ, Brazil. The wet fresh blood, thick smears, thin smears and modified Knott’s test were used to detect microfilariae. ELISA (Snap™ 4Dx ® IDEXX) was used to detect antigens and the polymerase chain reaction (PCR) was used to detect DNA and enable sequencing for species differentiation and confirmation. 19.4% of samples were positive according to microscopy. Through PCR, 15.5% of the total were positive. Using ELISA, the positivity rate was 29.1%. Occult heartworm infection was detected in 11.6% of the samples. ELISA sensitivity was shown to be higher than PCR or microscopy (P = 0.001). Sequencing of samples confirmed the presence of Dirofilaria immitis and Acanthocheilonema reconditum . ELISA was more effective for serological diagnosis canine heartworm and should be used in clinical and epidemiological studies.

Resumo Gatos são menos susceptíveis à infecção por Dirofilaria immitis do que cães. Apesar de rara, a doença nos gatos pode ser fatal mesmo com baixas cargas parasitárias. Muitas vezes, a doença é assintomática ou apresenta sintomas inespecíficos, principalmente associados com a morte de formas parasitárias imaturas. Microfilaremia é rara e transitória. Normalmente, quando ocorre microfilaremia, ela permanece por, no máximo, 33 dias. Este estudo descreve o caso de um felino que apresentava sinais inespecíficos e microfilaremia prolongada: um gato sem raça definida, portador de infecção pelo vírus da leucemia felina (FeLV) que foi diagnosticado como microfilaremico ao acaso. A infecção foi detectada pela presença de microfilárias em esfregaço sanguíneo e, posteriormente, confirmada pelo teste de antígenos (SNAP Feline Triple Test, Idexx®) e por ecocardiograma.

Abstract Cats are less susceptible to Dirofilaria immitis infection than dogs. Although rare, the feline disease can be fatal even with low parasitic loads. The infection is often asymptomatic or has non-specific symptoms that are mainly associated with the death of immature worms. Microfilaremia is rare and transient. Normally, microfilaremia, when present, lasts for not more than 33 days. This study describes a feline case presenting with non-specific clinical signs and prolonged microfilaremia. Case: a random bred cat infected by feline leukemia virus (FeLV) that was found to be microfilaremic by chance. The infection was detected by the presence of microfilariae in a blood smear and was confirmed by antigen test (SNAP Feline Triple Test, Idexx®) and echocardiogram.

Abstract INTRODUCTION: Visceral leishmaniasis is a disease that affects humans, wildlife, and domestic species. Since dogs play a key role in urban Leishmania spp. transmission, the Brazilian government maintains the Monitoring and Control Program of Visceral Leishmaniasis (VLMCP) in endemic regions, which promotes awareness campaigns aiming to enhance the control of the infection. The VLMCP recommends the Dual Path Platform (DPP®) canine visceral leishmaniasis test (Bio-Manguinhos, Brazil) for screening and enzyme-linked immunosorbent assay to confirm the infection. The DPP® test is produced and distributed by the Health Ministry to the Municipal Health Centers responsible for the local VLMCP. The test is not available to all the clinics, forcing some veterinarians to use other rapid tests for screening and diagnosis of this disease in their daily routine. METHODS: The present study was conducted to compare the performance of the DPP® and SNAP® tests using sera from the dogs with confirmed infections of L. infantum and from the dogs with no previous testing, residing in areas with a low Leishmania infection. RESULTS: There was 97.0% agreement between the two tests. Sensitivity and specificity of the SNAP® test were 96.3% and 100%, respectively. Agreement between both the antibody tests and the parasitological detection methods was 96.8%. The DPP® test had 95.8% sensitivity and 100% specificity. CONCLUSIONS: The SNAP® and the DPP® tests were virtually equivalent in terms of detection of canine antibodies against L. infantum, and both the tests demonstrated high and similar levels of sensitivity and specificity.

RESUMO: O presente estudo avaliou o uso de um dispositivo de coleta salivar (Salivette®) para mensurar o cortisol salivar de gatos domésticos e avaliar o efeito do análogo sintético do feromônio facial felino - fração F3 (Feliway®) sobre seus níveis de cortisol. Um total de 28 gatos domésticos mantidos em gatil particular tiveram amostras de saliva coletadas antes da exposição ao feromônio facial felino e após 35 dias de exposição. Dois difusores de feromônio foram instalados na área onde os animais se alimentavam, a fim de garantir que todos os gatos fossem expostos. Os dispositivos de coleta salivar permitiram a coleta de volume salivar suficiente (≥0,20 mL) para a mensuração do cortisol. Os níveis de cortisol salivar variaram de 0,02g/dL a 0,16ug/dL, com coeficiente de variação de 42,1% antes e de 62,6% após à exposição ao feromônio (F=3.2351; p≤ 0.0002). Não foi verificada diferença entre os níveis de cortisol salivar nas amostras obtidas antes (x =0,078μg/dL) e após (x =0,066μg/dL) (t=1,79; p=0,08) à exposição. Entretanto, os níveis de cortisol salivar diminuíram em 75% (21/28) dos gatos expostos ao feromônio (x 2=12,07; p=0,0005), sugerindo que os animais apresentam susceptibilidade diferente ao feromônio facial sintético ou que passaram períodos de tempo distintos na área onde os difusores foram instalados.

ABSTRACT: This study aimed to evaluate the ability of a saliva collection device (Salivette®) to measure cortisol levels in saliva samples of domestic cats and to assess the effect of a synthetic analogue of the feline facial pheromone fraction F3 (Feliway®) on cortisol levels. A total of 28 domestic cats from a private high-quality sanctuary were sampled before exposure to the facial pheromone and after 35 days of exposure. Two pheromone devices were placed in the area where the animals ate to guarantee the exposure of all cats. The collecting device yielded a sufficient volume of saliva (≥0.20mL) to allow cortisol measurement. Cortisol measurements ranged from 0.02g/dL to 0.16μg/dL, with a difference between before (42.1%) and after (62.6%) exposure to the pheromone (F=3.2351; p≤0.0002). No difference in cortisol levels was observed between before (x =0.078μg/dL) and after (x =0.066μg/dL) (t=1.79; p=0.08) exposure. However, salivary cortisol levels decreased in 75% (21/28) of the cats after exposure (x 2=12.07; p=0.0005), suggesting that the animals have different susceptibilities to the pheromone or that they spent different lengths of time in the area where the pheromone devices were installed.

Os gatos, como hospedeiros definitivos, apresentam um papel fundamental na transmissão do Toxoplasma gondii. Nosso estudo teve como objetivo determinar a presença de imunoglobulinas G e M anti-T. gondii, e a frequência de oocistos nas fezes de uma população de gatos domésticos do Rio de Janeiro, Brasil. Objetivou-se ainda estudar a associação da infecção por T. gondii com a idade, sexo, raça, estilo de vida, dieta e infecção por retrovírus. Um total de 108 gatos foi incluído no estudo e 54 amostras de fezes foram obtidas desses animais. Somente 5,6% dos gatos foram sororreagentes para T. gondii, utilizando-se o teste de hemaglutinação indireta. Nenhum dos 54 gatos apresentou oocistos em suas amostras fecais. Embora sem comprovação estatística, machos, sem raça definida, com acesso livre às ruas e gatos com mais de dois anos de idade tenderam a ser mais expostos ao parasito. Idade, estilo de vida e uso de caixa de areia foram considerados importantes fatores de risco. Anemia e infecção por retrovírus não apresentaram relação com infecção por T. gondii. Não foram detectados anticorpos na maioria dos gatos (94,4%), indicando que esses gatos nunca foram expostos ao parasito e que, se infectados, poderão eliminar grande número de oocistos no ambiente.

Cats, as definitive host, play an important role in the transmission of Toxoplasma gondii. This study aimed to establish the seroprevalence of anti-T. gondii immunoglobulins G and M, and determine the frequency of oocysts in the feces of the domestic cat population in Rio de Janeiro, Brazil. We also aimed to study the association between T. gondii infection and age, sex, breed, lifestyle, diet and retroviral infection. A total of 108 cats were included in the study and fecal samples of 54 of those cats were obtained. Only 5.6% of the cats were seropositive for anti-T. gondii immunoglobulins using the indirect hemagglutination test. None of the 54 cats presented oocysts in their fecal samples. Although not statistically significant, males, mixed-breed, free-roaming and cats aged two years and older were found to be more exposed. Age, lifestyle and the use of litter boxes were found to play an important role as risk factors. Anemia and retroviral infections were independent of T. gondii infection. No antibodies were detected in the majority of cats (94.4%), indicating that those cats had never been exposed to the parasite and, therefore, once infected, they could present the risk of shedding large numbers of oocysts into the environment.

O objetivo do presente trabalho foi determinar a frequência sorológica e coproparasitológica da toxoplasmose em gatos atendidos em clínicas veterinárias na região metropolitana de Lima, Peru. Foram analisadas 154 amostras de soros de gatos e 50 amostras de fezes de gato, independentemente da idade, gênero ou raça. Paralelamente ao ato da coleta, os proprietários responderam a um questionário epidemiológico onde foram tratadas as seguintes variáveis: faixa etária, gênero, estilo de vida (animais confinados, semiconfinados o de vida livre), hábitos alimentares e de caça. Os soros e amostras fecais foram analisados pelos testes de hemaglutinação indireta (HAI) e coproparasitológico, respectivamente. A frequência de gatos expostos foi 11,0%, segundo HAI. As variáveis de faixa etária e sexo não mostraram associação com a exposição ao parasito. A exposição dos animais mostrou associação com hábitos de caçar (x2 = 4.98, p = 0.016) e alimentação (x2 = 13.34, p = 0.001), sendo aqueles alimentados com carne crua os mais expostos, quando comparados aos alimentados com ração (x2 = 9.50, p = 0.004) ou com comida caseira (x2 = 4.1, p = 0.027). A frequência de gatos na fase crônica da infecção por T. gondii foi 88% (15/17). Não foram achados oocistos de Toxoplasma gondiiem nenhuma das 50 amostras de fezes.

The aim of this study was to determine the frequency of infection by T. gondii in cats, by examining serum and fecal samples from animals attended at veterinary clinics in the Metropolitan Region of Lima, Peru. We collected and analyzed 154 cat serum samples and 50 fecal samples, regardless of the age, gender or breed. In parallel with the sample collections, the owners answered an epidemiological questionnaire that investigated the following variables: age group, gender, lifestyle (confined, semi-confined or free-living animals), feeding and hunting habits. The serum and fecal samples were analyzed using indirect hemagglutination (IHA) and coproparasitological tests, respectively. IHA showed that the frequency of cat exposure to T. gondii was 11%. Age and gender showed no association with exposure to the parasite. Exposure among the cats was associated with hunting (x2 = 4.98, p = 0.016) and feeding habits (x2 = 13.34, p = 0.001): those fed with raw meat were more exposed than those fed with commercial cat food (x2 = 9.50, p = 0.004) or with homemade food (x2 = 4.1, p = 0.027). The frequency of cats diagnosed in the chronic phase of T. gondii infection was 88% (15/17). No T. gondii oocysts were found in any of the 50 fecal samples examined.

A espécie Dirofilaria immitis parasita principalmente canídeos, podendo infectar também gatos embora raramente (Felis catus Linnaeus, 1758). Seus hospedeiros intermediários podem ser diferentes espécies de culicídeos. Motivados por um caso autóctone de dirofilariose felina, encontrado na região do Engenho Novo - RJ (S 22º 90' e WO 43º 27'), estudou-se: 1) a fauna culicídica local e; 2) a ocorrência de cães portadores de microfilárias. Os estudos sobre a fauna culicídica foram conduzidos, de abril de 2003 a maio de 2004, realizando-se capturas peridomiciliares e intradomiciliares. Foram capturados 672 indivíduos (fêmeas) das espécies Aedes aegypti (Linnaeus) (561/672 - 83,5%), Culex quinquefasciatus Say (96/672 - 14,3%), Aedes (Oc.) scapularis (Rondani) (12/672 - 1,8%) e Aedes albopictus (Skuse) (3/672 - 0,4%). Apenas as espécies A. aegypti e C. quinquefasciatus foram capturadas em todos os meses do ano. Foram coletadas 235 amostras de sangue canino, durante a Campanha de Vacinação "Rio Sem Raiva", em outubro de 2003. Nenhuma amostra examinada continha microfilárias (técnicas de Knott e NAN), apesar de apenas 3,4% dos cães receberem quimioprofilaxia. A presença de culicídeos vetores, associada à possibilidade de mobilidade dos cães, pode ter favorecido o aparecimento da infecção felina na região.

Dirofilaria immitis preferably infects canids and can, in lower frequency, be found in cats (Felis catus Linnaeus, 1758). The parasite may be transmitted by various Culicidae species. Motivated by an autochtone feline heartworm case diagnosed in the region of Engenho Novo - RJ (S 22º 90' e WO 43º 27'), the following issues were surveyed: 1) the local Culicidae fauna and; 2) the ocurrence of microfilaremic dogs. The mosquito study was conducted from April 2003 through May 2004, by peri-domiciliary and intra-domiciliary captures. A total of 672 mosquitoes (female) of the following species were caught: Aedes aegypti (Linnaeus) (561/672 - 83.5%); Culex quinquefasciatus Say (96/672 - 14.3%); Aedes (Oc.) scapularis (Rondani) (12/672 - 1.8%) and Aedes albopictus (Skuse) (3/672 - 0.4%). Only A. aegypti and C. quinquefasciatus specimens were captured year-round. A total of 235 canine blood samples were collected during the Campanha de Vacinação "Rio Sem Raiva" in October 2003. No microfilaria could be found in any of the examined samples (Knott´s and NAN techniques), even though only 3.4% of the dogs received chemoprophylaxis and the majority of them frequently traveled to endemic areas. The presence of vectors associated to a possible dog mobility, may have provided the right scenario for the feline D. immitis infection to happen in the studied area.

O diagnóstico coproparasitológico é o recurso laboratorial mais utilizado para detecção de infecções parasitárias gastrintestinais. A técnica de centrífugo-flutuação com sulfato de zinco (FAUST et al., 1938) é técnica de eleição no diagnóstico de estruturas parasitárias leves, podendo também ser usada para detecção de estruturas pesadas. Para avaliar o desempenho dessa técnica no diagnóstico das helmintoses gastrintestinais de gatos domésticos, foram realizados exames coproparasitológicos de 13 gatos domésticos 15 dias antes de suas mortes. À necropsia, os helmintos adultos encontrados no tubo digestivo e fígado foram contados, fixados e identificados. A técnica utilizada mostrou-se capaz de recuperar ovos de todos os nematóides e trematódeos encontrados, mas não foi adequada para a recuperação de cestóides.

Fecal examinations are the most commonly used laboratory tool for diagnosis of gastrointestinal nematode parasitic infections. Zinc centrifugal floatation technique (FAUST et al., 1938) is not only considered to be efficient in detecting light parasitic structures, but it is also used for recovery of heavy structures. In order to evaluate the performance of that technique in the diagnosis of domestic cats' gastrointestinal helminthiasis, fecal examinations of 13 animals were carried out 15 days prior to the animals' deaths. At necropsy, adult gastrointestinal parasites found were collected, counted, fixed and identified. The studied technique was able to recover eggs from all detected nematodes and trematodes, but appeared inappropriate for recovery of cestode eggs.

As condições sanitárias e composição populacional de uma colônia de gatos urbanos, errantes, habitantes do zoológico do Rio de Janeiro foram estudadas, objetivando-se um programa de controle populacional e sanitário. Capturou-se o maior número de indivíduos possível durante dois meses (47 animais). Os animais capturados foram examinados quanto ao gênero, idade, peso, pelagem, inspeção geral e presença de ectoparasitas e eram encaminhados a um programa de esterilização cirúrgica. Cada animal teve uma amostra de sangue colhida para realização de hemograma completo, plaquetometria, pesquisa de hemoparasitas e de retrovírus. As marcações e cores de pelagem encontradas foram "tabby" (70%), preta (17%); bicolor (11%) e arlequim (2%). A presença de pulgas foi observada em 28% dos animais. Os hemoparasitas encontrados foram Haemobartonella felis (38%) e piroplasmas indistinguíveis entre Cytauxzoon spp. e Babesia spp. (47%). Nenhum dos gatos foi encontrado com antigenemia de Dirofilaria immitis ou do vírus da leucemia felina (FeLV), embora anticorpos contra o vírus da imunodeficiência felina (FIV) tenham sido detectados (21%). Não houve correlação entre infecção por FIV e hemoparasitas. A população total estimada (método de captura-recaptura) foi de 59 gatos, sendo 68% fêmeas e 32% machos. Os resultados sugerem que um programa de esterilização cirúrgica é de fato necessário.

The colony of urban stray cats living in the Rio de Janeiro zoological garden was studied in order to develop a population and health control program. As many cats as possible were captured during two months (47 animals) and were classified according to gender, age, weight and coat markings. They were submitted to a general health evaluation, examined for the presence of ectoparasites and sent to a surgical neutering program. All animals had a blood sample drawn for CBC, platelet count, heartworm and retroviruses detection. Capillary blood smears were made for hemoparasites detection. Coat marking and colors were tabby (59.7%), followed by solid black (17%); torbie (10.6%); bicolor (10.6%) and harlequin (2.1%). The only ectoparasites found were fleas, which infested 28% of the animals. The hemoparasites found were Haemobartonella felis (38%) and piroplasmas that could not be differentiated between Cytauxzoon spp. and Babesia spp. (47%). No cat was found infected by Dirofilaria immitis or FeLV (Feline Leukemia Virus), although FIV (Feline Immunodeficiency Virus) antibodies could be detected (21%). There was no correlation between hemoparasites and FIV infections. The estimated total cat population (mark-recapture method) was 59; 68% female and 32% male, suggesting that a neutering program is in fact needed.

The identification of arthropod bloodmeals is important in many epidemiological studies, as, the understanding of the life cycle of vectors and the patogens they transmit, as well as helping to define arthropods' control strategies. The precipitin test has been used for decades, but ELISA is slowly becoming more popular. To compare the two tests for sensitivity, specificity and accuracy to detect small insect bloodmeals, Aedes aegypti or Ae. fluviatilis mosquitoes were fed either on feline, canine or human hosts. Mosquitoes were frozen at 6, 12, 24, 48 or 72 h after feeding. Precipitin test showed better specificity and accuracy and ELISA test showed higher sensitivity. Better results with both tests were achieved when mosquitoes were frozen within 48 h from feeding.

Dirofilaria immitis (Leidy 1856), a nematode parasite, is the etiologic agent of canine heartworm disease and mosquitoes are essential intermediate hosts. Mosquito susceptibility to the worms differ with species, strains and also among individuals of the same strain. To evaluate the degree of susceptibility of Rio de Janeiro laboratory raised strain of Aedes aegypti, we fed mosquitoes on canine blood with different densities of microfilariae (mf). There was no significant difference in the rate of development among the three different densities of mf. Infective larvae were found in the head and proboscis of all mosquitoes provided bloodmeals with different densities of mf after the 11th day post-infection. The infection rate of mosquitoes after ingestion of blood containing 3,000 mf/ml, 5,000 mf/ml and 7,000 mf/ml were 55.3%, 66.7% and 100%, respectively. The vector efficiency indices ranged from 1.6 to 9.3. The finding of L3 stage larvae, high infection rates and vector efficiency indices suggest that Ae. aegypti, Rio de Janeiro laboratory strain, is a potential vector of D. immitis, although of low efficiency.

Amostras fecais de cães com gastrenterite, até 6 meses de idade, foram testadas para a presença do parvovírus canino (CPV-2) pela reação de hemaglutinação (HA) e confirmadas pela reação de inibição da hemaglutinação (HI). Quarenta das 79 amostras, recebidas no período de abril de 1995 a junho de 1997, foram consideradas positivas. Aproximadamente 70% destas amostras foram obtidas de animais entre 2 e 4 meses de idade, época em que o risco de contraírem a infecção pelo CPV-2 é alto apesar da vacinação. Nenhuma variação sazonal da infecção pelo parvovírus canino pôde ser observada, e um estudo retrospectivo realizado na PolVet - UFF mostrou que em um período de 6 anos (1991-97), casos de gastrenterite ocorreram durante todos os anos em Niterói, sem uma sazonalidade definida.

Fecal samples from puppies with gastroenteritis less than 7 months old were examined for canine parvovirus infection (CPV-2) by hemagglutination (HA) and subsequent hemagglutination-inhibition (HI) tests. Forty of the 79 samples collected from April 1995 to June 1997 were found to be positive. About 70% of these samples were from 2 to 4 months old puppies, age in which they are at increased risk of developing CPV-2 infection, despite of vaccination. No seasonal distribution of canine parvovirus cases was found and it was supported by the results of a retrospective study realized at PolVet-UFF, which showed that gastroenteritis cases occurred throughout the year, for a six-year period (1991-97) in Niterói, Rio de Janeiro.

Six hundred and eleven random-source dogs (338 male, 273 female) one year of age or older, from six sections of the city of Recife, Pernambuco, were examined antemortem for circulating microfilariae Dirofilaria immitis and Dipetalonema reconditum adult heartworm (D. immitis) antigen, and examined postmortem for adult heartworms. The prevalence of heartworm infection was 2.3% (14/611), as determined by necropsy for adult worms, and 1% (6/611) had circulating microfilariae of D. immitis; thus, 57.1% of the heartworm-infected dogs had occult infections. The results of serological testing indicated that 1.3% (8/611) of the dogs were positive for adult heartworm antigen. A total of 42 (6.9%) of the dogs had microfilariae of D. reconditum; 40 of these had only D. reconditum and two additional dogs had microfilariae of both species, D. immitis and D. reconditum.

Dirofilaria immitis is a widespread mosquito-borne parasite that causes dirofilariasis, a commonly diagnosed disease of dogs that is rarely reported in cats and humans. A mosquito survey was conducted in Itacoatiara in the State of Rio de Janeiro, from March 1995 to February 1996, using canine, feline and human baits. A total of 3,667 mosquitoes were dissected for D. immitis larvae, representing 19 species and 10 genera. From those, Ae. scapularis, Ae taeniorhynchus, Cx. quinquefasciatus, Cx. declarator, Cx. saltanensis and Wy. bourrouli were found infected with D. immitis parasites, and among those, only the first three harbored infective larvae. The majority of larvae were found in the Malpighian tubules (889/936), and larval melanization was observed in the two Aedes species. In descending order, the best vectors were Ae. scapularis, Ae. taeniorhynchus, and Cx. quinquefasciatus which alternate seasonally in importance. Cx. quinquefasciatus is suggested to be a vector to cats. The potential transmission of D. immitis parasites by these three vectors to man is discussed.