Lethal effect of Goniozus legneri on Cactoblastis cactorum: A potential biocontrol agent for inundative releases.

Cactoblastis cactorum (Berg) (Lepidoptera: Pyralidae), the cactus moth, is native to South America with a widespread distribution in Argentina. The larvae consume the interior of Opuntia spp. (Cactaceae) plants. The moth was used as a biocontrol agent against invasive non-native Opuntia spp. in many countries around the world. The cactus moth arrived unintentionally in Florida, USA, expanded its range and threatened Opuntia-based agriculture and natural ecosystems in southern North America. The insect is also a pest of cultivated O. ficus-indica L. in Argentina. An endemic South American parasitoid, Goniozus legneri Gordth (Hymenoptera: Bethylidae), is used in inundative biological control programmes against lepidopteran pests. The goal of this work was to evaluate G. legneri as a biocontrol agent to be used in inundative releases against C. cactorum. Mortality of C. cactorum by G. legneri was assessed at different spatial scales, as well as the interactions with Apanteles opuntiarum Martínez & Berta (Hymenoptera: Braconidae), a common Argentine natural enemy of C. cactorum. The ability of G. legneri to paralyse, parasitise and kill C. cactorum was confirmed. The paralysis inflicted on C. cactorum larvae reduced larval damage to the plants by 85%. Using two parasitoid species increased the mortality of C. cactorum larvae, but it was highly dependent on the order of their arrival. The combined mortality caused by both parasitoids was higher than a single one, in particular when G. legneri arrived first (56 ± 1%), suggesting asymmetric competition due to the preference of G. legneri attacking previously parasitised larvae. Goniozus legneri has potential as an inundative biocontrol agent of C. cactorum, but its interaction with the classical biocontrol agent A. opuntiarum needs to be considered.

Improved vector control of Triatoma infestans limited by emerging pyrethroid resistance across an urban-to-rural gradient in the Argentine Chaco.

BACKGROUND: The sustainable elimination of Triatoma infestans in the Gran Chaco region represents an enduring challenge. Following the limited effects of a routine pyrethroid insecticide spraying campaign conducted over 2011-2013 (first period) in Avia Terai, an endemic municipality with approximately 2300 houses, we implemented a rapid-impact intervention package to suppress house infestation across the urban-to-rural gradient over 2015-2019 (second period). Here, we assess their impacts and whether persisting infestations were associated with pyrethroid resistance. METHODS: The 2011-2013 campaign achieved a limited detection and spray coverage across settings (< 68%), more so during the surveillance phase. Following community mobilization and school-based interventions, the 2015-2019 program assessed baseline house infestation using a stratified sampling strategy; sprayed all rural houses with suspension concentrate beta-cypermethrin, and selectively sprayed infested and adjacent houses in urban and peri-urban settings; and monitored house infestation and performed selective treatments over the follow-up. RESULTS: Over the first period, house infestation returned to pre-intervention levels within 3-4 years. The adjusted relative odds of house infestation between 2011-2013 and 2015-2016 differed very little (adj. OR: 1.17, 95% CI 0.91-1.51). Over the second period, infestation decreased significantly between 0 and 1 year post-spraying (YPS) (adj. OR: 0.36, 95% CI 0.28-0.46), with heterogeneous effects across the gradient. Mean bug abundance also dropped between 0 and 1 YPS and thereafter remained stable in rural and peri-urban areas. Using multiple regression models, house infestation and bug abundance at 1 YPS were 3-4 times higher if the house had been infested before treatment, or was scored as high-risk or non-participating. No low-risk house was ever infested. Persistent foci over two successive surveys increased from 30.0 to 59.3% across the gradient. Infestation was more concentrated in peridomestic rather than domestic habitats. Discriminating-dose bioassays showed incipient or moderate pyrethroid resistance in 7% of 28 triatomine populations collected over 2015-2016 and in 83% of 52 post-spraying populations. CONCLUSIONS: The intervention package was substantially more effective than the routine insecticide spraying campaign, though the effects were lower than predicted due to unexpected incipient or moderate pyrethroid resistance. Increased awareness and diagnosis of vector control failures in the Gran Chaco, including appropriate remedial actions, are greatly needed.

Improved detection of house infestations with triatomines using sticky traps: a paired-comparison trial in the Argentine Chaco.

BACKGROUND: We conducted a matched-pairs trial of three methods for detecting house infestation with triatominae bugs in a well-defined endemic rural area in the Argentine Chaco. METHODS: The three methods included a simple double-sided adhesive tape (ST) installed near host resting sites; timed-manual collections with a dislodging aerosol (TMC, the reference method used by vector control programmes), and householders' bug notifications (HN). Triatomine infestations were evaluated in 103 sites of 54 houses, including domiciles, kitchens and storerooms. RESULTS: In domiciles where Triatoma infestans was collected, sensitivity of each single method decreased from 79% by ST and 77% by HN, to 57% by TMC, and increased to 92% when ST was combined with HN. In peridomestic kitchens and storerooms, TMC was relatively as sensitive as ST and significantly more sensitive than HN. On average, the number of bugs recovered by ST was 0.94 times that collected by TMC. The ST mainly collected early-instar nymphs whereas TMC yielded late (larger) stages. Triatomines caught by ST had significantly lower mean weight-to-length ratios and lower blood-feeding rates than those caught by TMC, suggesting the ST intercepted and trapped vectors seeking a blood meal host. CONCLUSIONS: The ST may effectively replace TMC for detecting T. infestans in domiciles, and is especially apt for early detection of low-density domestic infestations in the frame of community-based surveillance or elimination programmes; decision making on whether an area should be targeted for full-coverage insecticide spraying, and to corroborate that extant conditions are compatible with the interruption of vector-borne transmission.

Community-based surveillance and control of chagas disease vectors in remote rural areas of the Argentine Chaco: A five-year follow-up.

Prevention of Chagas disease vector-borne transmission mostly relies on the residual application of pyrethroid insecticide. Persistent or recurrent house infestation after insecticide spraying remains a serious challenge in remote, resource-poor rural areas where public health services face substantial constraints. Here we use generalized estimating equations and multimodel inference to model the fine-scale, time-lagged effects of a community-based vector surveillance-and-response strategy on house infestation and abundance of Triatoma infestans in four rural communities of the Argentine Chaco over a five-year period. Householders and community leaders were trained to detect triatomines and spray with insecticides their premises if infested. House infestation and vector abundance were consistently higher in peridomestic habitats than in human habitations (domiciles). Householders supplemented with sensor boxes detected infested domiciles (67%) more frequently than timed-manual searches (49%). Of all houses ever found to be infested by timed-manual searches, 76% were sprayed within six months upon detection. Domestic triatomine abundance was significantly related to house-level insecticide spraying during the previous year (inversely) and current peridomestic abundance (positively). Peridomestic triatomine abundance significantly increased with current domestic bug abundance and maximum peridomestic abundance during the previous year, and was unaffected by insecticide spraying. Our study provides new empirical evidence of the interconnection and flow between domestic and peridomestic populations of T. infestans under recurrent insecticide treatments, and supports targeting both habitats with appropriate tactics for longer-lasting, improved vector control. Community-directed efforts succeeded in controlling domestic infestations and interrupting domestic transmission, whereas persistent peridomestic infestations demand sustained control efforts to address domestic reinvasions.

Chagas disease vector control and Taylor's law.

BACKGROUND: Large spatial and temporal fluctuations in the population density of living organisms have profound consequences for biodiversity conservation, food production, pest control and disease control, especially vector-borne disease control. Chagas disease vector control based on insecticide spraying could benefit from improved concepts and methods to deal with spatial variations in vector population density. METHODOLOGY/PRINCIPAL FINDINGS: We show that Taylor's law (TL) of fluctuation scaling describes accurately the mean and variance over space of relative abundance, by habitat, of four insect vectors of Chagas disease (Triatoma infestans, Triatoma guasayana, Triatoma garciabesi and Triatoma sordida) in 33,908 searches of people's dwellings and associated habitats in 79 field surveys in four districts in the Argentine Chaco region, before and after insecticide spraying. As TL predicts, the logarithm of the sample variance of bug relative abundance closely approximates a linear function of the logarithm of the sample mean of abundance in different habitats. Slopes of TL indicate spatial aggregation or variation in habitat suitability. Predictions of new mathematical models of the effect of vector control measures on TL agree overall with field data before and after community-wide spraying of insecticide. CONCLUSIONS/SIGNIFICANCE: A spatial Taylor's law identifies key habitats with high average infestation and spatially highly variable infestation, providing a new instrument for the control and elimination of the vectors of a major human disease.

Body size and hosts of Triatoma infestans populations affect the size of bloodmeal contents and female fecundity in rural northwestern Argentina.

Human sleeping quarters (domiciles) and chicken coops are key source habitats of Triatoma infestans-the principal vector of the infection that causes Chagas disease-in rural communities in northern Argentina. Here we investigated the links among individual bug bloodmeal contents (BMC, mg), female fecundity, body length (L, mm), host blood sources and habitats. We tested whether L, habitat and host blood conferred relative fitness advantages using generalized linear mixed-effects models and a multimodel inference approach with model averaging. The data analyzed include 769 late-stage triatomines collected in 120 sites from six habitats in 87 houses in Figueroa, Santiago del Estero, during austral spring. L correlated positively with other body-size surrogates and was modified by habitat type, bug stage and recent feeding. Bugs from chicken coops were significantly larger than pig-corral and kitchen bugs. The best-fitting model of log BMC included habitat, a recent feeding, bug stage, log Lc (mean-centered log L) and all two-way interactions including log Lc. Human- and chicken-fed bugs had significantly larger BMC than bugs fed on other hosts whereas goat-fed bugs ranked last, in consistency with average blood-feeding rates. Fecundity was maximal in chicken-fed bugs from chicken coops, submaximal in human- and pig-fed bugs, and minimal in goat-fed bugs. This study is the first to reveal the allometric effects of body-size surrogates on BMC and female fecundity in a large set of triatomine populations occupying multiple habitats, and discloses the links between body size, microsite temperatures and various fitness components that affect the risks of transmission of Trypanosoma cruzi.

Temporal variations of fluctuating asymmetry in wing size and shape of Triatoma infestans populations from northwest Argentina.

Environmentally-induced developmental instability has frequently been assessed using fluctuating asymmetry (FA) methods. For Triatoma infestans, the major vector of Chagas disease, the combined effects of host-feeding sources and habitats affected wing developmental instability depending on sex in Figueroa villages, northwest Argentina. Here we investigated whether habitat, sex, season/year and insecticide applications affected wing developmental instability in T. infestans populations from Amamá and other rural villages of northwest Argentina over a four-year period. We measured the occurrence and amount of wing size and shape FA in 423 adult triatomines collected in domiciles, goat corrals, pig corrals, storerooms and wood piles. Significant wing size and wing shape FA occurred in females and males from all habitats as determined by two-way mixed ANOVA and Procrustes ANOVA, respectively. For wing size and shape, the highest corrected indices of FA (FAI) for females occurred in wood piles, goat corrals and domiciles in late summer or early autumn, whereas for males, the largest FAIs consistently appeared in domiciles and storerooms. Wing size FAIs were significantly higher in recently infested goat corrals rather than in persistently infested goat corrals. The follow-up of four infested peridomestic sites showed that FA patterns were not stable over time or sites. Temporal variation of FA among habitats appears to be modified by the history of insecticide spraying, either through direct effects on insect development or through indirect effects related to flight dispersal and house invasion. Whether FA may provide another marker to identify the sources of reinfestant triatomines requires further investigation.

Morphometric Wings Similarity among Sylvatic and Domestic Populations of Triatoma infestans (Hemiptera: Reduviidae) from the Gran Chaco Region of Paraguay.

Despite sustained efforts for eliminating Triatoma infestans, reinfestation still persists in large part of the endemic area of Chagas disease from the Gran Chaco region. Sylvatic T. infestans populations seem to threat success of control programs of domestic T. infestans. In this study, we analyze whether T. infestans collected after a community-wide spraying were survivors or were immigrants from elsewhere using geometric morphometric tools. We used 101 right wings of female T. infestans captured before and after intervention program carried out in 12 de Junio and Casuarina, villages from Paraguayan Chaco, and in Puerto Casado during presprayed collection. There were no significant differences in wing size of domestic T. infestans between pre- and postspraying populations, and between domestic and sylvatic ones. When shape variables originating from postintervention individuals from 12 de Junio were introduced one by one into a discriminant analysis, the greatest weight (53%) was allocated to the sylvatic group. Furthermore, from the prespraying population, 25% were reallocated as postintervention individuals. Only 11% of the insects were reassigned to other groups Puerto Casado and Casuarina. These results suggest that postspraying individuals appear to have different origins. Half of the postspraying individuals from 12 de Junio were similar to the sylvatic ones and 25% of these were similar to those captured in the prespraying period. This remarkable morphometric wings similarity between sylvatic and domestic populations is new evidence suggesting that they could be highly related to each other in the Paraguayan Chaco; human-fed bugs from sylvatic area also support this.

Key source habitats and potential dispersal of triatoma infestans populations in Northwestern Argentina: implications for vector control.

BACKGROUND: Triatoma infestans -the principal vector of the infection that causes Chagas disease- defies elimination efforts in the Gran Chaco region. This study identifies the types of human-made or -used structures that are key sources of these bugs in the initial stages of house reinfestation after an insecticide spraying campaign. METHODOLOGY AND PRINCIPAL FINDINGS: We measured demographic and blood-feeding parameters at two geographic scales in 11 rural communities in Figueroa, northwest Argentina. Of 1,297 sites searched in spring, 279 (21.5%) were infested. Bug abundance per site and female fecundity differed significantly among habitat types (ecotopes) and were highly aggregated. Domiciles (human sleeping quarters) had maximum infestation prevalence (38.7%), human-feeding bugs and total egg production, with submaximal values for other demographic and blood-feeding attributes. Taken collectively peridomestic sites were three times more often infested than domiciles. Chicken coops had greater bug abundance, blood-feeding rates, engorgement status, and female fecundity than pig and goat corrals. The host-feeding patterns were spatially structured yet there was strong evidence of active dispersal of late-stage bugs between ecotopes. Two flight indices predicted that female fliers were more likely to originate from kitchens and domiciles, rejecting our initial hypothesis that goat and pig corrals would dominate. CONCLUSIONS AND SIGNIFICANCE: Chicken coops and domiciles were key source habitats fueling rapid house reinfestation. Focusing control efforts on ecotopes with human-fed bugs (domiciles, storerooms, goat corrals) would neither eliminate the substantial contributions to bug population growth from kitchens, chicken coops, and pig corrals nor stop dispersal of adult female bugs from kitchens. Rather, comprehensive control of the linked network of ecotopes is required to prevent feeding on humans, bug population growth, and bug dispersal simultaneously. Our study illustrates a demographic approach that may be applied to other regions and triatomine species for the design of innovative, improved vector control strategies.

Geographic variation of Trypanosoma cruzi discrete typing units from Triatoma infestans at different spatial scales.

We assessed the diversity and distribution of Trypanosoma cruzi discrete typing units (DTU) in Triatoma infestans populations and its association with local vector-borne transmission levels at various geographic scales. At a local scale, we found high predominance (92.4%) of TcVI over TcV in 68 microscope-positive T. infestans collected in rural communities in Santiago del Estero province in northern Argentina. TcV was more often found in communities with higher house infestation prevalence compatible with active vector-borne transmission. Humans and dogs were the main bloodmeal sources of the TcV- and TcVI-infected bugs. At a broader scale, the greatest variation in DTU diversity was found within the Argentine Chaco (227 microscope-positive bugs), mainly related to differences in equitability between TcVI and TcV among study areas. At a country-wide level, a meta-analysis of published data revealed clear geographic variations in the distribution of DTUs across countries. A correspondence analysis showed that DTU distributions in domestic T. infestans were more similar within Argentina (dominated by TcVI) and within Bolivia (where TcI and TcV had similar relative frequencies), whereas large heterogeneity was found within Chile. DTU diversity was lower in the western Argentine Chaco region and Paraguay (D=0.14-0.22) than in the eastern Argentine Chaco, Bolivia and Chile (D=0.20-0.68). Simultaneous DTU identifications of T. cruzi-infected hosts and triatomines across areas differing in epidemiological status are needed to shed new light on the structure and dynamics of parasite transmission cycles.

Domestic animal hosts strongly influence human-feeding rates of the Chagas disease vector Triatoma infestans in Argentina.

BACKGROUND: The host species composition in a household and their relative availability affect the host-feeding choices of blood-sucking insects and parasite transmission risks. We investigated four hypotheses regarding factors that affect blood-feeding rates, proportion of human-fed bugs (human blood index), and daily human-feeding rates of Triatoma infestans, the main vector of Chagas disease. METHODS: A cross-sectional survey collected triatomines in human sleeping quarters (domiciles) of 49 of 270 rural houses in northwestern Argentina. We developed an improved way of estimating the human-feeding rate of domestic T. infestans populations. We fitted generalized linear mixed-effects models to a global model with six explanatory variables (chicken blood index, dog blood index, bug stage, numbers of human residents, bug abundance, and maximum temperature during the night preceding bug catch) and three response variables (daily blood-feeding rate, human blood index, and daily human-feeding rate). Coefficients were estimated via multimodel inference with model averaging. FINDINGS: Median blood-feeding intervals per late-stage bug were 4.1 days, with large variations among households. The main bloodmeal sources were humans (68%), chickens (22%), and dogs (9%). Blood-feeding rates decreased with increases in the chicken blood index. Both the human blood index and daily human-feeding rate decreased substantially with increasing proportions of chicken- or dog-fed bugs, or the presence of chickens indoors. Improved calculations estimated the mean daily human-feeding rate per late-stage bug at 0.231 (95% confidence interval, 0.157-0.305). CONCLUSIONS AND SIGNIFICANCE: Based on the changing availability of chickens in domiciles during spring-summer and the much larger infectivity of dogs compared with humans, we infer that the net effects of chickens in the presence of transmission-competent hosts may be more adequately described by zoopotentiation than by zooprophylaxis. Domestic animals in domiciles profoundly affect the host-feeding choices, human-vector contact rates and parasite transmission predicted by a model based on these estimates.

Improved chemical control of Chagas disease vectors in the dry Chaco region.

The effectiveness of two doses of suspension concentrate (SC) pyrethroid insecticides in suppressing peridomestic populations of Triatoma infestans (Klug) was evaluated in 28 rural communities located in Santiago del Estero province, northwestern Argentina, including 388 houses and 1,516 identified sites. Four treatments were randomly assigned to peridomiciles within each community: 5% SC beta-cypermethrin at standard (S, 50 mg active ingredient [AI]/m2) and double dose (2S), and 2.5% SC deltamethrin at standard (D, 25 mg [AI]/m2) and double dose (2D). Simultaneously, we assessed the effects of both pyrethroids applied at standard doses against domestic infestations. Bug infestation at the site level was assessed by timed manual collections with a dislodging agent at baseline, 13 and 21 mo postspraying (MPS). In domiciles, D and S nearly suppressed all T. infestans infestations up to 21 MPS. In peridomestic sites infested before interventions, multiple logistic regression analysis showed that site-level reinfestation at 13 MPS was significantly lower for treatment 2D (1%) than for other treatments, whereas 2S (6%), D (5%), and S (14%) did not differ significantly between them. The risk of reinfestation after spraying was significantly greater in goat or pig corrals than in other peridomestic ecotopes (in which treatments did not differ significantly), and in sites infested before interventions than in uninfested sites. The application of SC deltamethrin at double dose in goat or pig corrals may suppress T. infestans foci and achieve more sustained effects in the dry Chaco.

Impact of genetic variation and geographic distribution of porcine reproductive and respiratory syndrome virus on infectivity and pig growth.

BACKGROUND: The porcine reproductive and respiratory syndrome (PRRS) is a devastating disease for the pig industry. In this study, we analysed the genetic variability of PRRS virus (PRRSV) as well as the relationship between the genetic variability, the geographical and temporal distribution of the PRRSV strains. Moreover, we investigated the association between the glycosylation patterns in PRRSV sequences and pigs growth. RESULTS: The data highlight that PRRSV strains evolve rapidly on individual farms, and temporal evolution of PRRSV is an important factor of genetic variability. Analysis of glycosylation sites in the glycoprotein 5 (GP5) ectodomain revealed that PRRSV isolates had seven combinations of putative N-linked glycosylation sites of which the N37/46/53 sites was found in 79% of the sequences. No significant relationship was found between the genetic variation of the PRRSV strains and the geographic distance. A significant relationship was found between the genetic variation and time of sampling when farm was considered as a factor in the analysis. Furthermore, the commercial semen from artificial insemination centres was not a source of PRRS transmission.The PRRSV having the glycosylation site at position N46 (N46+) were observed to have higher burden on pigs and accordingly the corresponding infected pigs had lower average daily gain (ADG) compared with those infected with PRRSV lacking the glycosylation at N46 (N46-) position site. This study showed that the number of piglets by litter infected by PRRSV was lower for the Landrace breed than for the other studied breeds (Large White, Duroc and Pietrain). CONCLUSIONS: The PRRSV genetic variability which is determined by a local and temporal evolution at the farm level could be considered in a perspective of prevention. Moreover, the association between the PRRSV glycosylation patterns and its virulence could be of interest for vaccine development. The differences of resistance to PRRSV infections among pig breeds might open new horizons for the genetic selection of robustness against PRRSV infection.

Co-expression of host and viral microRNAs in porcine dendritic cells infected by the pseudorabies virus.

MicroRNAs are small non-coding RNAs approximately 22 nt long that modulate gene expression in animals and plants. It has been recently demonstrated that herpesviruses encode miRNAs to control the post-transcriptional regulation of expression from their own genomes and possibly that of their host, thus adding an additional layer of complexity to the physiological cross-talk between host and pathogen. The present study focussed on the interactions between porcine dendritic cells (DCs) and the Pseudorabies virus (PRV), an alpha-herpesvirus causing Aujeszky's disease in pigs. A catalogue of porcine and viral miRNAs, expressed eight hours post-infection, was established by deep sequencing. An average of 2 million reads per sample with a size of 21-24 nucleotides was obtained from six libraries representing three biological replicates of infected and mock-infected DCs. Almost 95% of reads mapped to the draft pig genome sequence and pig miRNAs previously annotated in dedicated databases were detected by sequence alignment. In silico prediction allowed the identification of unknown porcine as well as of five miRNAs transcribed by the Large Latency Transcript (LLT) of PRV. The gene target prediction of the viral miRNAs and the Ingenuity Pathway Analysis of differentially expressed pig miRNAs were conducted to contextualize the identified small RNA molecules and functionally characterize their involvement in the post-transcriptional regulation of gene expression. The results support a role for PRV miRNAs in the maintenance of the host cell latency state through the down-regulation of immediate-early viral genes which is similar to other herpesviruses. The differentially expressed swine miRNAs identified a unique network of target genes with highly significant functions in the development and function of the nervous system and in infectious mechanisms, suggesting that the modulation of both host and viral miRNAs is necessary for the establishment of PRV latency.

Spatial re-establishment dynamics of local populations of vectors of Chagas disease.

BACKGROUND: Prevention of Chagas disease depends mainly on control of the insect vectors that transmit infection. Unfortunately, the vectors have been resurgent in some areas. It is important to understand the dynamics of reinfestation where it occurs. Here we show how continuous- and discrete-time models fitted to patch-level infestation states can elucidate different aspects of re-establishment. Triatoma infestans, the main vector of Chagas disease, reinfested sites in three villages in northwest Argentina after community-wide insecticide spraying in October 1992. METHODOLOGY/PRINCIPAL FINDINGS: Different methods of estimating the probabilities of bug establishment on each site were compared. The results confirmed previous results showing a 6-month time lag between detection of a new infestation and dispersal events. The analysis showed that more new bug populations become established from May to November than from November to May. This seasonal increase in bug establishment coincides with a seasonal increase in dispersal distance. In the fitted models, the probability of new bug establishment increases with increasing time since last detected infestation. CONCLUSIONS/SIGNIFICANCE: These effects of season and previous infestation on bug establishment challenge our current understanding of T. infestans ecology and highlight important gaps in knowledge. Experiments necessary to close these gaps are discussed.

Genome-wide transcriptional response of primary alveolar macrophages following infection with porcine reproductive and respiratory syndrome virus.

Porcine reproductive and respiratory syndrome is a major cause of economic loss for the swine industry worldwide. Porcine reproductive and respiratory syndrome virus (PRRSV) triggers weak and atypical innate immune responses, but key genes and mechanisms by which the virus interferes with the host innate immunity have not yet been elucidated. In this study, genes that control the response of the main target of PRRSV, porcine alveolar macrophages (PAMs), were profiled in vitro with a time-course experiment spanning the first round of virus replication. PAMs were obtained from six piglets and challenged with the Lelystad PRRSV strain, and gene expression was investigated using Affymetrix microarrays and real-time PCR. Of the 1409 differentially expressed transcripts identified by analysis of variance, two, five, 25, 16 and 100 differed from controls by a minimum of 1.5-fold at 1, 3, 6, 9 and 12 h post-infection (p.i.), respectively. A PRRSV infection effect was detectable between 3 and 6 h p.i., and was characterized by a consistent downregulation of gene expression, followed by the start of the host innate immune response at 9 h p.i. The expression of beta interferon 1 (IFN-beta), but not of IFN-alpha, was strongly upregulated, whilst few genes commonly expressed in response to viral infections and/or induced by interferons were found to be differentially expressed. A predominance of anti-apoptotic transcripts (e.g. interleukin-10), a shift towards a T-helper cell type 2 response and a weak upregulation of tumour necrosis factor-alpha expression were observed within 12 h p.i., reinforcing the hypotheses that PRRSV has developed sophisticated mechanisms to escape the host defence.

Re-establishment of local populations of vectors of Chagas disease after insecticide spraying.

1. Prevention of Chagas disease is mainly dependent on control of the insect vectors that transmit infection. Unfortunately, this control is not wholly successful and the vectors have been resurgent in some areas. Where re-infestation has occurred, it is important to understand the dynamics of the process. We investigated how a metapopulation framework can elucidate key aspects of re-infestation and thereby contribute to more efficient disease control.2.Triatoma infestans, the main vector of Chagas disease, re-infested sites in three villages in north-west Argentina after community-wide insecticide spraying in October 1992. Ten surveys were carried out at 6-monthly intervals from November 1994 to May 1999.3. Comparisons were made of different methods of estimating the sources of dispersal and the number of sites in which bug infestations became established.4. The results indicated that (i) the number of dispersing Triatoma infestans from a given site was proportional to the number of bugs found at the site; (ii) there was a 6-month time lag between detection of a new infestation and dispersal events; (iii) the relationship between infestations and new establishments varied by season.5. Three of 156 sites at which bugs were found were estimated to be the source of more than 50% of establishment events. These three sites were the only ones with large, persistent bug populations.6.Synthesis and applications. To reduce the risk of human Chagas disease, identifying those few sites infested with large, persistent bug populations and targeting control measures at those sites should greatly improve the efficiency of vector control. The appropriate seasonal timing of vector control could also greatly increase its efficiency. Specific recommendations for the timing of insecticide spraying require further research to establish how the observed temporal pattern of bug establishment is associated with the seasonality of bug dispersal.

Comparative trial of effectiveness of pyrethroid insecticides against peridomestic populations of Triatoma infestans in northwestern Argentina.

The effects of different pyrethroid insecticides, formulations, and doses on peridomestic populations of Triatoma infestans (Klug) were evaluated in 128 houses with 148 identified infested peridomestic sites in northwestern Argentina between October 2003 and March 2005. Four treatments were randomly assigned within each community: two doses of 5% suspension concentrate beta-cypermethrin in water applied with manual compression sprayers, the standard dose (S) at 50 mg and a double dose (2S) at 100 mg active ingredient (AI)/m2; and two emulsifiable concentrates diluted in diesel fuel and applied with power sprayers, 25% cypermethrin (100 mg [AI] /m2) (CF) and 10% permethrin (170 mg [AI]/m2) (DF). Infestation was assessed by timed manual collections with a dislodging agent at baseline, 5, 12, and 17 mo postspraying, and the sites found to be reinfested at 5 mo postspraying were selectively resprayed. Only 2S eliminated T. infestans from all peridomestic sites up to 12 mo postspraying, and it was significantly more effective than all other treatments. At 5 mo postspraying, more sites treated with CF or DF rather than S had bug colonies that probably represented residual foci, which they also failed in eliminating after a second spray. At 17 mo postspraying, the prevalence of reinfested peridomestic sites was 5% for 2S, 29% for S, 43% for CF, and 54% for DF. The application of suspension concentrate pyrethroids in dose twice as large as that currently in use in the attack phase produces a greater initial impact and may eliminate peridomestic populations of T. infestans.

A prospective study of the effects of sustained vector surveillance following community-wide insecticide application on Trypanosoma cruzi infection of dogs and cats in rural Northwestern Argentina.

Domestic dogs were used as natural sentinels to assess prospectively the long-term impact of selective, community-based spraying with pyrethroid insecticides after community-wide spraying on transmission of Trypanosoma cruzi in rural villages under surveillance between 1992 and 2002. In 2000 and 2002 light infestations by Triatoma infestans were recorded, and 523 dogs and cats were examined serologically or by xenodiagnosis. The prevalence of T. cruzi infection in dogs decreased from 65% at baseline to 8.9% and 4.7% at 7.5 and 10 years after sustained vector surveillance, respectively. The average annual force of infection dropped 260-fold from 72.7 per 100 dog-years at baseline to <0.3% in 2002, as determined prospectively and retrospectively from the age-prevalence curve of native dogs born during surveillance. Multiple logistic regression analysis showed that prevalent cases in dogs in 2000 and 2002 were associated positively and significantly with the peak number of T. infestans caught in domestic areas at the dog's compound during its lifetime. The sustained decline in T. cruzi infections in dogs and cats is the result of selective, community-based insecticide spraying that kept the abundance of infected T. infestans at marginal levels, fast host population turnover, and low immigration rates from areas with active transmission.

Reinfestation sources for Chagas disease vector, Triatoma infestans, Argentina.

Reinfestation by Triatoma infestans after insecticide spraying has caused elimination efforts in the dry Chaco region to fail repeatedly. The sources and spatial extent that need to be considered to understand the reinfestation pattern and to plan a comprehensive control program were studied in 2 adjacent rural communities in northwestern Argentina from 1993 to 1997. The effects of external, residual, and primary sources on the reinfestation pattern were evaluated by using geographic information systems, satellite imagery, spatial statistics, and 5-year retrospective data for 1,881 sites. The reinfestation process depended on primary internal sources and on surrounding infested communities. In the dry Chaco, successfully reducing the risk for reinfestation in a community depends on treating all communities and isolated sites within 1,500 m of the target community. In addition, during the surveillance phase, spraying all sites within 500 m of new foci will delay reinfestation.