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.

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.

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.

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.

Chemoprophylaxis of Dirofilaria immitis (Leidy 1856) infection at a high challenge environment.

BACKGROUND: The frequency of canine heartworm infection in the state of Rio de Janeiro, Brazil was high before chemoprophylactic treatment was available, with one of the highest rates of infection (52.5 %) found among dogs living on the eastern shore of the state. Following the launch of a chemoprophylactic product, the rate of infection gradually decreased, and new infections were rarely reported. After 2005, outbreaks reported at the eastern shore as well as for new infections in other areas of high infection frequency were considered to possibly be related to reduced efficacy of macrocyclic lactones. Therefore, this study aimed to evaluate the efficacy of topical heartworm preventatives from different drug families at the high challenge area of the state of Rio de Janeiro. METHODS: A total of 46 dogs, including animals negative for Dirofilaria immitis microfilariae and antigen (Snap 4 Dx, IDEXX Laboratories, USA) at the initial screening were randomly allocated to two monthly treatment groups. Dogs in one group received topical moxidectin + imidacloprid and dogs in the other group received topical selamectin for eight consecutive months. Blood samples were obtained for microfilariae and antigen detection until the eleventh month after the first treatment. Dogs becoming microfilaremic or antigenemic on or before day 180 were considered to be infected prior to the first dose and were excluded from the study. RESULTS: A total of 29 dogs completed the study, including 14 treated with moxidectin + imidacloprid and 15 treated with selamectin. No dogs treated with moxidectin + imidacloprid (0/14) became infected during the treatment period, whereas four dogs of the selamectin group (4/15) became infected. CONCLUSION: Topical moxidectin + imidacloprid is 100 % effective in preventing D. immitis infections in dogs living in a high challenge natural environment.

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.

Characterization of ectoparasites in an urban cat (Felis catus Linnaeus, 1758) population of Rio de Janeiro, Brazil.

Ectoparasites are capable of transmitting infectious diseases and, therefore, are of zoonotic concern. Cats submitted to a spay/neuter program in the city of Rio de Janeiro were examined to determine the distribution of ectoparasites in cats from a city with a tropical climate. Independent of gender, breed, or age, 292 cats were combed and subjected to otoscopic examination. Ectoparasites were collected, and blood samples were taken to determine packed cell volume. The majority of the 292 cats were female (71%), and most of them were categorized as domestic short hair cats (92%). Different species of ectoparasites that produced both single agent and multi-agent infestations were detected in these cats. Most cats were infested by Ctenocephalides felis (60%); however, other ectoparasites were found to a lesser degree in the sampled population (Felicola subrostratus (5.4%), Rhipicephalus sanguineus (1.4%), Lynxacarus radovskyi (1%), and Otodectes cynotis (6.2%)). Within the infested cats, 16.3% were also anemic, and there was a significant association between the occurrence of anemia and flea infestation. Of all sampled cats, 65% were infested by at least one species of ectoparasites, which suggests both a heavy infestation of the environment and owner negligence. Ectoparasites are not only nuisances to both cats and owners, but they are also significant carriers of disease. Therefore, the implementation of rigorous, safe preventive measures is of great importance.