Cytotype-specific ISSR profiles and karyotypes in the Neotropical genus <Emphasis Type="Italic">Eigenmannia</Emphasis> (Teleostei: Gymnotiformes)

The genus Eigenmannia (Teleostei: Gymnotiformes), a widely distributed fish genus from the Neotropical region, presents very complex morphological patterns and many taxonomic problems. It is suggested that this genus harbors a species complex that is hard to differentiate using only morphological characteristics. As a result, many species of Eigenmannia may be currently gathered under a common name. With the objective of providing new tools for species characterization in this group, an analysis of the polymorphism of DNA inter-simple sequence repeats (ISSR), obtained by single primer amplification reaction (SPAR), combined with karyotype identification, was carried out in specimens sampled from populations of the Upper Paraná, São Francisco and Amazon river basins (Brazil). Specific ISSR patterns generated by primers (AAGC)₄ and (GGAC)₄ were found to characterize the ten cytotypes analyzed, even though the cytotypes 2n = 38 and 2n = 38 XX:XY, from the Upper Paraná basin, share some ISSR amplification patterns. The geographical distribution of all Eigenmannia specimens sampled was inferred, showing the cytotype 2n = 31/2n = 32 as the most frequent and largely distributed in the Upper Paraná basin. The cytotype 2n = 34 was reported for the first time in the genus Eigenmania, restricted to the São Francisco basin. Polymorphic ISSR patterns were also detected for each cytotype. Considering our results and the data reported previously in the literature, it is suggested that many of the forms of Eigenmannia herein analyzed might be regarded as different species. This work reinforces the importance of employing diverse approaches, such as molecular and cytogenetic characterization, to address taxonomic and evolutionary issues.     

Banding and FISH in three species of <Emphasis Type="Italic">Vernonia</Emphasis>, subsection <Emphasis Type="Italic">Macrocephalae</Emphasis> (Asteraceae,Vernonieae)

To complement our knowledge about the karyotypes of the genus Vernonia Schreb., different techniques of chromosome banding, including AgNOR, triple staining with fluorochromes CMA/DA/DAPI (CDD), and fluorescence in situ hybridization (FISH) for the 45S rDNA probe, were applied to three species of subsection Macrocephalae. Vernonia bardanoides was collected from an area of cerrado (savanna) vegetation in Itirapina, São Paulo State, Brazil, and V. linearifolia and V. tomentella were collected from areas of rocky, open altitudinal vegetation in Joaquim Felicio and Diamantina, respectively, in Minas Gerais State. All species showed two terminal CMA⁺ and NOR bands. FISH indicated two terminal 45S rDNA sites in V. linearifolia and V. tomentella, and six in V. bardanoides.     

Conservation genetics of the highly endangered Azorean endemics <Emphasis Type="Italic">Euphrasia azorica</Emphasis> and <Emphasis Type="Italic">Euphrasia grandiflora</Emphasis> using new SSR data

In the Azores Islands, two Euphrasia L. (Orobanchaceae) endemic species are recognized: Euphrasia azorica H.C.Watson, an annual herb, in Flores and Corvo, and Euphrasia grandiflora Hochst. ex Seub., a semi-shrub, in Pico, São Jorge and Terceira. Both species are highly endangered and protected by the Bern Convention and Habitats Directive. A population genetics study was conducted with new microsatellite primer pairs in 159 individuals of E. azorica and E. grandifolia, sampled from populations in Flores, Corvo, Pico and São Jorge. Allele sizing suggested that E. azorica is a diploid while E. grandiflora is a tetraploid. Euphrasia grandiflora revealed higher genetic diversity then E. azorica. The E. grandiflora population of Morro Pelado in São Jorge, displayed higher genetic diversity when compared with all others, while the E. azorica population of Madeira Seca in Corvo, showed the lowest. Private and less common bands were also overall higher in E. grandiflora populations. Population genetic structure analysis confirmed a distinctiveness between the two Azorean endemic Euphrasia, in addition to island-specific genetic patterns in E. azorica. The genetic structure obtained for E. grandiflora was complex with the populations of Cabeço do Mistério in Pico Island and of Pico da Esperança in São Jorge sharing the same genetic group, while a putative spatial barrier to gene flow was still retrieved between both islands. Although some populations of both species might benefit from propagation actions, studies are needed on plant host species and translocations between islands or between some populations of a same island should be avoided, due to the occurrence of putative ESUs. Eradication of invasive species and control of grazing will be fundamental to promote in situ restauration.     

Chromosomal studies in <Emphasis Type="Italic">Crenicichla lepidota</Emphasis> and <Emphasis Type="Italic">Australoheros facetus</Emphasis> (Cichlidae,Perciformes) from extreme Southern Brazil

Fishes of the family Cichlidae generally show low karyotype variability. Nevertheless, karyotype variants have been identified within some genera, providing information about their evolutionary history. In the present study, karyotype characteristics of Crenicichla lepidota and Australoheros facetus, two sympatric species found in the São Gonçalo-Mangueira basin, were studied. Besides conventional procedures, double fluorochromes staining chromomycin A₃/DAPI and fluorescent in situ hybridization (FISH) with rDNA probes were also used. Both species presented 2n = 48 chromosomes, but karyotypes were differentiated by fundamental number, which was equal to 70 in A. facetus and 56 in C. lepidota. Similar heterochromatin distribution patterns were also observed on the pericentromeric region of most chromosomes, although C. lepidota presented an additional heterochromatic block in the first pair. The Ag-NORs, 18S rDNA probe and CMA₃/DAPI were coincident in location on the first and second pairs of C. lepidota and A. facetus, respectively. The minor rDNA loci (5S rDNA) were found in four sites located on two distinct chromosomal pairs in C. lepidota. Although the data obtained here to C. lepidota and to A. facetus show chromosomal characteristics considered ancestral to the family, new data are presented to both species. Additionally, this study corroborates the hypothesis in which evolutionary processes like non-Robertsonian rearrangements are involved in the diversification of the major groups of Neotropical Cichlidae. Thus, the karyotype diversification observed in A. facetus have the high fundamental number pathway while C. lepidota has others evolutionary chromosomal mechanisms.     

Neotropical Monogenoidea. 55. Dactylogyrids parasitising the pintado-amarelo <Emphasis Type="Italic">Pimelodus maculatus</Emphasis> Lacépède (Actinopterygii: Pimelodidae) from the Rio São Francisco,Brazil

Eight species of Dactylogyridae were collected from the gills of the pintado-amarelo Pimelodus maculatus Lacépède in the Rio São Francisco in Brazil: Ameloblastella paranaensis (França, Isaac, Pavanelli &; Takemoto, 2003) Mendoza-Franco &; Scholz, 2009, A. satoi n. sp., Ameloblastella sp., Demidospermus armostus Kritsky &; Gutiérrez, 1998, D. cf. bidiverticulatum (Suriano &; Incorvaia, 1995) Kritsky &; Gutiérrez, 1998, D. ichthyocercus n. sp., D. paravalenciennesi Gutiérrez &; Suriano, 1992 and D. uncusvalidus Gutiérrez &; Suriano, 1992. Two new species, A. satoi n. sp. and D. ichthyocercus n. sp., are described, and A. paranaensis is redescribed. The Rio São Francisco represents new geographical records for the five previously described dactylogyrid species.     

Communities of arbuscular mycorrhizal fungi under <Emphasis Type="Italic">Picconia azorica</Emphasis> in native forests of Azores

Arbuscular mycorrhizal fungi (AMF) from the rhizosphere of the endemic Laurisilva tree, Picconia azorica, were characterised at two sites in each of two Azorean islands (Terceira and São Miguel). Forty-six spore morphotypes were found, and DNA extraction was attempted from individual spores of each of these. DNA was obtained from 18 of the morphotypes, from which a 1.5 kb long fragment of the nuclear ribosomal RNA gene (SSU-ITS-LSU) was sequenced. A total of 125 AMF sequences were obtained and assigned to 18 phylotypes. Phylogenetic analysis revealed sequences belonging to the families, Acaulosporaceae, Archaeosporaceae, Claroideoglomeraceae, Gigasporaceae and Glomeraceae. Phylotype richness changed between islands and between sampling sites at both islands suggesting that geographical and historical factors are determinant in shaping AMF communities in native forest of Azores. Ecological analysis of the molecular data revealed differences in AMF community composition between islands. In Terceira, the rhizosphere of P. azorica was dominated by species belonging to Acaulosporaceae and Glomeraceae, while São Miguel was dominated by members of Glomeraceae and Gigasporaceae. This is the first molecular study of AMF associated with P. azorica in native forest of the Azores. These symbiont fungi are key components of the ecosystem. Further research is needed to develop their use as promoters of plant establishment in conservation and restoration of such sites.     

mtDNA analysis of <Emphasis Type="Italic">Mytilopsis</Emphasis> (Bivalvia,Dreissenidae) invasion in Brazil reveals the existence of two species

The present study sought to identify general patterns of genetic variability and structure of fish stocks (migratory and non-migratory species) along one of the largest Brazilian rivers, the São Francisco. Given that genetic variability of populations of all organisms is governed by both present and past influences, herein we investigate both these aspects by evaluating the current genetic differences between populations of five species (Leporinus piau, Megaleporinus reinhardti, Pimelodus maculatus, Prochilodus argenteus, and Pygocentrus piraya) along the entire extension of the river, as well as their demographic history. Analyses were done through sequences of two mitochondrial fragments and microsatellite data. In general, the data showed no support for recent fragmentation of stocks by the dams present in this river, and that all species show signs of past population expansion. We discuss the possible reasons for the common patterns found between these species, including the influence of the river’s topography and history.     

Clonal Transmission of ESBL-Producing <Emphasis Type="Italic">Klebsiella</Emphasis> spp. at a University Hospital in Brazil

The present study was designed to determine the prevalence and extended-spectrum β-lactamase (ESBL) types in clinical isolates of Klebsiella spp. at a university hospital located in the Brazilian southern region (Ribeirão Preto, São Paulo) as well as their antibiotic susceptibility and genetic profiles. This study included 147 non-repeat Klebsiella spp. isolates collected from January to June 2000, of which 23 K. pneumoniae and 8 K. oxytoca were selected as ESBL producers by using the Oxoid combination disk method and Etest ESBL strip. β-lactamases were characterized by IEF, PCR and sequencing assays using primers for ESBL genes. Antibiotic susceptibility was evaluated by MicroScan system. Dissemination of two major clones of ESBL-producing Klebsiella spp. occurred in the hospital. According to the results obtained in this study there was a clonal spread of CTX-M-producing K. oxytoca in five clinics and dissemination of ESBL-producing K. pneumoniae in the nursery and pediatrics wards.     

Phylogenetic relationships of sculpin <Emphasis Type="Italic">Cottus volki</Emphasis> Taranetz, 1933 (Scorpaeniformes,Cottidae) according to the results of analysis of control region in mitochondrial DNA

Sequencing of the fragment of control region in mitochondrial DNA in sculpin Cottus volki and the comparison of obtained data with homologous nucleotide sequences in the other species from genus Cottus demonstrated that C. volki occupies the basal position in the “poecilopus” group, which includes also typical sculpin C. poecilopus Heckel, 1837 from water bodies of Europe, sculpin C. szanaga Dybowski, 1869 from the Amur River basin, and sculpin Cottus cf. poecilopus from the Lena River basin. Early separation of C. volki line from common trunk of “poecilopus” group explains the presence in C. volki of several primitive (plesiomorphic for this group) morphological characters: long interior ray of ventral fin, practically complete lateral body line, and well-developed dentition of palatine bones.     

Genetic analysis of forest species <Emphasis Type="Italic">Eugenia uniflora</Emphasis> L. through of newly developed SSR markers

Nine microsatellite loci for genetic analysis of three populations of the tropical tree Eugenia uniflora L. (pitanga or Brazilian cherry) from fragments of semideciduous forest were developed. We used the technique of building a (GA) _n and (CA) _n microsatellite-enriched library by capture with streptavidin-coated magnetic beads. We assessed the polymorphism of seven microsatellites in 84 mature trees found in three areas (Ribeirão Preto, Tambaú and São José do Rio Pardo), highly impacted by the agricultural practices, in a large region among Pardo river and Mogi-Guaçu river basins, in state of São Paulo, Brazil. All loci were polymorphic, and the number of alleles was high, ranging from 6 to 24, with a mean of 14.4. All stands showed the same high level of genetic diversity (mean H _E  = 0.83) and a low genetic differentiation (mean F _ST = 0.031), indicating that genetic diversity was higher within rather than among populations. Seven of the nine loci were highly variable, and sufficiently informative for E. uniflora. It was concluded that these new SSR markers can be efficiently used for gene flow studies.     

The fitness of the Brazilian damsel <Emphasis Type="Italic">Stegastes fuscus</Emphasis> is increased by sharing the territory with the dusky grouper <Emphasis Type="Italic">Epinephelus marginatus</Emphasis>

Stegastes fuscus and Epinephelus marginatus are known for co-habiting shelters. The damselfish S. fuscus uses the territory for nesting and must protect its eggs from grazers; the grouper E. marginatus is an omnivorous sit-and-wait predator. This study aims to evaluate the effect of juvenile groupers on the reproductive success of the Brazilian damsel. Twenty-five hours of underwater observations were done in São Sebastião and Ilhabela, Northern shore of São Paulo, Brazil. Fitness increase was measured by the egg-clutch area and number of contributing females in 130 nests shared by groupers and another 130 where damselfishes stood alone. An egg predator crab was placed into the damselfish territory, and behavioural responses during 2 min were recorded for nests with or without E. marginatus, 80 replicates each. Nests shared by the dusky groupers had more eggs and received eggs from more females too. While fathers who were alone in the territory had to deal with the egg predator crab, in shared nests, the grouper would take care of the intruder, sometimes feeding on it. Therefore, the Brazilian damsel may benefit from the presence of the dusky grouper by increasing the fitness and diminishing the costs of parental care.     

Differentiation of petroleum hydrocarbon-degrading <Emphasis Type="Italic">Pseudomonas</Emphasis> spp. based on PCR-RFLP of the 16S-23S rDNA intergenic spacer region

Genetic diversity among 43 petroleum hydrocarbon-degrading Pseudomonas belonging to four different species and the type strain Pseudomonas aeruginosa MTCC1034 was assessed by using restriction fragment length polymorphism (RFLP) of polymerase chain reaction (PCR)-amplified 16S–23S rDNA intergenic spacer regions (ISRs) polymorphism. PCR amplification from all Pseudomonas species yielded almost identical ISR amplicons of “?” 800 bp and in nested PCR of “?” 550 bp. The RFLP analysis with MboI and AluI revealed considerable intraspecific variations within the Pseudomonas species. The dendrogram constructed on the basis of the PCR-RFLP patterns of 16S–23S rDNA intergenic spacer regions differentiated all the species into seven different clusters.     

New species of <Emphasis Type="Italic">Jakutoproductus</Emphasis> from the Lower Permian of Verkhoyansk Region

Two new brachiopod species of Jakutoproductus, J. lenensis sp. nov. and J. talchanensis sp. nov., are described. J. lenensis occurs in the upper part of the J. verkhoyanicus Zone of the Khorokytian Horizon (Asselian–Sakmarian deposits) of the lower reaches of the Lena River and in the Sobolokh-Mayan and Allara-Khadaryn’ya river basins. J. talchanensis is identified from the Khorokytian Horizon of the Talchan River (Baraiy River basin). A new subzone, the J. lenensis Subzone, is proposed for the upper part of the J. verkhoyanicus Zone of the Khorokytian Horizon of the Verkhoyansk Region.     

New host species and new distribution areas of <Emphasis Type="Italic">Salmincola lavaretus</Emphasis> (Copepoda: Lernaeopodidae): A parasite of coregonid fishes of Siberia

Data on the infestation of certain fish species by the parasitic copepod Salmincola lavaretus are presented for the first time. The infested fish species included Bauntovsky whitefish (Coregonus baunti), an endemic species from the Maloye and Bolshoye Kapylyushi lakes; Siberian whitefish (C. pidschian) and Siberian cisco (C. sardinella) from Bolshoye Kapylyushi Lake and Baunt Lake, which belong to the Tsypo-Tsypikan lake system (the Lena River basin, Transbaikalia); and Teletsky whitefish (C. lavaretus natio smitti) from Teletskoe Lake (the Ob River basin). Previously, S. lavaretus had been described as a parasite of Baikal omul (C. migratorius) and Baikal whitefish (C. baicalensis) from Baikal. The taxons of S. longimanus complex (S. longimanus, S. l. sibirica, S. svetlanovi, and S. lavaretus)—parasites of nasal fossae of grayling and coregonid fishes—were registered in the lakes of the Lake Baikal basin (Lake Baikal and Khovsgol Lake), the Lena River, the Yenisei and the Ob rivers (the largest rivers of the Arctic zoogeographic province), and the Kobdo River (the Western Mongolian province). It was assumed that S. longimanus complex is more widely distributed in the water bodies of the Arctic zoogeographic province of the Palearctic.     

Loaches of the genus <Emphasis Type="Italic">Barbatula</Emphasis> (Nemacheilinae) of the Zavkhan River basin (Western Mongolia)

Two sympatric species of Siberian loaches are recorded in the Zavkhan River basin (Great Lakes Depression, Western Mongolia) and described as new for science: Barbatula conilobus and B. dsapchynensis spp. n. The former species, most likely, is widely distributed over the entire river basin, and the latter species, apparently, occurs locally, but in sympatry with B. conilobus. Morphological differences of the new species from other representatives of the genus Barbatula and their relationships are analyzed. Ecological distinctions between the species of the genera Barbatula and Triplophysa in the Zavkhan River basin are described. Possible mechanisms of development of the present fauna of loaches in Mongolia and adjacent territories with an appearance of sympatric species pairs in a single water basin are discussed.     

Population genetics of a Chinese endemic, <Emphasis Type="Italic">Gymnocypris potanini</Emphasis> Herzenstein,threatened by population isolation: conflicting patterns between microsatellites and mitochondrial DNA

The arrival of the rains and the hydrological changes they cause are essential in the reproductive cycle of Neotropical migratory fishes, but their association with the onset of migration is still poorly understood. This study aimed to determine the role played by rainfall, hydrological changes, and the lunar cycle as triggers for migration of Prochilodus costatus (Prochilodontidae) in the upper São Francisco River, Brazil. In total, 132 individuals were fitted with radiotelemetry transmitters over three consecutive spawning seasons. Spawning migration began with the onset of the rains from late September to mid-December. Individuals exhibited a strong preference for initiating migration at the beginning of the rainy season, when river discharge is low, in days with positive changes in water level, and at times of new or waxing moon. The fraction of the population that initiates migration each year appears to be dependent on rainfall, indicating that P. costatus may be a partial migrant. The results show that P. costatus and, possibly, other migratory Neotropical fish, rely on different but interconnected environmental cues to trigger their spawning migration, ensuring that individuals migrate in time to reach their spawning grounds and find appropriate environmental conditions for spawning.     

Conservation genetics of the threatened catfish <Emphasis Type="Italic">Conorhynchos conirostris</Emphasis> (Siluriformes: <Emphasis Type="Italic">incertae sedis</Emphasis>), an evolutionary relict endemic to the São Francisco River Basin,Brazil

Analysis of genetic datasets can be particularly useful in providing guidelines for conservation management of understudied species targeted by commercial activities. Here we used population genetic approaches to inform on the conservation status of the Neotropical long-nose pirá catfish, Conorhynchos conirostris. Pirá is a large migratory fish endemic to the São Francisco River Basin (SFRB). It is an evolutionarily divergent and relict species, being the sole representative of an incertae sedis family. The species is considered locally extinct in the upper and lower SFRB, listed as vulnerable on the IUCN Red List and as endangered on the Brazilian Red List (ICMBIO). Fishing prohibition has received severe criticism from middle SFRB fisheries that claim that this understudied species is relatively abundant in that region. We used information from 13 microsatellite markers and COI mitochondrial sequences to clarify the genetic diversity of this enigmatic species in the middle SFRB, to estimate contemporary effective population size (N_e), and to assess its conservation status. Results from bottleneck analyses indicated that the species has experienced recent reductions in population size, which is consistent with small estimates of contemporary N_e. The predicted amount of heterozygosity loss (H_t) in t generations ranged from 0.1152 (for an estimated N_e of 26.4; t?=?100) to 0.7573 (for an estimated N_e of 169.9; t?=?10). Our study supports the conservation status proposed by the ICMBIO to the remaining pirá population. Moreover, we highlight the need for demographic data and the re-assessment of the current IUCN classification for this evolutionary relict lineage.     

Redescription of <Emphasis Type="Italic">Cottus sibiricus</Emphasis> and an estimation of differences between this species and <Emphasis Type="Italic">Cottus dzungaricus</Emphasis> (Cottidae)

The type specimens of Cottus sibiricus and C. dzungaricus as well as individuals from the Olkha River (Yenisei River basin) are studied. The diagnostic features of C. sibiricus are established. Comparative morphological and molecular analysis of divergence between C. sibiricus and C. dzungaricus demonstrates that the differences lie within the limits of one polymorphous species. The rank of Dzungarian sculpin is lowered to the subspecies, C. sibiricus dzungaricus.     

The recent occurrence,establishment and potential impact of <Emphasis Type="Italic">Geophagus proximus</Emphasis> (Cichlidae: Perciformes) in the Tietê River reservoirs: an Amazonian fish species introduced in the Paraná Basin (Brazil)

The introduction of allochthonous fish species happens constantly in large bodies of freshwater, like as the reservoirs of Paraná Basin, located in Brazilian southeast, representing a threat for local biodiversity. The fish species Plagioscion squamosissimus and Cichla ocellaris were introduced from the 1970s in several water bodies of this basin and had successfully established themselves in all six reservoirs located in the middle and lower Tietê River (SP, Brazil), particularly. After six decades from the first recorded species introduction, this hydrographic system remains open to the invasion of further fish species, owing to widespread fish-farming activity and by the channels opened between this system and other reservoirs and river basin. This study was an effort to confirm the Geophagus proximus occurrence in the six Tietê River reservoirs, verifying the actual introduction status and analyzing its potential environmental impacts on local species by the analysis of the population structure (abundance, body dimensions and feeding habits). By the results, this species was confirmed in the Ibitinga, Nova Avanhandava and Três Irmãos reservoirs. The abundance and feeding analysis shows, respectively, it is successfully established in the Três Irmãos reservoir with the same feeding habitats of local species, such as Geophagus brasiliensis. It was further shown to be very likely that G. proximus would spread throughout the reservoir system of the middle and lower Tietê River, in the manner of P. squamosissimus and C. ocellaris, and the competition pressure for food resources between G. proximus and the local species which represents a potential environmental impact system. These scientific evidences fortifies the knowledge basin for the implantation of a fish management system, to control and reduce the abundance of the invader and to prevent its becoming established in all the Tietê River Basin, avoiding the disastrous consequences for the native species of Paraná River Basin.