Are Virginia opossums really ecological traps for ticks? Groundtruthing laboratory observations.

Virginia opossums (Didelphis virginiana) are a common synanthrope in North America, and serve as host to many species of ectoparasites. Research on captive Virginia opossums estimated that opossums eat, on average, 5500 larval ticks (Acari: Ixodidae) per week. To investigate this apparent preference exhibited by opossums for ingesting ticks, we comprehensively analyzed stomach contents of 32 Virginia opossums from central Illinois. Using a dissecting microscope, we searched the contents exhaustively for ticks and tick body parts, without sieving or pre-rinsing the stomach contents. We did not locate any ticks or tick parts in the stomach contents of Virginia opossums. We also performed a vigorous literature search for corroborating evidence of tick ingestion. Our search revealed 23 manuscripts that describe diet analyses of Virginia opossums, 19 of which were conducted on stomach or digestive tract contents and four of which were scat-based analyses. None of the studies identified ticks in their analyses of diet items. We conclude that ticks are not a preferred diet item for Virginia opossums. Considering that wildlife unconditioned to laboratory conditions may exhibit non-typical behaviors, we recommend that lab-based studies of wildlife behavior be groundtruthed with studies based in natural conditions.

Resistance of South American opossums to vWF-binding venom C-type lectins.

Opossums in the clade Didelphini are well known to be resistant to snake venom due to endogenous circulating inhibitors which target metalloproteinases and phospholipases. However, the mechanisms through which these opossums cope with a variety of other damaging venom proteins are unknown. A protein involved in blood clotting (von Willebrand Factor) has been found to have undergone rapid adaptive evolution in venom-resistant opossums. This protein is a known target for a subset of snake venom C-type lectins (CTLs), which bind it and then induce it to bind platelets, causing hemostatic disruption. Several amino acid changes in vWF unique to these opossums could explain their resistance; however, experimental evidence that these changes disrupt venom CTL binding was lacking. We used platelet aggregation assays to quantify resistance to a venom-induced platelet response in two species of venom-resistant opossums (Didelphis virginiana, Didelphis aurita), and one venom-sensitive opossum (Monodelphis domestica). We found that all three species have lost nearly all their aggregation response to the venom CTLs tested. Using washed platelet assays we showed that this loss of aggregation response is not due to inhibitors in the plasma, but rather to the failure of either vWF or platelets (or both) to respond to venom CTLs. These results demonstrate the potential adaptive function of a trait previously shown to be evolving under positive selection. Surprisingly, these findings also expand the list of potentially venom tolerant species to include Monodelphis domestica and suggest that an ecological relationship between opossums and vipers may be a broader driver of adaptive evolution across South American marsupials than previously thought.

Land management strategies can increase oil palm plantation use by some terrestrial mammals in Colombia.

While the conservation role of remaining natural habitats in anthropogenic landscapes is clear, the degree to which agricultural matrices impose limitations to animal use is not well understood, but vital to assess species' resilience to land use change. Using an occupancy framework, we evaluated how oil palm plantations affect the occurrence and habitat use of terrestrial mammals in the Colombian Llanos. Further, we evaluated the effect of undergrowth vegetation and proximity to forest on habitat use within plantations. Most species exhibited restricted distributions across the study area, especially in oil palm plantations. Habitat type strongly influenced habitat use of four of the 12 more widely distributed species with oil palm negatively affecting species such as capybara and naked-tailed armadillo. The remaining species showed no apparent effect of habitat type, but oil palm and forest use probabilities varied among species. Overall, generalist mesocarnivores, white-tailed deer, and giant anteater were more likely to use oil palm while the remaining species, including ocelot and lesser anteater, showed preferences for forest. Distance to nearest forest had mixed effects on species habitat use, while understory vegetation facilitated the presence of species using oil palm. Our findings suggest that allowing undergrowth vegetation inside plantations and maintaining nearby riparian corridors would increase the likelihood of terrestrial mammals' occurrence within oil palm landscapes.

PHYSIOLOGICAL BASAL PARAMETERS OF FREE-RANGING OPOSSUMS ( DIDELPHIS VIRGINIANA) IN THE YUCATAN PENINSULA, MEXICO.

The aim of the present study is to determine the basal parameters in opossums ( Didelphis virginiana) during physical examination in Yucatan, Mexico. Sixty-six opossums were captured and manually handled for physical examinations. Sex, age, cardiac and respiratory rate, body temperature, pulse, mucous membranes, capillary refill time, hydration, size of superficial lymph nodes, mental status, and body condition were evaluated. Also, comparisons between rectal and auricular temperatures were performed. The means of physiological parameters obtained were: cardiac frequency 146 beats per minute (95% confidence interval [CI]: 138.91-155.24), respiratory frequency 27.33 breaths per minute (95% CI: 25.15-29.64), and body temperature 34.01°C (95% CI: 33.71-34.31). In 54% of animals, a sinus arrhythmia was present. A significant difference was found between rectal and auricular temperature measurements. A variation of -1.64 to 1.14°C was obtained via the auricular thermometer compared with the rectal device.

Camera trap arrays improve detection probability of wildlife: Investigating study design considerations using an empirical dataset.

Camera trapping is a standard tool in ecological research and wildlife conservation. Study designs, particularly for small-bodied or cryptic wildlife species often attempt to boost low detection probabilities by using non-random camera placement or baited cameras, which may bias data, or incorrectly estimate detection and occupancy. We investigated the ability of non-baited, multi-camera arrays to increase detection probabilities of wildlife. Study design components were evaluated for their influence on wildlife detectability by iteratively parsing an empirical dataset (1) by different sizes of camera arrays deployed (1-10 cameras), and (2) by total season length (1-365 days). Four species from our dataset that represented a range of body sizes and differing degrees of presumed detectability based on life history traits were investigated: white-tailed deer (Odocoileus virginianus), bobcat (Lynx rufus), raccoon (Procyon lotor), and Virginia opossum (Didelphis virginiana). For all species, increasing from a single camera to a multi-camera array significantly improved detection probability across the range of season lengths and number of study sites evaluated. The use of a two camera array increased survey detection an average of 80% (range 40-128%) from the detection probability of a single camera across the four species. Species that were detected infrequently benefited most from a multiple-camera array, where the addition of up to eight cameras produced significant increases in detectability. However, for species detected at high frequencies, single cameras produced a season-long (i.e, the length of time over which cameras are deployed and actively monitored) detectability greater than 0.75. These results highlight the need for researchers to be critical about camera trap study designs based on their intended target species, as detectability for each focal species responded differently to array size and season length. We suggest that researchers a priori identify target species for which inference will be made, and then design camera trapping studies around the most difficult to detect of those species.

Which Factors Determine Spatial Segregation in the South American Opossums (Didelphis aurita and D. albiventris)? An Ecological Niche Modelling and Geometric Morphometrics Approach.

Didelphis albiventris and D. aurita are Neotropical marsupials that share a unique evolutionary history and both are largely distributed throughout South America, being primarily allopatric throughout their ranges. In the Araucaria moist forest of Southern Brazil these species are sympatric and they might potentially compete having similar ecology. For this reason, they are ideal biological models to address questions about ecological character displacement and how closely related species might share their geographic space. Little is known about how two morphologically similar species of marsupials may affect each other through competition, if by competitive exclusion and competitive release. We combined ecological niche modeling and geometric morphometrics to explore the possible effects of competition on their distributional ranges and skull morphology. Ecological niche modeling was used to predict their potential distribution and this method enabled us to identify a case of biotic exclusion where the habit generalist D. albiventris is excluded by the presence of the specialist D. aurita. The morphometric analyses show that a degree of shape discrimination occurs between the species, strengthened by allometric differences, which possibly allowed them to occupy marginally different feeding niches supplemented by behavioral shift in contact areas. Overlap in skull morphology is shown between sympatric and allopatric specimens and a significant, but weak, shift in shape occurs only in D. aurita in sympatric areas. This could be a residual evidence of a higher past competition between both species, when contact zones were possibly larger than today. Therefore, the specialist D. aurita acts a biotic barrier to D. albiventris when niche diversity is not available for coexistence. On the other hand, when there is niche diversification (e.g. habitat mosaic), both species are capable to coexist with a minimal competitive effect on the morphology of D. aurita.

Detecting and modelling delayed density-dependence in abundance time series of a small mammal (Didelphis aurita).

We study the population size time series of a Neotropical small mammal with the intent of detecting and modelling population regulation processes generated by density-dependent factors and their possible delayed effects. The application of analysis tools based on principles of statistical generality are nowadays a common practice for describing these phenomena, but, in general, they are more capable of generating clear diagnosis rather than granting valuable modelling. For this reason, in our approach, we detect the principal temporal structures on the bases of different correlation measures, and from these results we build an ad-hoc minimalist autoregressive model that incorporates the main drivers of the dynamics. Surprisingly our model is capable of reproducing very well the time patterns of the empirical series and, for the first time, clearly outlines the importance of the time of attaining sexual maturity as a central temporal scale for the dynamics of this species. In fact, an important advantage of this analysis scheme is that all the model parameters are directly biologically interpretable and potentially measurable, allowing a consistency check between model outputs and independent measurements.

Femoral loading mechanics in the Virginia opossum, Didelphis virginiana: torsion and mediolateral bending in mammalian locomotion.

Studies of limb bone loading in terrestrial mammals have typically found anteroposterior bending to be the primary loading regime, with torsion contributing minimally. However, previous studies have focused on large, cursorial eutherian species in which the limbs are held essentially upright. Recent in vivo strain data from the Virginia opossum (Didelphis virginiana), a marsupial that uses a crouched rather than an upright limb posture, have indicated that its femur experiences appreciable torsion during locomotion as well as strong mediolateral bending. The elevated femoral torsion and strong mediolateral bending observed in D. virginiana might result from external forces such as a medial inclination of the ground reaction force (GRF), internal forces deriving from a crouched limb posture, or a combination of these factors. To evaluate the mechanism underlying the loading regime of opossum femora, we filmed D. virginiana running over a force platform, allowing us to measure the magnitude of the GRF and its three-dimensional orientation relative to the limb, facilitating estimates of limb bone stresses. This three-dimensional analysis also allows evaluations of muscular forces, particularly those of hip adductor muscles, in the appropriate anatomical plane to a greater degree than previous two-dimensional analyses. At peak GRF and stress magnitudes, the GRF is oriented nearly vertically, inducing a strong abductor moment at the hip that is countered by adductor muscles on the medial aspect of the femur that place this surface in compression and induce mediolateral bending, corroborating and explaining loading patterns that were identified in strain analyses. The crouched orientation of the femur during stance in opossums also contributes to levels of femoral torsion as high as those seen in many reptilian taxa. Femoral safety factors were as high as those of non-avian reptiles and greater than those of upright, cursorial mammals, primarily because the load magnitudes experienced by opossums are lower than those of most mammals. Thus, the evolutionary transition from crouched to upright posture in mammalian ancestors may have been accompanied by an increase in limb bone load magnitudes.

In vivo strains in the femur of the Virginia opossum (Didelphis virginiana) during terrestrial locomotion: testing hypotheses of evolutionary shifts in mammalian bone loading and design.

Terrestrial locomotion can impose substantial loads on vertebrate limbs. Previous studies have shown that limb bones from cursorial species of eutherian mammals experience high bending loads with minimal torsion, whereas the limb bones of non-avian reptiles (and amphibians) exhibit considerable torsion in addition to bending. It has been hypothesized that these differences in loading regime are related to the difference in limb posture between upright mammals and sprawling reptiles, and that the loading patterns observed in non-avian reptiles may be ancestral for tetrapod vertebrates. To evaluate whether non-cursorial mammals show loading patterns more similar to those of sprawling lineages, we measured in vivo strains in the femur during terrestrial locomotion of the Virginia opossum (Didelphis virginiana), a marsupial that uses more crouched limb posture than most mammals from which bone strains have been recorded, and which belongs to a clade phylogenetically between reptiles and the eutherian mammals studied previously. The presence of substantial torsion in the femur of opossums, similar to non-avian reptiles, would suggest that this loading regime likely reflects an ancestral condition for tetrapod limb bone design. Strain recordings indicate the presence of both bending and appreciable torsion (shear strain: 419.1 ± 212.8 µÎµ) in the opossum femur, with planar strain analyses showing neutral axis orientations that placed the lateral aspect of the femur in tension at the time of peak strains. Such mediolateral bending was unexpected for a mammal running with near-parasagittal limb kinematics. Shear strains were similar in magnitude to peak compressive axial strains, with opossum femora experiencing similar bending loads but higher levels of torsion compared with most previously studied mammals. Analyses of peak femoral strains led to estimated safety factor ranges of 5.1-7.2 in bending and 5.5-7.3 in torsion, somewhat higher than typical mammalian values for bending, but approaching typical reptilian values for shear. Loading patterns of opossum limb bones therefore appear intermediate in some respects between those of eutherian mammals and non-avian reptiles, providing further support for hypotheses that high torsion and elevated limb bone safety factors may represent persistent ancestral conditions in the evolution of tetrapod limb bone loading and design.

Invasive plant species alters consumer behavior by providing refuge from predation.

Understanding the effects of invasive plants on native consumers is important because consumer-mediated indirect effects have the potential to alter the dynamics of coexistence in native communities. Invasive plants may promote changes in consumer pressure due to changes in protective cover (i.e., the architectural complexity of the invaded habitat) and in food availability (i.e., subsidies of fruits and seeds). No experimental studies have evaluated the relative interplay of these two effects. In a factorial experiment, we manipulated cover and food provided by the invasive shrub Amur honeysuckle (Lonicera maackii) to evaluate whether this plant alters the foraging activity of native mammals. Using tracking plates to quantify mammalian foraging activity, we found that removal of honeysuckle cover, rather than changes in the fruit resources it provides, reduced the activity of important seed consumers, mice in the genus Peromyscus. Two mesopredators, Procyon lotor and Didelphis virginiana, were also affected. Moreover, we found rodents used L. maackii for cover only on cloudless nights, indicating that the effect of honeysuckle was weather-dependent. Our work provides experimental evidence that this invasive plant species changes habitat characteristics, and in so doing alters the behavior of small- and medium-sized mammals. Changes in seed predator behavior may lead to cascading effects on the seeds that mice consume.

Cone pigments in a North American marsupial, the opossum (Didelphis virginiana).

Only two of the four cone opsin gene families found in vertebrates are represented in contemporary eutherian and marsupial species. Recent genetic studies of two species of South American marsupial detected the presence of representatives from two of the classes of cone opsin genes and the structures of these genes predicted cone pigments with respective peaks in the ultraviolet and long-wavelength portions of the spectrum. The Virginia opossum (Didelphis virginiana), a profoundly nocturnal animal, is the only marsupial species found in North America. The prospects for cone-based vision in this species were examined through recordings of the electroretinogram (ERG), a commonly examined retinal response to photic stimulation. Recorded under flickering-light conditions that elicit signals from cone photoreceptors, the spectral sensitivity of the opossum eye is well accounted for by contributions from the presence of a single cone pigment having peak absorption at 561-562 nm. A series of additional experiments that employed various chromatic adaptation paradigms were conducted in a search for possible contributions from a second (short-wavelength sensitive) cone pigment. We found no evidence that such a mechanism contributes to the ERG in this marsupial.

A genetic analysis of the Virginia opossum mating system: evidence of multiple paternity in a highly fragmented landscape.

Using molecular techniques, we examined patterns of paternity in Virginia opossums occupying a highly fragmented agricultural landscape in northern Indiana. During 2008, we collected tissue from 64 females and their pouch young in 34 forest patches distributed over a 1100-km(2) region. Using genotypes from 10 microsatellite loci, we determined the minimum number of fathers contributing to each litter using GERUD 1.0. Genotyped offspring with known mothers were then analyzed using CERVUS 3.0, incorporating genotypes from 317 males sampled from 2007-2008 to identify potential fathers. Our analyses revealed that promiscuity was common among females, with 26 (41%) litters having > or = 2 sires. Despite the fact that we intensively sampled forest patches for potential fathers, we only were able to identify 13 fathers contributing to 14 litters, with an average Euclidean distance of 18.7 km between father-offspring pairs found in disparate patches (N = 6). Our inability to identify most (85%) fathers of sampled litters, coupled with the extensive distances observed between putative father-offspring pairs, suggests that opossums may not maintain explicit home ranges in highly fragmented landscapes.

Seasonal source-sink dynamics at the edge of a species' range.

The roles of dispersal and population dynamics in determining species' range boundaries recently have received theoretical attention but little empirical work. Here we provide data on survival, reproduction, and movement for a Virginia opossum (Didelphis virginiana) population at a local distributional edge in central Massachusetts (USA). Most juvenile females that apparently exploited anthropogenic resources survived their first winter, whereas those using adjacent natural resources died of starvation. In spring, adult females recolonized natural areas. A life-table model suggests that a population exploiting anthropogenic resources may grow, acting as source to a geographically interlaced sink of opossums using only natural resources, and also providing emigrants for further range expansion to new human-dominated landscapes. In a geographical model, this source-sink dynamic is consistent with the local distribution identified through road-kill surveys. The Virginia opossum's exploitation of human resources likely ameliorates energetically restrictive winters and may explain both their local distribution and their northward expansion in unsuitable natural climatic regimes. Landscape heterogeneity, such as created by urbanization, may result in source-sink dynamics at highly localized scales. Differential fitness and individual dispersal movements within local populations are key to generating regional distributions, and thus species ranges, that exceed expectations.

Behavioral and nutritional aspects of the Virginian opossum (Didelphis virginiana).

Virginia opossums are widely distributed throughout the United States, except in the most arid regions, and wild individuals are commonly brought to practitioners for medical attention. Opossums' popularity as pets seems to be growing, and it is likely that pet opossums will be more common in veterinary practice. Clinicians must be aware of natural opossum behaviors so that thorough physical examination and diagnostic procedures can be performed on injured patients. For animals kept captive long-term or as pets, veterinarians must understand proper nutrition and nutritional disorders, such as secondary nutritional hyperparathyroidism, obesity, and dental disease, to properly treat this species.

Breathing with your belly: abdominal exhalation, loco-ventilatory integration and size constraints on locomotion in small mammals.

In mammals, diaphragmatic contractions control inhalation while contraction of some thoracic hypaxial muscles and the transversus abdominis muscle contribute to exhalation. Additional thoracic hypaxial muscles are recruited as accessory ventilatory muscles to improve inhalation and exhalation during locomotion. However, the contribution of abdominal hypaxial muscles to resting and locomotor ventilation is little understood in mammals and loco-ventilatory integration has not been studied in small basal mammals. We show for the first time that all of the abdominal hypaxial muscles actively contribute to both resting and locomotory ventilation in mammals but in a size-dependent manner. In large opossums (Didelphis), hypaxial muscles exhibit uniform mild tonus during resting ventilation (pressurizing the gut to aid in exhalation) and shift to phasic bursts of activity during each exhalation during locomotion. Smaller opossums (Monodelphis) actively exhale by firing the abdominal hypaxial muscles at approximately 10Hz at both rest and at preferred locomotor speeds. Furthermore, the large opossums entrained ventilation to limb cycling as speed increased while the small opossums entrained limb cycling to the resting ventilation rate during locomotion. Differences in these species are related to size effects on the natural frequency of the ventilatory system and increasing resting ventilation rates at small size. Large mammals, with lower resting ventilation rates, can increase ventilatory rates during locomotion, while the high resting ventilation rates of small mammals limits their ability to increase ventilation rates during locomotion. We propose that increase in mammalian body size during the Cenozoic may have been an adaptation or exaptation to overcome size effects on ventilation frequency.

Dental eruption in afrotherian mammals.

BACKGROUND: Afrotheria comprises a newly recognized clade of mammals with strong molecular evidence for its monophyly. In contrast, morphological data uniting its diverse constituents, including elephants, sea cows, hyraxes, aardvarks, sengis, tenrecs and golden moles, have been difficult to identify. Here, we suggest relatively late eruption of the permanent dentition as a shared characteristic of afrotherian mammals. This characteristic and other features (such as vertebral anomalies and testicondy) recall the phenotype of a human genetic pathology (cleidocranial dysplasia), correlations with which have not been explored previously in the context of character evolution within the recently established phylogeny of living mammalian clades. RESULTS: Although data on the absolute timing of eruption in sengis, golden moles and tenrecs are still unknown, craniometric comparisons for ontogenetic series of these taxa show that considerable skull growth takes place prior to the complete eruption of the permanent cheek teeth. Specimens showing less than half (sengis, golden moles) or two-thirds (tenrecs, hyraxes) of their permanent cheek teeth reach or exceed the median jaw length of conspecifics with a complete dentition. With few exceptions, afrotherians are closer to median adult jaw length with fewer erupted, permanent cheek teeth than comparable stages of non-afrotherians. Manatees (but not dugongs), elephants and hyraxes with known age data show eruption of permanent teeth late in ontogeny relative to other mammals. While the occurrence of delayed eruption, vertebral anomalies and other potential afrotherian synapomorphies resemble some symptoms of a human genetic pathology, these characteristics do not appear to covary significantly among mammalian clades. CONCLUSION: Morphological characteristics shared by such physically disparate animals such as elephants and golden moles are not easy to recognize, but are now known to include late eruption of permanent teeth, in addition to vertebral anomalies, testicondy and other features. Awareness of their possible genetic correlates promises insight into the developmental basis of shared morphological features of afrotherians and other vertebrates.

Role of BMP-4 during tooth development in a model with complete dentition.

There are no reports in literature about roles of bone morphogenetic protein 4 (BMP-4) in tooth development in mammals with complete dentition (with all dental groups). The classical model of study is the mouse, which has distinctive incisor and molar patterns. The opossum Didelphis albiventris with five upper and four lower incisors, one canine, three premolars and four molars, on each side of the jaw, seems to be a convenient model for odontogenesis study. This investigation searched for similarities and differences in BMP-4 expression pattern between the opossum and the mouse. BMP-4 cDNA was obtained by RT-PCR and the expression pattern during molar tooth development was investigated by the immunoperoxidase method. Opossum BMP-4 mature protein has 95% of sequence similarity in relation to mouse and 94% to human. The BMP-4 expression pattern during opossum tooth development was suggestive of a role in dental organ initiation and morphogenesis.

Dental development of Didelphis albiventris (Marsupialia): I--incisors and canines.

The formation of incisors and canines in marsupials of D. albiventris was studied at various stages of development. Seventy-six specimens, with ages varying from 0 to 100 days, were used in this investigation. Serial sections of the maxilla were obtained in the transverse plane and stained with hematoxylin and eosin. Histological analyses were made to verify the pattern of teeth development, as well as their chronology of eruption. The period of time from birth to 100 days comprised the entire process of teeth development, from epithelial bud formation to early eruption of the teeth. Oral epithelium thickening gave rise to the functional incisors and canines. In addition, a secondary dental lamina emerged in different phases of development in the outer epithelium of incisors and canines, which degenerated when it reached the bud stage. No evidence of deciduous dentition was observed. The results of this investigation suggest that secondary dental lamina represents remnants of a primitive condition in which secondary dentition used to be present.

Dental development of Didelphis albiventris (Marsupialia): I - incisors and canines

Estudou-se o desenvolvimento dos dentes incisivos e caninos em 76 amostras de Didelphis albiventris com idade entre 0 e 100 dias. Cortes transversais, seriados de 6 µm de espessura foram obtidos da região da maxila, corados com Hematoxilina e Eosina e analisados ao microscópio de luz. Verificou-se que o período estudado abrange todo o desenvolvimento dental, desde a fase de iniciação da interação epitélio/mesenquima até a completa formação e erupção dos incisivos e caninos. O espessamento do epitélio oral dá origem aos incisivos e caninos funcionais, enquanto o epitélio dental externo do órgão dental origina uma lâmina dental secundária, a qual sofre degeneração, quando o dente alcança o estágio de botão. Não há vestígios de dentição decídua. Sugere-se que a lâmina dental secundária é remanescente de uma condição primitiva na qual ocorria dentição secundária.

Fibroblast growth factor 9: cloning and immunolocalisation during tooth development in Didelphis albiventris.

There are no reports in literature about functional roles of fibroblast growth factor 9 (FGF-9) in tooth development in animals with complete tooth pattern. The classical model for studying tooth development is the mouse, which has small number of teeth and distinctive incisor and molar patterns. The opossum Didelphis albiventris with five upper and four lower incisors, one canine, three premolars, and four molars, on each side of the jaw, seems to be a convenient model to test results obtained in the mouse. Molecular expression studies indicate that FGF-9 participates in murine tooth initiation and regulation of morphogenesis. Searching for similarities and differences in FGF-9 expression between the opossum and the mouse, amino acid sequence and expression pattern of FGF-9 in the developing first molars of D. albiventris were characterised. FGF-9 cDNA sequence was obtained using RT-PCR and expressed in bacterial system for recombinant protein production and analysis of immunoreactivity. FGF-9 expression during tooth development was investigated by immunoperoxidase method. FGF-9 protein consists in a 209-residue polypeptide with a predicted molecular mass of 23.5 kDa. FGF-9 amino acid sequence has 98% of sequence identity to human and 97% to rodents. During tooth development, epithelial FGF-9 expression was seen at the dental lamina stage. Mesenchymal expression was seen at the bud stage and at the cap stage. No significant expression was found in the enamel knot. While in rodents FGF-9 is involved in initiation and regulation of tooth shape, it is suggested that it is only involved in tooth initiation in D. albiventris.