Enteric parasites in free-living Mediterranean pond turtle (Mauremys leprosa leprosa) in contrasted areas (natural vs polluted) from central-western of Morocco.

The objective of this study is to assess the occurrence of intestinal parasites in Mediterranean pond turtle Mauremys leprosa leprosa collected from three contrasting environments in Morocco. Stool samples from 92 turtles were examined for parasite detection and enumeration. The identified intestinal parasites belong to helminths (oxyurids and ascarid) and protozoa (Entamoebidae). A total of 25 turtles (27.17%) were found to be infected by helminths and/or protozoan parasites. No adult form of these parasites was detected. Eggs of oxyurid and ascarid were detected in individuals of populations studied from Oued Ksob (23.07% and 30.76% of n = 13 turtles) and Oued Zat (34.14% and 24.39% of n = 41 turtles), respectively. For protozoa, Entamoeba cysts were present in turtles in Oued Ksob (15.38% of n = 13 turtles), Oued Zat (12.19% of n = 41 turtles), and Oued Tensift (5.26% of n = 38 turtles) localities. Oxyurid eggs showed the highest intensity at Oued Zat reaching 29.30 ± 59.59 eggs per gram (EPG), versus 12 ± 0.38 EPG for ascaris eggs in Oued Ksob. Entamoeba cysts were detected in lower levels with a maximum of 1.66 ± 1.50 cysts per gram (CPG), in Oued Zat. The prevalence of turtles eliminating eggs was statistically significant between localities for different parasite groups. This study reports for the first time a parasitological characterization of gastrointestinal parasites in wild populations of M. leprosa leprosa from contrasting environments, suggesting a relationship between turtles' infestation and the quality of their habitat.

Molecular Phylogenies of Leeches and Haemoparasites Infecting Freshwater Turtles in Aquatic Ecosystems of Northern Africa Suggest Phylogenetic Congruence between Placobdella costata Sensu Lato and Haemogregarina stepanowi Sensu Lato.

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.

Molecular Survey and Microscopic Examination of Haemoparasites Infecting Lizards from Morocco.

PURPOSE: Understanding the factors influencing parasite diversification, their distribution and infection patterns among hosts provides valuable information on the impact of parasitism on natural populations and the dynamics and evolution of disease outbreaks. In this study, we investigated prevalence and intensity of infection by blood parasites, the diversity, distribution, host specificity and infection patterns of apicomplexan parasites in lizards of Morocco. METHODS: Samples from lizard specimens were collected between 2019 and 2021 from different prospected localities in Morocco. In total, 1500 lizards were collected, and their blood and tail tips were screened for blood parasites. RESULTS: Using a PCR approach, we detected two distinct apicomplexan parasite groups (haemogregarines and haemococcidians) and also Stramenopiles (Proteromonas). A significant difference was identified between hosts families, with no agamids or skinks infected and with particularly high infection rates in lacertids. Twenty-four samples were sequenced and included in a phylogenetic assessment. Two samples of Atlantolacerta andreanskyi were infected with unidentified species of Proteromonas. Three samples were infected with haemococcidians, including two samples with Schellackia and another sample with Lankesterella, more closely related to others isolated from bird hosts. These are the first reports of Lankesterella and Proteromonas in these lizards from Morocco. Nineteen different isolates of the complex of Hepatozoon/Karyolysus were identical or very similar to previously sequenced parasites from North Africa and Iberian Peninsula and included new host species. CONCLUSION: Our results highlight the need for future research on the vectors involved in transmission that may influence the ecology and distribution patterns of these parasites. Given that Morocco is a hot spot of endemicity and reptile richness, our results are relevant not only in the context of their taxonomy, but can also have implications for conservation.

New host and locality records of helminths' infection of seven lizards from Morocco.

Morocco has a great diversity of reptiles; more than 26% of the herpetofauna species are considered to be endemic. Nevertheless, there is little information available on helminth parasites of Moroccan lizards. The purpose of this article is to establish a helminth list using non-invasive methods for natural populations of seven lizard species: Acanthodactylus maculatus, Chalcides mionecton, Chalcides montanus, Chalcides polylepis, Quedenfeldtia moerens, Quedenfeldtia trachyblepharus, and Tarentola mauritanica. For each species, prevalence and intensity of infection were given as well as their relationship to some determinant parameters (sex, age class, size, altitude, and seasons). Four species of nematodes, Parapharyngodon micipsae, Pharyngodon mamillatus, Spauligodon auziensis, Thelandros alatus, and unassigned Spauligodon were found. Mean helminth intensity for the seven lizard species was 7.3 ± 16.2 SD (range = 1-92 in infected lizards), with total infection prevalence of 5.6%. Mixed infections were observed in Chalcides montanus and Chalcides polylepis co-infected by Pharyngodon mamillatus and Spauligodon sp. Furthermore, the intensity of infection by nematodes were significantly different among the seven studied species. The nematode infection in Chalcides mionecton were related to sex, host size, and altitude; in Chalcides polylepis to the host age; in Quedenfeldtia trachyblepharus to elevation, age, and host size; and in Tarentola Mauritanica to age, body size, seasons, and altitude. In conclusion, our study provides data about nine new host and locality records and the effect of some determinant factors on host parasitism.

Geographical Variation of Honey Bee (Apis mellifera L. 1758) Populations in South-Eastern Morocco: A Geometric Morphometric Analysis.

In Morocco, there are two well-recognised honey bee (Apis mellifera L.) subspecies: A. m. intermissa in the north and A. m. sahariensis in the south-east. The latter subspecies is found in the arid and semiarid climates of the Sahara Desert. In this study, we used honey bees from four areas of south-eastern Morocco which are, to some degree, isolated by arid zones. We analysed the shape and size of the forewings, using the method of geometric morphometrics. The bees from the four areas of south-eastern Morocco differed significantly in terms of wing shape. Moreover, bees from traditional hives were smaller than those from modern hives. The bees from south-eastern Morocco were clearly different from the reference samples obtained from the Morphometric Bee Data Bank in Oberursel, Germany, representing most of the global variation in honey bees. Surprisingly, the bees were also different from A. m. sahariensis, which should occur in the study area, according to earlier studies. This difference could have been caused by introgression with non-native subspecies imported by beekeepers. The distinct honey bees from south-eastern Morocco deserve to be protected. We provide a method for identifying them, which can help protect them.

Characterization and identification of haemogregarine hemoparasites (Apicomplexa: Adeleina: Hepatozoidae) in natural populations of Mauremys leprosa leprosa and M. leprosa saharica from Morocco.

Distribution, prevalence and parasitaemia of apicomplexan parasites of the genus Haemogregarina were studied in 858 freshwater turtles (735 Mauremys leprosa leprosa and 123 Mauremys leprosa saharica) throughout 30 localities from Morocco. Blood smears were collected from the turtles and partial 18S rRNA sequence data used to infer genetic diversity and phylogenetic relationships. Of the 858 individuals analyzed by microscopy 22.7% were infected, from 16 M. leprosa leprosa localities. Individuals of M. leprosa saharica sampled south of the High Atlas Mountains were not infected, probably due to the absence or rarity of the leech vectors in these localities. Within M. leprosa leprosa, we did not identify any patterns between geography and prevalence, which varied between 10% (Oued Nfiss) to 100% (Oued Tassaout). Five distinct genetic lineages were identified, indicating the likely presence of multiple species of haemogregarines, one of which probably corresponds to Haemogregarina stepanowi. Mixed infections were also detected. Additional studies are needed to better understand the ecology and epidemiology of this parasite in turtles, as well as the host-parasite relationship with their definitive hosts, leeches.

Determinants of prevalence and co-infestation by ecto- and endoparasites in the Atlas day gecko, Quedenfeldtia trachyblepharus, an endemic species of Morocco.

The present work was undertaken to investigate the occurrence of ectoparasites (mites and ticks) and endoparasites (haemogregarines and helminths) in the Atlas day gecko, Quedenfeldtia trachyblepharus (Boettger, 1874), a high-altitude Moroccan endemic lizard. The study examinated also the effect of some host parameters (age, sex, size, body condition), in addition to the season and altitude on the prevalence and intensity of parasite infestations. The study was conducted in three localities from May to September 2019. The results indicated that 35% of juvenile geckos were found to be parasited by one type of parasite. Contrarily, up to three types of parasites were detected in the adults. The prevalence of mite infestations was 48.02%, with a mean intensity of 11.80 ± 15.69. The patterns of mite infestations was found to be mainly related to the altitude, while the prevalence and intensity of infestations were linked to the host size, and to the sex and season, respectively. Larvae and nymphs of Ixodes ricinus (Linnaeus, 1758) were the only life stages infesting geckos, with a prevalence and mean intensity of infestations of 4.41% and 2.2 ± 1.48, respectively. The tick infestations observed were mainly related to the season. The prevalence and intensity of haemogregarine infections were, respectively, 7.92% and 0.24 ± 0.15. The altitude was found to be the only factor associated with this infection. This study also revealed the presence of one helminth genus, Spauligodon sp., with a prevalence and mean intensity of 12.33% and 1.46 ± 0.88, respectively. Spauligodon infestations was significantly associated with age, host size and altitude. This finding represents the first citation of this parasite in Quedenfeldtia genus. Our study indicated that there was no significant relationship between parasite load and geckos body condition, which suggested a stable interaction between the gecko and its parasites. However, it showed a difference of infection between the localities, which could be in relation with habitat conditions.

Absence of Hemolivia mauritanica (Apicomplexa: Haemogregarinidae) in natural populations of Testudo graeca in Morocco.

During spring 2018, we captured 101 spur-thighed tortoises, Testudo graeca, from seven localities in central Morocco. All tortoises were examined for the presence of blood parasites Hemolivia mauritanica and Hyalomma aegyptium ticks, the known vectors. We looked for H. mauritanica infection by examination of blood smears and by genetic analysis with PCR using extractions from both tortoises and ticks. On all tortoises collected, 71.29% were infested with at least one tick, with a mean infestation intensity of 7.12 ticks/tortoise and maximum of 15.55 ticks/tortoises in Had Draa locality (Essaouira region). Although the definitive host is present and abundant in all tortoise populations, all blood samples were Hemolivia-negative. Our results support and confirm the finding of studies previously conducted in other populations of Morocco and indicate that H. mauritanica has a narrower distribution range than its tick vector.

Host-parasite association of Placobdella costata (Glossiphoniidae: Hirudinea) and Mauremys leprosa (Geoemydidae: Testudinoidea) in aquatic ecosystems of Morocco.

Emys orbicularis (Linnaeus, 1758) was considered as a specific host of Placobdella costata (Fr. Mûller, 1846). However, since the parasite was recorded from outside the distribution area of its host, some authors suggested a possible relationship with other hosts. Although two accidental associations were found with another turtle, Mauremys leprosa (Schweigger, 1812), the obtained data remain insufficient to better understand this discovered host-parasite ecological system. In this context, the present study was carried out to evaluate the relationship between the Mediterranean pond turtle, M. leprosa, and the freshwater rhynchobdellid leech, P. costata (Hirudinida: Glossiphoniidae), in aquatic ecosystems of Morocco. During the period from April to June 2018, we found leeches attached to turtles in five out of 30 populations sampled with a prevalence of infection significantly higher in adult than that in juvenile turtles. Moreover, the males are the most infested with 51% of the total, followed by females (33.3%) and juveniles (15.7%). The obtained results indicated that 51 turtles were infested by 139 leeches with a mean intensity of infestation of 4.17 ± 0.47 leeches/turtle (up to 10 leeches/turtle). It was higher in males than that in females in almost all sites. The posterior limbs are the most preferred attachment site, and the body condition of turtles was not affected by the intensity of infestation but it is rather a function of altitude. Our findings proved that M. leprosa-P. costata association is more than accidental and that M. leprosa is rather the main host of P. costata in aquatic ecosystems of Morocco.

Large-scale geographic patterns of mercury contamination in Morocco revealed by freshwater turtles.

Mercury (Hg) is a toxic contaminant present in most aquatic ecosystems. High concentrations pose serious threats to organisms and to human health. Because previous studies focused on few countries, environmental hazard due to Hg contamination remains obscure in many geographic areas, and for example limited information is available in North Africa. We examined total Hg contamination in 13 sites in Morocco (12 rivers and one lake) spread over a large area, 400 km north-south and 350 km west-east, that encompasses different biogeographic zones separated by the Atlas Mountains. Due to their longevity and sedentary habits, we used freshwater turtles as biological probes to monitor Hg exposure. Keratinized tissues reflect long-term Hg exposure; thus, we assayed Hg concentration in the claws of > 200 individuals and supplemented these data with blood Hg concentrations of > 60 individuals (a tissue that provides shorter term Hg exposure integration). The results provide the first large-scale picture of Hg contamination in the aquatic freshwater systems of Morocco. Comparisons with previous studies revealed that some of the sites were highly contaminated (e.g. mean Hg concentrations were above 5 µg g-1, a very high level in keratinized tissues) whereas other sites presented moderate or baseline levels. Unexpectedly, all highly contaminated sites were found in less densely populated areas, while more densely urbanized northern sites, even the sewers of large cities, were not highly contaminated. We hypothesize that silver mining activities in the southern High Atlas and in the Anti-Atlas contaminate rivers of the catchment basins over long distances. These findings indicate that fish, water consumption and contamination levels in local people should be further scrutinized.

The mitochondrial genomes of Atlas Geckos (Quedenfeldtia): mitogenome assembly from transcriptomes and anchored hybrid enrichment datasets.

The nearly complete mitogenomes of the two species of North African Atlas geckos, Quedenfeldtia moerens and Q. trachyblepharus were assembled from anchored hybrid enrichment data and RNAseq data. Congruent assemblies were obtained for four samples included in both datasets. We recovered the 13 protein-coding genes, 22 tRNA genes, and two rRNA genes for both species, including partial control region. The order of genes agrees with that of other geckos.

Review of the systematics, distribution, biogeography and natural history of Moroccan amphibians.

The amphibian fauna of the Kingdom of Morocco was traditionally regarded as poor and closely related to its European counterpart. However, an increase in research during the last decades revealed a considerable degree of endemism amongst Moroccan amphibians, as well as phenotypic and genotypic inter- and intraspecific divergence. Despite this increase in knowledge, a comprehensible overview is lacking while several systematic issues have remained unresolved. We herein present a contemporary overview of the distribution, taxonomy and biogeography of Moroccan amphibians. Fourteen fieldtrips were made by the authors and colleagues between 2000 and 2012, which produced a total of 292 new distribution records. Furthermore, based on the results of the present work, we (i) review the systematics of the genus Salamandra in Morocco, including the description of a new subspecies from the Rif- and Middle Atlas Mountains, Salamandra algira splendens ssp. nov.; (ii) present data on intraspecific morphological variability of Pelobates varaldiiand Pleurodeles waltl in Morocco; (iii) attempt to resolve the phylogenetic position of Bufo brongersmai and erect a new genus for this species, Barbarophryne gen. nov.; (iv) summarize and assess the availability of tadpole-specific characteristics and bioacoustical data, and (v) summarize natural history data.

Does testosterone influence activity budget in the male Greek tortoise (Testudo graeca graeca)?

The stimulatory effect of testosterone on male sexual activity is one of the clearest examples linking hormones and behaviors. However, this relationship is complex in Chelonians. We experimentally studied the influence of testosterone levels on the activity budget and space use in male Greek tortoises (Testudo graeca graeca) during the spring mating season. We first described the annual pattern of changes in plasma testosterone levels in free-ranging animals in Morocco. Two peaks, one in winter and one in summer, corresponded to periods of inactivity; whereas mating periods in spring and to a lesser extent in autumn were associated with low plasma testosterone levels. Second, we experimentally manipulated plasma testosterone levels in free-ranging males, and analyzed the behavioral consequences. The strong contrasts in plasma hormone levels induced by the experimental treatments did not result in changes in activity budget or space use, both in the short-term or more than one month after the beginning of the hormonal treatment. Our results suggest that testosterone levels did not influence directly behavioral activity in this species, either immediately or after a time delay of one month.