Platynosomum illiciens infection in domestic cats: insights from a sanctuary.

Platynosomum illiciens, a trematode parasite known for its intricate life cycle, predominantly infests the liver, gallbladder, and bile ducts of domestic cats. In this study, we examined feline fecal samples from a cat sanctuary in Metropolitan Rio de Janeiro, Brazil, aiming to determine whether, even in such environments, cats retain their atavistic hunting habits, as evidenced by the presence of P. illiciens in fecal samples. The infection diagnosis utilized centrifugal sedimentation in the formalin-ethyl acetate test. Out of 72 fecal samples collected from various cats, four tested positive for P. illiciens eggs. This study serves as a reminder that even in environments where they are well-fed, cats exhibit predatory behavior, engaging in hunting and consuming prey, thus exposing themselves to parasites. It emphasizes the importance of veterinarians and cat caretakers being cognizant of the widespread presence of P. illiciens in Brazil and considering it in the differential diagnosis for cats presenting with liver or gallbladder issues. In conclusion, our findings underscore that the ancestral instinct for hunting and predation, preserved in domestic cats, persists despite fulfilling their nutritional needs.

Platynosomum illiciens, um parasito trematódeo conhecido por seu ciclo de vida complexo, infesta predominantemente o fígado, a vesícula biliar e os ductos biliares de gatos domésticos. Neste estudo, examinamos amostras fecais de felinos de um santuário de gatos na região metropolitana do Rio de Janeiro, Brasil, com o objetivo de determinar se, mesmo nesses ambientes, os gatos mantêm seus hábitos de caça, evidenciado pela presença de P. illiciens em amostras fecais. O diagnóstico da infecção foi realizado por sedimentação centrífuga no teste de formalina-acetato de etila. Foram obtidas 72 amostras fecais de diferentes gatos e quatro testaram positivo para ovos de P. illiciens. Este estudo é um lembrete de que os gatos terão comportamento predatório, caçando e consumindo presas mesmo quando bem alimentados, ficando expostos ao parasito. Veterinários e cuidadores de gatos devem estar atentos à ampla presença de P. illiciens no Brasil e considerá-la no diagnóstico diferencial de gatos com problemas de fígado ou vesícula biliar. Concluindo, nossos resultados mostram que o instinto atávico dos gatos que se mantiveram conservados durante a domesticação não pode ser anulado quando as necessidades nutricionais dos gatos são satisfeitas.

Dogs infected by Dirofilaria immitis: a threat to the health of human and non-human animals in Rio de Janeiro, Brazil.

This study aimed to investigate the presence of Dirofilaria immitis microfilaremia in dogs from two regions of the state of Rio de Janeiro, Brazil, where heartworm infections are highly prevalent. Blood samples were collected from dogs aged > 12 months, independent of the use of preventatives. All samples obtained and analyzed using Knott's modified test by the investigators. A total of 133 blood samples were tested, and D. immitis microfilariae were detected in 29 of them, resulting in an occurrence of 21.8%. The percentage of dogs with microfilaremia detected raises concerns for pet families, one health professionals, and small animal practitioners. Microfilaremic dogs are the richest source of infection for the mosquitoes, increasing the risk of transmission. Therefore, the stakeholders in One Health must raise concerns regarding the health of wild animals, as wild canids and other species of wild animals are exposed to the risk of D. immitis infection. In addition, humans can get infected and develop human pulmonary dirofilariasis. In conclusion, the presence of dogs with microfilaremia potentiates opportunities for D. immitis transmission, exposing all animals, wild or domestic, human or non-human to the disease.

Este estudo objetivou investigar a presença de cães portadores de microfilemia da espécie Dirofilaria immitis em duas áreas do estado do Rio de Janeiro, Brasil onde a infecções por vermes do coração são altamente prevalescentes. Amostras de sangue foram obtidas de cães com mais de 12 meses de idade, independentemente do uso de preventivos. Todas as amostras foram obtidas e analisadas pelo grupo do estudo utilizando-se o teste de Knott modificado. Um total de 133 amostras foi testado e microfilárias de D. immitis foram detectadas em 29 amostras, resultando em uma ocorrencia de 21,8%. A porcentagem de cães com microfilaremia detectada gera preocupações para as famílias de animais de estimação, para profissionais da saúde única e para clínicos de pequenos animais. Cães microfilarêmicos são a mais rica fonte de infecção para mosquitos, aumentando o perigo de transmissão. Assim, atores de todos os segmentos sociais com interesse na saúde única devem incluir a saude de animais selvagens em suas preocupações, uma vez que canideos selvagens e outras espécies de animais selvagens são expostos ao desafio da infecção por D.immitis. Além disso, seres humanos podem se tornar infectados e desevolver dirofilariose pulmonar humana. Em conclusão, a presença de cães microfilarêmicos potencializa as oportunidades de trasmissão de D. immitis, expondo todos os animais, selvagens ou domesticos, humanos ou não humanos à doença.

Genetics of randomly bred cats support the cradle of cat domestication being in the Near East.

Cat domestication likely initiated as a symbiotic relationship between wildcats (Felis silvestris subspecies) and the peoples of developing agrarian societies in the Fertile Crescent. As humans transitioned from hunter-gatherers to farmers ~12,000 years ago, bold wildcats likely capitalized on increased prey density (i.e., rodents). Humans benefited from the cats' predation on these vermin. To refine the site(s) of cat domestication, over 1000 random-bred cats of primarily Eurasian descent were genotyped for single-nucleotide variants and short tandem repeats. The overall cat population structure suggested a single worldwide population with significant isolation by the distance of peripheral subpopulations. The cat population heterozygosity decreased as genetic distance from the proposed cat progenitor's (F.s. lybica) natural habitat increased. Domestic cat origins are focused in the eastern Mediterranean Basin, spreading to nearby islands, and southernly via the Levantine coast into the Nile Valley. Cat population diversity supports the migration patterns of humans and other symbiotic species.

How does Dirofilaria immitis infection impact the health of dogs referred to cardiology care.

Dirofilaria immitis is a nematode that can cause a disease that may present clinical signs from severe to absent. When dogs are symptomatic, the clinical signs are cardiorespiratory and nonspecific, which may be misleading. This study aimed to demonstrate the clinical presentations to cardiac care by evaluating 26 dogs subjected to clinical examination, complete blood count (CBC), specific tests for D. immitis infection, chest radiography, and echocardiography. Among them, 11 (42.3%) dogs were infected and 15 (57.7%) were non-infected. Most dogs presented with coughing (65.4%) and abnormal lung sounds (81%) independent of infection. Murmur at the tricuspid focus was present in 26.9% of the dogs, of which 57.1% were infected. Echocardiography revealed tricuspid regurgitation in 30.8% of the dogs and pulmonary regurgitation in 46.1%, of which 37.5% and 50% were infected, respectively. Worms were detected by echocardiography in 45.5% of the infected dogs. The x-rays showed that the bronchial pattern was present in 45.5% of the infected dogs and in 46.7% of the non-infected dogs. The interstitial pattern was present in 18.2% of the infected animals, in contrast to 6.7% of the non-infected dogs. The CBC results for all dogs were within the reference range, except for platelets. Although similar, the percentage of dogs with thrombocytopenia was higher among infected dogs (36.4%) than among the non-infected (6.7%). These results reinforce that due to the non-specific signs of infection, it is mandatory to perform parasitological assays when evaluating dogs presenting with cardiopulmonary signs.

Dirofilaria immitis, é um nematoide que pode causar sinais clínicos de graves a ausentes. Quando os cães são sintomáticos, os sinais clínicos são cardiorrespiratórios e inespecíficos, o que pode confundir com outras doenças. Com o objetivo de descrever a apresentação clínica de cães atendidos em um serviço de cardiologia e doenças respiratórias, após consentimento dos tutores, 26 pacientes foram submetidos a exame clínico, hemograma, exames específicos para infecção por D. immitis, radiografia de tórax e ecocardiografia. Entre esses cães, 11 estavam infectados (42,3%) e 15 não infectados (57,7%). A maioria dos cães apresentou tosse (65,4%) e sons pulmonares anormais (81%) independentemente da infecção. O sopro no foco tricúspide estava presente em 26,9% dos cães, sendo 57,1% infectados. A ecocardiografia mostrou insuficiência tricúspide em 30,8% dos cães e insuficiência pulmonar em 46,1%, sendo 37,5% e 50% infectados, respectivamente. Parasitos foram detectados pela ecocardiografia em 45,5% dos cães infectados. As radiografias mostraram que o padrão brônquico estava presente em 45,5% dos cães infectados e em 46,7% dos não infectados. O padrão intersticial esteve presente em 18,2% dos infectados e em 6,7% dos não infectados. Os resultados do hemograma de todos os cães estavam dentro do intervalo de referência para todas as células, exceto plaquetas. Embora semelhante, a porcentagem de cães com trombocitopenia foi maior entre os cães infectados (36,4%), do que entre os livres de infecção (6,7%). Esses resultados reforçam que, devido aos sinais inespecíficos da infecção, é obrigatória a realização de ensaios parasitológicos na avaliação de cães que apresentem sinais cardiopulmonares.

Clinical and blood count findings in dogs naturally infected with Dirofilaria immitis.

Dirofilaria immitis is a nematode that infects canids worldwide as well as other mammalian species, including humans. Worms and dogs are well adapted to one another, making dogs the best urban host for the parasite. Nevertheless, 30% of dogs do not sufficiently present microfilaremia, that is, the low larval load impairs transmission by mosquitoes and diagnosis by its detection in the blood samples. Therefore, the canine diagnosis must always include a microfilaria test and serological tests to detect adult worm antigens. To describe the clinical findings in naturally infected dogs in Rio de Janeiro, 34 dogs were included in the study. All dogs were evaluated for history, anamnesis, physical examination, complete blood count (CBC), D. immitis testing for antigens (ELISA test SNAP 4Dx Plus®), and microfilarial burden. The most frequent complaint from the owners was coughing (14.7%, 5/34). The most common CBC finding was eosinophilia (29.4%), followed by thrombocytopenia (26.5%) and neutrophilia (14.7%). Of the 34 animals, 91.2% were microfilaremic, with a mean count of 11.939 microfilaria/mL. Veterinarians working in areas endemic to D. immitis should always undergo screening tests and pulmonary auscultation, and increased expiratory sounds, even in the absence of coughing, can be considered a sign of the disease, along with eosinophilia, thrombocytopenia, and neutrophilia.

Dirofilaria immitis, é um nematoide que infecta canídeos em todo o mundo, bem como outras espécies de mamíferos, incluindo humanos. Os vermes e os cães estão bem adaptados um ao outro, tornando os cães o melhor hospedeiro urbano para o parasita. Contudo, 30% dos cães não apresentam microfilaremia, prejudicando a transmissão pelos mosquitos e o diagnóstico por detecção de larvas em amostras de sangue. Portanto, o diagnóstico canino deve incluir sempre a pesquisa de microfilárias e sorologia para detecção do antígeno do verme adulto. Com o objetivo de descrever os achados clínicos de cães naturalmente infectados no Rio de Janeiro, 34 cães foram incluídos. Todos os cães foram avaliados por histórico, anamnese, exame físico, hemograma completo (CBC), teste de antígenos, pesquisa e contagem de microfilárias de D. immitis. A queixa mais frequente dos responsáveis foi a tosse (14,7% - 5/34). O achado de hemograma mais comum foi eosinofilia (29,4%), seguido de trombocitopenia (26,5%) e neutrofilia (14,7%). Dos 34 animais, 91,2% eram microfilarêmicos com contagem média de 11.939 microfilárias/mL. Os veterinários que atuam em áreas endêmicas de D. immitis devem sempre realizar exames de triagem e ausculta pulmonar, pois mesmo na ausência de tosse, sons expiratórios aumentados podem ser considerados um sinal da doença, assim como eosinofilia, trombocitopenia e neutrofilia.

Feline Heartworm in Clinical Settings in a High Canine Prevalence Area.

Heartworm (HTW) infection in cats is associated with persistent pulmonary pathology, even when clinical signs are absent. Treatment options for cats are limited once infected, making prevention an important topic for discussion with cat owners. In Brazil, tests to detect feline HTW infections are unavailable, likely leading to an underestimation of its impact on the wellbeing of cats. The present study investigated the seroprevalence of HTW antigen in cats living in an area with high canine HTW prevalence and investigated risk factors and clinical signs associated with HTW disease in cats from Rio de Janeiro, Brazil. Clinical examinations were conducted, and serological evaluations performed with a point-of-care test (SNAP® Feline Triple® Test, IDEXX Laboratories, Inc.). A total of 586 cats were examined. The HTW antigen seroprevalence was 1.2%. Heartworm positive results were significantly associated with vomiting and abnormal lung sounds. Results from this study indicate that cats residing in areas of high canine HTW prevalence should have HTW disease as a differential diagnosis when presenting with compatible clinical signs. Veterinarians should prioritize client education and promote regular use of effective prophylaxis to protect feline health.

Molecular characterization of canine filarioids in a previously non-endemic area of Rio de Janeiro State, Brazil.

Dirofilaria immitis is the causative agent of canine heartworm disease, a severe health problem in dogs, especially in coastal areas of tropical and subtropical regions of the world. We employed molecular methods to investigate the occurrence of canine infection by filarioids in five municipalities of Baixada Fluminense (Magé, Duque de Caxias, Guapimirim, Nova Iguaçu, and São João de Meriti), a non-endemic area of Rio de Janeiro State, Southeast Brazil. A total of 110 canine blood samples collected from 2017 to 2018 and positive for microfilariae at the modified Knott's test were screened by cPCR targeting DNA fragments of the 12S rDNA gene for filarial nematodes. Seventy-seven samples (70%) tested positive at the molecular analysis. Of these, 72 were identified as D. immitis and 5 (4.5%) as Acanthocheilonema reconditum. Dirofilaria repens was not detected in the studied municipalities of Baixada Fluminense. This is the first record of D. immitis and A. reconditum in the Baixada Fluminense region, Rio de Janeiro State, Brazil. The prevalence of D. immitis cases in the five municipalities suggests the establishment and maintenance of its enzootic cycle in the studied region, which indicate vulnerability to the occurrence of epidemic cycles and, possibly, human cases.

Infection with Dirofilaria immitis and Other Infections in Cats and Dogs from Rio de Janeiro, Brazil: The Need for Prophylactic Enforcement.

PURPOSE: Dirofilaria immitis, a mosquito-borne nematode that primarily infects dogs, can equally infect cats. Although there have been numerous studies on canine heartworm prevalence in Brazil, there have been few studies on feline infections. Feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) are both life-threatening retroviruses transmitted directly between cats. Infections with Ehrlichia spp. and Anaplasma spp. are highly prevalent among dogs in Brazil, with Rhipicephalus sanguineus being the main vector for both bacteria. This study aimed to gather information on these infections among dogs and cats in the metropolitan area of Rio de Janeiro by performing rapid point-of-care tests for prophylactic enforcement. METHODS: Surplus samples of serum or plasma from private laboratories were tested by enzyme-linked immunosorbent assay (ELISA) (SNAP Feline Triple Test or SNAP 4Dx Plus Test). RESULTS: The prevalence of heartworm disease was 7% among dogs and 0.9% among cats, the latter being 12.9% of the former. The prevalence of FIV and FeLV was 4.3 and 11.9%, respectively. Among dogs, the seroprevalence of Ehrlichia spp. and Anaplasma spp. was 27.1 and 9.8%, respectively, and Borrelia burgdorferi was not detected. CONCLUSION: Given that such infections circulate among pets, prophylactic measures should be encouraged by small animal practitioners.

Efficacy of semi-annual therapy of an extended-release injectable moxidectin suspension and oral doxycycline in Dirofilaria immitis naturally infected dogs.

BACKGROUND: Dirofilaria immitis is a life-threatening nematode spreading globally. Arsenical treatment is currently recommended for removal of adult worms. However, arsenical treatment is not available in some countries, and there are dogs that cannot tolerate the rapid kill of adult worms; therefore, alternative adulticide slow-kill treatments are needed. Criticisms against the use of these alternative protocols include the potential for allowing disease to progress and for the development of ML-resistant worms. METHODS: The efficacy of a protocol that includes semi-annual doses (i.e. every 6 months) of commercially available extended-release injectable moxidectin suspension (ProHeart® SR-12) with 30-day oral administration of doxycycline was studied in 20 dogs with naturally occurring D. immitis infections. Each dog received treatment with ProHeart® SR-12 (0.5 mg moxidectin/kg) by subcutaneous injection and oral doxycycline (10 mg/kg/bid × 30 days) every 6 months until two consecutive negative antigen test results were obtained. Pulmonary and cardiac evaluations were performed by radiographic and echocardiographic parameters. Physical examinations, complete blood counts, clinical chemistry profiles, microfilariae and antigen tests were performed periodically. RESULTS: At enrollment, all dogs were positive for D. immitis antigen and 18 were microfilaremic. On day 30, microfilaremia counts decreased, and all dogs became amicrofilaremic by day 150. On day 180, 11 dogs were antigen-negative, and 7 more became negative by day 360. The two remaining antigen-positive dogs converted to negative by day 540 or 810. All antigen tests performed 180 days after the first negative test were negative. There was no decline in cardiac performance of the dogs throughout the study. Overall, pulmonary clinical conditions, presence of worms by echocardiography, and enlargement of caudal and main pulmonary arteries improved after treatment. Physical examinations, complete blood count results, and clinical chemistry profiles were within normal reference values. Respiratory conditions were improved, no damage to the heart was observed, and the treatment protocol was well tolerated by the animals. CONCLUSIONS: This alternative adulticide treatment was efficacious and well tolerated in naturally infected dogs. The injectable formulation provides the advantage of having veterinarians able to administer, monitor, and assess the efficacy and condition of the dog throughout the treatment and post-treatment periods.

Unusual feline Dirofilaria immitis infection: a case report.

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.

Detection of Dirofilaria immitis using microscopic, serological and molecular techniques among dogs in Cabo Frio, RJ, Brazil.

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.

Detection of Dirofilaria immitis using microscopic, serological and molecular techniques among dogs in Cabo Frio, RJ, Brazil / Detecção de Dirofilaria immitis utilizando técnicas microscópicas, imunológicas e moleculares em cães de Cabo Frio, RJ

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 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.

Mosquito abundance in a Dirofilaria immitis hotspot in the eastern state of Rio de Janeiro, Brazil.

Coastal lowlands in the state of Rio de Janeiro, Brazil, support high numbers of heartworm-infected dogs. Microfilariae of heartworm need to be ingested by a potencial mosquito vector in order to develop into infective larvae and infect a new host. Ochlerotatus taeniorhynchus and Ochlerotatus scapularis are the primary vector species in the coastal lowlands of metropolitan Rio de Janeiro; thus, the aim of this study was to investigate whether these two species were abundant enough at the heartworm hotspot in the eastern area of the state to be important to the local parasite's life cycle. The study was conducted at the Massambaba sandbank (22°55'45″S;42°18'51″W), where canine heartworm prevalence was 53.1%. Mosquitoes were captured monthly using two traps with CO2, as well as eventual human landing. A total of 6118 mosquitoes of 16 species were collected. Ochlerotatus taeniorhynchus (37.5%), Culex quinquefasciatus (19.3%), Oc. scapularis (18.2%), and Coquillettidia sp. (10.5%) were the most abundant species. The monthly variation in frequency was marked for Oc. scapularis and Cx. quinquefasciatus and steadier for Oc. taeniorhynchus and Coquillettidia sp. The abundance of the two Ochlerotatus species reaffirms that they are the primary natural vectors for D. immitis in the coastal Rio de Janeiro state lowlands.

Evaluation of DPP® and SNAP® Rapid Tests for diagnosis of Leishmania infantum canine infections.

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.

Use of Slow-Release Injectable Moxidectin for Treatment of Dirofilaria immitis Infection During Pregnancy.

Canine heartworm disease is a life-threatening disease caused by Dirofilaria immitis and is prevalent in Brazil. The standard drug for its treatment, melarsomine dihydrochloride, is a fast-killing organic arsenical chemotherapeutic agent not approved in Brazil. Therefore, an alternative strategy, such as macrocyclic lactone in combination with a tetracycline antibiotic, has to be used. The alternative method is a long-term therapy that could lead to compliance issues during treatment. The aim of this case report is to present a preliminary assessment on the efficacy and safety of an off-label biannual administration of slow-release moxidectin (0.5 mg/kg every 6 months), which is formulated for annual administration (0.5 mg/kg annually). This overdose was chosen to test if moxidectin serum levels could be maintained high enough to harm the worms. It was administered to a 4-year-old female dog in combination with a 30-day doxycycline course. The second dose of moxidectin was administered approximately a week before she gave birth to three healthy puppies. Microfilariae were not detected on day 180 of treatment. Serological tests showed that the worms were eliminated, as two negative antigen tests were obtained 6 months apart (at day 180 and day 360 of treatment). Therefore, the off-label biannual use of moxidectin in combination with doxycycline was effective in eliminating D. immitis in 360 days and was harmless for the pregnant dog and her offspring, suggesting that this strategy is promising. Although these results are encouraging, further studies are needed to confirm safety and efficacy issues.

Evaluation of DPP® and SNAP® Rapid Tests for diagnosis of Leishmania infantum canine infections

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.

Can P-glycoprotein and ß-tubulin polymorphisms be used as genetic markers of resistance in Dirofilaria immitis from Rio de Janeiro, Brazil?

OBJECTIVE: Dirofilaria immitis, the causative agent of canine heartworm infection, is worldwide the most important filarid to affect domestic dogs. Prevention of this infection is done by macrocyclic lactones, but some reports on the lack of efficacy have been published. Although the actual cause of resistance is unknown, single nucleotide polymorphisms (SNPs) on a P-glycoprotein ABC transporter and ß-tubulin genes have been pointed out as candidates for genetic markers of resistance. We conducted a survey to verify the presence of these suggested genetic markers in microfilariae from 30 naturally infected dogs under macrocyclic lactones treatment living in an endemic area in the state of Rio de Janeiro. RESULTS: The analysis of these specific SNPs demonstrated no sign of polymorphism on the P-glycoprotein loci, while 72 and 48% of the samples were polymorphic to the first and second SNPs on ß-tubulin loci, respectively. This work demonstrates that the P-glycoprotein position 11 and 618 were not polymorphic and, therefore, not suitable as a genetic marker of resistance in Rio de Janeiro whereas both ß-tubulin loci were polimorphic. This work points out the difficulty of finding a universal genetic marker for resistance.

Heat pretreatment of canine samples to evaluate efficacy of imidacloprid + moxidectin and doxycycline in heartworm treatment.

BACKGROUND: Considering the recent information on the increase of Dirofilaria immitis antigen detection by rapid assays in canine blood samples after heat treatment, the proposal that immune complexes block D. immitis antigen detection and that macrocyclic lactone + doxycycline (alternative protocol) might lead to increased production of those immune complexes, resulting in the erroneous diagnosis of adult worm elimination, and that there is no recommended adulticide marketed in Brazil, a study was performed to evaluate the interference of moxidectin + doxycycline (moxi-doxy) on diagnostic procedures when heartworm positive dogs are treated with this alternative protocol. Twenty-two naturally infected pet dogs were treated monthly with topical 10% imidacloprid + 2.5% moxidectin and with oral doxycycline (10 mg/kg BID/30 days) (moxi-doxy). All the dogs had their microfilaremia level determined prior to the first day of treatment, and were tested every 6 months for microfilariae (Mf) detection prior to heating, and for antigen detection prior to and after heating, the sample. RESULTS: The results indicate that the treatment protocol can eliminate adult heartworms as early as 6 months after the first dose, especially in low microfilaremic dogs (< 300 Mf/ml). In this study, all dogs were free of heartworm antigen after 18-24 months of treatment. In a comparison of pre-heated samples and non-heated samples, sample pre-heating increased antigen detection sensitivity, and non-heated samples tended to be antigen-negative earlier than the pre-heated samples, especially when dogs had low microfilaremia levels. These discrepancies were not present in a subsequent sample of the same dog 6 months later. CONCLUSIONS: Two negative antigen test results 6 months apart can be recommended as the criterion to consider when a dog has been cleared of infection. The initial microfilaremia level of a dog can be used to estimate the necessary time frame to end the treatment period.

Effect of a synthetic analogue of the feline facial pheromone on salivary cortisol levels in the domestic cat / Efeito do análogo sintético do feromônio facial felino sobre o nível de cortisol salivar de gatos domésticos

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.(AU)

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.(AU)

Serological evidence of canine exposure to arthropod-borne pathogens in different landscapes in Rio de Janeiro, Brazil.

Arthropod-borne infections are dependent on environmental conditions, and several combinations of natural and human-related variables play an important role in vector populations as well as the life cycle of agents carried by the arthropods. The top 5 canine arthropod-transmitted agents, Dirofilaria immitis, Leishmania spp., Ehrlichia canis, Anaplasma phagocytophilum, and Borrelia burgdorferi infect unprotected animals without propensity. The purpose of this study was to determine the prevalence of these parasite species in three different landscape settings (sandbanks, plains and mountains) along a 60-km line. During a 6-month period, blood samples were collected from dogs (>12months old) living in the different settings. Prevalence of D. immitis was determined by modified Knott test and ELISA. Prevalence of E. canis, A. phagocytophilum, and B. burgdorferi was determined by ELISA, and Leishmania spp. by ELISA, indirect immunofluorescence, and immunocromatographic assays. D. immitis was most prevalent in the sandbank (68.9%) as well as Leishmania spp. (34.5%), and tick-transmitted agents, A. phagocytophilum and E. canis in the plains (61.7%). B. burgdorferi was not detected. Depending on the resources for arthropods present in regions, dogs are likely to be exposed to different arthropod-borne parasites and should receive preventives tailored to the risk of infection in the region in which the dog resides.