Seroepidemiological Studies of Arboviruses in Africa.

The literature on sero-epidemiological studies of flaviviral infections in the African continent is quite scarce. Much of the viral epidemiology studies have been focussing on diseases such as HIV/AIDS because of their sheer magnitude and impact on the lives of people in the various affected countries. Increasingly disease outbreaks caused by arboviruses such as the recent cases of chikungunya virus, dengue virus and yellow fever virus have prompted renewed interest in studying these viruses. International agencies from the US, several EU nations and China are starting to build collaborations to build capacity in many African countries together with established institutions to conduct these studies. The Tofo Advanced Study Week (TASW) was established to bring the best scientists from the world to the tiny seaside town of Praia do Tofo to rub shoulders with African virologists and discuss cutting-edge science and listen to the work of researchers in the field. In 2015 the 1st TASW focussed on Ebola virus. The collections of abstracts from participants at the 2nd TASW which focused on Dengue and Zika virus as well as presentations on other arboviruses are collated in this chapter.

Predicting West Nile Virus Infection Risk From the Synergistic Effects of Rainfall and Temperature.

Mosquito-based surveillance is a practical way to estimate the risk of transmission of West Nile virus (WNV) to people. Variations in temperature and precipitation play a role in driving mosquito infection rates and transmission of WNV, motivating efforts to predict infection rates based on prior weather conditions. Weather conditions and sequential patterns of meteorological events can have particularly important, but regionally distinctive, consequences for WNV transmission, with high temperatures and low precipitation often increasing WNV mosquito infection. Predictive models that incorporate weather can thus be used to provide early indications of the risk of WNV infection. The purpose of this study was first, to assess the ability of a previously published model of WNV mosquito infection to predict infection for an area within the region for which it was developed, and second, to improve the predictive ability of this model by incorporating new weather factors that may affect mosquito development. The legacy model captured the primary trends in mosquito infection, but it was improved considerably when calibrated with local mosquito infection rates. The use of interaction terms between precipitation and temperature improved model performance. Specifically, temperature had a stronger influence than rainfall, so that lower than average temperature greatly reduced the effect of low rainfall on increased infection rates. When rainfall was lower, high temperature had an even stronger positive impact on infection rates. The final model is practical, stable, and operationally valid for predicting West Nile virus infection rates in future weeks when calibrated with local data.

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.

Microgeographic spatial structuring of Triatoma infestans (Hemiptera: Reduviidae) populations using wing geometric morphometry in the Argentine Chaco.

We investigated the occurrence of spatial structuring in Triatoma infestans (KLug) (Hemiptera: Reduviidae) populations 12 yr after the last community-wide insecticide spraying campaign in rural Pampa del Indio, in the Gran Chaco of northeastern Argentina. In total, 172 male and 149 female right wings collected at 16 georeferenced sites with at least 10 individuals of the same sex were analyzed using geometric morphometry. Mean female body length and wing centroid size (CS) were significantly larger than for males. Log-transformed CS and length were significantly and positively correlated both for males and females. Males collected in domiciles had significantly smaller CS than those collected in peridomestic structures both closed (kitchens or storerooms) or open (chicken coops), in agreement with our previous results elsewhere in the dry Argentine Chaco. Female wing CS was not significantly different between ecotopes. Wing shape analyses showed the occurrence of significant geographic structuring in males and females combined and in males only. Male wings showed a strong association between Mahalanobis distance and geographic distance. In general, Mahalanobis distances were significantly different between collection sites located > 4 km apart. For collection sites located < 4 km apart, the greater the geographic distance the larger the difference in wing shape variables. Among females, only a partial correspondence between geographic groups and Mahalanobis distances was recorded. The strong spatial structuring found in T. infestans populations may be useful for the identification of putative reinfestation sources after vector control interventions.

Genetic variability, phylogenetic relationships and gene flow in Triatoma infestans dark morphs from the Argentinean Chaco.

The recent discovery of sylvatic populations of Triatoma infestans outside the Andean Valleys of Bolivia prompted an evolutionary question about the putative ancestral area of origin and dispersal of the species, and an epidemiological question regarding the possible role of these sylvatic populations in the recolonization process of insecticide-treated houses. The finding of a population of sylvatic melanic T. infestans (dark morphs) in the Argentinean dry Chaco at 7 km from a peridomestic bug population of typical coloration gave us the opportunity to test both questions simultaneously by employing phylogenetic and population genetic approaches. For this purpose we analyzed sylvatic and peridomestic bugs using sequence-based mitochondrial and nuclear markers (mtCOI and ITS-1) and microsatellites. Sylvatic bugs were confirmed to be T. infestans and not hybrids, and showed high levels of genetic variability and departures from neutral expectations for mtCOI variation. New ITS-1 and mtCOI haplotypes were recorded, as well as haplotypes shared with peridomestic and/or domestic bugs from previous records. The peridomestic population was invariant for ITS-1 and mtCOI, but showed variability for microsatellites and signatures of a population bottleneck, probably due to a limited number of founders. Phylogenetic analyses were consistent with the presence of ancestral haplotypes in sylvatic bugs. According to F-statistics and assignment methods there was a significant differentiation between sylvatic and peridomestic bugs and gene flow was low and asymmetric, with more bugs moving from the peridomicile to the sylvatic environment. These results support the hypothesis of the Chaco region as the area of origin of T. infestans, and a limited role of sylvatic melanic T. infestans in peridomestic infestation in the Argentinean Chaco.

Weather variability impacts on oviposition dynamics of the southern house mosquito at intermediate time scales.

Oviposition is a major event in the life history of mosquitoes, shaping both individual fitness and vectorial capacity. Several exogenous factors have been shown as important for the dynamic forcing of oviposition at finely (hourly) and coarsely (monthly or season to season) grained temporal scales. However, field studies addressing the interplay of weather factors on oviposition dynamics at the intermediate (days to weeks) time scale are missing. Here, we present the results from a field study that showed the oviposition dynamics of the southern house mosquito, Culex quinquefasciatus Say (Diptera: Culicidae), to be: (i) primarily dictated by relative humidity; and (ii) disrupted by rainfall events that resulted in a modified sensitivity to relative humidity. Rainfall changed the concentration of ammonia, a major limiting resource of microbes used as food by mosquito larvae. Following major rainfall events, the importance of relative humidity in forcing the oviposition dynamics also changed. Finally, our results indicate that qualitative changes in oviposition habitats modify the importance of weather variables as predictors of mosquito oviposition dynamics.

Geographical information systems and tropical medicine.

In terms of their applicability to the field of tropical medicine, geographical information systems (GIS) have developed enormously in the last two decades. This article reviews some of the pertinent and representative applications of GIS, including the use of such systems and remote sensing for the mapping of Chagas disease and human helminthiases, the use of GIS in vaccine trials, and the global applications of GIS for health-information management, disease epidemiology, and pandemic planning. The future use of GIS as a decision-making tool and some barriers to the widespread implementation of such systems in developing settings are also discussed.

Detection of "Candidatus Rickettsia sp. strain Argentina"and Rickettsia bellii in Amblyomma ticks (Acari: Ixodidae) from Northern Argentina.

Ixodid ticks were collected from vegetation and from humans, wild and domestic mammals in a rural area in the semi-arid Argentine Chaco in late spring 2006 to evaluate their potential role as vectors of Spotted Fever Group (SFG) rickettsiae. A total of 233 adult ticks, identified as Amblyomma parvum, Amblyomma tigrinum and Amblyomma pseudoconcolor, was examined for Rickettsia spp. We identified an SFG rickettsia of unknown pathogenicity, "Candidatus Rickettsia sp. strain Argentina", in A. parvum and A. pseudoconcolor by PCR assays targeting gltA, ompA, ompB and 17-kDa outer membrane antigen rickettsial genes. Rickettsia bellii was detected in a host-seeking male of A. tigrinum. Amblyomma parvum is widespread in the study area and is a potential threat to human health.

First finding of melanic sylvatic Triatoma infestans (Hemiptera: Reduviidae) colonies in the Argentine Chaco.

Triatoma infestans (Klug), the most important vector of Chagas disease in southern South America, is a highly domiciliated species with well-known sylvatic foci only in the Bolivian Andean valleys and in the Bolivian Chaco, where melanic insects designated as "dark morphs" were found. After the tentative identification of two melanic bugs collected from parrot nests in a forest reserve in the Argentine Chaco as T. infestans, we conducted an intensive search there using mouse-baited sticky traps in summer 2006 and 2007. Four live T. infestans bugs were collected in trees without parrot nests in 288 trap-nights, whereas no bug was collected from inside trees with active parrot nests in 51 trap-nights. To increase bug captures, hollow tree trunks that recently had had Amazona aestiva (Berlepsch) and Aratinga acuticaudata (Vieillot) parrot nests were treated with insecticide fumigant canisters exhibiting strong knockdown power. Four (22%) of 18 trees were positive for T. infestans with a dark phenotype. A fragment of the mitochondrial gene COI of 8 of the 14 triatomine bugs collected was successfully sequenced and confirmed as T. infestans. Most of the bugs were captured from Aspidosperma quebracho-blanco (Schlechter) hollow tree trunks harboring parrot nests. All of the T. infestans collected from the nearest house located at 10 km from the sylvatic foci displayed normal chromatic characters. The repeated finding of T. infestans in sylvatic habitats, albeit at very low density, shows that this species is capable of maintaining viable sylvatic foci in the absence of human hosts and immigration from domestic populations. These are the first confirmed findings of sylvatic T. infestans colonies in Argentina and of dark morphs in the Argentine Chaco.

Molecular population genetics and phylogeography of the Chagas disease vector Triatoma infestans in South America.

Knowledge of the genetic variability, population structure, and evolutionary history of Triatoma infestans may be useful for developing rational vector control strategies. A 661-bp fragment of the mitochondrial gene cytochrome oxidase I (COI) was sequenced and analyzed in bugs from Argentina, Uruguay, Peru, and Bolivia, including peridomestic, domestic, Andean, and Chaco sylvatic bugs. A total of 48 polymorphic sites among 37 haplotypes were described. Nucleotide variation fluctuated among samples, with the highest nucleotide diversity observed in seven Argentinean provinces. Within this group, some populations showed patterns of variability compatible with population expansions and/or fine-scale population structure, whereas others suggested population bottlenecks and/or population admixture processes. A maximum parsimony analysis of the haplotypes showed the presence of a Bolivian/Peruvian and an Argentinean/Uruguayan clade. Bolivian sequences were further divided in Chaco sylvatic and Andean domestic and sylvatic. Two different nested clades were found within the Argentinean/Uruguayan cluster. Analysis of molecular variance (AMOVA) and K(ST)* analysis supported a strong population structure in Argentina, where genetic differentiation was correlated with geographic distance. Departures from neutrality expectations and a nested cladistic analysis suggest a recent population expansion of T. infestans in Argentina, followed by restricted gene flow and patterns of isolation by distance. This expansion could have taken place as a two-wave process, as was shown by the phylogenetic analysis and signatures of population admixture in the southern most Argentinean populations.

Genetic structure of Triatoma infestans populations in rural communities of Santiago del Estero, northern Argentina.

To gain an understanding of the genetic structure and dispersal dynamics of Triatoma infestans populations, we analyzed the multilocus genotype of 10 microsatellite loci for 352 T. infestans collected in 21 houses of 11 rural communities in October 2002. Genetic structure was analyzed at the community and house compound levels. Analysis revealed that vector control actions affected the genetic structure of T. infestans populations. Bug populations from communities under sustained vector control (core area) were highly structured and genetic differentiation between neighboring house compounds was significant. In contrast, bug populations from communities with sporadic vector control actions were more homogeneous and lacked defined genetic clusters. Genetic differentiation between population pairs did not fit a model of isolation by distance at the microgeographical level. Evidence consistent with flight or walking bug dispersal was detected within and among communities, dispersal was more female-biased in the core area and results suggested that houses received immigrants from more than one source. Putative sources and mechanisms of re-infestation are described. These data may be use to design improved vector control strategies.

Environmental and demographic factors determining the spatial distribution of Triatoma guasayana in peridomestic and semi-sylvatic habitats of rural northwestern Argentina.

Triatoma guasayana (Wygodzinsky & Abalos), a sylvatic vector of Chagas' disease, occurs in natural and peridomestic habitats of the dry Chaco region of Argentina, Bolivia and Paraguay. Ten-year retrospective spatial analyses of peridomestic T. guasayana abundance in the rural community of Amamá were expanded to the neighbouring community of Trinidad in northwestern Argentina. The distribution of T. guasayana in domiciles, peridomiciles (storerooms, chicken coops and corrals) and natural habitats (bromeliads, dry cacti and logs) around houses (i.e. 'semi-sylvatic' habitats) was analysed. The distribution of the 316 T. guasayana specimens collected in domestic and peridomestic sites during 1993-2002 was significantly clustered in both communities. Searches confirmed that the spatial distribution of semi-sylvatic and peridomestic T. guasayana was determined by the joint effects of the local abundance of goats and the density of semi-sylvatic habitats. The integration of detailed entomological and demographic longitudinal data with geographic information system data, high-resolution satellite imagery, appropriate spatial and temporal analyses and field observations allowed us to infer the underlying processes determining the distribution of T. guasayana in rural communities. This approach may be applied to other sylvatic and peridomestic vectors of Chagas' disease in order to identify high-risk areas for targeted control or environmental management.

Impact of community-based vector control on house infestation and Trypanosoma cruzi infection in Triatoma infestans, dogs and cats in the Argentine Chaco.

The relative impact of two community-based vector control strategies on house infestation by Triatoma infestans and Trypanosoma cruzi infection in bugs, domestic dogs and cats was assessed in two neighboring rural areas comprising 40 small villages and 323 houses in one of the regions most endemic for Chagas disease in northern Argentina. The prevalence and abundance of domestic infestation were 1.5- and 6.5-fold higher, respectively, in the area under pulsed, non-supervised control actions operating under the guidelines of the National Vector Control Program (NCVP) than in the area under sustained, supervised surveillance carried out jointly by the UBA research team and NCVP. The prevalence of infestation and infection varied widely among village groups within each area. In the pulsed control area, the prevalence of infection in bugs, dogs and cats was two- to three-fold higher than in the area under sustained surveillance, most of the infected animals qualified as autochthonous cases, and evidence of recent transmission was observed. Infection was highly aggregated at the household level and fell close to the 80/20 rule. Using multiple logistic regression analysis clustered by household, infection in dogs was associated positively and significantly with variables reflecting local exposure to infected T. infestans, thus demonstrating weak performance of the vector surveillance system. For high-risk areas in the Gran Chaco region, interruption of vector-mediated domestic transmission of T. cruzi requires residual insecticide spraying that is more intense, of a higher quality and sustained in time, combined with community participation and environmental management measures.

Spatial analysis of the 1999-2000 highly pathogenic avian influenza (H7N1) epidemic in northern Italy.

The effect of proximity on infected premises was evaluated during the highly pathogenic avian influenza (HPAI) epidemic that struck northern Italy in 1999-2000 by quantifying the spatial and temporal clustering of cases. The epidemic was caused by an H7N1 subtype of type A influenza virus that originated from a low-pathogenic AI virus that spread among poultry farms in northeastern Italy in 1999 and eventually became virulent by mutation. More than 90% of 413 infected premises were located in Lombardy and Veneto regions; of 382 outbreaks, 60% occurred in the Lombardy region and 40% in the Veneto region. Global and local spatial statistics were used to estimate the location and degree of clustering of cases with respect to the population at risk. Outbreaks were spatially clustered primarily in Lombardy, with a large cluster in Brescia province and another in Mantua province, on the border of Veneto. Time series analysis was used to assess the temporal clustering of outbreaks. Temporal aggregation increased during the first 5 wk and decreased thereafter (probably as a result of eradication measures enforced in the Veneto region). Spatio-temporal clustering was assessed considering the Temporal Risk Window (TRW), the time period during which premises remain infectious and infection can spread to neighboring premises. The clustering pattern was similar to the one detected when considering spatial clustering (i.e., the larger clusters were identified in the Brescia and Mantua provinces of Lombardy). These results highlight the role of proximity in the spread of AI virus and, when considering the TRW, indicate the possible direction of virus spread.

Hydrological modelling of snail dispersal patterns in Msambweni, Kenya and potential resurgence of Schistosoma haematobium transmission.

Urinary schistosomiasis is an important source of human morbidity in Msambweni, Kenya, where the intermediate host snail, Bulinus nasutus is found in ponds and water pools. In the past, aquatic habitats in the area have been studied separately; however, recent collections of B. nasutus snails and shells indicated that many of these ponds are in fact connected during and following sufficient rains. Satellite imagery and a geographical information system (GIS) were used to survey the main water courses and potential drainage routes, to locate potential source populations of snails and to determine probable snail dispersal routes. The 2 water bodies implicated as being the most important Schistosoma haematobium transmission foci in the area were found to differ in their degree of connectivity to other B. nasutus source habitats. One pond becomes connected even after normal rains, while the other pond requires prolonged rains or flooding to become connected with source habitats. Consequently, the transmission foci differ in their susceptibility to snail population control measures. Spatially explicit dispersal models that consider the spatial and temporal patterns of connectivity between aquatic habitats will contribute to improved snail surveillance and more focused control for urinary schistosomiasis at a local level.

Domestic dogs and cats as sources of Trypanosoma cruzi infection in rural northwestern Argentina.

The reservoir capacity of domestic cats and dogs for Trypanosoma cruzi infection and the host-feeding patterns of domestic Triatoma infestans were assessed longitudinally in 2 infested rural villages in north-western Argentina. A total of 86 dogs and 38 cats was repeatedly examined for T. cruzi infection by serology and/or xenodiagnosis. The composite prevalence of infection in dogs (60%), but not in cats, increased significantly with age and with the domiciliary density of infected T. infestans. Dogs and cats had similarly high forces of infection, prevalence of infectious hosts (41-42%), and infectiousness to bugs at a wide range of infected bug densities. The infectiousness to bugs of seropositive dogs declined significantly with increasing dog age and was highly aggregated. Individual dog infectiousness to bugs was significantly autocorrelated over time. Domestic T. infestans fed on dogs showed higher infection prevalence (49%) than those fed on cats (39%), humans (38%) or chickens (29%) among 1085 bugs examined. The basic reproduction number of T. cruzi in dogs was at least 8.2. Both cats and dogs are epidemiologically important sources of infection for bugs and householders, dogs nearly 3 times more than cats.

Seasonal variations in active dispersal of natural populations of Triatoma infestans in rural north-western Argentina.

The flight dispersal of Triatoma infestans Klug (Hemiptera: Reduviidae) is one of the main mechanisms determining community re-infestation after control interventions. An empirical model of flight initiation coupled with data from a longitudinal study predicted that the flight dispersal of T. infestans would peak in summer. To test this prediction, longitudinal light trap collections were conducted during 3-8 nights in March (late summer), July (winter) and November (spring) 2003, and in March 2004 in a rural community in north-west Argentina. Following each light-trapping collection date, all peridomestic sites around light traps were inspected to assess the relative abundance and nutritional status of T. infestans at each site. A total of 21 adult and five nymph T. infestans, six Triatoma guasayana Wygodzinsky & Abalos, and nine Triatoma garciabesi Carcavallo et al. were collected in 96 light-trapping nights, whereas 696 T. infestans were collected from the peridomestic sites that surrounded the light traps. The arrival of T. infestans in the light traps occurred in 64% of catch stations and peaked in the summer surveys (10-14 bugs) compared with spring and winter surveys. When winds were < 5 km/h, the arrival of adult T. infestans at the light traps was significantly associated with maximum temperature and relative humidity. This is the first field report of seasonal variations in the flight dispersal activity of T. infestans.

Differential detection of Blastocrithidia triatomae and Trypanosoma cruzi by amplification of 24salpha ribosomal RNA genes in faeces of sylvatic triatomine species from rural northwestern Argentina.

Flagellates indistinguishable from Trypanosoma cruzi were detected by microscopy in faecal samples of 2/110 Triatoma guasayana and 2/283 Triatoma garciabesi captured in a rural area of northwestern Argentina. Inoculation of faecal homogenates to mice followed by xenodiagnosis, haemoculture, histopathology and culture from cardiac homogenates, and PCR based on T. cruzi minicircle and nuclear sequences failed to detect T. cruzi infection, pointing to another trypanosomatidean. A PCR strategy targeted to the D7 domain of 24salpha ribosomal DNA genes amplified a 250 bp sequence from one T. guasayana and one T. garciabesi faecal lysate. Sequence analysis revealed 100% identity with 24salpha rDNA amplicons from Blastocrithidia triatomae obtained from faeces of reared Triatoma infestans bugs. Phylogenetic analysis clustered this sequence with C. fasciculata and L. major, separated from the Trypanosoma branch (bootstrap: 968/1000), in concordance with a Neighbour-joining dendrogram based on 18s rDNA sequences. This PCR procedure provides a rapid sensitive tool for differential diagnosis of morphologically similar trypanosomatids in field surveys of Chagas disease vectors and laboratory-reared triatomines used for xenodiagnosis.

Long-term reduction of Trypanosoma cruzi infection in sylvatic mammals following deforestation and sustained vector surveillance in northwestern Argentina.

Long-term variations in the dynamics and intensity of sylvatic transmission of Trypanosoma cruzi were investigated around eight rural villages in the semiarid Argentine Chaco in 2002-2004 and compared to data collected locally in 1984-1991. Of 501 wild mammals from 13 identified species examined by xenodiagnosis, only 3 (7.9%) of 38 Didelphis albiventris opossums and 1 (1.1%) of 91 Conepatus chinga skunks were infected by T. cruzi. The period prevalence in opossums was four-fold lower in 2002-2004 than in 1984-1991 (32-36%). The infection prevalence of skunks also decreased five-fold from 4.1-5.6% in 1984-1991 to 1.1% in 2002-2004. Infection in opossums increased with age and from summer to spring in both study periods. The force of infection per 100 opossum-months after weaning declined more than six-fold from 8.2 in 1988-1991 to 1.2 in 2002-2004. Opossums were mainly infected by T. cruzi lineage I and secondarily by lineage IId in 1984-1991, and only by T. cruzi I in 2002-2004; skunks were infected by T. cruzi IId in 1984-1991 and by IIc in 2002-2004. The striking decline of T. cruzi infection in opossums and skunks occurred in parallel to community-wide insecticide spraying followed by selective sprays leading to very low densities of infected Triatoma infestans in domestic and peridomestic habitats since 1992; to massive deforestation around one of the villages or selective extraction of older trees, and apparent reductions in opossum abundance jointly with increases in foxes and skunks. These factors may underlie the dramatic decrease of T. cruzi infection in wild reservoir hosts.

Spatiotemporal patterns of host-seeking Ixodes scapularis nymphs (Acari: Ixodidae) in the United States.

The risk of Lyme disease for humans in the eastern United States is dependent on the density of host-seeking Ixodes scapularis Say nymphal stage ticks infected with Borrelia burgdorferi. Although many local and regional studies have estimated Lyme disease risk using these parameters, this is the first large-scale study using a standardized methodology. Density of host-seeking I. scapularis nymphs was measured by drag sampling of closed canopy deciduous forest habitats in 95 locations spaced among 2 degrees quadrants covering the entire United States east of the 100th meridian. Sampling was done in five standardized transects at each site and repeated three to six times during the summer of 2004. The total number of adults and nymphs of the seven tick species collected was 17,972, with 1,405 nymphal I. scapularis collected in 31 of the 95 sites. Peak global spatial autocorrelation values were found at the smallest lag distance (300 km) and decreased significantly after 1,000 km. Local auto-correlation statistics identified two significant high-density clusters around endemic areas in the northeast and upper Midwest and a low-density cluster in sites south of the 39th parallel, where only 21 nymphs were collected. Peak nymphal host-seeking density occurred earlier in the southern than in the most northern sites. Spatiotemporal density patterns will be combined with Borrelia prevalence data as part of a 4-yr survey to generate a nationwide spatial risk model for I. scapularis-borne Borrelia, which will improve targeting of disease prevention efforts.