Americanum Populations Research Articles

Feeding and reproductive parameters of adult female Ixodes scapularis (Acari: Ixodidae) and Amblyomma americanum parasitizing white-tailed deer (Odocoileus virginianus).

White-tailed deer Odocoileus virginianus (Zimmermann) (Artiodactyla: Cervidae) are the main host for adult Ixodes scapularis Say (Acari: Ixodidae) (blacklegged tick) and all stages of Amblyomma americanum Linnaeus (Acari: Ixodidae) (lone star tick). However, literature describing the feeding and reproductive parameters of these tick species when feeding on this host is limited. We experimentally infested white-tailed deer with adult pairs of either I. scapularis or A. americanum to improve our understanding of these tick-host relationships. Our study used tick-naïve white-tailed deer and restricted host grooming throughout the infestation. For I. scapularis, the days to repletion (mean ± SE, 6.04 ± 0.07), engorgement weight of replete females (0.20 ± 0.0032 g), duration of oviposition (32 ± 0.45 d), egg mass weight (0.10 ± 0.0027 g), and number of eggs laid per tick (1,803.00 ± 49.00) were recorded. Data from A. americanum were also recorded, including days to repletion (11.00 ± 0.063), engorgement weight of replete females (0.63 ± 0.025 g), duration of oviposition (37.00 ± 1.30 d), egg mass weight (0.34 ± 0.017 g), and number of eggs laid per tick (5,873.00 ± 291.00). These biological parameter data could be used as variables in models (e.g., LYMESIM 2.0) to determine how white-tailed deer influence I. scapularis and A. americanum populations in nature, and to evaluate the protective efficacy of tick-antigen-based antitick vaccines.

Identification and reproduction of dagger nematode, Xiphinema americanum, in potato

Summary Plant-parasitic nematodes limit potato production by feeding on roots, reducing tuber yield and quality, and resulting in poor growth. Dagger nematodes (Xiphinema spp.) can pose a significant threat to crop production even in low numbers. Dagger nematodes have been reported in North Dakota, USA, potato fields. In this study, a dagger nematode species was identified and its reproduction potential in 21 potato cultivars was evaluated. The dagger nematode was identified as X. americanum via morphometric measurements and DNA sequence analysis. At the molecular level, D2-D3 of 28S ribosomal RNA (rRNA) and internal transcribed spacer (ITS)-regions of rDNA were targeted. Sequencing data of D2-D3 (749 bp) showed 98.18% similarity with three isolates of X. americanum. ITS sequence (860 bp) analysis showed 97.68% similarity with an isolate of X. americanum but lower similarity with other species of Xiphinema. The phylogenetic analysis of ITS region further confirmed the species of dagger nematode as X. americanum. Using naturally infested soil, two glasshouse experiments were conducted to assess the reproduction rate of X. americanum on 21 potato cultivars belonging to four distinct market potato classes (yellow, red, white and russet). The reproduction rates were found to vary among the potato classes and cultivars. Numerically, X. americanum population density was higher in ‘Soraya’, whereas ‘Ranger Russet’, ‘Manistee’, ‘Kennebec’, ‘Russet Norkotah 278’, ‘Modoc’, ‘Pomerelle Russet’ and ‘Dakota Rose’ reduced nematode reproduction in both trials. This is the first study demonstrating the reproduction of X. americanum in potato cultivars and could be helpful to manage dagger nematodes.

Establishment of Amblyomma americanum populations and new records of Borrelia burgdorferi-infected Ixodes scapularis in South Dakota.

Tick-borne diseases are an emerging public health threat in the United States, but surveillance is lacking in some regions. To advance current knowledge of the ecology of ticks and tick-borne diseases in South Dakota, we conducted a survey in the summer of 2019, focusing on the eastern counties of the state. We collected and identified 266 ticks and a subset were tested for the presence of Borrelia burgdorferi by polymerase chain reaction (PCR). Dermacentor variabilis, a ubiquitous species in the state, was the most commonly identified tick, present in all counties surveyed. However, we also identified 15 Amblyomma americanum from three different locations, providing the first evidence of established populations in the state and expanding the range of this species. In addition, we identified 22 Ixodes scapularis from five different locations, confirming a previous report of an established population in the state. Two adult I. scapularis from two different sites were found to harbor B. burgdorferi, including an individual from Lincoln County, suggesting the ongoing presence of the pathogen in tick populations in the state and representing its southwestern-most detection in the midwest United States. These findings provide important information for assessing and monitoring the public health risk from tick-borne diseases in an area where surveillance is lacking.

Effective control of the motile stages of Amblyomma americanum and reduced Ehrlichia spp. prevalence in adults via permethrin treatment of white-tailed deer in coastal Connecticut, USA

The lone star tick, Amblyomma americanum, is a common human-biting species whose range has been largely restricted to the southeastern United States, until recent detections of established populations on Long Island, New York and throughout coastal southern New England. We evaluated the effectiveness of topical treatment of 10 % permethrin delivered via 4-poster devices to white-tailed deer, Odocoileus virginianus, in the management of a newly discovered A. americanum population in Norwalk, Connecticut. Using a high-density deployment of one 4-poster device/12.7 ha, we were successful in significantly reducing densities of host-seeking adults (93 % reduction), nymphs (92 %), and larvae (96 %) from 2018 to 2020. We also documented a significant reduction (87 %) in parasitizing adults and nymphs on white-tailed deer from 2018 to 2019. The prevalence of Ehrlichia chaffeensis and Ehrlichia ewingii combined in host-seeking adults declined significantly from 47 % at the time the A. americanum population was discovered in 2017 to 7% in 2020. However, the prevalence in nymphs remained static (∼9%) throughout the study period. These data demonstrate that, when properly deployed in a density-dependent manner in terms of deer abundance, 4-poster devices can effectively manage parasitizing and host-seeking A. americanum populations and reduce the prevalence of two ehrlichial species of public health importance.

Field Performance of Winegrape Rootstocks and Fumigation during Establishment of a Chardonnay Vineyard in Washington

In Washington, most winegrapes are own-rooted Vitis vinifera, which is susceptible to the plant-parasitic nematodes Meloidogyne hapla and Xiphinema americanum. Using resistant rootstocks to manage nematodes has not been evaluated in Washington vineyards. A long-term vineyard trial was established to evaluate the effects of soil fumigation and rootstock genotype on M. hapla and X. americanum population dynamics and vine growth during vineyard establishment (first three years) in a replant scenario. Vines in an existing V. vinifera ‘Chardonnay’ vineyard were first treated with foliar glyphosate in fall 2014. Randomized areas within the vineyard were then either fumigated or not with drip-applied metam sodium. Following fumigation, vines were removed. In spring 2015, the vineyard was replanted to Chardonnay on the following rootstocks: 1103 Paulsen, 101-14 Millardet et de Grasset, Teleki 5C, and Harmony. Self-grafted and own-rooted Chardonnay were included. Fumigation reduced M. hapla soil second-stage juvenile (J2) population densities on own-rooted and self-grafted vines for only the first year after fumigation. One year after fumigation, the self-grafted and own-rooted vines had higher population densities of M. hapla J2 than rootstocks. All rootstocks supported measurable densities of M. hapla J2 but were poor hosts relative to V. vinifera. Fumigation effectively reduced population densities of X. americanum for up to 3.5 years. Fumigation also reduced early establishment pruning weights. Vines grown in fumigated areas had lower pruning weights through year 2; but rootstock was the bigger influence on pruning weights by year 3. This trial demonstrates that rootstocks have a more sustained impact on nematode re-establishment and subsequent vine health in a vineyard replant scenario than that of pre-plant fumigation.

Comparative population genetics of Amblyomma maculatum and Amblyomma americanum in the mid-Atlantic United States

The Gulf Coast tick, Amblyomma maculatum, is undergoing a northward expansion along the United States East Coast, most recently establishing populations in Virginia, Maryland, and Delaware. This expansion has human health implications, as A. maculatum is the primary natural vector of the bacterium Rickettsia parkeri, which causes a spotted fever-type rickettsiosis. Newly established populations of A. maculatum in Virginia tend to have high prevalence of R. parkeri, compared to lower infection rates in the historical range. The factors contributing to high R. parkeri prevalence in Virginia are not known. Investigating connectivity between sites colonized with A. maculatum can help determine whether sites with higher prevalence are isolated or well-connected through migration, thus serving as a source of infected individuals. We characterized 16S rRNA haplotypes of A. maculatum and, for comparison, the congeneric Amblyomma americanum collected from sites where these species co-occur. We then explored connectivity and genetic structure among Virginia populations using pairwise ΦST and AMOVA analyses. Our study identified one recently restored native grassland site with low A. maculatum haplotype diversity and strong evidence of a founder effect, whereas most sites are haplotypically diverse but with no clear genetic structure or connectivity between sites. These findings contrast with high connectivity and a slight mainland/island structure among A. americanum populations. Our results suggest that A. maculatum populations occasionally arise following long-distance drop-offs of few individual ticks in suitable habitat, but no clear migration patterns were observed. The distinct population genetic patterns between species might result from differences in host utilization.

Evaluation of Fipronil Susceptibility in the Lone Star Tick (Acari: Ixodidae).

The lone star tick, Amblyomma americanum Linnaeus, is ubiquitously present in the southeastern United States and will readily parasitize humans and companion animals. Fipronil is the active ingredient in several topically applied products used to manage ticks and fleas on companion animals. Reducing ectoparasite infestations on companion animals decreases the risk that pathogens vectored by these pests are transmitted to these animals and their owners. However, dependence on acaricides can promote the development of resistance. In this study, the Food and Agriculture Organization of the United Nations larval packet test was used to determine the lethal concentration (LC) values and discriminating concentration (DC) for fipronil in the lone star tick. The DC was calculated as 0.02080%. The low magnitude of the DC value suggests that fipronil is an effective active ingredient for A. americanum management. With the LC and DC values determined, emergent resistance can be monitored, potentially allowing for intervention should tolerance develop in A. americanum populations that are in frequent contact with humans and their companion animals.

Predicted Northward Expansion of the Geographic Range of the Tick Vector Amblyomma americanum in North America under Future Climate Conditions.

Background:The geographic range of the tick Amblyomma americanum, a vector of diseases of public health significance such as ehrlichiosis, has expanded from the southeast of the United States northward during the 20th century. Recently, populations of this tick have been reported to be present close to the Canadian border in Michigan and New York states, but established populations are not known in Canada. Previous research suggests that changing temperature patterns with climate change may influence tick life cycles and permit northward range expansion of ticks in the northern hemisphere.Objectives:We aimed to estimate minimal temperature conditions for survival of A. americanum populations at the northern edge of the tick’s range and to investigate the possibility of range expansion of A. americanum into northern U.S. states and southern Canada in the coming decades.Methods:A simulation model of the tick A. americanum was used, via simulations using climate data from meteorological stations in the United States and Canada, to estimate minimal temperature conditions for survival of A. americanum populations at the northern edge of the tick’s range.Results:The predicted geographic scope of temperature suitability [ annual cumulative degree days (DD) ] included most of the central and eastern U.S. states east of longitude 110°W, which is consistent with current surveillance data for the presence of the tick in this region, as well as parts of southern Quebec and Ontario in Canada. Regional climate model output raises the possibility of northward range expansion into all provinces of Canada from Alberta to Newfoundland and Labrador during the coming decades, with the greatest northward range expansion (up to by the year 2100) occurring under the greenhouse gas (GHG) emissions of Representative Concentration Pathway (RCP) 8.5. Predicted northward range expansion was reduced by approximately half under the reduced GHG emissions of RCP4.5.Discussion:Our results raise the possibility of range expansion of A. americanum into northern U.S. states and southern Canada in the coming decades, and conclude that surveillance for this tick, and the diseases it transmits, would be prudent. https://doi.org/10.1289/EHP5668

Discovery of filarial nematode DNA in Amblyomma americanum in Northern Virginia

Ticks collected in 2011 were screened for the presence of filarial nematode genetic material, and positive samples were sequenced for analysis. Monanema-like filarial nematode DNA was recently discovered in Amblyomma americanum in northern Virginia, marking the first time genetic material from this parasite has been discovered in ticks in the state. Phylogenetic analysis revealed that this material was directly related to a previously discovered filarial nematode in A. americanum populations in Maryland as well as recently identified parasites in Ixodes scapularis from southern Connecticut. Further study is warranted to visually confirm the presence of these nematodes, characterize their distribution, and determine if these ticks are intermediate hosts.

A Dynamic Population Model to Investigate Effects of Climate and Climate-Independent Factors on the Lifecycle of Amblyomma americanum (Acari: Ixodidae).

The lone star tick, Amblyomma americanum, is a disease vector of significance for human and animal health throughout much of the eastern United States. To model the potential effects of climate change on this tick, a better understanding is needed of the relative roles of temperature-dependent and temperature-independent (day-length-dependent behavioral or morphogenetic diapause) processes acting on the tick lifecycle. In this study, we explored the roles of these processes by simulating seasonal activity patterns using models with site-specific temperature and day-length-dependent processes. We first modeled the transitions from engorged larvae to feeding nymphs, engorged nymphs to feeding adults, and engorged adult females to feeding larvae. The simulated seasonal patterns were compared against field observations at three locations in United States. Simulations suggested that 1) during the larva-to-nymph transition, some larvae undergo no diapause while others undergo morphogenetic diapause of engorged larvae; 2) molted adults undergo behavioral diapause during the transition from nymph-to-adult; and 3) there is no diapause during the adult-to-larva transition. A model constructed to simulate the full lifecycle of A. americanum successfully predicted observed tick activity at the three U.S. study locations. Some differences between observed and simulated seasonality patterns were observed, however, identifying the need for research to refine some model parameters. In simulations run using temperature data for Montreal, deterministic die-out of A. americanum populations did not occur, suggesting the possibility that current climate in parts of southern Canada is suitable for survival and reproduction of this tick.

The mitochondrial genome of the lone star tick (Amblyomma americanum)

Amblyomma americanum is an abundant tick in the southeastern, midwestern, and northeastern United States. It is a vector of multiple diseases, but limited genomic resources are available for it. We sequenced the complete mitochondrial genome of a single female A. americanum collected in Georgia using the Illumina platform. The consensus sequence was 14,709bp long, and the mean coverage across the assembly was >12,000×. All expected tick genomic features were present, including two “Tick-Box” motifs, and in the expected order for the Metastriata. Heteroplasmy rates were low compared to the most closely related tick for which data are available, Amblyomma cajennense. The phylogeny derived from the concatenated protein coding and rRNA genes from the 33 available tick mitochondrial genomes was consistent with those previously proposed for the Acari. This is the first complete mitochondrial sequence for A. americanum, which provides a useful reference for future studies of A. americanum population genetics and tick phylogeny.

Microsymbionts of Phaseolus vulgaris in acid and alkaline soils of Mexico

In order to investigate bean-nodulating rhizobia in different types of soil, 41 nodule isolates from acid and alkaline soils in Mexico were characterized. Based upon the phylogenetic studies of 16S rRNA, atpD, glnII, recA, rpoB, gyrB, nifH and nodC genes, the isolates originating from acid soils were identified as the phaseoli symbiovar of the Rhizobium leguminosarum-like group and Rhizobium grahamii, whereas the isolates from alkaline soils were defined as Ensifer americanum sv. mediterranense and Rhizobium radiobacter. The isolates of “R. leguminosarum” and E. americanum harbored nodC and nifH genes, but the symbiotic genes were not detected in the four isolates of the other two species. It was the first time that “R. leguminosarum” and E. americanum have been reported as bean-nodulating bacteria in Mexico. The high similarity of symbiotic genes in the Rhizobium and Ensifer populations showed that these genes had the same origin and have diversified recently in different rhizobial species. Phenotypic characterization revealed that the “R. leguminosarum” population was more adapted to the acid and low salinity conditions, while the E. americanum population preferred alkaline conditions. The findings of this study have improved the knowledge of the diversity, geographic distribution and evolution of bean-nodulating rhizobia in Mexico.

Prevalence of Ehrlichia chaffeensis and Ehrlichia ewingii in Amblyomma americanum and Dermacentor variabilis collected from southeastern Virginia, 2010–2011

Amblyomma americanum is the most commonly encountered tick species in southeastern Virginia, representing approximately 95% of the human-biting tick population in this area. Here we investigated the prevalence of Ehrlichia chaffeensis and Ehrlichia ewingii in questing Amblyomma americanum and Dermacentor variabilis ticks collected from multiple sites in southeastern Virginia from 2010 to 2011. Although both Ehrlichia species were detected in Amblyomma americanum, no evidence of either pathogen was found in Dermacentor variabilis. Prevalence of E. chaffeensis varied by location, ranging from 0 to 5.08% among Amblyomma americanum populations. Ehrlichia ewingii prevalence was slightly higher, ranging from 0 to 8.20% among A. americanum populations. We conclude that both pathogens are established in southeastern Virginia A. americanum populations, and that although there are no apparent temporal trends in Ehrlichia prevalence, there is variation among locations, suggesting the potential for disease hotspots.

Relationship between habitat type, fire frequency, andAmblyomma americanumpopulations in east-central Alabama

Ticks were collected from 20 sites in the Calhoun, Cherokee, and Cleburne Counties in east-central Alabama areas to determine the relationship between plant physiognomy, environmental variables, and tick populations. Sites investigated included various burning regimes, wildland-urban-interface (WUI), a college campus, and an unmanaged area. Amblyomma americanum (L.) (Acari: Ixodidae) dominated the tick population while Ixodes scapularis Say was not encountered. There were complex differences in tick populations among site conditions. After prescribed burning, the tick population size was small but was larger in subsequent 2- and 5-year post-burn sites. An increase in Odocoileus virginianus foraging in recently burned sites is likely responsible for this phenomenon. WUI areas had the largest tick populations likely due to Odocoileus virginianus activity in an area that provides cover, forage, and a connection to a wildlife refuge. It is possible that the likelihood of humans coming in contact with ticks and tick-borne diseases is greater in WUI areas than in unbroken contiguous forest. A. americanum showed a positive correlation with percent cover of grass and leaf litter mass and a negative relationship with pine sapling density. Variables expected to be strongly correlated with A. americanum populations such as soil moisture, canopy closure, and tree density were found to have weak correlations.

Distribution and infection frequency of ‘Candidatus Rickettsia amblyommii’ in Maryland populations of the lone star tick (Amblyomma americanum) and culture in an Anopheles gambiae mosquito cell line

Amblyomma americanum (the lone star tick) is a broadly distributed tick that transmits multiple pathogens of humans and domestic animals. ‘Candidatus Rickettsia amblyommii’ is a spotted-fever group rickettsial species that is potentially associated with human disease. In 2008 and 2009, we assayed over 500 unfed adult ticks from 19 Maryland populations for the presence of ‘Candidatus R. amblyommii’. Infection frequencies ranged from 33% to 100%, with an average infection rate of 60% in 2008 and 69% in 2009. Infection frequencies did not differ statistically between sexes. To develop a system in which to study ‘Candidatus R. amblyommii’ in the laboratory, we used a cell line developed from Anopheles gambiae mosquitoes (Sua5B) to isolate and culture ‘Candidatus R. amblyommii’ from field-collected A. americanum ticks from 2 localities in Maryland. After infection, Sua5B cells were infected for more than 40 passages. Infection was confirmed by Rickettsia-specific PCR, gene sequencing, and Rickettsia-specific fluorescence in situ hybridization (FISH). These data show that ‘Candidatus R. amblyommii’ is widespread in Maryland A. americanum populations and that Sua5B cells are a useful tool for culturing Rickettsia infections from wild ticks.

Distribution and molecular characterization of Wolbachia endosymbionts and filarial nematodes in Maryland populations of the lone star tick (Amblyomma americanum)

The lone star tick Amblyomma americanum is host to a wide diversity of endosymbiotic bacteria. We identified a novel Wolbachia symbiont infecting A. americanum. Multilocus sequence typing phylogenetically placed the endosymbiont in the increasingly diverse F supergroup. We assayed a total of 1031 ticks (119 females, 78 males and 834 nymphs in 89 pools) from 16 Maryland populations for infection. Infection frequencies in the natural populations were approximately 5% in females and <2% (minimum infection rate) in nymphs; infection was not detected in males. Infected populations were only observed in southern Maryland, suggesting the possibility that Wolbachia is currently invading Maryland A. americanum populations. Because F supergroup Wolbachia have been detected previously in filarial nematodes, tick samples were assayed for nematodes by PCR. Filarial nematodes were detected in 70% and 9% of Wolbachia-positive and Wolbachia-negative tick samples, respectively. While nematodes were more common in Wolbachia-positive tick samples, the lack of a strict infection concordance (Wolbachia-positive, nematode-negative and Wolbachia-negative, nematode-positive ticks) suggests that Wolbachia prevalence in ticks is not due to nematode infection. Supporting this hypothesis, phylogenetic analysis indicated that the nematodes were likely a novel species within the genus Acanthocheilonema, which has been previously shown to be Wolbachia-free.

Spatial distribution of Crinum americanum L. in tropical blind estuary: Hydrologic, edaphic and biotic drivers

The competitive abilities of a given species are inversely proportional to its tolerance to environmental stress. Thus, in estuaries, vegetation is generally controlled by salinity and flooding in their lower limits, and by biotic drivers in their upper limits. Crinum americanum L. is vastly distributed over flooded regions of American seacoast, frequently associated with stressful habitats. We aimed to explain the role of hydrologic, edaphic, and biotic drivers in the distribution of this species on the Massaguaçu River estuary, Southeastern Brazil (23°37′20″S and 54°21′25″W). We sampled randomly 400 plots in the estuary, and registered covering of all species, the height of the C. americanum individuals, and the relative height of the plots. We collected soil samples from every five plots. We measured the estuary level daily for two years. We used Correspondence Analysis, Simple and Canonical, and graphic analysis. The salinity has explained the major part of the observed pattern, and the C. americanum population was positively related to it. The estuary level was also important. C. americanum has presented higher densities in intermediary flooding classes than in the extremes of the gradient. Species reduction in regions of low salinity or its absence has probably been due to the highly competitive environment, and not to the lack of salt per se.

Population Genetics ofAmblyomma americanum(Acari: Ixodidae) Collected From Arkansas

Lone star ticks, Amblyomma americanum L. (Acari: Ixodidae), infest multiple hosts such as birds, and mammals of various sizes (rodents to white-tailed deer) and can harbor human pathogens such as Borrelia lonestari and Ehrlichiosis chaffeensis. The population structure of 251 A. americanum ticks, collected from canines and two white-tailed deer in six Arkansas ecoregions, was examined using DNA sequences of a 247-bp region of the mitochondrial DNA ribosomal RNA 16S gene. Of the 247 nucleotide characters, 26 were variable. Thirty-three haplotypes were identified of which 25 haplotypes occurred once (10%). The most common haplotype was Aa25, occurring in 60% of the samples and found in all six ecoregions. The excess of low frequency haplotypes combined with the overall negative Tajima’s D and Fu and Li statistics suggests population expansion. Phylogenetic relationships of the 33 A. americanum haplotypes were constructed with other Amblyomma species and identified A. americanum as a monophyletic species with two groups. The patterns of high nucleotide and haplotype diversity found in this study suggests that the A. americanum population is expanding perhaps due to its ability to survive in a variety of habitats and feed on multiple hosts. Given the gene flow in Arkansas, the spread of acaricide resistance and pathogens may be rapid.

Population Genetics of &lt;I&gt;Amblyomma americanum&lt;/I&gt; (Acari: Ixodidae) Collected From Arkansas

Lone star ticks, Amblyomma americanum L. (Acari: Ixodidae),infest multiple hosts such as birds, and mammals of various sizes (rodents to white-tailed deer) and can harbor human pathogens such as Borrelia lonestari and Ehrlichiosis chaffeensis. The population structure of 251 A. americanum ticks, collected from canines and two white-tailed deer in six Arkansas ecoregions, was examined using DNA sequences of a 247-bp region of the mitochondrial DNA ribosomal RNA 16S gene. Of the 247 nucleotide characters, 26 were variable. Thirty-three haplotypes were identified of which 25 haplotypes occurred once (10%). The most common haplotype was Aa25, occurring in 60% of the samples and found in all six ecoregions. The excess of low frequency haplotypes combined with the overall negative Tajima's D and Fu and Li statistics suggests population expansion. Phylogenetic relationships of the 33 A. americanum haplotypes were constructed with other Amblyomma species and identified A. americanum as a monophyletic species with two groups. The patterns of high nucleotide and haplotype diversity found in this study suggests that the A. americanum population is expanding perhaps due to its ability to survive in a variety of habitats and feed on multiple hosts. Given the gene flow in Arkansas, the spread of acaricide resistance and pathogens may be rapid.

Chronoclinal variation and sexual dimorphism in Mammut americanum (American mastodon) from the Pleistocene of Florida

This study quantitatively and statistically analyzes chronoclinal variation in Mammut americanum fossils over a broad temporal range (~1. 5 Ma to 10,000 year BP). A large sample of elements (third molars, mandibles, limb bones) from the Irvingtonian and Rancholabrean of Florida were measured and statistically compared for significant differences. Rancholabrean elements are generally found to be significantly larger than Irvingtonian elements. Florida M. americanum shows directional selection for increased body mass from the Irvingtonian to the Rancholabrean. Anterior mandibular height is the only measurement that demonstrates significant sexual dimorphism in Rancholabrean individuals, with males having a larger anterior mandibular height than females. Mandibular tusk presence/absence ratios. in Florida M. americanum show directional selection against these small tusks from the Irvingtonian to the Rancholabrean. M. americanum demonstrates a heterochronic increase in growth rate at the onset of puberty from the Irvingtonian to the Rancholabrean. In addition, Rancholabrean M. americanum populations also exhibit holobiotic heteromorphism, with males growing at a faster rate than females. The results of this study demonstrate that throughout its ~1.5 Ma range in Florida, M. americanum was not static in its evolution.