Genetic and morphological analyses reveal a complex biogeographic pattern in the endemic barbel populations of the southern Italian peninsula

The Italian peninsula is a biodiversity hotspot, with its freshwater fish fauna characterized by high levels of local endemism. Two endemic fluvio‐lacustrine fishes of the genus Barbus (barbel, family Cyprinidae) have allopatric distributions in the Tyrrhenian and Adriatic basins of Italy. Barbus plebejus inhabits the mid‐ to northern Adriatic basins, while B. tyberinus is widespread in all central‐northern basins draining into the Tyrrhenian Sea. For basins in Southern Italy draining into the southern parts of these seas, there remains a knowledge gap on their barbel populations due to no previous genetic and morphological studies, despite their apparent biogeographic isolation. Correspondingly, this study quantified the presence and distribution of barbels in the Adriatic and Tyrrhenian basins of Southern Italy through genetic and morphological analyses of 197 fish sampled across eight populations. Testing of how local isolation has influenced the evolution and persistence of these populations was completed by examining sequence variation at two mitochondrial loci (cytochrome b and D‐loop) and performing geometric morphometric analyses of body shape, plus measuring 11 morphometric and meristic characters. Phylogenetic and morphological analyses revealed the presence of two genetically distinct lineages that differed significantly from adjacent B. tyberinus and B. plebejus populations. These two new taxa, here described as SI1 and SI2 Barbus lineages, are highly structured and reflect a complex mosaic biogeographic pattern that is strongly associated with the underlying hydrographical scenarios of the basins. The geographic isolation of these basins thus has high evolutionary importance that has to be considered for maintaining endemism.     

Morphologic and genetic variability in the Barbus fishes (Teleostei,Cyprinidae) of Central Italy

Italian freshwaters are highly biodiverse, with species present including the native fishes Barbus plebejus and Barbus tyberinus that are threatened by habitat alteration, fish stocking and invasive fishes, especially European barbel Barbus barbus. In central Italy, native fluvio‐lacustrine barbels are mainly allopatric and so provide an excellent natural system to evaluate the permeability of the Apennine Mountains. Here, the morphologic and genetic distinctiveness was determined for 611 Barbus fishes collected along the Padany–Venetian (Adriatic basins; PV) and Tuscany–Latium (Tyrrhenian basins; TL) districts. Analyses of morphological traits and mitochondrial DNA sequence data explored the natural and anthropogenic factors that have shaped their distribution ranges. Over 100 alien B. barbus were recorded in the Tiber basin (TL district) and Metauro basin (PV district). Comparisons of genetic and morphometric data revealed that morphometric data could identify alien B. barbus from native Barbus, but could not differentiate between B. tyberinus and B. plebejus. Genetic analyses revealed ~50 D‐loop mtDNA haplotypes and identified a distinct Barbus lineage present only in the Vomano River at the southern boundary of PV district. Demographic expansion and molecular variance analyses revealed a lack of geographic structuring across the sampling regions. While the contemporary B. plebejus distribution has been driven primarily by anthropogenic fish translocations, the dispersal of B. tyberinus has been via natural dispersion, including their crossing of the Apennine Mountains via temporary river connectivity. The results also revealed that the Barbus fishes of the mid‐Adriatic region of Europe have a complex pattern of local endemism. To conserve these patterns of genetic uniqueness, especially in the mid‐Adriatic basins, Barbus fishes should be managed by treating them as unique evolutionary units and ceasing translocations of all Barbus fishes between river basins.     

Combining geometric morphometrics with molecular genetics to investigate a putative hybrid complex: a case study with barbels Barbus spp. (Teleostei: Cyprinidae)

This integrative study examined the morphological and genetic affinities of three endemic barbel species from Italy (brook barbel Barbus caninus, Italian barbel Barbus plebejus and horse barbel Barbus tyberinus) and of putative hybrid specimens to their species of origin. Two of the species frequently occur together with the non‐native barbel Barbus barbus. DNA barcoding indicates that mitochondrial (mt) haplotypes often do not match the species expected from morphology. Linear distance measurements and meristics are not informative for discrimination of the species and putative hybrids, but a discriminant analysis of principal components (DAPC) of geometric landmark data produces reassignments largely in congruence with mt and nuclear genetic data. Cyto‐nuclear conflicts confirm the presence of hybridization in B. plebejus and B. tyberinus and identify additional introgressed specimens. A comparison between mixed genotypes and their morphology‐based assignment reveals no predictable pattern. The finding that most individuals of the morphologically similar B. plebejus and B. tyberinus have very high assignment probabilities to their respective species suggests that the presented approach may serve as a valuable tool to distinguish morphologically very similar taxa.     

Population dynamics of Barbus barbus plebejus (Valenciennes) and Leuciscus cephalus cabeda (Risso) in the middle River Po (Italy)

Scales from 427 Barbus barbus plebejus and 406 Leuciscus cephalus cabeda from the middle River Po were examined. The last annulus was formed between April and May 1977, except the 0–1 age-group of L.c. cabeda which developed its first year-ring between March and April 1977. The survival (S) and annual mortality (A) rates were equivalent in both fish populations; the same was true of both sexes of Leuciscus, but S in Barbus males was lower than in females. The sex ratio showed a preponderance of females in both species.Growth rate from June to September 1976 was higher than in the rest of the year. Annual growth in Barbus females was higher than in males, whereas both sexes in Leuciscus showed equivalent growth rates; growth rate was higher in Barbus than in Leuciscus. The P/B ratio of both species was higher in the young age-groups.Presented at the XLVII Congress of the Italian Zoological Union, Bergamo (Italy), September 1979.Presented at the XLVII Congress of the Italian Zoological Union, Bergamo (Italy), September 1979.     

Genetic structure of Barbus spp. populations in the Marches Region of central Italy and its relevance to conservation actions

A genetic survey of Barbus spp. populations in the Marches Region (Adriatic River basins), central Italy, was carried out using mitochondrial and nuclear markers (partial D‐loop, cyt b sequences and microsatellite loci) in order to ascertain their systematic position and to address their genetic structure which is key to conservation action planning. Analyses were conducted on sequences obtained from 91 individuals collected from eight sampling sites in five different rivers, from two specimens provided by the Ichthyological Centre of Rome and mitochondrial sequences of Barbus spp. retrieved from GenBank. Presumptive classification based on external morphological characters was not confirmed by genetic analysis, by means of which all specimens collected in the Marches Region were ascribed to Barbus plebejus. Genetic diversity values (h and π) of sampling groups were all different from 0 except the one sample collected from the upper reaches of the River Tenna, above a hydroelectric dam. Population connectivity and colonization patterns of the studied area were inferred from an analysis of molecular variance distribution and evolutionary relationships among haplotypes. The results point to different levels of isolation among sampling groups due to ecological and anthropogenic factors and the effect of an artificial barrier on genetic variability and conservation status of the population. Finally, this study confirms the uncertainty associated with systematic classification of Barbus spp. based on morphological characters due to the phenotypic plasticity of the species.     

Massive invasion of exotic Barbus barbus and introgressive hybridization with endemic Barbus plebejus in Northern Italy: where,how and why?

Biological invasions and introgressive hybridization are major drivers for the decline of native freshwater fish. However, the magnitude of the problem across a native species range, the mechanisms shaping introgression as well as invader's dispersal and the relative role of biological invasions in the light of multiple environmental stressors are rarely described. Here, we report extensive (N = 665) mtDNA sequence and (N = 692) microsatellite genotypic data of 32 Northern Adriatic sites aimed to unravel the invasion of the European Barbus barbus in Italy and the hybridization and decline of the endemic B. plebejus. We highlight an exceptionally fast breakthrough of B. barbus within the Po River basin, leading to widespread introgressive hybridization with the endemic B. plebejus within few generations. In contrast, adjacent drainage systems are still unaffected from B. barbus invasion. We show that barriers to migration are inefficient to halt the invasion process and that propagule pressure, and not environmental quality, is the major driver responsible for B. barbus success. Both introgressive hybridization and invader's dispersal are facilitated by ongoing fisheries management practices. Therefore, immediate changes in fisheries management (i.e. stocking and translocation measures) and a detailed conservation plan, focussed on remnant purebred B. plebejus populations, are urgently needed.     

A taxonomic revision of western Eupholidoptera bush crickets (Orthoptera: Tettigoniidae): testing the discrimination power of DNA barcode

The genus Eupholidoptera includes 46 Mediterranean species distributed from Turkey to Greece, Italy and southern France. In the eastern part of its range, Eupholidoptera has been considered to consist of several distinct species, while in the Balkans and Italian peninsula only E. chabrieri has been recognized. However, the status of some Italian populations, confined to particular geographic areas, remains uncertain. To investigate the delimitation of the Italian taxa of Eupholidoptera, we performed both morphological and molecular analyses. Morphological analysis was carried out by considering diagnostic characters usually used to distinguish different taxa, such as the shape of titillators in males and the subgenital plate in females. Molecular analysis was performed by sequencing three mitochondrial genes: 12S rRNA, 16S rRNA, partially sequenced and the entire gene of cox1. Molecular markers were used to infer phylogenetic relationships among the Italian Eupholidoptera species and to reconstruct the historical processes that shaped their current geographic distribution. Results from both morphological and molecular analyses were used to revise the taxonomic arrangement of species. On the whole we were able to distinguish nine lineages of Italian Eupholidoptera, of which E. tyrrhenica sp.n. from Corsica is described as a new species. This published work has been registered in ZooBank, http://zoobank.org/urn:lsid:zoobank.org:pub:EBD181A0-5263-4880-AC80-66F624506E3A .     

Barbus xanthos,a new barbel from the Southern Aegean basin (Teleostei: Cyprinidae)

Barbus xanthos, a new species, is described from the Eşen, Dalaman, Tersakan and Büyük Menderes rivers in south-western Anatolia. It differs from other Barbus species in the adjacent basins by having 53–60 lateral line scales, a weakly ossified last unbranched dorsal-fin ray (about 33–50%), numerous small irregular-shaped black or dark-brown spots smaller than scales, often forming large, black or dark-brown blotches on back and flank in juveniles and adults, and a straight or slightly convex posterior dorsal-fin margin. B. xanthos differs from its most closely related congener, B. pergamonensis, by nine nucleotide substitution sites in the mitochondrial DNA cytochrome oxidase I barcode region.     

Karyological study of some cyprinid species from Ethiopia. The ploidy differences between large and small Barbus of Africa

Chromosome sets of nine species of Barbus and a species of Varicorhinus from Ethiopia were investigated. All large Barbus ( B. bynni, B. intermedius, B. ethiopicus ) and V. beso are shown to have 2N =150, whereas all small Barbus ( B. anema, B. kerstenii, B. paludinosus , and three unidentified (probably undescribed) species have 2N = 50. Based on the karyological data and on the published morphological data, an independent origin of small and large Barbus of Africa is suggested. The authors believe that large Barbus of Africa constitute a monophyletic group and that this group has no sister-group relationships with any of the small Barbus of Africa.     

Cardamine apennina: a new endemic diploid species of the C. pratensis group (Brassicaceae) from Italy

Italian representatives of the Cardamine pratensis group were investigated using morphometric, karyological, and molecular (AFLP-fingerprinting) analyses. Focus was on resolving the taxonomic and evolutionary position of C. granulosa from Piemonte (NW Italy) and of populations from central Italy, previously included in C. granulosa or C. pratensis s. str. Results show that C. granulosa is a morphologically and genetically well defined diploid species, but is extremely rare and restricted to a small area in Piemonte. Central Italian populations differ from both C. granulosa and C. pratensis s. str., and, although the morphological differentiation is not strongly pronounced, they show marked molecular divergence, suggesting recognition as a new species, Cardamine apennina. Because of their restricted distribution and diploid chromosomal level, C. apennina and C. granulosa may represent basal and quite isolated lineages, whereas reticulation on diploid and polyploid level has affected remaining lineages of the C. pratensis group. Both Italian endemics are endangered due to rarity and habitat loss. A key for identification of the Italian taxa of the C. pratensis group is also provided.     

Integrative taxonomy at work: DNA barcoding of taeniids harboured by wild and domestic cats

In modern taxonomy, DNA barcoding is particularly useful where biometric parameters are difficult to determine or useless owing to the poor quality of samples. These situations are frequent in parasitology. Here, we present an integrated study, based on both DNA barcoding and morphological analysis, on cestodes belonging to the genus Taenia, for which biodiversity is still largely underestimated. In particular, we characterized cestodes from Italian wildcats (Felis silvestris silvestris), free‐ranging domestic cats (Felis silvestris catus) and hybrids populations. Adult taeniids were collected by post‐mortem examinations of the hosts and morphologically identified as Taenia taeniaeformis. We produced cox1 barcode sequences for all the analysed specimens, and we compared them with reference sequences of individuals belonging to the genus Taenia retrieved from GenBank. In order to evaluate the performance of a DNA barcoding approach to discriminate these parasites, the strength of correlation between species identification based on classical morphology and the molecular divergence of cox1 sequences was measured. Our study provides clear evidence that DNA barcoding is highly efficient to reveal the presence of cryptic lineages within already‐described taeniid species. Indeed, we detected three well‐defined molecular lineages within the whole panel of specimens morphologically identified as T. taeniaeformis. Two of these molecular groups were already identified by other authors and should be ranked at species level. The third molecular group encompasses only samples collected in Italy during this study, and it represents a third candidate species, still morphologically undescribed.     

Abundance and distribution of fish in the lower Mnembo River, Malawi-Mozambique

Endorheic Lake Chilwa is one of the most productive lakes for fisheries in Africa. High human population densities and agricultural practices in the Chilwa catchment have been linked to observed declines of the annual fish catch in the lake. The Mnembo River is a major inflow into Lake Chilwa which has received little scientific study to date. In 2003/2004, fish diversity, abundance and distribution were monitored monthly at three sites in the Mnembo River. Twelve fish species were collected, a subset of the fourteen fish species present in Lake Chilwa. Although Barbus spp. were the most abundant species sampled in the Mnembo, total biomass was higher for Labeo cylindricus (6709 and 10,434 g, respectively). Total biomass of Pareutropius longifilis (1741 g) and Brycinus imberi (1174 g) were also high in the river. Catches of Barbus and Labeo were highly correlated (r = 0.763). Compared with other inflows into Lake Chilwa (Likangala and Domasi), the Mnembo River appears to be less adversely influenced by human populations, with high abundance of Barbus and strong presence of the riverine species Labeo, Pareutropius and Brycinus.     

SPECIATIONAL HISTORY IN A DIVERSE CLADE OF HABITAT-SPECIALIZED SPIDERS (ARANEAE: NESTICIDAE: NESTICUS): INFERENCES FROM GEOGRAPHIC-BASED SAMPLING

This paper summarizes the results of an initial effort to reconstruct the speciational history of cave spiders (Nesticus) from the southern Appalachian Mountains of eastern North America. The Appalachian Nesticus fauna includes a large series of about 30 species distributed across islandlike cave and montane habitats. Many of the species are geographically restricted; all of the species are found in allopatry. Observed patterns of morphological variation and biogeographic evidence suggest that species diversification in this lineage may have occurred recently, perhaps in response to Pleistocene climatic fluctuations. To address questions about the spatial and temporal dynamics of Nesticus speciation, while accounting for potential phylogenetic difficulties, I have gathered nuclear and mitochondrial DNA sequences for a sample of individuals from 81 populations representing 28 Nesticus species. Analyses of these data indicate that considerable genetic divergence exists within and among currently recognized morphological species. Consistent with relatively deep species divergences, most of which likely predate the Pleistocene, is a prevailing pattern of phylogenetic concordance between taxonomic species and monophyletic gene tree lineages. The few deviations from monophyly detected can be tentatively attributed to a peripatric mode of speciation. Although species limits as inferred by the molecular data are generally concordant with patterns of morphological continuity and discontinuity in genitalia, there is evidence to suggest that cryptic phylogenetic lineages exist within some morphologically continuous units. This observation, in combination with the general depth of species lineages, makes any argument about rapid evolution in Nesticus genitalic characteristics unnecessary.     

Molecular phylogeny and biogeography of African diploid barbs, ‘Barbus’, and allies in Africa and Asia (Teleostei: Cypriniformes)

African diploid barbs (‘Barbus’, Clypeobarbus, Barboides, etc.) are a group of small cyprinids with a body size less than 20 cm and widely distributed in drainages across Africa. These species constitute a significant component of African freshwater fish fauna. This study is the first to focus on the molecular systematics and biogeography of African diploid barbs ‘Barbus’ and its African and Asian allies using both mitochondrial and nuclear genes. We test for monophyly of groups, determine interspecific relationships and estimate the time of divergence of 52 species of ‘Barbus’ and allies using two mitochondrial and four nuclear genes. Resulting trees demonstrate that ‘Barbus’ and allies (Systomus, Barboides, Clypeobarbus and African tetraploid barbs) form a strongly supported clade; however, ‘Barbus’ is not resolved as monophyletic. Divergence time analyses identify the separation between Systomus and ‘Barbus’ plus African allies may have occurred around 26 MYA. In addition to the phylogenetic results, these findings highlight the need for more thorough taxonomic and systematic studies on ‘Barbus’ and allies using morphological and additional molecular data and greater taxon sampling, including the type species of the genus Enteromius, ‘Barbus’ potamogalis.     

Monogènes Dactylogyridae parasites de Cyprinidae du genreBarbus d'Afrique du Nord

Resumé Quinze nouvelles espèces de Monogènes Dactylogyridae sont décrites chez quinze espèces deBarbus (Teleostei, Cyprinidae) appartenant aux sous-genresB. (Barbus) etB. (Labeobarbus) en Afrique du Nord. Les barbeaux examinés proviennent des différents bassins hydrographiques du Maroc et d'une localité nommée Hamman Bourgiba en Tunisie. Dans cette dernière région, le genreBarbus n'est représenté que par une seule espèce:Barbus (B.) callensis. Au Maroc, on en dénombre actuellement quatorze dont quatre appartiennent au sous-genreLabeobarbus: il s'agit deBarbus (L.) fritschii; B. (L.) harteti; B. (L.) paytonii etB. (L.) reinii. Les dix espèces appartenant au sous-genreBarbus sont:Barbus (B.) figuiensis; B. (B.) ksibi; B. (B.) lepineyi; B. (B.) magniatlantis; B. (B.) massaensis; B. (B.) moulouyensis; B. (B.) nasus; B. (B.) pallaryi; B. (B.) setivemensis etB. (B.) issenensis.Chaque sous-genre possède son propre pool parasitaire, à l'exception deDactylogyrus marocanus n. sp., recontré sur des espèces appartenant aux deux sous-genres (B. (L.) fritschii, B. (L.) paytonii, B. (L.) harteti, B. (L.) reinii, B. (L.) nasus, B. (B.) setivimensis, B. (B.) ksibi). Sur les cinqDactylogyrus parasitant lesLabeobarbus, trois présentent une spécificité stricte vis à vis de leur hôte. Il s'agit deDactylogyrus reinii n. sp. surB. (L.) reinii; D. volutus n. sp. etD. zatensis n. sp. surB. (L.) fritschii. Les espècesD. oumiensis n. sp. etD. kulindrii n. sp. présentent une spécifité stenoxène et parasitent respectivementB. (L.) harteti, B. (L.) paytonii, B. (L.) reinii etB. (L.) fritschii, B. (L.) reinii.Nous avons recontré neufDactylogyrus chez les poisson-hôtes appartenent au sous-genreBarbus. Six d'entre eux ont une spécificité oïoxène; ce sont:D. guirensis n. sp.,D. atlasensis n. sp. etD. draaensis n. sp. surB. (B.) pallaryi; D. borjensis n. sp. surB. (B.) nasus etD. heteromorphus n. sp. etD. tunisiensis n. sp. surB. (B.) callensis. Les trois autres parasites ont un spectre d'hôtes plus large. Il s'agit deD. ksibii n. sp. recontré chezB. (B.) ksibi, B. (B.) setivimensis etB. (B.) magniatlantis; D. ksibioïdes n. sp. recontré chezB. (B.) setivimensis etB. (B.) moulouyensis. L'espèceD. fimbriphallus n. sp. stenoxène, se recontre chez les poisson-hôtes du versant Sud de l'Atlas et de la façade méditerranéenne à savoir:B. (B.) figuiensis, B. (B.) lepineyi, B. (B.) massaensis, B. (B.) moulouyensis, B. (B.) pallaryi etB. (B.) issenensis.Le rôle des Dactylogyridae en tant que marqueurs biogéographiques, phylogénétiques et taxonomiques est discuté à partir de la composition spécifique des communautés de Monogènes rencontrés et de leurs différents types morphologiques.

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Fifteen new species of the Dactylogyridae (Monogenea) parasitic on fifteen species of barbels (Barbus) from North Africa (Teleostei, Cyprinidae) are described. The fishes studied belong to two subgenera,B. (Labeobarbus) andB. (Barbus), collected from various hydrographical basins of Morocco and from the Hamman Bourgiba locality in Tunisia. In the latter area, the genusBarbus is represented by onlyBarbus (Barbus) callensis. In Morocco, fourteen species are listed, four of which belong to the subgenusLabeobarbus; these areBarbus (L.) fritschii; B. (L.) harteti; B. (L.) paytonii andB. (L.) reinii. The other ten species belong to the subgenusBarbus: these areBarbus (B.) figuiensis; B. (B.) ksibi; B. (B.) lepineyi; B. (B.) magniatlantis; B. (B.) massaensis; B. (B.) moulouyensis; B. (B.) nasus; B. (B.) pallaryi; B. (B.) setivimensis andB. (B.) issenensis. Each of the two subgenera has its unique parasitic fauna, except forDactylogyrus marocanus n. sp. collected on species belonging to both subgenera (B. (L.) fritschii, B. (L.) paytonii, B. (L.) harteti, B. (L.) reinii, B. (B.) nasus, B. (B.) setivimensis andB. (B.) ksibi). Of the five monogeneans found onLabeobarbus, three appear to be specific to one host: they areDactylogyrus reinii n. sp. onB. (L.) reinii, andD. volutus n. sp. andD. zatensis n. sp. onB. (L.) fritschii. D. kulindrii n. sp. parasitisedB. (L.) reinii andB. (L.) fritschii; andD. oumiensis n. sp. occurred onB. (L.) reinii, B. (L.) paytonii andB. (L.) harteti. NineDactylogyrus species were found in fishes belonging to the subgenusBarbus. Six of them have an oïoxenous specificity: these areD. guirensis n. sp.,D. atlasensis n. sp. andD. draaensis n. sp. onB. (B.) pallaryi; D. borjensis n. sp. onB. (B.) nasus andD. heteromorphus n. sp. andD. tunisiensis n. sp. on(B.) callensis. These other three have a wider range of hosts: they areD. ksibii n. sp. collected fromB. (B.) ksibi, B. (B.) setivimensis andB. (B.) magniatlantis, andD. ksibioïdes n. sp. found onB. (B.) setivimensis andB. (B.) moulouyensis. D. fimbriphallus n. sp. is a characteristic parasite of fishes from the southern side of the Atlas mountains and the Mediterranean coast (B. (B.) figuiensis, B. (B.) lepineyi, B. (B.) massaensis, B. (B.) moulouyensis, B. (B.) pallaryi andB. (B.) issenensis).The role of dactylogyrids as biogeographical phylogenetic and taxonomic indicators is discussed in relation to the specific structure of the monogenean communities and the different morphological types found.