Abstract: Background: Glossina fuscipes fuscipes is the major vector of human African trypanosomiasis, commonly referred to as sleeping sickness, in Uganda. In western and eastern Africa, the disease has distinct clinical manifestations and is caused by two different parasites: Trypanosoma brucei rhodesiense and T. b. gambiense. Uganda is exceptional in that it harbors both parasites, which are separated by a narrow 160-km belt. This separation is puzzling considering there are no restrictions on the movement of people and animals across this region. xD;Methodology and Results: We investigated whether genetic heterogeneity of G. f. fuscipes vector populations can provide an explanation for this disjunct distribution of the Trypanosoma parasites. Therefore, we examined genetic structuring of G. f. fuscipes populations across Uganda using newly developed microsatellite markers, as well as mtDNA. Our data show that G. f. fuscipes populations are highly structured, with two clearly defined clusters that are separated by Lake Kyoga, located in central Uganda. Interestingly, we did not find a correlation between genetic heterogeneity and the type of Trypanosoma parasite transmitted. xD;Conclusions: The lack of a correlation between genetic structuring of G. f. fuscipes populations and the distribution of T. b. gambiense and T. b. rhodesiense indicates that it is unlikely that genetic heterogeneity of G. f. fuscipes populations explains the disjunct distribution of the parasites. These results have important epidemiological implications, suggesting that a fusion of the two disease distributions is unlikely to be prevented by an incompatibility between vector populations and parasite.
Abstract: The snake-eyed lizards of the genus Ophisops (Lacertidae) have been through a series of taxonomical revisions, but still their phylogenetic relationships remain uncertain. In the present study we estimate the phylogeographic structure of O. elegans across its distributional range and we evaluate the relationships between O. elegans and the sympatric, in North Africa, species O. occidentalis, using partial mtDNA sequences (16S rRNA, CO1, and cyt b). All phylogenetic analyses produced topologically identical trees where extant populations of O. elegans and O. occidentalis were found polyphyletic. Taking into account all the potential causes of polyphyly (introgressive hybridization, incomplete lineage sorting, and imperfect taxonomy) we suggest the inaccurate taxonomy as the most likely explanation for the observed pattern. Our results stress the need for re-evaluation of the current taxonomical status of these species and their subspecies. Furthermore, Our biogeographic analyses and the estimated time of divergences suggest a late Miocene diversification within these species, where the present distribution of O. elegans and O. occidentalis was the result of several dispersal and vicariant events, which are associated with climatic oscillations (the late Miocene aridification of Asia and northern Africa) and paleogeographic barriers of late Miocene and Pliocene period. (C) 2008 Published by Elsevier Inc.
Abstract: Background: If the insect innate immune system is to be used as a potential blocking step in transmission of malaria, then it will require targeting one or a few genes with highest relevance and ease of manipulation. The problem is to identify and manipulate those of most importance to malaria infection without the risk of decreasing the mosquito's ability to stave off infections by microbes in general. Molecular evolution methodologies and concepts can help identify such genes. Within the setting of a comparative molecular population genetic and phylogenetic framework, involving six species of the Anopheles gambiae complex, we investigated whether a set of four preselected immunity genes (gambicin, NOS, Rel2 and FBN9) might have evolved under selection pressure imposed by the malaria parasite. xD;Results: We document varying levels of polymorphism within and divergence between the species, in all four genes. Introgression and the sharing of ancestral polymorphisms, two processes that have been documented in the past, were verified in this study in all four studied genes. These processes appear to affect each gene in different ways and to different degrees. However, there is no evidence of positive selection acting on these genes. xD;Conclusion: Considering the results presented here in concert with previous studies, genes that interact directly with the Plasmodium parasite, and play little or no role in defense against other microbes, are probably the most likely candidates for a specific adaptive response against P. falciparum. Furthermore, since it is hard to establish direct evidence linking the adaptation of any candidate gene to P. falciparum infection, a comparative framework allowing at least an indirect link should be provided. Such a framework could be achieved, if a similar approach like the one involved here, was applied to all other anopheline complexes that transmit P. falciparum malaria.
Abstract: Attributed to human-mediated dispersal, a species of the Anopheles gambiae complex invaded northeastern Brazil in 1930. This event is considered unique among the intercontinental introductions of disease vectors and the most serious one: "Few threats to the future health of the Americas have equalled that inherent in the invasion of Brazil, in 1930, by Anopheles gambiae." Because it was only in the 1960s that An. gambiae was recognized as a species complex now including seven species, the precise species identity of the Brazilian invader remains a mystery. Here we used historical DNA analysis of museum specimens, collected at the time of invasion from Brazil, and aimed at the identification of the Brazilian invader. Our results identify the arid-adapted Anopheles arabiensis as being the actual invading species. Establishing the identity of the species, in addition to being of intrinsic historical interest, can inform future threats of this sort especially in a changing environment. Furthermore, these results highlight the potential danger of human-mediated range expansions of insect disease vectors and the importance of museum collections in retrieving historical information.
Abstract: Mesalina is a widespread lacertid genus occurring throughout the Saharo-Sindian region from North Africa to Pakistan. It has been through a series of taxonomic revisions, but the phylogenetic relationships among the species remain unclear. In this study we estimate the phylogeographic structure of M. guttulata across most of its distributional range and we evaluate the relationships between M. guttulata and the sympatric species M. brevirostris and M. bahaeldini using partial mitochondrial DNA (mtDNA) sequences (cyt b and 16S). M, guttulata and M. brevirostris represent species complexes, whereas M. bahaeldini considered before as M. guttulata is a recently described species with very restricted distribution. Here we present the first evidence that M. guttulata is a paraphyletic taxon with respect to M. bahaeldini, while M. brevirostris proves to be a polytypic species or even a species complex, confirming previous studies. Although mtDNA markers have several properties that make them suitable for phylogeographic studies, they are not free of difficulties. Phylogeographic inferences within and between closely related species can be mislead by introgression and retention of ancestral polymorphism (incomplete lineage sorting). However, the present distribution pattern, the estimated times of divergence and the significant variation in morphology within M. guttulata led us to accept that the paraphyletic pattern observed, is most likely due to inaccurate taxonomy. Our hypothesis is that what has hitherto been considered as intraspecific variation, actually reflects species-level variation. Furthermore, our biogeographic analyses and the estimated time of divergences suggest that the present distribution of M. guttulata was the result of several dispersal and vicariant events, which are associated with historical changes (climatic oscillations and paleogeographic barriers) of late Miocene and Pliocene period. (C) 2008 Elsevier Inc. All rights reserved.
Abstract: In the present study, we employed three mitochondrial DNA genetic markers in a phylogenetic analysis aiming at the delineation of the relationships amongst nominal Trachelipus kytherensis populations, as well as between populations of this species and of Trachelipus aegaeus and a new form, occurring syntopically with the latter. Both the phylogenetic analysis and the genetic distances separating populations, show the presence of several distinct and well differentiated clades that undermine the monophyly of T kytherensis. On the other hand, despite the insular distribution of T aegaeus populations, their divergence is low and the monophyly of this taxon can be rescued by the inclusion of two more insular populations previously assigned to T kytherensis. The patterns of genetic divergence among clades are only partially congruent with the geographic distribution of populations. The validity of taxonomic characters used so far in the genus appears to be questionable; therefore, a more comprehensive phylogenetic study at a population level is deemed necessary for understanding the divergence of Trachelipus lineages. (C) 2008 The Linnean Society of London, Biological Journal of the Linnean Society, 2008, 95, 361-370.
Abstract: Sequence data derived from two mitochondrial markers, 16S rRNA and COI genes, were used to infer the evolutionary history of 47 insular and mainland populations covering most of the distributional range of the northeastern Mediterranean scorpion species Mesobuthus gibbosus. Based on the estimated divergence times of Mesobuthus lineages, the temporal frame of the genus differentiation in the northeastern Mediterranean region is placed in middle Miocene (15 million years ago). The biogeographic affinities of M. gibbosus populations point towards a mainly vicariant pattern of differentiation of the species which is consistent with the geological events that transformed the Aegean region during the period from 12 to 5 million years ago. M. gibbosus is an old northeastern Mediterranean species that has retained valuable bits of genetic information, reflecting some of the oldest vicariant events that have occurred in the area. Most importantly, the history witnessed by M. gibbosus has not been obscured by more recent palaeoevents of the region. Therefore, the case of M. gibbosus is in favour of a taxon-oriented 'perception' of the natural history of a given area.
Abstract: Aim This study uses molecular data in conjunction with palaeogeography to infer the most plausible biogeographical scenario accounting for the current distributional pattern of Iurus dufoureius. Location North-eastern Mediterranean region. Methods Sequencing of a 441-bp segment of the mitochondrial 16S rRNA gene in seven populations covering the whole distributional range of the species. Phylogenetic analyses performed included neighbour joining, maximum parsimony and Bayesian inference. Results The molecular phylogeny showed that two Iurus clades are strongly supported. These clades correspond to the two subspecies Iurus dufoureius dufoureius and Iurus dufoureius asiaticus, currently recognized within the genus. The assumption of a clock-like evolution could not be rejected, and this enabled us to estimate an approximation of the local rate of evolution for the I. dufoureius lineages. Based on the estimated evolutionary rate (0.79 +/- 0.17 Myr(-1)), the split between the two Iurus clades occurred c. 8 Ma. Main conclusions Contrary to what was believed in the past, the genus Iurus is an old north-eastern Mediterranean genus that has been differentiating in southern Greece and south-west Turkey at least from the middle Miocene. According to the phylogenetic trees obtained and the dating of the divergence times of lineages, the genus dispersed into the Aegean Archipelago when the Aegean was still a uniform land mass. Although the phylogenetic relationships of I. d. dufoureius populations have been shaped by the most recent vicariant events, the phylogenetic relationships of I. d. asiaticus populations are mostly attributable to older palaeoevents occurring in the area.
Abstract: The phylogeny of Greek populations of the terrestrial isopod genus Ligidium is reconstructed based on three mtDNA gene segments: 12S rRNA, 16S rRNA and COI. Two widely distributed European species, as well as three outgroups belonging to different isopod genera, were also included in the analyses. The samples used represent almost all Ligidium species known to occur in Greece, as well as several populations of unknown specific status plus some new records. Phylogenetic analyses of the combined data set were performed using Bayesian inference and maximum parsimony. The two main sister clades with good support indicate the sympatric differentiation of two lineages in southern continental Greece (Peloponnisos), where Ligidium populations exhibit a mosaic distribution of sibling species. The insular populations of the Aegean Islands show increased genetic divergence and form separate clades. The presence of a third lineage of Asiatic origin is strongly suggested by both the molecular phylogeny and morphology. The only presumably valid diagnostic morphological character exhibits only partial correspondence to well supported clades of the molecular phylogeny. Genetic differentiation between populations is very high, a fact that can be attributed to the strict ecological specialization of these animals that leads to increased levels of isolation even between populations that are in close proximity. As a consequence, Greek Ligidium populations, especially those present on islands, are unique genetic pools and extremely vulnerable to extinction.
Abstract: During the Pleistocene pygmy elephantids, some only a quarter of their ancestors' size, were present on Mediterranean islands until about 10 000 years ago (y.a.). Using a new methodology for ancient DNA (aDNA) studies, the whole genomic multiple displacement amplification method, we were able to retrieve cytochrome b (cytb) DNA fragments from 4200 to 800 000 y.a. specimens from island and mainland samples, including pygmy and normal-sized forms. The short DNA sequence (43 bp) retrieved from the 800 000 y.a. sample is one of the oldest DNA fragment ever retrieved. Duplication of the experiments in two laboratories, the occurrence of three diagnostic sites and the results of the phylogenetic analyses strongly support its authenticity. Our results challenge the prevailing view that pygmy elephantids of the eastern Mediterranean originated exclusively from Elephas, suggesting independent histories of dwarfism and the presence of both pygmy mammoths and elephant-like taxa on these islands. Based on our molecular data, the origin of the Tilos and Cyprus elephantids from a lineage within the genus Elephas is confirmed, while the DNA sequence from the Cretan sample falls clearly within the mammoth clade. Thus, the name Mammuthus creticus rather than Elephas creticus, seems to be justified for this form. Our findings also suggest a need to re-evaluate the evolutionary history of the Sicilian/Maltese species, traditionally included in the genus Elephas.
Abstract: The Mediterranean land snail genus Mastus (Beck, 1837) is highly divergent. Thirty-two Mastus species have been recorded throughout the genus range, and 23 of them are endemic to the islands of the Aegean Sea and mainland Greece. Of these, all 16 Mastus species reported from Crete are endemic to this island. A robust molecular phylogenetic framework based on mitochondrial and nuclear genes (1623 bp) allowed us to explore the temporal diversification pattern of lineages, using molecular clock approaches. Our results showed an initial radiation in the evolutionary history of the Cretan lineage, followed by a subsequent slowdown of lineage splitting rate. Using a dated major vicariant event of the Aegean area, we estimated the absolute time of the radiation event and proposed a biogeographic scenario accounting for the observed pattern. Additionally, we tried to infer the processes that led to the divergence of the Cretan Mastus species, by applying comparative methods in phylogenetically informated context. Overall, our results favoured a nonecological radiation scenario in the Cretan Mastus species due to an allopatric divergence of secondary sexual characters.
Abstract: We studied the population dynamics of Mastus by investigating the effects of population structure, spatial ecology and biology of reproduction on the genetic diversity of two sympatric Mastus species endemic to the island of Crete. Over a period of 27 months, we carried out both mark-recapture and random quadrat sampling techniques in order to assess the dispersal trends, the aggregation patterns, the neighbourhood size and the habitat preferences of these species. There were 154 recorded movements for M. butoti and 114 for M. cretensis. Mean monthly dispersal was estimated at d = 0.5 m for M. butoti and d = 1 m for M. cretensis. Both species showed a random dispersal pattern but tended to aggregate in the field. Their populations were found to be highly structured owing to their highly parsimonious dispersal behaviour and the very low population densities, estimated at D = 2.07 +/- 0.16 and D = 0.73 +/- 0.16 individuals m(-2) for M. butoti and M. cretensis, respectively. The neighbourhood size did not exceed 150 individuals for either species. The habitat occupied by each species changed during the active season, but both the immature and the adult individuals of each species seemed to prefer the same habitats throughout the active season. Partial population activation during the active season was observed in both species. We conclude that the population structure, the partial population activation and the species-specific reproductive strategies have a profound effect on maintaining the genetic diversity of the genus' populations. (C) 2004 The Linnean Society of London.
Abstract: The land mollusc fauna of 1 km(2) of Cretan maquis was surveyed by sampling fourteen 400-m(2) plots in May 2001, and by resampling six of these in February 2002. Sampling methods were designed to resemble those used in similar surveys of 1-km(2) sites in tropical rainforests in Cameroon and Sabah. A total of 27 species were recorded for the site. Slugs and a semi-slug were found only in the 2002 survey. Individual plots were very similar in species richness and composition; the richest plot contained about 85% of the fauna recorded for the whole site. Overall densities were very high. This local homogeneity contrasts with marked heterogeneity over Crete as a whole. These results are contrasted with those from tropical forests, where individual plots vary considerably in richness and composition, where densities are lower and where the total site faunas are larger. Although there are problems associated with differing amounts of sampling error between studies, this contrast is striking and some possible causes are discussed. Further work in tropical forest on limestone may elucidate this.
Abstract: Three endemic Cretan land snail species of the genus Mastus (Beck, 1837) from the island group of Koufonisi (southeast Crete) and the eastern part of Crete, were studied by multivariate analysis of shell morphology and analysis of mtDNA sequences. The phylogeny of the populations studied and the processes effecting the genetic and morphological diversity of the species were investigated. Extremely high mtDNA sequence divergence was observed both within and between populations. The Cretan populations could not be distinguished morphologically, while the populations of the islets were more distinct. We argue that the active geological past of the area (including sea level changes) and the long-term presence of humans has produced a mixing up of Mastus populations leading to the accumulation of high divergence of mtDNA sequences on a small spatial scale. The limited morphological diversity and the distinct shell 'identity' of the islets' populations can be attributed to the selective pressures of the island group. (C) 2003 The Linnean Society of London.
Abstract: The present study reports an investigation into the reproductive biology and ecology of two sympatric species of Mastus endemic to the island of Crete (southern Greece). The study lasted 6 months, from September 1997 to March 1998, and involved field observations on single and copulating individuals, and the study of the spermatophores within the genital tract. The two species have different aestivation habits and emergence times. Mastus olivaceus, which aestivates closer to the surface of the soil or in the litter, emerges soon after the first rains, while Mastus cretensis emerges later. Additionally, M. olivaceus starts aestivating over a month earlier than M. cretensis. M. olivaceus reproduces from late September till early January, while M. cretensis starts in early November and ends in early March. The bursa-like diverticulum of M. cretensis bears only one spermatophore, but in M. olivaceus from one to three spermatophores were found. The 91 spermatophores of M. olivaceus examined indicate intrapopulation variability in the number of transverse ridges and number of spines in the ornamentation of its distal part. These types of difference in spermatophore morphology should not, therefore, be considered species-specific.
