Sperm packages are widespread in the order Scorpiones, but absent in the family Buthidae. The morphology of sperm packages is diverse and apparently has phylogenetic information. The objectives of this work were to show diversity of sperm packages and to provide a quantitative basis for using sperm packages' morphology as a taxonomic character. For this, we conducted a morphological analysis and comparison of the different sperm packages of species of the family Bothriuridae. The seminal content from males of species of Bothriuridae was studied. Specimens from Iuridae, Buthidae, Euscorpiidae, Liochelidae, Scorpionidae, Vaejovidae, Chaerilidae, and Chactidae were used for comparison. Digital images of sperm packages were measured and statistically analysed based on the following variables: total length, head width, head-body angle, total area, and head length. Pairs of variables were also contrasted, and all the variables were correlated with the current phylogenetic hypothesis for Bothriuridae. High morphological diversity and variability in measures was observed. In general, measurements were similar within each genus, but differed amongst genera. Cane-like sperm packages are very common in species of the family Bothriuridae. Species from Bothriurus show a wide range of sperm package shapes, some of them shared with Timogenes and Vachonia species, supporting the idea of nonmonophyly of the genus. Many species showed sperm package dimorphism inside a single male. Some of the analysed features fit well with the phylogenetic hypothesis in Bothriuridae, and the general package shape shows high correlation with scorpion phylogeny in other families. Bent and round packages are the most common amongst the different families. Sperm packages are not developed in Chaerilidae, as in Buthidae. This is the first morphological and comparative analysis of sperm packages in scorpions, and reveals much greater diversity in this trait than previously known. Our results reinforce the idea that the study of morphology of sperm packages would contribute characters for scorpion phylogeny at different levels.

Many environmental factors may influence the activity of scorpions in arid ecosystems. In this study, we examined the effects of such factors on scorpion activity at a mature and a secondary forest site in the Chancaní Reserve (Arid Chaco ecoregion in Córdoba, Argentina). Scorpions were collected using pitfall traps, and their nocturnal activity was observed by means of UV light during 14 nights. Temperature, air humidity, atmospheric pressure, and wind velocity were measured, and the percentage of visible moon was estimated. A total of 440 scorpions, representing seven species belonging to two families (Bothriuridae and Buthidae), were observed in the study area. Brachistosternus ferrugineus was the species most commonly observed using both methods. Mature and secondary forest shared slightly more than 50% of species. Surface activity of scorpions observed with the two methods differed significantly between the two areas. Surface activity of all scorpions and of B. ferrugineus were negatively related to the phase of the moon and positively related to air temperature. Physical factors thus have important effects on scorpion activity.

There is strong evidence that polyandrous taxa have evolved relatively larger testes than monogamous relatives. Sperm size may either increase or decrease across species with the risk or intensity of sperm competition. Scorpions represent an ancient direct mode with spermatophore-mediated sperm transfer and are particularly well suited for studies in sperm competition. This work aims to analyze for the first time the variables affecting testes mass, ejaculate volume and sperm length, according with their levels of polyandry, in species belonging to the Neotropical family Bothriuridae. Variables influencing testes mass and sperm length were obtained by model selection analysis using corrected Akaike Information Criterion. Testes mass varied greatly among the seven species analyzed, ranging from 1.6±1.1 mg in Timogenes dorbignyi to 16.3±4.5 mg in Brachistosternus pentheri with an average of 8.4±5.0 mg in all the species. The relationship between testes mass and body mass was not significant. Body allocation in testes mass, taken as Gonadosomatic Index, was high in Bothriurus cordubensis and Brachistosternus ferrugineus and low in Timogenes species. The best-fitting model for testes mass considered only polyandry as predictor with a positive influence. Model selection showed that body mass influenced sperm length negatively but after correcting for body mass, none of the variables analyzed explained sperm length. Both body mass and testes mass influenced spermatophore volume positively. There was a strong phylogenetic effect on the model containing testes mass. As predicted by the sperm competition theory and according to what happens in other arthropods, testes mass increased in species with higher levels of sperm competition, and influenced positively spermatophore volume, but data was not conclusive for sperm length.

After mating, females may experience a decline in sexual receptivity and attractiveness that may be associated with changes in the production and emission of sex pheromones. In some cases, these changes are produced by chemical substances or structures (e.g., mating plugs) produced by males as a strategy to avoid or reduce sperm competition. In scorpions, sex pheromones may be involved in finding potential mates and starting courtship. Here, we tested the hypothesis that the males of Urophonius brachycentrus, a species that produces a mating plug, use chemical communication (sex pheromones) to detect, localize, and discriminate females according to their mating status (virgin or inseminated), aided by chemical signaling. We also explored the effect of extracting of the mating plug on chemical communication and mating acceptance. We used Y‐maze olfactometers with different stimuli to analyze male choice and exploration time. To evaluate mating acceptance, we measured the attractiveness and receptivity of females of different mating status. We found that chemical communication occurs through volatile pheromones, but not contact pheromones. Males equally preferred sites with virgin or inseminated females with removed mating plug. In turn, females with these mating statuses were more attractive and receptive for males than inseminated females. This study suggests that the mating plug significantly affects female chemical attractiveness with an effect on volatile pheromones and decreasing sexual mating acceptance of females. The decline in the female's sexual receptivity is a complex process that may respond to several non‐exclusive mechanisms imposed by males and strategically modulated by females.