Polyparasitism and zoonotic parasites in dogs from a rural area of the Argentine Chaco.

Dogs play an important role as reservoirs and hosts of multiple pathogens shared with humans and wildlife, which contribute significantly to the global burden of disease. Here, we assessed the occurrence of a broad range of zoonotic and non-zoonotic parasites in dogs from a rural area in the humid Chaco; determined the occurrence of polyparasitism; and explored its association with selected risk factors. In total, 212 dogs were examined serologically to determine Trypanosoma cruzi infection and 152 of them also were examined for Ehrlichia canis, Borrelia bugderfori, Anaplasma phagocitophylum, Dirofilaria immitis and Toxoplasma gondii. Fecal samples from 85 dogs were examined for intestinal parasites. Seventeen parasite species were seen, 77% of which are zoonotic. The most prevalent parasites were Ancylostoma caninum (68.2%), T. gondii (55.3%, first report for dogs in Argentina), Giardia sp. (25.9%), Cryptosporidium sp. (20.0%), T. cruzi (16.5%), trematodes (15.3%) and Toxocara canis (14.1%). Polyparasitism was found in 96% of the dogs, with up to six parasite species in a single dog, and was significantly associated with age of dog but not with host body condition or sex. The most frequent pair of parasites found together were T. gondii-A. caninum (46%), A. caninum-T. cruzi (34%) and T. gondii-T. cruzi (27%). The prevalence of anemia and leukocytosis was significantly higher in dogs showing the worst body condition. Our findings likely reflect structural poverty, poor sanitation and lack of a safe water supply. Importantly, many of the prevalent parasites seen are threats to human health. 243 words.

An oral dose of Fluralaner administered to dogs kills pyrethroid-resistant and susceptible Chagas disease vectors for at least four months.

New vector control tools that can fit into a broader integrated vector management strategy are notably lacking. We conducted a seven-month randomized trial to assess the efficacy of a single oral dose of Fluralaner (Bravecto®) administered to dogs on the blood-feeding success, engorgement levels and mortality of pyrethroid-resistant and -susceptible Triatoma infestans third- and fifth-instar nymphs. The trial included 10 Fluralaner-treated and 10 placebo-treated (control) outbred healthy dogs residing in rural houses of the Argentine Chaco. Most (92.7%) of the 3017 triatomines exposed were able to blood-feed. Generalized linear models showed that blood-feeding success was not significantly modified by Fluralaner treatment, time posttreatment and their interaction. However, pyrethroid-susceptible fifth instars blood-fed significantly more frequently than susceptible third instars, and no significant differences were observed between the latter and resistant fifth instars. Engorgement levels were not significantly modified by Fluralaner treatment, time posttreatment and their interaction. Nearly all the triatomines that blood-fed on treated dogs up to 60 days posttreatment (DPT) died within 24 h regardless of pyrethroid susceptibility status combined with bug stage. Cumulative bug mortality over 4 days postexposure remained high over 90-120 DPT (70-81% in susceptible third and fifth instars, and 47-49% in resistant fifth instars), and was virtually nil at 210 DPT. Triatomines that fed on control dogs suffered marginal mortality (0-4%) except at 4 and 30 DPT. Fluralaner and xenointoxication are eligible for Phase III efficacy trials alone or combined with other methods in the frame of an integrated vector management strategy in areas with or without pyrethroid resistance.

Is the infectiousness of dogs naturally infected with Trypanosoma cruzi associated with poly-parasitism?

Interactions among different species of parasites co-infecting the same host could be synergistic or antagonistic. These interactions may modify both the frequency of infected hosts and their infectiousness, and therefore impact on transmission dynamics. This study determined the infectiousness of Trypanosoma cruzi-seropositive dogs (using xenodiagnosis) and their parasite load (quantified by qPCR), and tested the association between both variables and the presence of concomitant endoparasites. A cross-sectional serosurvey conducted in eight rural villages from Pampa del Indio and neighboring municipalities (northeastern Argentina) detected 32 T. cruzi-seropositive dogs out of 217 individuals examined for infection. Both the infectiousness to the vector Triatoma infestans and parasite load of T. cruzi-seropositive dogs examined were heterogeneous. A statistically significant, nine-fold higher mean infectiousness was registered in T. cruzi-seropositive dogs co-infected with Ancylostoma caninum and a trematode than in T. cruzi-seropositive dogs without these infections. The median parasite load of T. cruzi was also significantly higher in dogs co-infected with these helminths. An opposite trend was observed in T. cruzi-seropositive dogs that were serologically positive to Toxoplasma gondii or Neospora caninum relative to dogs seronegative for these parasites. Using multiple logistic regression analysis with random effects, we found a positive and significant association between the infectiousness of T. cruzi-seropositive dogs and co-infections with A. caninum and a trematode. Our results suggest that co-infections may be a modifier of host infectiousness in dogs naturally infected with T. cruzi.

A comparative study of Trypanosoma cruzi infection in sylvatic mammals from a protected and a disturbed area in the Argentine Chaco.

Understanding the complex epidemiology of Trypanosoma cruzi transmission cycles requires comparative studies in widely different environments. We assessed the occurrence of T. cruzi infection in sylvatic mammals, their infectiousness to the vector, and parasite genotypes in a protected area of the Argentine Chaco, and compared them with information obtained similarly in a nearby disturbed area. A total of 278 mammals from >23 species in the protected area were diagnosed for T. cruzi infection using xenodiagnosis, kDNA-PCR and nuclear satellite DNA-PCR (SAT) from blood samples. The relative abundance and species composition differed substantially between areas. Didelphis albiventris opossums were less abundant in the protected area; had a significantly lower body mass index, and a stage structure biased toward earlier stages. The capture of armadillos was lower in the protected area. The composite prevalence of T. cruzi infection across host species was significantly lower in the protected area (11.1%) than in the disturbed area (22.1%), and heterogeneous across species groups. The prevalence of infection in D. albiventris and Thylamys pusilla opossums was significantly lower in the protected area (nil for D. albiventris), whereas infection in sigmodontine rodents was three times higher in the protected area (17.5 versus 5.7%). Parasite isolates from the two xenodiagnosis-positive mammals (1 Dasypus novemcinctus and 1 Conepatus chinga) were typed as TcIII; both specimens were highly infectious to Triatoma infestans. Fat-tailed opossums, bats and rodents were kDNA-PCR-positive and xenodiagnosis-negative. Desmodus rotundus and Myotis bats were found infected with T. cruzi for the first time in the Gran Chaco.

High levels of Trypanosoma cruzi DNA determined by qPCR and infectiousness to Triatoma infestans support dogs and cats are major sources of parasites for domestic transmission.

The competence of reservoir hosts of vector-borne pathogens is directly linked to its capacity to infect the vector. Domestic dogs and cats are major domestic reservoir hosts of Trypanosoma cruzi, and exhibit a much higher infectiousness to triatomines than seropositive humans. We quantified the concentration of T. cruzi DNA in the peripheral blood of naturally-infected dogs and cats (a surrogate of intensity of parasitemia), and evaluated its association with infectiousness to the vector in a high-risk area of the Argentinean Chaco. To measure infectiousness, 44 infected dogs and 15 infected cats were each exposed to xenodiagnosis with 10-20 uninfected, laboratory-reared Triatoma infestans that blood-fed to repletion and were later individually examined for infection by optical microscopy. Parasite DNA concentration (expressed as equivalent amounts of parasite DNA per mL, Pe/mL) was estimated by real-time PCR amplification of the nuclear satellite DNA. Infectiousness increased steeply with parasite DNA concentration both in dogs and cats. Neither the median parasite load nor the mean infectiousness differed significantly between dogs (8.1Pe/mL and 48%) and cats (9.7Pe/mL and 44%), respectively. The infectiousness of dogs was positively and significantly associated with parasite load and an index of the host's body condition, but not with dog's age, parasite discrete typing unit and exposure to infected bugs in a random-effects multiple logistic regression model. Real-time PCR was more sensitive and less time-consuming than xenodiagnosis, and in conjunction with the body condition index, may be used to identify highly infectious hosts and implement novel control strategies.

Detection of Trypanosoma cruzi infection in naturally infected dogs and cats using serological, parasitological and molecular methods.

Domestic dogs and cats are major domestic reservoir hosts of Trypanosoma cruzi and a risk factor for parasite transmission. In this study we assessed the relative performance of a polymerase chain reaction assay targeted to minicircle DNA (kDNA-PCR) in reference to conventional serological tests, a rapid dipstick test and xenodiagnosis to detect T. cruzi infection in dogs and cats from an endemic rural area in northeastern Argentina. A total of 43 dogs and 13 cats seropositive for T. cruzi by an immunosorbent assay (ELISA) and an indirect hemagglutination assay (IHA), which had been examined by xenodiagnosis, were also tested by kDNA-PCR. kDNA-PCR was nearly as sensitive as xenodiagnosis for detecting T. cruzi-infectious dogs and cats. kDNA-PCR was slightly more sensitive than xenodiagnosis in seropositive dogs (91% versus 86%, respectively) and cats (77% against 54%, respectively), but failed to detect all of the seropositive individuals. ELISA and IHA detected all xenodiagnosis-positive dogs and both outcomes largely agreed (kappa coefficient, κ=0.92), whereas both assays failed to detect all of the xenodiagnosis-positive cats and their agreement was moderate (κ=0.68). In dogs, the sensitivity of the dipstick test was 95% and agreed closely with the outcome of conventional serological tests (κ=0.82). The high sensitivity of kDNA-PCR to detect T. cruzi infections in naturally infected dogs and cats supports its application as a diagnostic tool complementary to serology and may replace the use of xenodiagnosis or hemoculture.

Discrete typing units of Trypanosoma cruzi identified in rural dogs and cats in the humid Argentinean Chaco.

The discrete typing units (DTUs) of Trypanosoma cruzi that infect domestic dogs and cats have rarely been studied. With this purpose we conducted a cross-sectional xenodiagnostic survey of dog and cat populations residing in 2 infested rural villages in Pampa del Indio, in the humid Argentine Chaco. Parasites were isolated by culture from 44 dogs and 12 cats with a positive xenodiagnosis. DTUs were identified from parasite culture samples using a strategy based on multiple polymerase-chain reactions. TcVI was identified in 37 of 44 dogs and in 10 of 12 cats, whereas TcV was identified in 5 dogs and in 2 cats -a new finding for cats. No mixed infections were detected. The occurrence of 2 dogs infected with TcIII -classically found in armadillos- suggests a probable link with the local sylvatic transmission cycle involving Dasypus novemcinctus armadillos and a potential risk of human infection with TcIII. Our study reinforces the importance of dogs and cats as domestic reservoir hosts and sources of various DTUs infecting humans, and suggests a link between dogs and the sylvatic transmission cycle of TcIII.

The sylvatic transmission cycle of Trypanosoma cruzi in a rural area in the humid Chaco of Argentina.

Little is known about the sylvatic transmission cycle of Trypanosoma cruzi in the Gran Chaco ecoregion. We conducted surveys to identify the main sylvatic hosts of T. cruzi, parasite discrete typing units and vector species involved in Pampa del Indio, a rural area in the humid Argentinean Chaco. A total of 44 mammals from 14 species were captured and examined for infection by xenodiagnosis and polymerase chain reaction amplification of the hyper-variable region of kinetoplast DNA minicircles of T. cruzi (kDNA-PCR). Ten (22.7%) mammals were positive by xenodiagnosis or kDNA-PCR. Four of 11 (36%) Didelphis albiventris (white-eared opossums) and six of nine (67%) Dasypus novemcinctus (nine-banded armadillos) were positive by xenodiagnosis and or kDNA-PCR. Rodents, other armadillo species, felids, crab-eating raccoons, hares and rabbits were not infected. Positive animals were highly infectious to the bugs that fed upon them as determined by xenodiagnosis. All positive opossums were infected with T. cruzi I and all positive nine-banded armadillos with T. cruzi III. Extensive searches in sylvatic habitats using 718 Noireau trap-nights only yielded Triatoma sordida whereas no bug was collected in 26 light-trap nights. Four armadillos or opossums fitted with a spool-and-line device were successfully tracked to their refuges; only one Panstrongylus geniculatus was found in an armadillo burrow. No sylvatic triatomine was infected with T. cruzi by microscopical examination or kDNA-PCR. Our results indicate that two independent sylvatic transmission cycles of T. cruzi occur in the humid Chaco. The putative vectors of both cycles need to be identified conclusively.