Genetic Characterization and Alternative Preservation Ways of Locally Adapted Sheep Breeds: Cases of Private and Public Sheep Sectors in Tunisia and Italy.

Non-commercialized sheep breeds known as local or native breeds are well adapted to their environmental constraints and constitute precious genetic resources that need prioritization for genetic diversity characterization and preservation. The aim of the present study was to assess the genetic diversity level and the related preservation decisions of very old and traditional native Mediterranean sheep breeds from Tunisia and Italy using 17 microsatellite markers. In total, 975 sheep were sampled from five Tunisian, one Center Italian, and four Venetian native breeds. Both private and publicly available farms were considered for each breed for breeding strategies' comparison purposes. The microsatellite set used was highly informative (PIC = 0.80 ± 0.08), with a total of 383 alleles. Moderate genetic differentiation was revealed between the native sheep of the two Mediterranean sides (global overall loci FST = 0.081). The genetic diversity level was higher in the case of the Tunisian native breeds compared to the Italian ones, as evidenced by higher mean allelic richness, higher expected and observed heterozygosities, and lower inbreeding levels. Priority for conservation suggestions was carried out for each private or public breed population based on the contribution of each population to the diversity of the whole data. The four Venetian breeds, already undergoing conservation, the Tunisian dairy breed, and the very ancient Maghrebian breed, would be favored for conservation. In conclusion, our results highlighted the importance of the analyzed Mediterranean native sheep breeds as valuable inherited genetic reservoirs and supported previous conservation decisions made for the threatened breeds.

Historical Westward Migration Phases of Ovis aries Inferred from the Population Structure and the Phylogeography of Occidental Mediterranean Native Sheep Breeds.

In this study, the genetic relationship and the population structure of western Mediterranean basin native sheep breeds are investigated, analyzing Maghrebian, Central Italian, and Venetian sheep with a highly informative microsatellite markers panel. The phylogeographical analysis, between breeds' differentiation level (Wright's fixation index), gene flow, ancestral relatedness measured by molecular coancestry, genetic distances, divergence times estimates and structure analyses, were revealed based on the assessment of 975 genotyped animals. The results unveiled the past introduction and migration history of sheep in the occidental Mediterranean basin since the early Neolithic. Our findings provided a scenario of three westward sheep migration phases fitting properly to the westward Neolithic expansion argued by zooarcheological, historical and human genetic studies.

Genetic Variability and Population Structure of the Tunisian Sicilo-Sarde Dairy Sheep Breed Inferred from Microsatellites Analysis.

This study analyzed the genetic variability, inbreeding and population structure of the Tunisian-North African dairy sheep breed, the Sicilo-Sarde (SS), created by crossing the Sarda and Comisana dairy breeds. The level of variability in the SS, considered as an endangered breed after a dramatic decrease, was assessed using 17 microsatellite markers by analyzing the two breed populations sampled from their respective cradles: SS of Beja (SSB, n = 27) and SS of Mateur (SSM, n = 25). High levels of genetic diversity in SS were revealed, with a total of 212 alleles, a high mean number of alleles (12.47 ± 4.17) and a high average polymorphism information content (PIC) (0.81 ± 0.10). The observed heterozygosity was considerable in SSB and SSM (0.795 and 0.785, respectively). The inbreeding level measured by the population inbreeding coefficient FIS is higher in the SSM population (0.121) than in the SSB population (0.090). The higher genetic diversity level detected in SSB reflected the effect of new Italian Sarda genes introduced by intra-uterine artificial insemination recently practiced in this population. The Wilcoxon test and the mode-shift distribution indicated that the SS breed is a non-bottlenecked population. The structural analysis reflected the historical miscegenation practiced during the breed creation and highlighted further ancient miscegenation, which could date back to the first waves of sheep introduction to the western Mediterranean region. Microsatellite markers were successfully applied in the assessment of the genetic variability of SS and should be used in monitoring this variability during the application of conservation strategies.

A comparative study of trace elements in Cymodocea nodosa from three semi-enclosed coastal areas in Tunisia.

The present study quantifies the levels of five trace elements (TEs) Zn, Cu, Ni, Pb, and Cd in the leaves and rhizomes of Cymodocea nodosa as well as the surficial sediments from three semi-enclosed coastal areas in Tunisia, in the south Mediterranean Sea. Samples were taken from the Bizerte and Ghar El Melh lagoons and from marina Cap Monastir. The TE ranking was found to be Zn > Cu > Pb > Ni > Cd in sediments and Zn > Cu = Ni = Pb = Cd in C. nodosa leaves and rhizomes. Except for Ni, levels of Zn, Cu, Pb, and Cd significantly differed between the sites. Translocation factors (TFs) were > 1 for all trace elements proving the high capacity of C. nodosa to accumulate TEs in its above-ground tissues. Results show that marina Cap Monastir's meadow exhibits higher TFs than the Bizerte and Ghar El Melh lagoons. This can be due to the presence of the non-indigenous species Halophila stipulacea. The present study highlights the need for further investigation on the effect of interspecific interaction on TE uptake by seagrasses.

Variation along the year of trace metal levels in the compartments of the seagrass Posidonia oceanica in Port El Kantaoui, Tunisia.

The accumulation of the five trace metals (TMs) cadmium, copper, lead, nickel and zinc was measured in Posidonia oceanica leaves. Shoots were seasonally sampled at 8-10-m depth from four stations located in Port El Kantaoui area, Tunisia, during four campaigns performed in 2012. Levels of the five TMs were analyzed using inductively coupled plasma atomic emission spectrometry (ICP-AES) in three compartments of P. oceanica shoots: blades and sheaths of adult leaves and intermediate leaves. Results showed a preferential accumulation of Cd, Pb, Ni and Zn in adult leaf blades. Therefore, we focus on the study of this compartment. TM levels of blades of adult leaves decreased in the following order: Zn > Ni > Cu > Pb > Cd, irrespective of the season. Levels of the five TMs significantly differed between seasons (p < 0.01). Levels of Cd and Cu showed a seasonal pattern: Cd decreased from spring to winter while Cu increased during that same period of time. A significant correlation (p < 0.01) was found between Cd-Cu and Cd-Pb. A significant correlation (p < 0.05) was also noted between Cd-Ni in the adult leaf blades. A relationship was recorded between the foliar surface of the adult leaf blades and Zn accumulation. This survey allowed to highlight the annual variation of TM accumulation in adult leaf blades of P. oceanica, in relation with ecophysiology of this seagrass. Therefore, this study reinforces the usefulness and the relevance of this compartment of P. oceanica, easy to sample without destruction of whole shoot, as a bioindicator of Zn, Ni, Cd and Pb contamination.

Abundance and diversity of amphipods (Crustacea: Peracarida) on shallow algae and seagrass in lagoonal ecosystem of the Mediterranean Tunisian coast.

BACKGROUND: Bizerte lagoon is a semi-enclosed marine ecosystem, where various types of human activities have been developed. To investigate the distribution and temporal variation of amphipod assemblage, monthly sampling was conducted at the Menzel Jemil site from October 2009 to September 2010. RESULTS: Atotal number of 3,620 specimens were collected from floating algae and seagrass allowing the identification of 10 amphipod species. Moreover, several indices, species richness, dominance, mean density, and diversity index were estimated to characterize the amphipode assemblage. Gammarusaequicauda wasthe most dominant species in all seasons. In addition, the minimum and maximum values of species richness of amphipod were observed in January (3 species) and April (8 species), respectively. The mean density and species richness were negatively correlated with plant biomass. Mean Shannon index (H')and evenness (J')were 1.62 ± 0.34 and 0.67 ± 0.16, respectively. Non-metric multidimensional scaling (MDS) analysis based on the mean species density showed three seasonal groups of samples. Therefore, canonical correspondence analysis (CCA) made it possible to summarize the overall situation for the species, monthly sampling, and environmental parameters on a single graph. CONCLUSIONS: Thus, the temperature, turbidity, and chlorophyll a content are the most often reported factors for the distribution and structure of amphipods in the Bizerte lagoon.

Widespread Wolbachia infection in terrestrial isopods and other crustaceans.

Wolbachia bacteria are obligate intracellular alpha-Proteobacteria of arthropods and nematodes. Although widespread among isopod crustaceans, they have seldom been found in non-isopod crustacean species. Here, we report Wolbachia infection in fourteen new crustacean species. Our results extend the range of Wolbachia infections in terrestrial isopods and amphipods (class Malacostraca). We report the occurrence of two different Wolbachia strains in two host species (a terrestrial isopod and an amphipod). Moreover, the discovery of Wolbachia in the goose barnacle Lepas anatifera (subclass Thecostraca) establishes Wolbachia infection in class Maxillopoda. The new bacterial strains are closely related to B-supergroup Wolbachia strains previously reported from crustacean hosts. Our results suggest that Wolbachia infection may be much more widespread in crustaceans than previously thought. The presence of related Wolbachia strains in highly divergent crustacean hosts suggests that Wolbachia endosymbionts can naturally adapt to a wide range of crustacean hosts. Given the ability of isopod Wolbachia strains to induce feminization of genetic males or cytoplasmic incompatibility, we speculate that manipulation of crustacean-borne Wolbachia bacteria might represent potential tools for controlling crustacean species of commercial interest and crustacean or insect disease vectors.

[Phenetic analysis of populations of Porcellionides pruinosus (Brandt, 1833) (Crustacea, Isopoda, Oniscidea)]. / Analyse phénétique des populations de Porcellionides pruinosus (Brandt, 1833) (Crustacea, Isopoda, Oniscidea).

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.

[Reproductive phenology of a natural population of Armadillidium pelagicum Arcangeli, 1955 (Isopoda, Oniscidea)]. / Phénologie de la reproduction d'une population naturelle de Armadillidium pelagicum Arcangeli, 1955 (Isopoda, Oniscidea).

Armadillidium pelagicum, the most abundant species of the genus in Tunisia is endemic to the North of Tunisia and the circumsicilian islands. In the laboratory and under the natural conditions of temperature and photoperiod, virgin females of A. pelagicum exhibit a spontaneous ovarian maturation, followed by a parturial moult. Nevertheless, the onset of reproduction is greatly accelerated by the presence of a male. In fact, mating shortened the lag time (from the beginning of experiment to the parturial moult) by reducing the number of normal moult preceding the first parturial moult and the duration of the preparturial intermoult. In this species, only one mating is enough to ensure the fertility for several egg layings. Otherwise, it has been shown that A. pelagicum has a reproductive period rather than a reproductive activity and that mating females have a longer reproductive period than virgins. Like several species of terrestrial Isopods, the duration of the gestation period is temperature-dependent.

Life cycle and population dynamic of Armadillidium pelagicum Arcangeli, 1955 (Isopoda, Oniscidea) at Aouina.

Armadillidium pelagicum Arcangeli, 1955 is a terrestrial isopod endemic to the circum-Sicilian islands and the North of Tunisia. The life cycle and the population structure of this species were studied on a natural population at Aouina, in the surroundings of Tunis, over 16 months from, January 2000 to April 2001, by monthly or semi-monthly samplings. The ovigerous females were present from March/April to the end of August and absent from September to February/March. These observations indicate that A. pelagicum at Aouina has a seasonal reproduction, followed by a sexual rest. The recruitment period is spread from April/May to mid-September. The fecundity, estimated by the number of eggs in the marsupium of ovigerous females, exhibited a great variability, which is related to the weight of these females. The sex ratio underwent fluctuations throughout the sampling period. It was female-biased in most samplings. Mass frequency distribution was analysed and nine cohorts were identified during the sampling period. The field growth rates are high in the first life phase, decrease during winter and increase during spring. The characteristics of the life cycle of A. pelagicum at Aouina may be summarized as follows: (i) Semi-annual species, since females appear to produce up to five broods per year, (ii) iteroparous females, since females seem to reproduce twice or more in life; (iii) bivoltine life cycle, since the population produces two generations per year; (iv) variability of cohorts' life span.

[Biology and population dynamics of Porcellionides sexfasciatus (Crustacea, Isopoda, Oniscidea)]. / Biologie et dynamique de population de Porcellionides sexfasciatus (Crustacea, Isopoda, Oniscidea).

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.