Elucidating the Mechanism of Trypanosoma cruzi Acquisition by Triatomine Insects: Evidence from a Large Field Survey of Triatoma infestans.

Blood-sucking triatomine bugs transmit the protozoan parasite Trypanosoma cruzi, the etiologic agent of Chagas disease. We measured the prevalence of T. cruzi infection in 58,519 Triatoma infestans captured in residences in and near Arequipa, Peru. Among bugs from infected colonies, T. cruzi prevalence increased with stage from 12% in second instars to 36% in adults. Regression models demonstrated that the probability of parasite acquisition was roughly the same for each developmental stage. Prevalence increased by 5.9% with each additional stage. We postulate that the probability of acquiring the parasite may be related to the number of feeding events. Transmission of the parasite does not appear to be correlated with the amount of blood ingested during feeding. Similarly, other hypothesized transmission routes such as coprophagy fail to explain the observed pattern of prevalence. Our results could have implications for the feasibility of late-acting control strategies that preferentially kill older insects.

Trypanosoma cruzi Infection Does Not Decrease Survival or Reproduction of the Common Bed Bug, Cimex lectularius.

Although not presently implicated as a vector of human pathogens, the common bed bug, Cimex lectularius, has been suspected of carrying human pathogens because of its close association with humans and its obligate hematophagy. Recently, we characterized the vectorial competence of C. lectularius for the parasite Trypanosoma cruzi, the causative agent of Chagas disease. We observed that C. lectularius can acquire T. cruzi infection when fed on T. cruzi-carrying mice, and subsequently transmit T. cruzi to uninfected mice. This led us to ask why has C. lectularius not been implicated in the transmission of T. cruzi outside of the laboratory? We hypothesized that T. cruzi reduces C. lectularius fitness (i.e., survival and/or reproduction) as an explanation for why C. lectularius does not to transmit T. cruzi in natural settings. We tested this hypothesis by comparing the survival and reproduction of uninfected and T. cruzi-infected C. lectularius. We observed that T. cruzi had a variable effect on C. lectularius survival and reproduction. There were negligible differences between treatments in juveniles. Infected adult females tended to live longer and produce more eggs. However, no effect was consistent, and infected bugs showed more variation in survival and reproduction metrics than control bugs. We did not observe any negative effects of T. cruzi infection on C. lectularius survival or reproduction, suggesting that decreased fitness in T. cruzi-infected C. lectularius is not why bed bugs have not been observed to transmit T. cruzi in natural settings.

Host-seeking behavior and dispersal of Triatoma infestans, a vector of Chagas disease, under semi-field conditions.

Chagas disease affects millions of people in Latin America. The control of this vector-borne disease focuses on halting transmission by reducing or eliminating insect vector populations. Most transmission of Trypanosoma cruzi, the causative agent of Chagas disease, involves insects living within or very close to households and feeding mostly on domestic animals. As animal hosts can be intermittently present it is important to understand how host availability can modify transmission risk to humans and to characterize the host-seeking dispersal of triatomine vectors on a very fine scale. We used a semi-field system with motion-detection cameras to characterize the dispersal of Triatoma infestans, and compare the behavior of vector populations in the constant presence of hosts (guinea pigs), and after the removal of the hosts. The emigration rate - net insect population decline in original refuge - following host removal was on average 19.7% of insects per 10 days compared to 10.2% in constant host populations (p = 0.029). However, dispersal of T. infestans occurred in both directions, towards and away from the initial location of the hosts. The majority of insects that moved towards the original location of guinea pigs remained there for 4 weeks. Oviposition and mortality were observed and analyzed in the context of insect dispersal, but only mortality was higher in the group where animal hosts were removed (p-value <0.01). We discuss different survival strategies associated with the observed behavior and its implications for vector control. Removing domestic animals in infested areas increases vector dispersal from the first day of host removal. The implications of these patterns of vector dispersal in a field setting are not yet known but could result in movement towards human rooms.

Bed bugs (Cimex lectularius) as vectors of Trypanosoma cruzi.

Populations of the common bed bug, Cimex lectularius, have recently undergone explosive growth. Bed bugs share many important traits with triatomine insects, but it remains unclear whether these similarities include the ability to transmit Trypanosoma cruzi, the etiologic agent of Chagas disease. Here, we show efficient and bidirectional transmission of T. cruzi between hosts and bed bugs in a laboratory environment. Most bed bugs that fed on experimentally infected mice acquired the parasite. A majority of previously uninfected mice became infected after a period of cohabitation with exposed bed bugs. T. cruzi was also transmitted to mice after the feces of infected bed bugs were applied directly to broken host skin. Quantitative bed bug defecation measures were similar to those of important triatomine vectors. Our findings suggest that the common bed bug may be a competent vector of T. cruzi and could pose a risk for vector-borne transmission of Chagas disease.

Residual infestation and recolonization during urban Triatoma infestans Bug Control Campaign, Peru.

Chagas disease vector control campaigns are being conducted in Latin America, but little is known about medium-term or long-term effectiveness of these efforts, especially in urban areas. After analyzing entomologic data for 56,491 households during the treatment phase of a Triatoma infestans bug control campaign in Arequipa, Peru, during 2003-2011, we estimated that 97.1% of residual infestations are attributable to untreated households. Multivariate models for the surveillance phase of the campaign obtained during 2009-2012 confirm that nonparticipation in the initial treatment phase is a major risk factor (odds ratio [OR] 21.5, 95% CI 3.35-138). Infestation during surveillance also increased over time (OR 1.55, 95% CI 1.15-2.09 per year). In addition, we observed a negative interaction between nonparticipation and time (OR 0.73, 95% CI 0.53-0.99), suggesting that recolonization by vectors progressively dilutes risk associated with nonparticipation. Although the treatment phase was effective, recolonization in untreated households threatens the long-term success of vector control.

Comparison of insecticidal paint and deltamethrin against Triatoma infestans (Hemiptera: Reduviidae) feeding and mortality in simulated natural conditions.

The vector of Chagas disease, Triatoma infestans, is largely controlled by the household application of pyrethroid insecticides. Because effective, large-scale insecticide application is costly and necessitates numerous trained personnel, alternative control techniques are badly needed. We compared the residual effect of organophosphate-based insecticidal paint (Inesfly 5A IGR™ (I5A)) to standard deltamethrin, and a negative control, against T. infestans in a simulated natural environment. We evaluated mortality, knockdown, and ability to take a blood meal among 5(th) instar nymphs. I5A paint caused significantly greater mortality at time points up to nine months compared to deltamethrin (Fisher's Exact Test, p < 0.01 in all instances). A year following application, mortality among nymphs in the I5A was similar to those in the deltamethrin (χ2 = 0.76, df=1, p < 0.76). At months 0 and 1 after application, fewer nymphs exposed to deltamethrin took a blood meal compared to insects exposed to paint (Fisher's Exact Tests, p < 0.01 and p < 0.01, respectively). Insecticidal paint may provide an easily-applied means of protection against vectors of Chagas disease.

Temporal differences in blood meal detection from the midguts of Triatoma infestans.

We used genus/species specific PCRs to determine the temporal persistence of host DNA in Triatoma infestans experimentally fed on blood from six common vertebrate species: humans, domestic dogs, guinea pigs, chickens, mice, and pigs. Twenty third or fourth instar nymphs per animal group were allowed to feed to engorgement, followed by fasting-maintenance in the insectary. At 7, 14, 21, or 28 days post-feeding, the midgut contents from five triatomines per group were tested with the respective PCR assay. DNA from all vertebrate species was detected in at least four of five study nymphs at seven and 14 days post-feeding. DNA of humans, domestic dogs, guinea pigs, pigs, and chickens were more successfully detected (80-100%) through day 21, and less successfully (20-100%) at day 28. Findings demonstrate that species-specific PCRs can consistently identify feeding sources of T. infestans within two weeks, a biologically relevant time interval.

Temporal differences in blood meal detection from the midguts of Triatoma infestans / Diferencias temporales en la detección de fuentes de alimentación en el contenido intestinal de Triatoma infestans

We used genus/species specific PCRs to determine the temporal persistence of host DNA in Triatoma infestans experimentally fed on blood from six common vertebrate species: humans, domestic dogs, guinea pigs, chickens, mice, and pigs. Twenty third or fourth instar nymphs per animal group were allowed to feed to engorgement, followed by fasting-maintenance in the insectary. At 7, 14, 21, or 28 days post-feeding, the midgut contents from five triatomines per group were tested with the respective PCR assay. DNA from all vertebrate species was detected in at least four of five study nymphs at seven and 14 days post-feeding. DNA of humans, domestic dogs, guinea pigs, pigs, and chickens were more successfully detected (80-100%) through day 21, and less successfully (20-100%) at day 28. Findings demonstrate that species-specific PCRs can consistently identify feeding sources of T. infestans within two weeks, a biologically relevant time interval.

Se utilizó pruebas PCR género o especie específicas para determinar la persistencia temporal de ADN del hospedero en el contenido intestinal de Triatoma infestans que fueron alimentados experimentalmente con sangre de seis vertebrados muy frecuentemente asociados a enfermedad de Chagas: humano, perro, cobayo, pollo, ratón, y cerdo. Se emplearon 20 ninfas de tercer y cuarto estadio por cada especie de hospedero. Fueron alimentados a saciedad y mantenidas en el insectario sin alimentación posterior. Se obtuvo el contenido intestinal de cinco triatominos por cada grupo a los 7, 14, 21 y 28 días post - alimentación, que fueron evaluados con los respectivos PCRs específicos. El ADN de todos los vertebrados fue detectado en al menos 4 de 5 ninfas evaluadas a los 7 y 14 días post - alimentación. El ADN de humano, perro, cobayo, cerdo y pollo fue detectado exitosamente (80-100%) hasta el día 21 y con menos éxito (20-100%) en el día 28. Estos resultados demuestran que PCRs específicos para cada especie de hospedero pueden identificar consistentemente la fuente de alimentación de T. infestans dentro de las dos semanas post - alimentación, siendo un intervalo de tiempo biológicamente relevante.

Secondary kill effect of deltamethrin on Triatoma infestans.

Control of the Chagas disease vector, Triatoma infestans, relies on the application of pyrethroid insecticides, especially deltamethrin. We performed laboratory studies to determine whether a T. infestans nymph that comes into contact with a deltamethrin-treated surface horizontally transfers the insecticide to subsequent triatomines. We found that a triatomine that walks on a deltamethrin-treated surface for a short period of time has the ability to transport the insecticide in concentrations sufficient to kill other triatomines with which it comes into contact. The effect was limited to high-density environments, and mortality as a result of secondary exposure was greater among second-instar nymphs compared with fifth-instar nymphs. Our results suggest that deltamethrin could be killing triatomines through both direct and indirect contact, although it remains unclear whether the phenomenon occurs in natural conditions.