We used a combination of culture-dependent and independent approaches to study in depth the microbial community associated with the digestive tract of the terrestrial isopod Porcellionides pruinosus (Brandt, 1833 ). Specimens from different sampling sites in Tunisia harbored distinct microbiota profiles indicating the impact of both host origin and environmental factors on shaping the microbial flora within P. pruinosus. Our results revealed unexpected bacterial diversity especially via metagenomic analysis; a total of 819 operational taxonomic units (OTUs) assigned to two major bacterial phyla; Proteobacteria and Bacteroidetes. We used Nutrient Agar to isolate the cultivable fraction of bacteria associated with the gut of three geographically distant populations of P. pruinosus. The isolated bacteria belong to Actinobacteria, Firmicutes and Proteobacteria. Enrichment cultures on carboxymethylcellulose (CMC) medium gave evidence that the gut of this Oniscidea harbors cellulolytic Firmicutes and Proteobacteria probably involved in the lignocellulose degradation and then in mediating the functional role of terrestrial isopods as litter decomposers and regulators of nutrient cycling in soil ecosystems.

Growth rate, survival, mean size of reproductive females and fertility were assessed in the terrestrial isopod Porcellionides pruinosus fed an artificial diet supplemented with potential sources of symbionts. The acquisition of symbionts was experimentally manipulated through feeding experiments to investigate the effect of the acquisition mechanism on isopod performances over 4 months. Five groups of isolated newborn were fed differently (basal diet or control; basal diet with leaf litter, faeces, gut; and basal diet mixed with antibiotic), weighed and counted at 0, 2 and 4 months. Growth rate differed significantly between diet groups at 2 and 4 months. No significant difference of survival was detected, but groups fed on a diet with supplements (leaf litter, faeces and gut) had a higher rate of survival. Size and fertility of reproductive females differed significantly across diet groups. Our findings showed the importance of the acquisition mode of symbionts on the life history of Oniscidea and the role of diet intake on the performance of P. pruinosus.

Haemogregarines are blood parasites with a life-cycle involving a vertebrate as the intermediate host and an invertebrate as the definitive host and vector. Extensive phylogenetic investigations based on 18S-rRNA gene sequences have shown that Haemogregarina stepanowi (Apicomplexa: Haemogregarinidae) is able to infest a large diversity of freshwater turtle species, including the European pond turtle Emys orbicularis, the Sicilian pond turtle Emys trinacris, the Caspian turtle Mauremys caspica, the Mediterranean pond turtle Mauremys leprosa, and the Western Caspian turtle Mauremys rivulata, among others. From the same molecular markers, H. stepanowi is further considered to be a complex of cryptic species predisposed to infect the same host species. While Placobdella costata is known to be the unique vector of H. stepanowi, it is only recently that independent lineages within P. costata have been illustrated—suggesting the presence of at least five unique leech species across Western Europe. The aims of our study were therefore to investigate from mitochondrial markers (COI) the genetic diversity within haemogregarines and leeches infecting freshwater turtles of the Maghreb, in order to identify processes of parasite speciation. We showed that H. stepanowi consists of at least five cryptic species in the Maghreb, while two Placobella species were identified in the same area. Although an Eastern–Western speciation pattern was apparent for both leeches and haemogregarines, we cannot make definitive conclusions regarding co-speciation patterns between parasites and vectors. However, we cannot reject the hypothesis of a very strict host–parasite specificity within leeches.

Iso-enzymatic polymorphism analysis was carried out on Tunisian populations of Porcellionides pruinosus to clarify their confused taxonomic status, previously revealed by reproductive and morphological approaches. Seventeen allozyme loci were studied, using horizontal starch gel electrophoresis, in 230 adult specimens belonging to nine populations of P. pruinosus located in north, central and south Tunisia, to assess the levels of their intra-genetic and inter-genetic diversity, and to analyse their genetic structure. Our results indicate a low genetic variability with a polymorphism ratio ranging from 29.4% in Tabarka to 47.1% in the Garat Nâam and Raccada populations, and the mean number of alleles per locus was estimated at 1.51 ± 0.15. The direct-count of heterozygosity ranged between 8.1% and 15%. Substantial estimates of demic structuring were found at the total population (F_ST = 0.198), with a generalized and significant heterozygote deficiency detected in every population (with a mean value for all populations of F_IS = 0.372). Genetic distances between the studied populations were low, indicating that they belong to the same species. Nevertheless, the genetic analyses showed the divergence of the population of Tabarka from all the other studied populations, with the lowest mean gene flow, 0.58, and the highest mean genetic distance, (0.182) obtained also by the morphological and reproductive approaches.