Infection Rate of Trypanosoma cruzi (Trypanosomatida: Trypanosomatidae) in Dipetalogaster maxima (Hemiptera: Reduviidae).

Chagas disease is caused by the infection of the parasite Trypanosoma cruzi (Chagas, 1909). Mexico is estimated to be among the countries with the highest rates of human infections. The southernmost region of the Baja California peninsula is home to the endemic, highly aggressive, and largest Triatominae vector, thus far described: Dipetalogaster maxima (Uhler 1894). Previous single-year studies have attempted to estimate the natural infection rate of T. cruzi in this species, none encompassing a multiyear sampling design nor a species-specific diagnostic tool. We report the infection rate based on more than 717 individuals examined via a PCR species-specific diagnosis. The infection rate of T. cruzi was of 4.4% (n = 5/112), 0.9% (n = 4/411), and 4.6% (n = 9/194) for 2016, 2017, and 2018, respectively, resulting in an infection rate of 2% across all sites and years (n = 18/717).

Distribution and natural infection status of synantrophic triatomines (Hemiptera: Reduviidae), vectors of Trypanosoma cruzi, reveals new epidemiological scenarios for chagas disease in the Highlands of Colombia.

INTRODUCTION: Updating the distribution and natural infection status of triatomine bugs is critical for planning, prioritizing, and implementing strategies to control Chagas disease (CD), especially after vector reduction programs. After carrying out a control program, the Department of Boyaca contains the highest number of Colombian municipalities certified by PAHO to be free of intradomiciliary transmission of Trypanosoma cruzi by Rhodnius prolixus. The present work describes the spatial distribution, natural infection (NI), and molecular characterization of T. cruzi in synanthropic triatomines from the Department of Boyaca in 2017 and 2018. MATERIALS AND METHODS: An entomological survey was conducted in 52 municipalities in Boyaca known to have had previous infestations of triatomine bugs. Insects were collected through active searches carried out by technical personnel from the Secretary of Health and community members using Triatomine Collection Stations (PITs-acronym in Spanish). For evaluation of natural infection, triatomines were identified morphologically and grouped in pools of one to five individuals of the same species collected in the same household. DNA derived from the feces of each pool of insects was analyzed by PCR for the presence of T. cruzi using primers flanking the satellite DNA of the parasite. SL-IR primers were used to differentiate TCI from the other DTUs and to identify different genotypes. The distribution of the collected triatomines was analyzed to determine any vector hotspots using spatial recreation. RESULTS: A total of 670 triatomine bugs was collected, belonging to five species: Triatoma dimidiata (73.2%), Triatoma venosa (16.7%), Panstrongylus geniculatus (5.7%), Rhodnius prolixus (4.4%), and Panstrongylus rufotuberculatus (0.4%), from 29 of the 52 municipalities. In total, 71.6% of the bugs were collected within houses (intradomiciliary) and the rest around the houses (peridomiciliary). Triatoma dimidiata was the most widely distributed species and had the highest natural infection index (37.8%), followed by T. venosa and P. geniculatus. TcI was the only DTU found, with the TcI Dom genotype identified in 80% of positive samples and TcI sylvatic in the other insects. Spatial analysis showed clusters of T. dimidiata and T. venosa in the northeast and southwest regions of Boyaca. CONCLUSIONS: After some municipalities were certified free of natural transmission within houses (intradomiciliary transmission) of T. cruzi by R. prolixus, T. dimidiata has become the most prevalent vector present, and represents a significant risk of resurgent CD transmission. However, T. venosa, P. geniculatus, and P. rufotuberculatus also contribute to the increased risk of transmission. The presence of residual R. prolixus may undo the successes achieved through vector elimination programs. The molecular and spatial analysis used here allows us to identify areas with an ongoing threat of parasite transmission and improve entomological surveillance strategies.

Evidencia molecular de la infección por Trypanosoma cruzi TcI en Meccus pallidipennis capturados en el municipio de Tepecuacuilco, Guerrero.

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Behavioral parameters of six populations of Meccus phyllosomus longipennis (Heteroptera: Reduviidae) from areas with high and low prevalences of Trypanosoma cruzi human infection.

Three behaviors of epidemiological importance, namely feeding latency, feeding duration and defecation latency, for six populations of Meccus phyllosomus longipennis (Usinger) from areas of central, western and north-central Mexico with high (HP) and low (LP) prevalence of Trypanosoma cruzi (Chagas) human infection were evaluated in this study. The median feeding latency (the time taken to begin feeding) was highly variable between instars. Within-instar comparisons showed that at least 65% of the LP populations (N3 to adult) started to feed significantly (P < 0.05) later than the HP population, with N1 showing no difference, and N2 from LP populations feeding sooner than those from HP populations. The six populations had similar median feeding durations within instars. A higher (P < 0.05) percentage of the instars from HP populations defecated faster than the respective instars from the three LP populations. Approximately 25% of the young nymphs (N1 to N3) and females in the HP populations defecated < 2 min postfeeding, compared with 4%-6% of the young nymphs and 1.3%-3% of females in the LP populations. Moreover, 17.7%-38.8% of the older nymphs (N4 to N5) in the HP populations and 6.8%-13.4% in the LP populations defecated during or immediately after feeding. Our results indicate that the HP populations have a greater potential than the LP populations to transmit T. cruzi infections, which may underlie the differences in the prevalence of T. cruzi infection in some areas where M. p. longipennis is currently distributed.

Effects of Trypanosoma cruzi on the phenoloxidase and prophenoloxidase activity in the vector Meccus pallidipennis (Hemiptera: Reduviidae).

BACKGROUND: Triatomine insects are vectors of Trypanosoma cruzi, the causal agent of Chagas disease. The insect-parasite interaction has been studied in relation to the transmission and prevalence of this disease. For most triatomines, however, several crucial aspects of the insect immune response are still unknown. For example, only for Rhodnius prolixus and Triatoma infestans has the activity of phenoloxidase (PO) and its zymogen prophenoloxidase (proPO) been reported in relation to the hemolymph and anterior midgut (AM). The aim of this study was to gain insight into the immune response to T. cruzi infection of an important triatomine in Mexico, Meccus pallidipennis. METHODS: Parasites were quantified in the rectal contents of infected M. pallidipennis groups. We examined some key factors in disease transmission, including the systemic (hemolymph) and local (gut) immune response. RESULTS: Parasites were present in the rectal contents at 4 days post-infection (pi) and reached their maximum density on day 7 pi. At 7 and 9 days pi mainly metacyclic trypomastigotes occurred. Compared to the control, the infected insects exhibited diminished PO activity in the hemolymph on days 9, 16 and 20 pi, and in the AM only on day 9. Additionally, infected insects displayed lower proPO activity in the hemolymph on day 1, but greater activity in the AM on day 28. CONCLUSIONS: The parasite strain originating from M. pallidipennis rapidly colonized the rectum of nymphs of this triatomine and developed high numbers of metacyclic trypomastigotes. Neither the changes of concentrations of PO and proPO in the hemolymph nor in the AM correlated with the changes in the population of T. cruzi.

Impact of climate change on vector transmission of Trypanosoma cruzi (Chagas, 1909) in North America.

Climate change can influence the geographical range of the ecological niche of pathogens by altering biotic interactions with vectors and reservoirs. The distributions of 20 epidemiologically important triatomine species in North America were modelled, comparing the genetic algorithm for rule-set prediction (GARP) and maximum entropy (MaxEnt), with or without topographical variables. Potential shifts in transmission niche for Trypanosoma cruzi (Trypanosomatida: Trypanosomatidae) (Chagas, 1909) were analysed for 2050 and 2070 in Representative Concentration Pathway (RCP) 4.5 and RCP 8.5. There were no significant quantitative range differences between the GARP and MaxEnt models, but GARP models best represented known distributions for most species [partial-receiver operating characteristic (ROC) > 1]; elevation was an important variable contributing to the ecological niche model (ENM). There was little difference between niche breadth projections for RCP 4.5 and RCP 8.5; the majority of species shifted significantly in both periods. Those species with the greatest current distribution range are expected to have the greatest shifts. Positional changes in the centroid, although reduced for most species, were associated with latitude. A significant increase or decrease in mean niche elevation is expected principally for Neotropical 1 species. The impact of climate change will be specific to each species, its biogeographical region and its latitude. North American triatomines with the greatest current distribution ranges (Nearctic 2 and Nearctic/Neotropical) will have the greatest future distribution shifts. Significant shifts (increases or decreases) in mean elevation over time are projected principally for the Neotropical species with the broadest current distributions. Changes in the vector exposure threat to the human population were significant for both future periods, with a 1.48% increase for urban populations and a 1.76% increase for rural populations in 2050.

Transmission Capacity of Trypanosoma cruzi (Trypanosomatida: Trypanosomatidae) by Three Subspecies of Meccus phyllosomus (Heteroptera: Reduviidae) and Their Hybrids.

Three behaviors of epidemiological importance: the time lapse for the onset of feeding, actual feeding, and defecation time for Meccus phyllosomus pallidipennis (Stål), Meccus phyllosomus longipennis (Usinger), Meccus phyllosomus picturatus (Usinger), and their laboratory hybrids were evaluated in this study. The mean time lapse for the beginning of feeding was between 0.5 and 8.3 min considering all instars in each cohort, with highly significant differences only among fifth-instar nymphs, females, and males of M. p. pallidipennis and M. p. longipennis relative to the hybrid cohorts. Four hybrid (LoPa [M. p. longipennis and M. p. pallidipennis] and LoPi [M. p. longipennis and M. p. picturatus] and their reciprocal experimental crosses) cohorts had similar mean feeding times to one of the parental subspecies, but longer than the other one. The remaining hybrid cohort (PaPi [M. p. pallidipennis and M. p. picturatus]) had longer feeding times than both of its parental subspecies. The specimens of the LoPa and LoPi hybrid cohorts defecated faster than the respective instars of the three parental cohorts. With exception of first- and fifth-instar nymphs, PaPi cohorts defecated faster than the remaining seven cohorts. More than 60% of defecation events occurred during feeding in the six hybrid cohorts. Our results indicate that hybrid cohorts have more potential to acquire infection and transmit Trypanosoma cruzi Chagas than their parental cohorts.

Geographic variation on biological parameters of Meccus picturatus (Usinger), 1939 (Hemiptera: Reduviidae).

The biological parameters related to the life cycles of three populations of Meccus picturatus (Usinger) (Hemiptera: Reduviidae), one of the main vectors of Trypanosoma cruzi Chagas (Trypanosomatida: Trypanosomatidae), were evaluated. A cohort of each of three populations from geographically isolated localities in western Mexico was maintained under similar laboratory conditions, after which all three populations were compared to each other. In each cohort, 50.9 to 72.1% of nymphs completed the cycle. The average time from N1 to adult was approximately 175 days and different for the three studied cohorts. The number of blood meals between molts varied from one to three. The hatching rates as well as the fecundity per female per day were different among the three studied populations. Our results show that the three isolated populations are statistically different in some parameters from each other, which points to the importance of studying the biological characteristics of local populations of triatomines for estimating their capacity of transmitting T. cruzi to reservoir hosts.

Differences on biological attributes of three populations of Meccus pallidipennis Stål (Hemiptera: Reduviidae).

BACKGROUND & OBJECTIVES: Meccus pallidipennis is one of the most epidemiologically important vectors of Trypanosoma cruzi to reservoir hosts in nine states of Mexico. Triatomines occurring in distinct locations normally adapt to local conditions. The aim of this study was to examine the biological attributes of three populations of M. pallidipennis from areas with different environmental characteristics as a factor influencing the triatomine capacity for T. cruzi transmission. METHODS: The values of biological parameters related to the life cycle, the number of blood meals to molt to next instar, fecundity and percentage of females after a biological cycle of three populations of M. pallidipennis were evaluated. A cohort of each of the three studied populations from different geographical areas of Mexico was maintained under similar laboratory conditions and then compared with each other. RESULTS: The life cycle was less than six months in all the studied cohorts, with differences among them. The number of blood meals to molt was lower for the cohort from Izϊcar de Matamoros. Laid eggs per day per female was lower for the cohort from Luvianos. In contrast, no important differences were recorded on the percentage of mortality, egg eclosion rate or percentages of obtained females. INTERPRETATION & CONCLUSION: It was found that an important level of heterogeneity exist between the three studied populations of M. pallidipennis, apparently influenced by the remarkable differences on environmental conditions on the localities where the founders were initially collected, that emphasizes the necessity of studies on local populations of triatomines.

Trypanosoma cruzi infection in the sylvatic kissing bug Mepraia gajardoi from the Chilean Southern Pacific Ocean coast.

The Southern Pacific Ocean coast has been traditionally considered a non-active transmission area for Chagas disease. In this report, we show evidence of Trypanosoma cruzi infection in the sylvatic kissing bug Mepraia gajardoi from the northern Chilean coast.

First record of Megaselia scalaris (Loew) (Diptera: Phoridae) infesting laboratory colonies of Triatoma brasiliensis Neiva (Hemiptera: Reduviidae)

Megaselia scalaris (Loew) is a cosmopolitan and synanthropic scuttle fly, eclectic in its feeding habits and acts as detritivore, parasite, facultative parasite, and parasitoid. Here we report for the first time M. scalaris infesting laboratory colonies of Triatoma brasiliensis Neiva, the most important Chagas disease vector in semiarid areas of Brazil. M. scalaris larvae were found feeding inside bugs; pupae were found in the esophagus and intestinal regions of T. brasiliensis through dissection. Other relevant information about this finding is also described in this note, including some preventive measures to avoid laboratory colonies infestations.

Megaselia scalaris (Loew) é uma mosca cosmopolita, sinantrópica e eclética quanto aos seus hábitos alimentares, tendo sido descrita como detritívora, parasita, parasita facultativa e parasitóide. Nesta nota, registra-se pela primeira vez, M. scalaris infestando colônias de laboratório de Triatoma brasiliensis Neiva, o mais importante vetor da doença de Chagas no semi-árido brasileiro. Larvas de M. scalaris foram encontradas alimentando-se dos órgãos internos dos triatomíneos; pupas foram encontradas na região intestinal e esofagiana de exemplares de T. brasiliensis dissecados. Outras informações relevantes sobre o achado são descritas nesta nota, bem como medidas preventivas para evitar infestações por M. scalaris em colônias de triatomíneos.

Susceptibility of Mepraia spinolai and Triatoma infestans to different Trypanosoma cruzi strains from naturally infected rodent hosts.

Trypanosoma cruzi is the causative agent of Chagas disease, a zoonosis involving domestic and sylvatic mammalian reservoirs. Since scarce information has been published about the susceptibility of T. cruzi lineages to other triatomine species besides Triatoma infestans, we evaluate the susceptibility of T. infestans and Mepraia spinolai to different T. cruzi lineages, originated from naturally infected Octodon degus rodents as mammal host. Xenodiagnosis-PCR methods to detect T. cruzi positive rodents and genotyping to differentiate T. cruzi lineages (TcI, TcIIb, TcIId and TcIIe) identified singly and mixed T. cruzi infections. More infections and nearly all mixed infections were identified using the wild vector M. spinolai than T. infestans.

Effect of ectoparasitic Pimeliaphilus plumifer mites (Acari: Pterygosomatidae) on Meccus pallidipennis (Hemiptera: Reduviidae) and several other Chagas' disease vectors under laboratory conditions.

Several biological parameters were evaluated to determine the capacity of Pimeliaphilus plumifer as biological control agent of Triatominae bugs. When P. plumifer and bugs of a variety of triatomine species were forced together in cages in the laboratory, the incidence of mite infestation was the following: Meccus pallidipennis > M. bassolsae > Triatoma rubida > M. longipennis > M. picturatus, and practically no mites were found on T. infestans and Rhodnius prolixus. Adults and hexapod larvae of P. plumifer were the only stages found to parasitize on M. pallidipennis. Fourth and fifth instar nymphs of this bug appeared most susceptible to mite infestation. P. plumifer mites located preferably on the coxae, ventral abdomen and pronotum of M. pallidipennis bugs. The number of blood meals, amount of ingested blood, and resistance to starvation of M. pallidipennis were similar in both the control and the mite infested groups. On the other hand, mite infection reduced molting rate in nymphs and longevity in adults, increased mortality in third-fifth instar nymphs, and fewer viable eggs were laid by females infected with P. plumifer. These effects could be related with nutritional deficiencies. Our results support the use of P. plumifer mites as control agents of host Triatominae species.

First record of Megaselia scalaris (Loew) (Diptera: Phoridae) infesting laboratory colonies of Triatoma brasiliensis Neiva (Hemiptera: Reduviidae).

Megaselia scalaris (Loew) is a cosmopolitan and synanthropic scuttle fly, eclectic in its feeding habits and acts as detritivore, parasite, facultative parasite, and parasitoid. Here we report for the first time M. scalaris infesting laboratory colonies of Triatoma brasiliensis Neiva, the most important Chagas disease vector in semiarid areas of Brazil. M. scalaris larvae were found feeding inside bugs; pupae were found in the esophagus and intestinal regions of T. brasiliensis through dissection. Other relevant information about this finding is also described in this note, including some preventive measures to avoid laboratory colonies infestations.

Methylene blue vital staining for Trypanosoma cruzi trypomastigotes and epimastigotes

The morphological identification of Trypanosoma cruzi is currently considered to have a high specificity, but its sensitivity, which depends on the volume of the sample examined, is rather low. Trypanosome developmental stages suspended in blood, reduviid feces, and culture media are routinely searched for by means of fresh film examination (about 2 æL). High speed centrifugation of blood samples separates the buffy coat, where most trypomastigotes concentrate. As the parasites are transparent and colorless, their detection is mostly dependent on their motility. The fluorescent vital stain acridine orange has been used to enhance image contrast, as exemplified by the QBC (Quantitative Buffy Coat) technique. Staining blood, buffy coat, reduviid feces, and culture media samples with methylene blue (also a vital dye) is a means of producing sharp, well contrasted images of motile or non-motile T. cruzi developmental stages, only standard laboratory microscopes being required. Slides previously coated with a thin layer of methylene blue are used to stain fresh blood films. Photomicrographs exemplify the results of methylene blue staining applied to living and fixed parasites.

A identificação morfológica de Trypanosoma cruzi tem alta especificidade, segundo é geralmente aceito; entretanto, sua sensibilidade, dependente do volume da amostra examinada, é baixa. Formas evolutivas de T. cruzi suspensas em sangue, fezes de reduviídeos e meios de cultura são rotineiramente pesquisadas em esfregaços a fresco (cerca de 2 æL). Centrifugação de amostras de sangue a altas velocidades produz a separação do creme leucocitário, onde se concentram as formas tripomastigotas. Em preparações a fresco, a motilidade das formas tripomastigotas e epimastigotas de T. cruzi, protozoário transparente e incolor, facilita sua detecção. Laranja de acridina, corante vital fluorescente, tem sido usada para acentuar o contraste das imagens de parasitas. Disto é exemplo a técnica QBC (Quantitative Buffy Coat). A coloração por meio de azul de metileno (também um corante vital), de amostras de sangue, de fezes de reduviídeos ou de meios de cultura permite obter imagens nítidas e contrastadas de formas evolutivas de T. cruzi com ou sem motilidade. Microscópios de uso geral em laboratórios permitem o exame dos parasitas corados. Uma camada bem delgada de azul de metileno colocada sobre a parte central da lâmina limpa (por meio da evaporação de solução diluída do corante) é usada para corar as preparações a fresco. O aspecto dos parasitas corados em materiais frescos ou previamente fixados pode ser observado em fotomicrografias.

Methylene blue vital staining for Trypanosoma cruzi trypomastigotes and epimastigotes.

The morphological identification of Trypanosoma cruzi is currently considered to have a high specificity, but its sensitivity, which depends on the volume of the sample examined, is rather low. Trypanosome developmental stages suspended in blood, reduviid feces, and culture media are routinely searched for by means of fresh film examination (about 2 microL). High speed centrifugation of blood samples separates the buffy coat, where most trypomastigotes concentrate. As the parasites are transparent and colorless, their detection is mostly dependent on their motility. The fluorescent vital stain acridine orange has been used to enhance image contrast, as exemplified by the QBC (Quantitative Buffy Coat) technique. Staining blood, buffy coat, reduviid feces, and culture media samples with methylene blue (also a vital dye) is a means of producing sharp, well contrasted images of motile or non-motile T. cruzi developmental stages, only standard laboratory microscopes being required. Slides previously coated with a thin layer of methylene blue are used to stain fresh blood films. Photomicrographs exemplify the results of methylene blue staining applied to living and fixed parasites.

Development of Triatoma rubida sonoriana, Triatoma barberi, and Meccus mazzottii (Heteroptera, Reduviidae) under laboratory conditions.

The objective of this study was to determine some of the most important bionomic parameters related to the capacity of transmission of Trypanosoma cruzi and to estimate the potential transmission capacity and role of 3 groups of recent colonized Mexican Triatoma barberi, T. rubida sonoriana, and Meccus (formerly Triatoma) mazzottii in the prevalence of Chagas disease cases based on the distribution area of each species. Among the studied Triatominae species, the life cycle was shorter in T. r. sonoriana and longer in T. barberi, by as much as 4 times. The 1st and 5th instars had the highest rates of mortality in most of the studied species. Statistically significant shorter duration of feeding of instars was observed for T. r. sonoriana and longer duration was found for M. mazzottii when the species were compared. The mean number of blood meals per nymphal stadium was statistically greater in T. barberi than in T. r. sonoriana and M. mazzottii. Triatoma r. sonoriana and M. mazzotti had shorter defecation delay than in T. barberi and the delay was uniform in all instars. Most of the studied parameters showed that T. r. sonoriana and M. mazzotti could have an important potential role in the prevalence of Chagas disease cases where these species are commonly found.

DNA evidence of Trypanosoma cruzi in the Chilean wild vector Mepraia spinolai (Hemiptera: Reduviidae).

Molecular evidence showed 46.2% of Trypanosoma cruzi infection in Mepraia spinolai insects from North-Central Chile, which is significantly higher than previous reports of up to 26% by microscopic observation. Our results show similar infection levels among nymphal stages, ranging from 38.3 to 54.1%, indicating that younger nymphs could be as important as older ones in parasite transmission. A cautionary note must be stressed to indicate the potential role of M. spinolai in transmitting T. cruzi in country areas due to the high infection level detected by molecular analysis.

DNA evidence of Trypanosoma cruzi in the Chilean wild vector Mepraia spinolai (Hemiptera: Reduviidae)

Molecular evidence showed 46.2 percent of Trypanosoma cruzi infection in Mepraia spinolai insects from North-Central Chile, which is significantly higher than previous reports of up to 26 percent by microscopic observation. Our results show similar infection levels among nymphal stages, ranging from 38.3 to 54.1 percent, indicating that younger nymphs could be as important as older ones in parasite transmission. A cautionary note must be stressed to indicate the potential role of M. spinolai in transmitting T. cruzi in country areas due to the high infection level detected by molecular analysis.