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.

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.

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.

Endoparasites of household and shelter cats in the city of Rio de Janeiro, Brazil.

Endoparasitic infections are associated with morbidity in cats. This study aimed to investigate the occurrence of endoparasites among cats of different life stages in the city of Rio de Janeiro, Brazil. The samples were analyzed individually by macroscopic exploration for proglottids and centrifugal-flotation. Stool samples were obtained from household cats (n = 57) and shelter cats (n = 336). Endoparasites were detected in 50.64% of the samples. Among household and shelter cats, 21.05% and 55.66% were infected with endoparasites, respectively. In household cats, the most prevalent endoparasites were Ancylostoma spp. (in 25.0%) and Strongyloides spp. (in 25.0%), followed by Toxocara spp. (in 16.67%), Dipylidium caninum (in 16.67%), Cystoisospora spp. (in 8.33%), and Uncinaria spp. + Ancylostoma spp. (in 8.33%). In shelter cats, the most prevalent endoparasite was Ancylostoma spp. (in 29.41%), followed by Cystoisospora spp. (in 26.20%) and Toxocara spp. (in 16.58%), as well as Cystoisospora spp. + Toxocara spp. (in 8.02%); Ancylostoma spp. + Toxocara spp. (in 11.76%); Cystoisospora spp. + Ancylostoma spp. (in 3.74%); Cystoisospora spp. + Toxocara spp. + Ancylostoma spp. (in 3.21%); and Dipylidium caninum + Ancylostoma spp. (in 0.53%). Endoparasitic infections in cats underscore the need for preventive veterinary care and routine coproparasitologic tests.

Investigation of Mycoplasma spp. in birds of the Rio de Janeiro Zoo by isolation and PCR / Investigação de Mycoplasma spp. em aves do Zoológico do Rio de Janeiro por isolamento e PCR

Brazil is one of the countries with the most abundant avifauna in the world. The confinement of birds associated with close contact with other animals and humans favor the spread of agents of respiratory diseases. Among them, mycoplasmas can cause asymptomatic or apparent disease that manifests in birds by coughing, sneezing, rales, conjunctivitis, ocular and nasal discharge. Several described mycoplasmas cause disease in birds, especially Mycoplasma gallisepticum(MG) andMycoplasma synoviae(MS). The diagnosis ofMycoplasmaspp. can be done by clinical observation and laboratory analysis. Molecular diagnosis by PCR was boosted by its speed, sensitivity, and low cost of agent isolation techniques that take up to 21 days to complete. This study aimed to verify the occurrence ofMycoplasmaspp. in birds of the Rio de Janeiro Zoo (Rio Zoo), by isolation and PCR. Of the total 635 birds from the Rio Zoo, 81 were studied for detection ofMycoplasmaspp., when taken for routine health assessment exams. These birds belonged to the following orders: Psittaciformes (45), Accipitriformes (18), Galliformes (7), Piciformes (5), Strigiformes (4), Falconiformes (1) and Cariamiformes (1), all individuals already identified by microchip or leg-ring. There was no isolation of mycoplasmas in any of the samples tested, whereas, in the PCR, 62.96% (51/81) were positive, with 1.96% (1/51) identified as MG and 19.61% (10/51) as MS, representing 1.23% (1/81) and 12.34% (10/81) of the total population studied. PCR was shown to be a more effective technique than isolation in the detection ofMycoplasmaspp. in birds. It was possible to detect mycoplasmas in birds from Riozoo with no clinical respiratory signs, with higher MS prevalence than MG. The positivities forMycoplasmaspp., MS, and MG were different among the orders studied, being the highest occurrence in birds of prey, followed by Galliformes and Piciformes. The presence of MG and MS in birds of Rio de Janeiro Zoo confirms the circulation of these agents and the need for further studies on the dissemination of mycoplasmas in zoos for the epidemiological analysis of these bacteria in these places.(AU)

O Brasil é um dos países com maior avifauna do mundo. O confinamento de aves associado ao contato próximo a outros animais e seres humanos favorece a disseminação de agentes etiológicos causadores de doenças respiratórias. Dentre eles, os micoplasmas podem causar doença assintomática ou aparente que se manifesta em aves por espirros, estertores, conjuntivite, corrimentos oculares e nasais. São diversos os micoplasmas descritos causadores de doença em aves, com destaque para Mycoplasma gallisepticum (MG) e Mycoplasma synoviae (MS). O diagnóstico de Mycoplasma spp. pode ser feito pela observação clínica e análises laboratoriais. O diagnóstico molecular pela Reação em Cadeia da Polimerase (PCR) ganhou impulso por sua rapidez, sensibilidade e baixo custo em relação às técnicas de isolamento do agente que levam até 21 dias para conclusão do gênero Mycoplasma. Objetivou-se verificar a ocorrência da infecção por Mycoplasma spp. em aves no Zoológico do Rio de Janeiro (Rio Zoo), por isolamento e PCR. Do plantel de 635 aves do Rio Zoo, foram estudadas 81 para detecção de Mycoplasma spp., quando contidas para exames rotineiros de avaliação da condição de saúde. Essas aves eram pertencentes às ordens Psittaciformes (45), Accipitriformes (18), Galliformes (7), Piciformes (5), Strigiformes (4), Falconiformes (1) e Cariamiformes (1), todas já identificadas por microchip ou por anilha. Não houve isolamento de micoplasmas em nenhuma das amostras testadas, enquanto na PCR, 62,96% (51/81) foram positivas, sendo 1,96% (1/51) identificadas como MG e 19,61% (10/51) como MS, representando 1,23% (1/81) e 12,34% (10/81) da população total estudada. A PCR demonstrou ser uma técnica mais efetiva que o isolamento na detecção de Mycoplasma spp. em aves. Foi possível detectar micoplasmas nas aves do Riozoo sem sinal clínico respiratório, tendo MS maior prevalência do que MG. As positividades para Mycoplasma spp., MG e MS foram diferentes entre as ordens de aves estudadas, sendo a maior ocorrência nas aves de rapina, seguida dos Galliformes e dos Piciformes. A presença de MG e MS nas aves do Rio de Janeiro Zoo confirma a circulação destes agentes e a necessidade de mais estudos sobre a disseminação de micoplasmas em zoológicos para análise epidemiológica dessas bactérias nesse local.(AU)

Endoparasites of household and shelter cats in the city of Rio de Janeiro, Brazil / Endoparasitos de gatos domiciliados e de abrigos da cidade do Rio de Janeiro, Brasil

Abstract Endoparasitic infections are associated with morbidity in cats. This study aimed to investigate the occurrence of endoparasites among cats of different life stages in the city of Rio de Janeiro, Brazil. The samples were analyzed individually by macroscopic exploration for proglottids and centrifugal-flotation. Stool samples were obtained from household cats (n = 57) and shelter cats (n = 336). Endoparasites were detected in 50.64% of the samples. Among household and shelter cats, 21.05% and 55.66% were infected with endoparasites, respectively. In household cats, the most prevalent endoparasites were Ancylostoma spp. (in 25.0%) and Strongyloides spp. (in 25.0%), followed by Toxocara spp. (in 16.67%), Dipylidium caninum (in 16.67%), Cystoisospora spp. (in 8.33%), and Uncinaria spp. + Ancylostoma spp. (in 8.33%). In shelter cats, the most prevalent endoparasite was Ancylostoma spp. (in 29.41%), followed by Cystoisospora spp. (in 26.20%) and Toxocara spp. (in 16.58%), as well as Cystoisospora spp. + Toxocara spp. (in 8.02%); Ancylostoma spp. + Toxocara spp. (in 11.76%); Cystoisospora spp. + Ancylostoma spp. (in 3.74%); Cystoisospora spp. + Toxocara spp. + Ancylostoma spp. (in 3.21%); and Dipylidium caninum + Ancylostoma spp. (in 0.53%). Endoparasitic infections in cats underscore the need for preventive veterinary care and routine coproparasitologic tests.

Resumo Endoparasitoses estão associadas à morbidade em gatos. Este estudo objetivou investigar a ocorrência de endoparasitos em gatos de diversas faixas etárias, na cidade do Rio de Janeiro. As amostras passaram por exploração macroscópica à procura de proglotes de cestódeos e analisadas individualmente por centrífugo-flutuação. Assim, amostras fecais foram obtidas de gatos domiciliados (n = 57) e de abrigos (n = 336). Endoparasitos foram detectados em 50,64% das amostras fecais. Nos gatos domiciliados e de abrigos, 21,05% e 55,66% estavam infectados por endoparasitos, respectivamente. Ancylostoma spp. (25%) e Strongyloides spp. (25%) foram os helmintos mais prevalentes encontrados nas amostras de fezes dos gatos domiciliados, seguido por Toxocara spp. (16,67%), Dypilidium caninum (16,67%), Cystoisospora spp. (8,33%) e a associação de Uncinaria spp. e Ancylostoma spp. (8,33%). Entre os gatos dos abrigos, Ancylostoma spp. estavam presentes em 29,41% das amostras, seguidos por Cystoisospora spp. (26,20%), Toxocara spp. (16,58%) e as associações de parasitos Cystoisospora spp. + Toxocara spp. (8,02%), Ancylostoma spp. + Toxocara spp. (11,76%), Cystoisospora spp. + Ancylostoma spp (3,74%), Cystoisospora spp. + Toxocaraspp. + Ancylostoma spp. (3,21%) e Dipylidium caninum + Ancylostoma spp. (0,53%). Cystoisospora spp. foi o único protozoário encontrado. A presença de endoparasitos reforça a necessidade de cuidados veterinários preventivos e testes coproparasitológicos de rotina para evitar sua disseminação.

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.

Pentatrichomonas hominis infection in two domestic cats with chronic diarrhea.

CASE SERIES SUMMARY: Chronic diarrhea is a common and recurring problem in feline medicine. Intestinal trichomonads have been reported as causative agents of diarrhea in cats. While Pentatrichomonas hominis is considered commensal, Tritrichomonas foetus has been found to cause feline large bowel diarrhea in cats. In our report, two young cats infected with the feline leukemia virus (FeLV) and presenting with chronic diarrhea were identified as having trichomonads in their feces, based on direct examination and fecal culture. Molecular assays (PCR and DNA sequencing) revealed that the parasite involved was P hominis, not T foetus, as was suspected. The animals had already been subjected to ineffective therapy with metronidazole, and after the use of ronidazole, their feces became dry and formed. RELEVANCE AND NOVEL INFORMATION: This case report describes P hominis infection as a possible cause of chronic diarrhea in two young cats also infected by FeLV. The parasite was probably resistant to metronidazole, the drug of choice in the literature, and sensitive to ronidazole. Although considered commensal, P hominis infection should be evaluated as a differential diagnosis in cats with chronic diarrhea, especially those that are immunocompromised. Moreover, a combination of techniques such as direct examination and/or fecal culture and PCR is essential for an accurate diagnosis of P hominis infection.

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.

Update on Dirofilaria immitis epidemiology in South America and Mexico: literature review / Atualização sobre a epidemiologia de Dirofilaria immitis na América do Sul e no México: revisão de literatura

Dirofilaria immitis (Leidy, 1856; Raillet & Henry 1911) is a parasite that is widely disseminated around the globe, with a higher prevalence in warm, humid climates. The first report of its occurrence in South America is from 1878 in Brazil. At that time, reports were scarce and difficult to retrieve ­ therefore, gathering them will facilitate record-keeping over time. Four databases were searched (Scopus, MEDLINE, LILACS, and PubMed) and the search keywords were "Dirofilaria" or "heartworm" and the countries' names. Four countries lacked reports (Bolivia, Ecuador, French Guiana, and Uruguay) and other three (Suriname, Guyana, and Paraguay) had only old reports. Chile was the only country in which studies were conducted over time, and no infected dogs were registered. For the other six countries (Mexico, Peru, Colombia, Venezuela, Argentina, and Brazil), reports showed that the infection frequency varied over time and with the surveyed area. Therefore, the information indicates that D. immitis is established, and veterinarians must institute preventive programs to optimally care for their patients and protect the health of their families.(AU)

Dirofilaria immitis é um nematoide de ampla distribuição geográfica, que ocorre com maior frequência em áreas quentes e úmidas do planeta. O primeiro registro de sua ocorrência na América do Sul foi realizado em 1878, no Brasil. Naquela época os registros eram poucos e raramente de fácil obtenção, razão pela qual reuni-los facilitará a recuperação da memória ao longo dos anos. Quatro bases de dados (Scopus, MEDLINE, LILACS e PubMed) foram estudadas utilizando-se as palavras-chave "Dirofilaria" ou "heartworm", os nomes dos países da América do Sul e o México. Nenhum registro foi encontrado para quatro países (Bolívia, Equador, Guiana Francesa e Uruguai) e para outros três (Suriname, Guiana e Paraguai) os registros eram antigos. Apenas o Chile é o território onde houve estudos registrados com ausência do parasita. Os outros países (México, Peru, Colômbia, Venezuela, Argentina e Brasil) apresentam registros com frequência variável no tempo ou no espaço. Assim, as informações reunidas indicam que infecções por D. immitis ocorrem na maior parte da América do Sul e no México e que os médicos veterinários devem instituir programas preventivos para garantir cuidados médicos de qualidade aos pacientes e para proteger a saúde destes e de suas famílias.(AU)

Selamectin for the prevention of canine Dirofilaria immitis infection: field efficacy in client-owned dogs in a high risk area.

BACKGROUND: Dog owners and veterinarians in small animal practices began to waive prevention of canine heartworm disease after heartworm infections seemed to have disappeared in Brazil. After 2013, infection rates rebounded, and an evaluation of the efficacy of chemoprophylactic drugs became necessary. Included in this re-evaluation was the efficacy of selamectin in client-owned dogs residing in a high infection-risk area. METHODS: The preventive efficacy of selamectin was evaluated by the topical application of selamectin to 24 client-owned dogs at the recommended rate (minimum of 6 mg/kg) by a veterinarian monthly for 36 months. Blood samples were collected before the first treatment and at the end of the study for testing to detect microfilariae by the modified Knott's test and Dirofilaria immitis antigens using a commercial antigen test. Exposure to risk of heartworm infection was confirmed by the presence of infection in dogs living in low-income communities within a 2 km radius from the homes of dogs in the study. The dogs were managed according to routine practice by the owners within each household throughout the study. RESULTS: All dogs tested negative by both tests after receiving topical treatment with selamectin monthly for 36 months. Testing of 204 dogs from the communities confirmed the presence of heartworm in the area by detection of microfilariae or D. immitis antigen in 44 dogs (21.6 %). CONCLUSIONS: Topical selamectin was 100 % effective for D. immitis prevention in 24 dogs that received monthly treatments by a veterinarian. Detection of heartworm infections in untreated dogs in the area suggests that clients need to be better informed regarding the prevalence of D. immitis and the importance of maintaining regular preventive treatments.

Updated canine infection rates for Dirofilaria immitis in areas of Brazil previously identified as having a high incidence of heartworm-infected dogs.

BACKGROUND: Canine heartworm infections were frequently diagnosed in Brazil before the new millennium. After the year 2000, the frequency of diagnosis showed a sharp decline; however, a few years later, new evidence indicated that the parasite was still present and that canine infection rates seemed to be increasing. Therefore, an updated survey of canine heartworm prevalence was conducted in several locations in south, southeast, and northeast Brazil. METHODS: Dogs from 15 locations having previously reported a high prevalence of heartworm infection were included in the survey according to defined criteria, including the absence of treatment with a macrocyclic lactone for at least 1 year. Blood samples from 1531 dogs were evaluated by an in-clinic immunochromatography test kit (Witness® Heartworm, Zoetis, USA) for detection of Dirofilaria immitis antigen. At each location, epidemiologic data, including physical characteristics and clinical signs reported by owners or observed by veterinarians, were recorded on prepared forms for tabulation of results by location, clinical signs, and physical characteristics. RESULTS: The overall prevalence of canine heartworm infection was 23.1%, with evidence of heartworm-infected dogs detected in all 15 locations studied. There was a tendency for higher prevalence rates in environmentally protected areas, despite some locations having less-than-ideal environmental temperatures for survival of vector mosquitoes. Among physical characteristics, it was noted that dogs with predominantly white hair coats and residing in areas with a high (≥20%) prevalence of heartworm were less likely to have heartworm infection detected by a commercial heartworm antigen test kit than were dogs with other coat colors. In general, dogs older than 2 years were more frequently positive for D. immitis antigen than were younger dogs. Clinical signs of heartworm infections were rare or owners were unable to detect them, and could not be used for reliable prediction of the presence of heartworm. CONCLUSIONS: These results indicate that the prevalence of D. immitis has increased in these areas of Brazil over the past few years. Small animal practitioners in these areas should include routine screening tests for heartworm infections in every dog's annual evaluation protocol and make sure to have uninfected dogs on prevention.

Seroprevalence of Toxoplasma gondii (Nicole & Manceaux, 1909) and retroviral status of client-owned pet cats (Felis catus, Linnaeus, 1758) in Rio de Janeiro, Brazil.

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.