Technical note: An in vitro study of dental microwear formation using the BITE Master II chewing machine

Dental microwear has been used for decades to reconstruct the diets of fossil hominins and bioarchaeological populations. The basic theory has been that hard‐brittle foods (e.g., nuts, bone) require crushing and leave pits as they are pressed between opposing cheek‐tooth surfaces, whereas soft‐tough foods (e.g., grass blades, meat) require shearing and leave scratches as they are dragged along opposing surfaces that slide past one another. However, recent studies have called into question the efficacy of microwear as an indicator of diet. One issue has been the limited number of in vitro studies providing empirical evidence for associations between microwear pattern and chewing behavior. We here describe a new study using a chewing simulator, the BITE Master II, to examine the effects of angle of approach between opposing teeth and food consistency on microwear surface texture. Results indicate that opposing teeth that approach one another: 1) perpendicular to the occlusal plane (crushing) result in pits; 2) parallel to the occlusal plane (shearing) result in striations in the direction of movement; and 3) oblique to the occlusal plane (45°) result in both striations and pits. Results further suggest that different food types and abrasive loads affect the propensity to accumulate microwear features independent of feature shapes. Am J Phys Anthropol 158:769–775, 2015. © 2015 Wiley Periodicals, Inc.     

Technical note: Dental microwear textures of "Phase I" and "Phase II" facets

The power stroke of mastication has been traditionally divided into two parts, one which precedes centric occlusion, and the other which follows it-"Phase I" and "Phase II," respectively. Recent studies of primate mastication have called into question the role of Phase II in food processing, as they have found little muscle activity or accompanying bone strain following centric occlusion. That said, many researchers today look to Phase II facets to relate diet to patterns of dental microwear. This suggests the need to reevaluate microwear patterns on Phase I facets. Here we use texture analysis to compare and contrast microwear on facets representing both phases in three primate species with differing diets (Alouatta palliata, Cebus apella, and Lophocebus albigena). Results reaffirm that microwear patterns on Phase II facets better distinguish taxa with differing diets than do those on Phase I facets. Further, differences in microwear textures between facet types for a given taxon may themselves reflect diet. Some possible explanations for differences in microwear textures between facet types are proposed.     

Dietary inferences from dental occlusal microwear at Mission San Luis de Apalachee

San Luis de Apalachee, one of a chain of Roman Catholic missions established in Spanish Florida (modern states of Georgia and Florida) in the sixteenth and seventeenth centuries, was the principal center of missionization of native populations in the Florida panhandle. Paleoethnobotanical remains yield evidence of production of various kinds of crops at the site, typical of nearly all mission period sites in the area. Stable isotope (carbon and nitrogen) analysis and dental caries evidence suggest that maize was not as important in the diet at San Luis as in other contemporary settings in the region. Ethnohistorical research indicates a heavier reliance on meat consumption compared to other mission settings. This study examines dental microwear of occlusal surfaces of maxillary molars from San Luis and five other Native American mission period sites in Spanish Florida. Epoxy casts of molar crushing facets were photographed under 500x magnification, using a scanning electron microscope. Photomicrographs were digitized using Microware 4.02 (Ungar [2002a]) and statistically evaluated using one-way ANOVA and post hoc Tukey's multiple comparisons tests. These analyses reveal that the frequency of pitting on the San Luis molars is significantly greater than for other mission period sites. Consistent with bioarchaeological, historical, and archaeological documentation, these findings suggest that diets were different in the San Luis natives in comparison with the other native populations in Spanish Florida. Various dietary factors likely came into play, resulting in these differences, and may have included significantly greater meat consumption at San Luis.     

A study of microwear on chimpanzee molars: Implications for dental microwear analysis

Recent investigations of dental microwear have shown that such analyses may ultimately provide valuable information about the diets of fossil species. However, no background information about intraspecific variability of microwear patterns has been available until now. This study presents the results of an SEM survey of microwear patterns found on occlusal enamel of chimpanzee molars. Methods of pattern analysis are described. Selected sites on the occlusal surface included shearing, grinding, and puncture-crushing surfaces formed by both phases of the power stroke of mastication. The microwear patterns found in this sample of chimpanzees showed a high degree of regularity. However, certain parameters such as relative pit-to-striation frequencies, feature density, striation length, and pit diameter were significantly affected by facet type and molar position. Sex and age of individuals also influenced some microwear parameters, but due to the small sample size these findings are considered to be preliminary. These results show that microwear within a single species may vary because of factors that are due more to biomechanics than to diet. The study also supplies some metrical estimates of “normal” pattern variability due to functional and morphological influences. These estimates should provide a useful baseline for assessing the significance of microwear pattern differences that may be found between species of differing diets.     

MicroWeaR: A new R package for dental microwear analysis

Mastication of dietary items with different mechanical properties leaves distinctive microscopic marks on the surface of tooth enamel. The inspection of such marks (dental microwear analysis) is informative about the dietary habitus in fossil as well as in modern species. Dental microwear analysis relies on the morphology, abundance, direction, and distribution of these microscopic marks. We present a new freely available software implementation, MicroWeaR, that, compared to traditional dental microwear tools, allows more rapid, observer error free, and inexpensive quantification and classification of all the microscopic marks (also including for the first time different subtypes of scars). Classification parameters and graphical rendering of the output are fully settable by the user. MicroWeaR includes functions to (a) sample the marks, (b) classify features into categories as pits or scratches and then into their respective subcategories (large pits, coarse scratches, etc.), (c) generate an output table with summary information, and (d) obtain a visual surface‐map where marks are highlighted. We provide a tutorial to reproduce the steps required to perform microwear analysis and to test tool functionalities. Then, we present two case studies to illustrate how MicroWeaR works. The first regards a Miocene great ape obtained from through environmental scanning electron microscope, and other a Pleistocene cervid acquired by a stereomicroscope.     

Technical note: Enhancement of Scott's molar wear scoring method

A method is described for orienting maxillary and mandibular molars in order to standardize the reporting of wear scores on quadrants of the occlusal surfaces (Scott: Am J Phys Anthropol 51 (1979) 213–217). The method, which was developed on an archeological sample from ancient Mendes, Egypt, further requires that quadrant scores be reported individually and sequentially for each tooth, rather than summed, in order to identify more easily differential and directional wear patterns. Intraobserver and interobserver error was found to be negligible when the appropriate diagrams and instructions were consulted. Thus, observer error does not add further to the potential for error associated with Scott's original scoring method. Am J Phys Anthropol, 2010. © 2010 Wiley‐Liss, Inc.     

Testing hypotheses of dietary reconstruction from buccal dental microwear in Australopithecus afarensis

A recent study of occlusal microwear in Australopithecus afarensis described this species as an opportunistic dweller, living in both forested and open environments and greatly relying on fallback resources and using fewer food-processing activities than previously suggested. In the present study, analysis of buccal microwear variability in a sample of A. afarensis specimens (n = 75 teeth) showed no significant correlations with the ecological shift that took place around 3.5 Ma in Africa. These results are consistent with the occlusal microwear data available. In fact, significant correlations between buccal and occlusal microwear variables were found. However, comparison of the buccal microwear patterns showed clear similarities between A. afarensis and those hominoid species living in somewhat open environments, especially the Cameroon gorillas. A diet based mainly on succulent fruits and seasonal fallback resources would be consistent with the buccal microwear patterns observed.     

Deciduous dental microwear of prehistoric juveniles from the lower Illinois River valley

Scanning electron microscopy was used to study age-related changes in the dental microwear of 36 prehistoric juveniles ranging from 6 to 27 months of age. Juveniles from horticultural (Middle Woodland) and agricultural (Mississippian) groups were studied to allow an investigation of the impact of diet on deciduous microwear. Inclusion of both molars and incisors in the sample permitted identification of age at earliest appearance of wear and comparisons between the age-related microwear characterizing different tooth types. Data on feature frequency and enamel surface characteristics were analyzed. Microwear feature frequencies generally increase with age and/or exposure to wear. Enamel surface characteristics show consistent qualitative changes associated with both age and exposure to wear. Molars and incisors differ for such surface characteristics in a way that make biomechanical sense, given the relative bite forces characterizing these teeth. Dietary reconstruction based on deciduous microwear is complex because of the effects of both age and exposure to wear on feature frequencies and enamel surface characteristics. Nonetheless, the present analyses suggest that 1) diets differed for younger and older juveniles within each cultural group and 2) the Middle Woodland juvenile diet was both harder and more varied in physical consistency than the Mississippian juvenile diet.     

Diet reconstruction in antebellum Baltimore: Insights from dental microwear analysis

Construction in the City of Baltimore during 1996 led to the recovery of human skeletal remains dating from 1792 to 1856. Historical research indicates that the skeletal remains come from two adjacent graveyards: Christ's Church Episcopalian Cemetery and the Potters Field East. The different socioeconomic status of the internees in each cemetery suggests the possibility of marked contrasts in lifestyle, health, and diet. To shed further light on these possibilities, analyses of microscopic wear patterns on teeth, or dental microwear analyses, were undertaken. A sample from Spanish Florida was used to help interpret the results. Epoxy casts of incisor and molar teeth were placed in an SEM and photomicrographs of clean wear facets were taken. The photomicrographs were digitized using the software package Microware 4.02. Statistical analyses of rank transformed data consisted of single‐factor ANOVA, followed by post hoc tests. No significant differences were found between Christ's Church and Potters Field East samples for any of the variables examined in either molar or incisor teeth. However, differences between each Baltimore sample and the La Florida samples give suggestions of possible diet differences in antebellum Baltimore. The mosaic of differences between the Baltimore and La Florida samples probably reflects the wide variety of foods available to antebellum Baltimoreans as well as the relative lack of abrasives in their diet. Am J Phys Anthropol, 2010. © 2009 Wiley‐Liss, Inc.     

Quantitative analysis of dental microwear in threespine stickleback: a new approach to analysis of trophic ecology in aquatic vertebrates

1. The threespine stickleback Gasterosteus aculeatus is an important model organism in studies of genomic and phenotypic evolution, adaptation and speciation. Fossil Gasterosteus offer the potential to test models derived from studies of extant fishes over true evolutionary time-scales. Competition for food resources, for example, plays an important part in stickleback speciation, causing divergence in food gathering traits and ecological character displacement, but it is not possible to test this model in fossils because evidence of diet is almost never preserved. 2. We demonstrate here that quantitative analysis of dental microwear, a technique previously applied only to mammals, provides a reliable guide to the dietary preferences of stickleback. Teeth from stickleback raised under laboratory conditions exhibit microwear patterns that vary systematically according to substrate coarseness and whether fishes feed on Daphnia within the water column, or on chironomid larvae from the bottom. Furthermore, microwear data exhibit a progressive shift in their distribution that tracks differences in experimental feeding treatments. 3. Microwear in wild populations also exhibits a relationship with feeding. In blind assessments of trophic niche based on microwear patterns we were able to correctly assign all but one equivocal population to trophic group. Microwear data from wild stickleback exhibit a shift in distribution comparable with that observed across the range of treatments in the laboratory and these allow populations to be ranked according to the degree to which they approach fully benthic or fully limnetic feeding. 4. Our results demonstrate that microwear has the potential to be a powerful tool in the analysis of fish trophic ecology, particularly in the analysis of species pairs and niche differentiation. It has advantages over the trophic snapshot provided by analysis of stomach contents in that microwear reflects feeding and food preferences over a longer period of time, and can be applied where these data are unavailable. Furthermore, it is applicable to extinct organisms and fossils, allowing the role of trophic ecology, niche partitioning and competition over evolutionary time-scales to be investigated for the first time.     

Molar microwear textures and the diets of Australopithecus anamensis and Australopithecus afarensis

Many researchers have suggested that Australopithecus anamensis and Australopithecus afarensis were among the earliest hominins to have diets that included hard, brittle items. Here we examine dental microwear textures of these hominins for evidence of this. The molars of three Au. anamensis and 19 Au. afarensis specimens examined preserve unobscured antemortem microwear. Microwear textures of these individuals closely resemble those of Paranthropus boisei, having lower complexity values than Australopithecus africanus and especially Paranthropus robustus. The microwear texture complexity values for Au. anamensis and Au. afarensis are similar to those of the grass-eating Theropithecus gelada and folivorous Alouatta palliata and Trachypithecus cristatus. This implies that these Au. anamensis and Au. afarensis individuals did not have diets dominated by hard, brittle foods shortly before their deaths. On the other hand, microwear texture anisotropy values for these taxa are lower on average than those of Theropithecus, Alouatta or Trachypithecus. This suggests that the fossil taxa did not have diets dominated by tough foods either, or if they did that directions of tooth–tooth movement were less constrained than in higher cusped and sharper crested extant primate grass eaters and folivores.     

Dust accumulation in the canopy: A potential cause of dental microwear in primates

Dental microwear researchers consider exogenous grit or dust to be an important cause of microscopic wear on primate teeth. No study to date has examined the accumulation of such abrasives on foods eaten by primates in the forest. This investigation introduces a method to collect dust at various heights in the canopy. Results from dust collection studies conducted at the primate research stations at Ketambe in Indonesia, and Hacienda La Pacifica in Costa Rica indicate that 1) grit collects throughout the canopy in both open country and tropical rain forest environments; and 2) the sizes and concentrations of dust particles accumulated over a fixed period of time differ depending on site location and season of investigation. These results may hold important implications for the interpretation of microwear on primate teeth. © 1995 Wiley-Liss, Inc.     

Quantitative differences in dental microwear between primate species with different diets and a comment on the presumed diet of Sivapithecus

Studies of dental microwear have been used to relate tooth form to function in a variety of recent and extinct mammals. Probably the most important aspect of microwear analysis is the possibility of using it to deduce the diet of extinct animals. Such deductions must be based on comparative studies of modern species with known diets, but to date, only qualitative studies have been attempted and all have been based on small samples. Here we report quantitative differences in dental microwear between primate species that are known to have different diets. Occlusal facets with different functions have previously been shown to exhibit different microwear patterns. However, the differences between facets of one species are shown to be far less than those between homologous facets of different species. Study of seven species of extant primates shows that enamel microwear can be used to distinguish between those with a mainly frugivorous diet and those with a mainly folivorous one. Microwear can also distinguish hard-object feeders from soft-fruit eaters. The microwear of Miocene Sivapithecus indicus cannot be distinguished statistically from that of the chimpanzee, but it is different from that of the other species. On this evidence S. indicus was not a hard-object feeder and the adaptive significance of its thick molar enamel is at present unknown.     

Human dental microwear caused by calcium oxalate phytoliths in prehistoric diet of the lower Pecos region,Texas

Recent research demonstrates that silica phytoliths of dietary origin are associated with microwear of human teeth. Previous research has shown that severe enamel microwear and dental wear characterizes Archaic hunter-gatherers in the lower Pecos region of west Texas. Calcium oxalate crystals are especially common in Archaic coprolites. The vast majority are derived from prickly pear and agave, which were the dietary staples in west Texas for 6,000 years. The calcium oxalate phytoliths are harder than enamel. Therefore, calcium oxalate crystals are the most likely source of previously documented dental microwear and wear in the lower Pecos region. Am J Phys Anthropol 107:297–304, 1998. © 1998 Wiley-Liss, Inc.     

Enamel structure and microwear: an experimental study of the response of enamel to shearing force.

The anisotropic fracturing and differential wear properties of enamel microstructure represent factors that can obscure the predictive relationship between dental microwear and diet. To assess the impact of enamel structure on microwear, this in vitro experimental study examines the relative contributions to wear of three factors: 1) species differences in microstructure, 2) direction of shearing force relative to enamel prisms and crystallites, and 3) size of abrasive particles. Teeth of Lemur, Ovis, Homo, and Crocodylus, representing, respectively, the structural categories of prismatic patterns 1, 2, and 3 and nonprismatic enamel, were abraded by shearing forces (forces having a component directed parallel to abraded surfaces) and examined by scanning electron microscopy. Striation width increased with particle size for nonprismatic, but not for prismatic, specimens. Direction of shear relative to prism and crystallite orientation had a significant influence on striation width in only some prismatic enamels. The different responses of prismatic and nonprismatic enamels to abrasion reflect the influence of structure, but at the level of organization of crystallites rather than prisms per se. Such interactions explain in part the inability of striation width to discriminate among animals with different dietary habits. Heteroscedasticity and deviations from normality also may confound parametric analyses of microwear variables. Variation in crystallite orientation in prismatic enamels may contribute to optimal dental function through the property of differential wear in functionally distinct regions of teeth.     

Inferring parrotfish (Teleostei: Scaridae) pharyngeal mill function from dental morphology, wear, and microstructure

Morphology, occlusal surface topography, macrowear, and microwear features of parrotfish pharyngeal teeth were investigated to relate microstructural characteristics to the function of the pharyngeal mill using scanning electron microscopy of whole and sectioned pharyngeal jaws and teeth. Pharyngeal tooth migration is anterior in the lower jaw (fifth ceratobranchial) and posterior in the upper jaw (paired third pharyngobranchials), making the interaction of occlusal surfaces and wear-generating forces complex. The extent of wear can be used to define three regions through which teeth migrate: a region containing newly erupted teeth showing little or no wear; a midregion in which the apical enameloid is swiftly worn; and a region containing teeth with only basal enameloid remaining, which shows low to moderate wear. The shape of the occlusal surface alters as the teeth progress along the pharyngeal jaw, generating conditions that appear suited to the reduction of coral particles. It is likely that the interaction between these particles and algal cells during the process of the rendering of the former is responsible for the rupture of the latter, with the consequent liberation of cell contents from which parrotfish obtain their nutrients.