Gastropods of the genus Bulimulus Leach, 1814 have a neotropical native distribution and are considered large in size because they measure between 20.01-40.00mm. The majority of its species were originally described based on their most widely used set of characters, conchological morphology. In some cases, information about the morphology of hard parts, such as the radula and jaw, was provided; however, data regarding their internal anatomy are scarce. Thus, of the 68 valid species of Bulimulus, according to the specialized database on molluscs, approximately 20% of the species have some anatomical data while; in terms of genetic approaches, only a few sequences corresponding to about 10% of the entities are currently available. As a result, for the majority of species, conchological characteristics are the only diagnostic tool available. Unfortunately, the length of the descriptions, the overlapping of conchological features, and the scarcity of anatomical information made it difficult to correctly identify numerous entities. The genus is represented in Argentina by eight species, three of which are found in the province of Buenos Aires.
Considering the on the one hand, B. bonariensis has little anatomical information, a wide distribution in the country, and has experienced population explosions with negative effects on crops on several occasions. And that, on the other hand, there is little or no anatomical information on B. rushii and B. vesicalis to complement the conchological data, as well as the fact that they were included on the list of priority species for conservation. Due to this, the objectives of this thesis are to conduct conchological and anatomical analyses with genetic support in order to complement the diagnosis of the species to contribute to elucidate certain confusing taxonomically situations as synonyms with other entities such as B. sporadicus and problems in the identification of specimens of B. rushii. On the other hand, this thesis focused on B. bonariensis to monitor an experimental population in order to gather information on its rate and pattern of growth, survival, mortality, age estimation and size at first reproduction, among other population parameters and characteristics of the life cycle. In addition to the analysis of gametic development, this study will provide us with valuable demographic information for conducting controls at the population level or comparisons with other members of the genus. Thus, using qualitative conchological analysis, we were able to identify specimens corresponding to the three species, with the sculptures of the protoconch being the most distinguishing feature of B. rushii, along with the length of the spire, length of the aperture, and general configuration, allowed the assignment of specimens to this species. As for B. vesicalis and B. bonariensis, the length of the spire, length of the aperture, and the low convexity of the whorls were the features that allowed such distinction.
In the morphometric analysis of the three species, a Principal Component Analysis (PCA) was first performed in order to explore the morphological variation of the shell shape, then the differences between the groups were deepened by means of a Discriminant Analysis (DA). In this way, the variables that allowed PCA discrimination were those associated with the penultimate whorl, length and width, and the smallest diameter. However, in the DA, the smallest diameter and the length of the spire were the variables of greatest contribution. Given the intraspecific morphological variability, morphometric tools were used to further analyze each species. Therefore, B. bonariensis displayed three distinct morphologies, with length penultimate whorl, the length spire, length total shell, length apertural, minor diameter and the largest diameter being the variables that contributed the most. B. vesicalis and B. bonariensis were qualitatively distinguishable from one another; yet, PCA and DA revealed a wide overlapping area. In B. rushii, variability at the conchological level was not observed within each population studied, but was found to be associated with geographic location. As a result, the length of the parietal space and aperture showed a significant difference in relation to altitude.
In terms of internal anatomy, differences that allowed complementing the diagnosis of each species were shown. At the level of the mantle organs, the location of the heart and pericardial cavity in relation to the kidney differed between the three species. In addition, B. rushii has a secondary collar vessel and a larger lung area. Regarding the digestive system, the main differences were observed in the buccal bulb, since B. rushii presented a greater number of muscles and a more quadrangular morphology of the jaw plates. On the other hand, there was only a subtle difference in the length of the cerebro-pleural connective tissue in the central nervous system between B. bonariensis and B. vesicalis, while in B. rushii there was extensive fusion of the pleural and pedial ganglia. There was also a significant difference at the reproductive level, which was considerably simpler compared to the other treated species, with a basic histological structure of the penial complex and no diverticula. When B. bonariensis and B. vesicalis were compared, differences in the thickness of the bursa copulatrix duct and the general and histological morphology of the penial complex were found. Moreover, when compared to samples from Buenos Aires, seven different penial complexes were present within the nominal species B. bonariensis. The molecular information through genetic distances, phylogenetic reconstructions and two species delimitation methods (ABGD and к/θ) allowed to consolidate what was observed from the anatomical point of view. So that, what was understood as a single species, is actually a complex of species formed by at least eight, which are considered cryptic based on the analysis of the conchological morphology.
Concerning the second part of the thesis, the laboratory study was carried out using B. bonariensis as the model species. In this way, it was possible to monitor a complete life cycle. The average number of eggs per clutch was calculated at 44, with a 14-day embryonic development time. The birth rate was nearly 42% and of this percentage, during the first 50 days mortality of 50% of the individuals born was observed. Thus, they presented a type III survival curve, with a high early mortality rate and a life expectancy of two to three years. The logistic growth model was the best fit. Through the histological study of the gonad, in a natural population and over the course of two years, biological peculiarities such as the location of the different cell types in the follicles could be identified. Furthermore, at the beginning of their development they are protogynous hermaphrodites and after reaching gonadal maturity, they become simultaneous hermaphrodites. It has also been determined that it is an iteroparous species. On the other hand, three reproductive periods were identified: pre-recruitment in the fall; recruitment, which begins in winter, but the main event occurs in spring; post-recruitment in summer, where the absence of gonadal rest was observed. In pre-recruitment resorption was observed in addition to abundant primary and secondary spermatocytes and growing vitellogenic oocytes. At the same time, it was possible to correlate the degree of spermoviduct development with the stage of gonadal development. By means of logistic regression, the size at which they reach gonadal maturity was calculated, which was established at 12 mm of total shell length. By relating this length with the laboratory study, it was possible to deduce that this size is reached after 200 days of life. Therefore, the first gametic maturation is achieved before one year of life, where a first decrease in the growth rate occurs. Finally, it was inferred that, since the reproductive period occurs the following spring, at one year of life during the recruitment period, the first reproduction takes place when they reach an average of 17 mm and there is a second decrease in the growth rate.