Using genetic comparisons of populations from Arizona, Mexico, and Texas to investigate fall armyworm migration in the American southwest.

Fall armyworm (FAW) is a global agricultural pest, causing substantial economic losses in corn and many other crops. Complicating efforts to control this pest is its capacity for long distance flights, which has been described in greatest detail for the central and eastern sections of the United States. FAW infestations are also routinely found in agricultural areas in southern Arizona, which lie beyond the western limits of the mapped migratory pathways. Climate suitability analysis found that the affected Arizona locations cannot support permanent FAW populations, indicating that these FAW most likely arise from annual migrations. A better understanding of this migration would provide insights into how large moth populations can move across desert habitats as well as the degree of gene flow occurring between FAW populations across the North American continent. In this study the Arizona populations were genetically characterized and compared to a selection of permanent and migratory FAW from multiple sites in the United States and Mexico. The results are consistent with migratory contributions from permanent populations in the states of Texas (United States) and Sinaloa (Mexico), while also providing evidence of significant barriers to gene flow between populations within Mexico. An unexpected finding was that two genetically distinct FAW subpopulations known as "host strains" have a differential distribution in the southwest that may indicate significant differences in their migration behavior in this region. These findings indicate that the combination of mitochondrial and Z-linked markers have advantages in comparing FAW populations that can complement and extend the findings from other methods.

Investigating the Migratory Behavior of Soybean Looper, a Major Pest of Soybean, through Comparisons with the Corn Pest Fall Armyworm Using Mitochondrial Haplotypes and a Sex-Linked Marker.

The Noctuid moth soybean looper (SBL), Chrysodeixis includens (Walker), is an economically important pest of soybean (Glycine max (Linnaeus) Merrill). Because it is not known to survive freezing winters, permanent populations in the United States are believed to be limited to the southern regions of Texas and Florida, yet its geographical range of infestations annually extend to Canada. This indicates annual migrations of thousands of kilometers during the spring and summer growing season. This behavior is like that of the fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith), also a Noctuid that is a major global pest of corn. SBL and FAW are projected to have very similar distributions of permanent populations in North America based on climate suitability modeling and the overlap in the distribution of their preferred host plants (corn and soybean). It therefore seems likely that the two species will display similar migratory behavior in the United States. This was tested by identifying genetic markers in SBL analogous to those successfully used to delineate FAW migratory pathways and comparing the distribution patterns of the markers from the two species. Contrary to expectations, the results indicate substantial differences in migratory behavior that appear to be related to differences in the timing of corn and soybean plantings. These findings underscore the importance of agricultural practices in influencing pest migration patterns, in particular the timing of host availability relative to mean seasonal air transport patterns.

Evidence for Two Soybean Looper Strains in the United States with Limited Capacity for Cross-Hybridization.

The noctuid moth soybean looper (SBL), Chrysodeixis includens (Walker) is an economically important pest of soybeans (Glycine max (L.) Merr.) in the southeastern United States. It has characteristics that are of particular concern for pest mitigation that include a broad host range, the capacity for annual long-distance flight, and resistance in some populations to important pesticides such as pyrethroids and chitin synthesis inhibitor. The biology of SBL in the United States resembles that of the fellow noctuid fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith), a major pest of corn and several other crops. FAW exhibits a population structure in that it can be divided into two groups (host strains) that differ in their host preferences but are broadly sympatric and exhibit incomplete reproductive isolation. In this paper, strategies used to characterize the FAW strains were applied to SBL to assess the likelihood of population structure in the United States. Evidence is presented for two SBL strains that were defined phylogenetically and display differences in the proportions of a small set of genetic markers. The populations exhibit evidence of reproductive barriers sufficient to allow persistent asymmetry in the distribution of mitochondrial haplotypes. The identified molecular markers will facilitate studies characterizing the behaviors of these two populations, with relevance to pest mitigation and efforts to prevent further dispersal of the resistance traits.

Screening for resistance alleles to Cry1 proteins through targeted sequencing in the native and invasive range of Spodoptera frugiperda (Lepidoptera: Noctuidae).

The fall armyworm, Spodoptera frugiperda (J. E. Smith), is a highly polyphagous pest native to the tropical Americas that has recently spread to become a global super-pest threatening food and fiber production. Transgenic crops producing insecticidal Cry and Vip3Aa proteins from Bacillus thuringiensis (Bt) are used for control of this pest in its native range. The evolution of practical resistance represents the greatest threat to sustainability of this technology and its potential efficacy in the S. frugiperda invasive range. Monitoring for resistance is vital to management approaches delaying S. frugiperda resistance to Bt crops. DNA-based resistance screening provides higher sensitivity and cost-effectiveness than currently used bioassay-based monitoring. So far, practical S. frugiperda resistance to Bt corn-producing Cry1F has been genetically linked to mutations in the SfABCC2 gene, providing a model to develop and test monitoring tools. In this study, we performed targeted SfABCC2 sequencing followed by Sanger sequencing to confirm the detection of known and candidate resistance alleles to Cry1F corn in field-collected S. frugiperda from continental USA, Puerto Rico, Africa (Ghana, Togo, and South Africa), and Southeast Asia (Myanmar). Results confirm that the distribution of a previously characterized resistance allele (SfABCC2mut) is limited to Puerto Rico and identify 2 new candidate SfABCC2 alleles for resistance to Cry1F, one of them potentially spreading along the S. frugiperda migratory route in North America. No candidate resistance alleles were found in samples from the invasive S. frugiperda range. These results provide support for the potential use of targeted sequencing in Bt resistance monitoring programs.

Population genomics of fall armyworm by genotyping-by-sequencing: Implications for pest management.

The fall armyworm (FAW), Spodoptera frugiperda, is a significant pest of many crops in the world and it is native to the Americas, where the species has shown the ability to rapidly evolve resistance to insecticides and transgenic plants. Despite the importance of this species, there is a gap in the knowledge regarding the genetic structure of FAW in South America. Here, we examined the genetic diversity of FAW populations across a wide agricultural area of Brazil and Argentina using a Genotyping-by-Sequencing (GBS) approach. We also characterized samples by their host strain based on mitochondrial and Z-linked genetic markers. The GBS methodology enabled us to discover 3309 SNPs, including neutral and outlier markers. Data showed significant genetic structure between Brazil and Argentina populations, and also among the Argentinian ecoregions. Populations inside Brazil showed little genetic differentiation indicating high gene flow among locations and confirming that structure is related to the presence of corn and rice strains. Outlier analysis indicated 456 loci putatively under selection, including genes possibly related to resistance evolution. This study provides clarification of the population genetic structure of FAW in South America and highlights the importance of genomic research to understand the risks of spread of resistance genes.

Revisiting fall armyworm population movement in the United States and Canada.

Introduction: Biophysical approaches validated against haplotype and trap catch patterns have modeled the migratory trajectory of fall armyworms at a semi-continental scale, from their natal origins in Texas or Florida through much of the United States east of the Rocky Mountains. However, unexplained variation in the validation analysis was present, and misalignments between the simulated movement patterns of fall armyworm populations and the haplotype ratios at several locations, especially in the northeastern US and Canada, have been reported. Methods: Using an expanded dataset extending into Canada, we assess the consistency of haplotype patterns that relate overwintered origins of fall armyworm populations to hypothesized dispersal trajectories in North America and compare the geographic distribution of these patterns with previous model projections. Results and discussion: We confirm the general accuracy of previous modeling efforts, except for late in the season where our data suggests a higher proportion of Texas populations invading the northeast, extending into eastern Canada. We delineate geographic limits to the range of both overwintering populations and show that substantial intermixing of the Texas and Florida migrants routinely occurs north of South Carolina. We discuss annual variation to these migratory trajectories and test the hypothesis that the Appalachian Mountains influence geographic patterns of haplotypes. We discuss how these results may limit gene flow between the Texas and Florida natal populations and limit the hereditary consequences of interbreeding between these populations.

The Spodoptera frugiperda Host Strains: What They Are and Why They Matter for Understanding and Controlling This Global Agricultural Pest.

Fall armyworm (Spodoptera frugiperda [J.E. Smith]) is a moth native to the Western Hemisphere where it is a major pest of corn (maize) and multiple other crops. It is now a global threat with its recent discovery and dissemination in the Eastern Hemisphere. Its broad host range is in part due to two subpopulations denoted as 'host strains' that differ in host plant specificity. Therefore, identifying the strains present in a location is critical to assessing what crops are at risk of infestation. However, much remains uncertain about how the strains differ and even on the fundamental issue of how they are identified. Complicating factors include the host strains are morphologically indistinguishable, the defining behavior of the strains (host plant specificity) is variable, and the existence of significant differences between geographical populations and laboratory colonies that are independent of strain identity. These factors contribute to substantial disagreements in the literature on presumptive strain differences. This paper presents a summary of strain characteristics and suggests the criteria that should be met before concluding a trait is 'strain-specific'. The intent is to facilitate a common understanding of what the strains represent and to develop a more consistent experimental framework for studies on strain phenotypes. Evidence is summarized that supports a primary role for Z-linked genes in strain identity, which has potential implications for genetic approaches to define the strains, and we discuss the possibility that the strains arose from allopatric (rather than sympatric) speciation processes.

Observations of genetic differentiation between the fall armyworm host strains.

The threat of invasive species is increasing with the expansion of global trade and habitat disruption. A recent example is the establishment of fall armyworm (FAW), a noctuid moth native to the Americas, into most of the Eastern Hemisphere with projections of significant economic losses on a global scale. The species has traditionally been subdivided into two populations that differ in their propensity to use different plant hosts, a phenotype with clear relevance for identifying crops at risk. However, inconsistencies in the genetic and phenotypic descriptions of these "host strains" has led to controversy about their composition and even existence. In this study, the locus for the Triosephosphate isomerase gene (Tpi) is used both as a host strain marker and for phylogenetic analysis. Association of the host choice phenotype with the Tpi-derived phylogenetic tree uncovered genetic differentiation between populations that supports the existence of the host strains and provided evidence that they are subject to different selection pressures. This correspondence of differential host use with Tpi was demonstrated for populations from a broad geographical range and supports the involvement of one or more Z-chromosome functions controlling the phenotype. Comparisons of collections from multiple locations identified significant differences in the efficacy of different molecular markers that implicate regional variations in host strain behavior.

Genetic studies of fall armyworm indicate a new introduction into Africa and identify limits to its migratory behavior.

The fall armyworm, Spodoptera frugiperda (J.E. Smith) is native to the Americas and a major pest of corn and several other crops of economic importance. The species has characteristics that make it of particular concern as an invasive pest, including broad host range, long-distance migration behavior, and a propensity for field-evolved pesticide resistance. The discovery of fall armyworm in western Africa in 2016 was followed by what was apparently a remarkably rapid spread throughout sub-Saharan Africa by 2018, causing economic damage estimated in the tens of billions USD and threatening the food security of the continent. Understanding the history of the fall armyworm invasion of Africa and the genetic composition of the African populations is critical to assessing the risk posed to different crop types, the development of effective mitigation strategies, and to make Africa less vulnerable to future invasions of migratory moth pests. This paper tested and expanded on previous studies by combining data from 22 sub-Saharan nations during the period from 2016 to 2019. The results support initial descriptions of the fall armyworm invasion, including the near absence of the strain that prefers rice, millet, and pasture grasses, while providing additional evidence that the magnitude and extent of FAW natural migration on the continent is more limited than expected. The results also show that a second entry of fall armyworm likely occurred in western Africa from a source different than that of the original introduction. These findings indicate that western Africa continues to be at high risk of future introductions of FAW, which could complicate mitigation efforts.

The fall armyworm strain associated with most rice, millet, and pasture infestations in the Western Hemisphere is rare or absent in Ghana and Togo.

The moth pest fall armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) is now present throughout much of the Eastern Hemisphere where it poses a significant economic threat to a number of crops. Native to the Western Hemisphere, fall armyworm is one of the primary pests of corn in the Americas and periodically causes significant economic damage to sorghum, millet, cotton, rice, and forage grasses. This broad host range is in part the result of two populations historically designated as host strains (C-strain and R-strain) that differ in their host plant preferences. Reports of infestations in Africa have to date mostly been limited to the C-strain preferred crops of corn and sorghum, with little evidence of an R-strain presence. However, this could reflect a bias in monitoring intensity, with the R-strain perhaps being more prevalent in other crop systems that have not been as routinely examined for the pest. Because knowledge of whether and to what extent both strains are present is critical to assessments of crops at immediate risk, we analyzed specimens obtained from a systematic survey of pasture grass and rice fields, habitats typically preferred by the R-strain, done contemporaneously with collections from corn fields in Ghana and Togo. Substantial larval infestations were only observed in corn, while pheromone trap capture numbers were high only in corn and rice habitats. Little to no fall armyworm were found in the pasture setting. Comparisons with a meta-analysis of studies from South America identified differences in the pattern of strain-specific markers typically found in fall armyworm collected from rice habitats between the two hemispheres. Genetic tests of specimens from rice and corn area traps failed to show evidence of differential mating between strains. These results are consistent with the R-strain being rare or even absent in Africa and, at least for the Ghana-Togo area, this R-strain lack does not appear to be due to limitations in pest monitoring. The implications of these results to the crops at risk in Africa and the accuracy of existing molecular markers of strain identity are discussed.

Trapping Spodoptera frugiperda (Lepidoptera: Noctuidae) Moths in Different Crop Habitats in Togo and Ghana.

The economic impact of the invasion of Spodoptera frugiperda (J.E. Smith, Lepidoptera: Noctuidae) into Africa has so far been limited to maize agriculture but could potentially impact many other crops. Trapping based on pheromone lures provides a cost-effective method for detecting this important pest (commonly known as fall armyworm) and will be essential for large-scale monitoring of populations to determine its geographical distribution and migration behavior as the species equilibrates to its new environment. However, the effective use of pheromone trapping requires optimization for a given location. An earlier report demonstrated that two commercial lures (one 3-component and the other 4-component) that were effective for trapping S. frugiperda in maize fields in Togo, Africa. The current study extends these findings to agricultural areas that differ in plant host composition (maize, pasture grasses, rice, and sorghum) in multiple locations in Ghana and Togo. In two seasons, significantly higher numbers of moths were found in maize, and in one season, higher numbers were found in rice than in sorghum and pasture grass systems. The results confirm the effectiveness of pheromone trapping and identify pheromone lures and trapping methods best suited for the different agroecosystems common to West Africa and that are at risk of infestation by S. frugiperda.

Comparisons of fall armyworm haplotypes between the Galápagos Islands and mainland Ecuador indicate limited migration to and between islands.

The migration of the fall armyworm (Spodoptera frugiperda) is of topical interest because of its recent introduction and rapid dissemination throughout the Eastern Hemisphere. This study compares fall armyworm from island and mainland locations in Ecuador to estimate migration behavior. The Galápagos Islands is a province of Ecuador whose mainland coast lies approximately 1000 km to the west and is the closest major land mass. Air transport modeling indicates that natural migration from the mainland to the Galápagos is unlikely, suggesting that most, if not all, the introgressions of mainland fall armyworm into the Galápagos are occurring through trade-assisted transport in contaminated cargo, which is offloaded at the Galápagos port of entry in San Cristóbal island. Haplotype studies are consistent with this limited migration and further show divergence in the fall armyworm from San Cristóbal with those from the neighboring island of Santa Cruz despite their close proximity (less than 100 km distance) and favorable winds for inter-island flights. These observations indicate that water poses a significant barrier for moth migration in this region, with human-assisted transport probably playing a more important role than natural migration.

The genetic characterization of fall armyworm populations in Ecuador and its implications to migration and pest management in the northern regions of South America.

The fall armyworm (Spodoptera frugiperda) is a moth pest native to the Western Hemisphere that has recently become a global problem, invading Africa, Asia, and Australia. The species has a broad host range, long-distance migration capability, and a propensity for the generation of pesticide resistance traits that make it a formidable invasive threat and a difficult pest to control. While fall armyworm migration has been extensively studied in North America, where annual migrations of thousands of kilometers are the norm, migration patterns in South America are less understood. As a first step to address this issue we have been genetically characterizing fall armyworm populations in Ecuador, a country in the northern portion of South America that has not been extensively surveyed for this pest. These studies confirm and extend past findings indicating similarities in the fall armyworm populations from Ecuador, Trinidad-Tobago, Peru, and Bolivia that suggest substantial migratory interactions. Specifically, we found that populations throughout Ecuador are genetically homogeneous, indicating that the Andes mountain range is not a long-term barrier to fall armyworm migration. Quantification of genetic variation in an intron sequence describe patterns of similarity between fall armyworm from different locations in South America with implications for how migration might be occurring. In addition, we unexpectedly found these observations only apply to one subset of fall armyworm (the C-strain), as the other group (R-strain) was not present in Ecuador. The results suggest differences in migration behavior between fall armyworm groups in South America that appear to be related to differences in host plant preferences.

Southeastern Asia fall armyworms are closely related to populations in Africa and India, consistent with common origin and recent migration.

The discovery of fall armyworm, a native of the Western Hemisphere, in western Africa in 2016 was rapidly followed by detections throughout sub-Saharan Africa, India, and most recently southeastern Asia. This moth pest has a broad host range that threatens such important crops as corn, rice, millet, and sorghum, creating concern for its potential impact on agriculture in the Eastern Hemisphere. Although genetic data suggest populations sampled in Africa and India originate from a recent common source, it is not known whether this is the case for populations in southeastern Asia, nor whether the subgroup with a preference for rice and millet is present in the region. This study found through comparisons of genetic markers that the fall armyworm from Myanmar and southern China are closely related to those from Africa and India, suggesting a common origin for these geographically distant populations. The results are consistent with a single recent introduction into the Eastern Hemisphere followed by rapid dispersion. The molecular similarities include discrepancies between the genetic markers that brings into question whether the subpopulation most likely to be a threat to rice and millet is present in significant numbers in Asia.

Genetic characterization of fall armyworm (Spodoptera frugiperda) in Ecuador and comparisons with regional populations identify likely migratory relationships.

Fall armyworm, Spodoptera frugiperda (J. E. Smith), is an important agricultural pest native to the Americas that has recently been introduced into the Eastern Hemisphere where it has spread rapidly through most of Africa and much of Asia. The long-term economic consequences of this invasion will depend on how the species and important subpopulations become distributed upon reaching equilibrium, which is expected to be influenced by a number of factors including climate, geography, agricultural practices, and seasonal winds, among others. Much of our understanding of fall armyworm movements have come from mapping genetically defined subpopulations in the Western Hemisphere, particularly in North America where annual long-distance migrations of thousands of kilometers have been documented and modeled. In contrast, fall armyworm mapping in much of the rest of the hemisphere is relatively incomplete, with the northern portion of South America particularly lacking despite its potential importance for understanding fall armyworm migration patterns. Here we describe the first genetic description of fall armyworm infesting corn in Ecuador, which lies near a likely migration conduit based on the location of regional trade winds. The results were compared with populations from corn habitats in select locations in the Caribbean and South America to investigate the possible migratory relationship between these populations and was further assessed with respect to prevailing wind patterns and the distribution of locations with climate favorable for fall armyworm population establishment and growth.

Genetic comparisons of fall armyworm populations from 11 countries spanning sub-Saharan Africa provide insights into strain composition and migratory behaviors.

The recent discovery of fall armyworm (Spodoptera frugiperda, J.E. Smith) in Africa presents a significant threat to that continent's food security. The species exhibits several traits in the Western Hemisphere that if transferred to Africa would significantly complicate control efforts. These include a broad host range, long-distance migratory behavior, and resistance to multiple pesticides that varies by regional population. Therefore, determining which fall armyworm subpopulations are present in Africa could have important implications for risk assessments and mitigation efforts. The current study is an extension of earlier surveys that together combine the collections from 11 nations to produce the first genetic description of fall armyworm populations spanning the sub-Saharan region. Comparisons of haplotype frequencies indicate significant differences between geographically distant populations. The haplotype profile from all locations continue to identify Florida and the Caribbean regions as the most likely Western Hemisphere origins of the African infestations. The current data confirm the uncertainty of fall armyworm strain identification in Africa by genetic methods, with the possibility discussed that the African infestation may represent a novel interstrain hybrid population of potentially uncertain behavioral characteristics.

Genetic characterization of fall armyworm infesting South Africa and India indicate recent introduction from a common source population.

The invasion of the Western Hemisphere native fall armyworm (Spodoptera frugiperda; J. E. Smith) (Lepidoptera: Noctuidae) into the Eastern Hemisphere has been notable for the rapidity and geographical breadth of new detections. In the year following the first discovery in western sub-Saharan Africa in 2016, infestations have been documented in most sub-Saharan maize growing regions and has now expanded beyond Africa with populations recently reported in India. These observations could indicate a remarkable capacity for rapid establishment and long-distance dissemination. However, while fall armyworm does exhibit extended migration in North America where it annually traverses thousands of kilometers, this behavior is known to be dependent on highly favorable wind patterns and so can't be assumed to occur in all locations. An alternative possibility is that the species has long been present in Africa, and perhaps the rest of the hemisphere, but was undetected until the enhanced monitoring that resulted after its initial discovery. Determining whether the fall armyworm in the Eastern Hemisphere is newly arrived or long pre-existing is important for assessing the risks of significant economic impacts, as the former indicates a change in pest composition while the latter does not. This study examined this issue by comparing collections from two geographically distant locations, South Africa and India. Sequence comparisons were used to quantify differences between the South Africa and India collections, assess the likelihood of their sharing a common source population, and their possible relationship with previously characterized fall armyworm from other regions of Africa. The results indicate genetic homogeneity between the South African and Indian fall armyworm populations tested and substantial similarities between these and collections from eastern Africa. The implications of these findings on fall armyworm population behavior and composition are discussed.

Evidence that a major subpopulation of fall armyworm found in the Western Hemisphere is rare or absent in Africa, which may limit the range of crops at risk of infestation.

The introduction and establishment of fall armyworm (Spodoptera frugiperda) in Africa presents a major threat to agriculture in that continent and potentially to the entire Eastern Hemisphere. The species is subdivided into two subpopulations called the R-strain and C-strain that differ in their distribution on different plant hosts. This means that the scope of the economic risk posed by invasive fall armyworm is influenced by whether one or both strains are present. Multiple studies have found mitochondrial markers diagnostic of the two strains throughout Africa but there is substantial disagreement with a nuclear strain marker that makes conclusions about strain composition uncertain. In this study the issue of whether both strains are present in Africa was tested by an assay that can detect strain-biased mating behaviors. Western Hemisphere fall armyworm consistently showed evidence of strain-specific assortative mating in the field that was not found in surveys from multiple locations in Africa. The absence of strain mating biases and the disagreements between the strain diagnostic genetic markers indicates that the R-strain is rare (<1% of the population) or absent in Africa. Instead, it appears that the African fall armyworm populations are dominated by two groups, the C-strain and the descendants of interstrain hybrids. These results suggest that plant hosts associated with the R-strain may not be at high risk of fall armyworm infestation in Africa.

Documenting Potential Sunn Hemp (Crotalaria juncea L.) (Fabaceae) Pollinators in Florida.

Sunn hemp, Crotalaria juncea L., is a warm-season legume that can be planted in rotation to cash crops to add nitrogen and organic matter to the soils, for weed growth prevention, and to suppress nematode populations. Sunn hemp flowers also provide nectar and pollen for pollinators and enhance biological control by furnishing habitat for natural enemies. Experiments were conducted in Northern and North Central Florida to evaluate bee populations that visited flowers within mixed plots of sunn hemp and sorghum-sudangrass and plots of two sunn hemp germplasm lines. Collections of bees that visited 'AU Golden' and Tillage Sunn flowers indicated that Xylocopa virginica (L.) (Hymenoptera: Apidae), Xylocopa micans Lepeletier (Hymenoptera: Apidae), Megachile sculpturalis Smith (Hymenoptera: Megachilidae), Megachile mendica (Cresson) (Hymenoptera: Megachilidae), and Megachile georgica Cresson (Hymenoptera: Megachilidae) were present in large numbers in May through July and then again in October. Although Tillage Sunn seeds planted in March flowered in May, percent bloom and number of bee visits were low. Compared with short day sunn hemp cultivars, 'AU Golden' plants produced flowers early in the season to provide food and habitat for pollinators and have the potential to produce an abundant seed crop in Northern and North Central Florida.

Analysis of strain distribution, migratory potential, and invasion history of fall armyworm populations in northern Sub-Saharan Africa.

Fall armyworm (Spodoptera frugiperda J.E. Smith) is a noctuid moth pest endemic throughout the Western Hemisphere that has recently become widespread in sub-Saharan Africa. There is a strong expectation of significant damage to African maize crop yield and a high likelihood of further dispersal, putting the rest of the Eastern Hemisphere at risk. Specimens from multiple locations in six countries spanning the northern portion of the infested region were analyzed for genetic markers. The similarity of haplotypes between the African collections was consistent with a common origin, but significant differences in the relative frequency of the haplotypes indicated limitations in migration. The mitochondrial marker frequently used to identify two host strains appears to be compromised, making uncertain previous reports that both strains are present in Africa. This more extensive study confirmed initial indications based on Togo populations that Florida and the Greater Antilles are the likely source of at least a subset of the African infestation and further suggest an entry point in western Africa. The origin of a second subgroup is less clear as it was rarely found in the collections and has a haplotype that has not yet been observed in the Western Hemisphere.