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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.

Species of Haemogregarina are apicomplexan blood parasites that use vertebrates as intermediate hosts. Due to limited interspecific morphological characters within the genus during the last decade, 18S rRNA gene sequences were widely used for species identification. As coinfection patterns were recently reported from nuclear molecular data for two sympatric freshwater turtles Mauremys leprosa and Emys orbicularis from Tunisia, our objectives were to design COI specific primers to confirm the presence of three distinct species in both host species. Blood samples were collected from 22 turtles, from which DNAs were extracted and used as templates for amplification. Following different rounds of PCR and nested PCR, we designed specific Haemogregarina COI primers that allowed the sequencing of nine distinct haplotypes. Phylogenetic Bayesian analysis revealed the occurrence of three well-differentiated sublineages that clustered together into a single clade. Based on pairwise genetic distances (p-distance), we confirmed the occurrence of three distinct but phylogenetically closely related species coinfecting M. leprosa and E. orbicularis in the same aquatic environments. Our results demonstrate that the use of fast evolving genes within Haemogregarina will help to investigate the parasite diversity within both intermediate vertebrate and definitive invertebrate hosts, and to assess the evolution, historical biogeography and specificity of haemogregarines., (Copyright © 2021. Published by Elsevier B.V.)

Several endemic species of Blaps occur in Tunisia, and the species Blaps nefrauensis nefrauensis has been reported in Moulares (urban zone in west-central Tunisia), where it lives and reproduces in home gardens and old buildings. The aim of this work is to study the life cycle of the darkling beetle, considering both field and laboratory rearing conditions. As a result, the beetle species has different developmental stages (egg, larva, prepupa, pupa, and adult) that last about 15 months. Each year during the same period, adults emerge (early summer) and expire (late autumn), larvae hatch (late summer) and pupate (early summer). There is only one generation per year. Females began laying eggs in late July. The eggs were ovoid, white, and about 2.7 mm in length and 1.5 mm in width. Embryogenesis took an average of nine days. The first instar larvae were at initially only 4.5 mm long and ivory white in color. A brief description of the newly egg hatched larva was provided; thus, the nerve fibers innervating the apical setae in the antennae and ligula were detected. Further light microscopic examination of the embryo before hatching from the egg pointed out that the antennal sensilla are protected during the embryogenesis stage.

Haemogregarina species are apicomplexan blood parasites infecting vertebrates such as fish, lizards, and turtles. Due to the high morphological similarity of the erythrocytic stages infecting host species, it has always been a challenge to identify the true diversity of these parasites. Therefore, taxonomic studies are presently based on the combination of morphological and molecular data. In Tunisia, two species of Haemogregarina have been reported within the freshwater turtle Mauremys leprosa (Geoemydidae) for more than 40 years. Since M. leprosa occurs in the same aquatic environments as Emys orbicularis (Emydidae) in Tunisia, our objectives were to assess parasite diversity and specificity on the basis of both morphological and molecular approaches. The turtles were surveyed and sampled across six aquatic areas of Tunisia. Among the 39 specimens of M. leprosa and seven of E. orbicularis that were trapped and investigated, the presence of haemogregarines was detected in the blood of turtles only at sites where leeches were observed. Three 18S variants were identified, which corresponded to three distinct Haemogregarina species, among which one was identified as Haemogregarina stepanowi. The two other species that were detected are likely new to science. Because we show the occurrence of more than one blood parasite species within a single host specimen, our study provides the first report of coinfection with molecularly distinct Haemogregarina spp.

While Liocranidae is a family encompassing 283 species distributed in 32 genera (World Spider Catalog 2019), the genus Agraecina Simon, 1932 currently counts only six described species: Agraecina canariensis Wunderlich, 1992 from the Canaries, A. cristriani (Georgescu, 1989) from Romania, A. hodna Bosmans, 1999 from Algeria, A. lineata (Simon, 1878) from Europe and Kazakhstan, A. rutilia (Simon, 1897) from Sierrra Leone, and A. scupiensis Deltshev, 2016 from Macedonia. During an ecological survey in the north of Tunisia, a new Agraecina species was discovered and is described here. Spiders were collected using pitfall traps in a salt marsh.

The cosmopolitan species, Porcellionides pruinosus (Brandt, 1833), exhibited a geographical variation of its morphological features and its reproduction pattern. In fact, some Tunisian populations had a seasonal reproductive period and other ones showed a reproductive activity. A phenetic analysis has been performed to compare populations belonging to P. pruinosus from different geographical localities. For this, nineteen quantitative characters in 800 specimens of P. pruinosus issued from nine populations located in the North, the Centre, and the South of Tunisia, and one population from Athens (Greece) were studied. These populations were characterized by different reproductive behaviours. The populations of Tabarka, Korba, and Tamerza showed reproductive activity, whereas those of Garat Nâam, Raccada, Sahline, Gafsa, Rdayef, Gabès, and Athens exhibited a seasonal reproductive period. The variability of the first three axes of the principal components analysis (PCA) and their significance showed an intraspecific variability structure. The graphical representation of the population's dispersion generated by the two first axes (63.55% of the total variability) revealed a great heterogeneity among females of the different populations of P. pruinosus. The Garat Nâam population was set apart from the other ones by its cephalic width and length and also by its apophysis length. Furthermore, cephalic and uropod length separated the Garat Nâam and Tabarka populations. The dendrogram, based on Euclidean coordinates, confirms an isolation of the populations of Tabarka and Garat Nâam, exhibiting respectively reproductive activity and seasonal reproduction. However, the other populations do not show any relationship with the reproduction behaviour. They were clustered in two groups. The first one is represented by the populations of the Southeast and the West of Tunisia. The second pooled the populations of the Northeast and the Centre of Tunisia with the population of Athens (Greece). Phenetic analysis of Tunisian populations exhibits an important heterogeneity, which is worth noting. Thus, we need to complete and deepen this study by genetic analysis and crossbreeding tests in order to define the taxonomic status of these populations.

The biology and the population dynamics of Porcellionides sexfasciatus Budde-Lund (1879) were studied on a field and carried out at Garat Nâam (Kasserine, Tunisia) from July 1996 to June 1998. The reproduction exhibited a seasonal pattern extending from February/March to October/November. The juveniles appeared in the population from April to November. Size frequency distributions were analysed and 14 cohorts were recognised during the sampling period. Six cohorts were identified at the first sampling and eight new ones on the other samplings. Among these latter cohorts, three were tracked till they disappeared. Minimum average length of new cohorts ranged from 3.07 +/- 0.35 mm to 3.47 +/- 0.2 mm. Maximum average length of cohorts was 10.42 mm. P. sexfasciatus is a semi-annual species (females producing two or three broods per year), with iteroparous and amphogenes females (females reproducing twice or more in their life and producing both males and females), and bivoltine life cycle (two generations per year). The females are able to produce two broods, in the laboratory, without new mating. Fecundity and fertility, corresponding to the number of eggs or embryos per brood, appeared positively correlated with females' size. Although oscillating throughout the year, the sex ratio was often in equilibrium.