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Heterakis gallinarum is one of the most recurrently diagnosed nematodes within the gastrointestinal tract of galliform birds. In the present study, we investigated the genetic diversity of 96 H. gallinarum collected from free-range chickens from different localities in Tunisia. We assessed phylogeny and genetic variability using the internal transcribed spacers (ITS) of the nuclear ribosomal DNA and the mitochondrial cytochrome c oxidase subunit I gene. Haplotype and nucleotide diversities indicate that H. gallinarum is a species with extremely low genetic diversity. Based on the networks and phylogenetic trees, there was strong support for the absence of significant geographical structuring among the H. gallinarum populations in different localities in Tunisia. Mismatch distributions and neutrality tests of both genetic markers suggest that at least one expansion event occurred in the population demographic history of H. gallinarum. Our data showed a lack of population structure using the pairwise fixation index (FST) and an extensive gene flow. It indicated that the most likely major contributor to the low genetic diversity and gene flow is the movement of the parasitized birds; in other words, the frequency of the poultry trade considerably affects patterns of worm gene flow. [ABSTRACT FROM AUTHOR]

Larval forms of the genus Hysterothylacium have been previously reported in teleost fish from the North African coasts of central Mediterranean Sea by morphological analysis. In the present study, samples identified morphologically as Hysterothylacium aduncum ( n = 62), from Merluccius merluccius, Trachurus mediterraneus and Pagellus erythrinus from different geographical locations of the Tunisian coasts, were genetically characterised by sequences of the first (ITS-1), the 5.8S and second (ITS-2) Internal Transcribed Spacers (ITS) of nuclear ribosomal DNA (rDNA). Comparison of the sequences obtained with those available in public gene databases confirmed that all the samples from the Tunisian coasts belong to a single species, namely H. aduncum. All specimens from the Tunisian coasts showed one indel in position 787 in ITS-2 sequences not reported by any of the previously published sequences from the Atlantic and Pacific Oceans, Adriatic Sea (Mediterranean Sea) and the East Greenland Sea, suggesting the existence of a population-specific pattern exhibiting a low differentiation of this parasite in this area. This is the first molecular characterization of H. aduncum from the Tunisian coasts using ITS rDNA sequences which allows the definition of genetic markers for their unequivocal identification, and provides further biological data on these nematodes in marine fish off the Tunisian coasts, improving the picture of the occurrence of these taxa in the North African coasts of central Mediterranean Sea. [ABSTRACT FROM AUTHOR]

Fasciolosis caused by Fasciola spp. (Platyhelminthes: Trematoda: Digenea) is considered the most important helminth infection of ruminants in tropical countries, causing considerable socioeconomic problems. Samples identified morphologically as Fasciola sp. in Equus caballus host species from the city of Tunis (north of Tunisia) were genetically characterized, for the first time, by sequences of the 1st (ITS-1), the 5.8S and 2nd (ITS-2) Internal Transcribed Spacers (ITS) of nuclear ribosomal DNA (rDNA). Comparison of the ITS of the Tunisian samples with sequences of Fasciola spp. from GenBank showed that the specimens examined had sequences identical to those of F. hepatica sequences selected as reference. The nucleotide sequencing of ITS rDNA showed no nucleotide variation in the ITS-1, 5.8S, or ITS-2 rDNA sequences among all F. hepatica samples parasitizing horses from Tunis, versus 2 ITS-2 haplotypes in standard F. hepatica, showing a substitution C/T in position 859, reported previously from Tunisia, Algeria, and Spain. The present study is the first demonstration of the existence of the most frequent haplotype (FhITS-H1) of F. hepatica species in horses from Tunis (north of Tunisia) by the genetic approach using ITS rDNA as genetic marker, providing a foundation for further studies on Fasciola sp. in North African countries. [ABSTRACT FROM AUTHOR]

Morphometric variation of Pelophylax saharicus was analysed using univariate and multivariate statistics, with both traditional and geometric morphometrics, based on 148 specimens from four different geographical localities in Tunisia and Algeria. The results show the existence of three morphotypes in Tunisia and one in Algeria, and indicate a significant degree of variation in morphometric characters between regions. Specimens from the southernmost region have the smallest body size and the greatest morphometric divergence from other populations. This pattern of morphometric variation probably results from phenotypic plasticity in response to local environmental factors. The results of our chromosomal study (C-, Ag-NOR-, endonuclease digestion, DAPI, CMA3 and Q-banding) reveal this species to exhibit the plesiomorphic Pelophylax karyotype of 2n = 26 biarmed chromosomes and NORs on the eleventh pair. Similarities and differences of the North African green frog are discussed in relation to the different forms of data collected (chromosomal, morphometric, ecological). [ABSTRACT FROM AUTHOR]

The intraspecific variation of mitochondrial cytochrome b sequence was analysed for two widely distributed Tunisian anuran species, Discoglossus pictus and Pelophylax saharicus. We observed low genetic variation within both species (D. p. auritus 0.6% uncorrected genetic distances, P. saharicus 0.2%) and the absence of genetic structure within both species in the studied region. There is no correspondence between previously described morphotypes of P saharicus in Tunisia and our genetic data. Our results show a lack of phylogeographic structure in both species despite their fragmented distribution across Tunisia. [ABSTRACT FROM AUTHOR]

Rana saharica is the most widely distributed anuran in Tunisia. We examined morphological variation in 124 specimens as a function of their geographical origin, using univariate and multivariate statistics with traditional morphometrics. Our results supported the existence of three morphotypes of this species, correctly classified 84.7% of specimens, and indicated significant degrees of variation in morphometric characteristics between regions. No significant differences were found between sexes among populations. We observed an increase in body size and a decrease in body mass as a function of latitude. Specimens from the southernmost region had the smallest body size and the greatest morphometric divergence. The pattern of morphometric variation probably results from phenotypic plasticity correlated with local environmental factors. [ABSTRACT FROM AUTHOR]

We present the results of a chromosomal study (C-, Ag-NOR-, endonuclease digestion, DAPI and CMA3 banding) carried out on four Tunisian species of amphibians: Burg mauritanicus, B. viridis, Rana saharica and Discoglossus pictus auritus. For comparative purposes, our cytogenetic analysis was also extended to Moroccan specimens of B. mauritanicus and D. pictus scovazzi. Tunisian specimens of B. viridis display chromosome and chromatin characters very similar to those found in other specimens, both North African and Euro-Asian. By contrast, chromosomes of Tunisian specimens of B. mauritanicus differed from the Moroccan specimens in presenting a different Q-banding phenotype. Tunisian specimens of R. saharica displayed the plesiomorphic Pelophylax karyotype of 2n = 26 biarmed chromosomes and NORs on the eleventh pair. The Tunisian specimens of D. pictus auritus had the typical D. pictus 2n = 28 karyotype with the 13th pair of telocentric elements, in contrast to the Spanish auritus specimens that display the chromosomes of this pair shaped as metacentric. Similarities and differences of the four studied Tunisians anuran taxa are discussed with references to published data on their relatives. [ABSTRACT FROM AUTHOR]

• Novel and common molecular markers support the monophyly of F. hepatica from Tunisia and Algeria. • Pepck multiplex PCR patterns could not differentiate between Fasciola species. • Sequence analysis of pepck revealed high polymorphism, length variation, within this intronic marker. • Nuclear and mitochondrial markers revealed an important gene flow between Tunisian and Algerian populations of F. hepatica. • The combination of nuclear and mitochondrial sequence analysis is still the best method to distinguish these taxa. Fasciolosis is a foodborne trematodosis characterised by a worldwide distribution. Various approaches have been developed for the study of the causative agents of this parasitic infection: Fasciola hepatica , Fasciola gigantica and the aspermic intermediated forms (hybrid and introgressed). In the present study, novel and common molecular markers (pepck and pold, ITS, CO1, ND1 and CO1-trnT-rrnL) were used to characterise Fasciola flukes from the Tunisian-Algerian border, to estimate the gene flow between these populations and to evaluate the reliability of different molecular markers. All nuclear and mitochondrial markers, apart from pepck, supported the monophyly of the studied flukes identified as F. hepatica. Multiplex PCR for pepck revealed three different genotypes corresponding to F. hepatica (pepck-Fh), F. gigantica (pepck-Fg) and the aspermic Fasciola flukes (pepck-Fh/Fg). Sequence analysis of pepck revealed high polymorphism, length variation, within this intronic marker. The observed inconsistencies were due to the position of the forward primer within the intronic region. Pepck sequences showed different level of heterozygosity and homozygosity with length polymorphisms in the introns. Pepck multiplex PCR patterns could not differentiate between Fasciola species. All studies based on only pepck multiplex PCR with mitochondrial markers should be revised. Nuclear and mitochondrial markers revealed an important gene flow between Tunisian and Algerian populations of F. hepatica. The combination of nuclear and mitochondrial sequence analysis is still the best method to distinguish these taxa. Effective measures are needed in order to better control cross-country illegal trade of vector. [ABSTRACT FROM AUTHOR]

Fasciolosis caused by Fasciola spp. (Platyhelminthes: Trematoda: Digenea) is considered the most important helminth infection of ruminants in tropical countries, causing considerable socioeconomic problems. In the present study, samples identified morphologically as Fasciola hepatica from sheep and cattle from different geographical locations of Tunisia and Algeria were genetically characterised by sequences of the first (ITS-1), the 5.8S and second (ITS-2) Internal Transcribed Spacers (ITS) of nuclear ribosomal DNA (rDNA) and mitochondrial Cytochrome c Oxidase subunit I (COI) gene. Comparison of the ITS and COI sequences of the North African samples with sequences of Fasciola spp. from GenBank confirmed that all samples from Tunisia and Algeria samples belong to a single species, namely F. hepatica. Several specimens from Tunisia and Algeria showed a substitution C/T in position 859 in the ITS-2 sequences, previously reported from Spain, suggesting that the above mentioned variant may have a common origin and spread recently throughout the three countries because of movement of infected animals. This is the first molecular characterization of F. hepatica in North Africa which provides a foundation for further studies on Fasciola spp. in Tunisia and Algeria.

Two gadiform species with a successive bathymetric and an ecological and economical importance in the Mediterranean fishing industry, Phycis blennoides and Phycis phycis, were selected for the present study. A total of 592 fresh specimens belonging to the Gadiformes genus were obtained from local commercial fisheries. The investigation was centred on anisakid parasites of 272 specimens of the greater forkbeard (P. blennoides) and 320 of the forkbeard (P. phycis) captured off the Mediterranean coasts of Tunisia (eastern Mediterranean Sea). Four species of nematodes were identified: Anisakis simplex s.1., Anisakis physeteris, Hysterothylacium aduncum and Hysterothylacium fabri. The total prevalence was 53.75% in the forkbeard and 51.47% in the greater forkbeard. The highest values of prevalence (38.75%, 2-14), mean intensity (6.74+/-3.4) and mean abundance (2.61) were all obtained for H. fabri L4 in the forkbeard. The most frequent parasite in the greater forkbeard was H. aduncum L3 with 32.35% (1-3) prevalence and values of 1.21+/-0.58 and 0.39 for mean intensity and mean abundance, respectively. The infestation parameters were also analysed according to the host length, and prevalence was highest in P. blennoides longer than 35 cm in respect of all anisakid species. Whilst in P. phycis, the highest prevalence, conditioned by H. fabri parasitisation, was found in fish with length reaching a maximum of more than 40 cm. When the data were grouped seasonally, clear patterns were observed for P. blennoides and P. phycis species, with prevalence and mean intensity of all the anisakid species peaking in spring and summer.