Isolation and characterization of polymorphic microsatellites for the natural populations of barker frog Physalaemus cuvieri

Ten polymorphic microsatellite loci were isolated for Physalaemus cuvieri from a GA—CA enriched library. In 160 P. cuvieri individuals, the number of alleles per locus ranged to 2–9 and the expected heterozygosity ranged from 0.30 to 0.85. The primers were successfully cross-amplified in the congeneric species P. albonotatus, P. ephippifer and Physalaemus cf. cuvieri, suggesting that these loci are potentially useful for studies on population genetic structure of Physalaemus sp.     

Genetic identification of larvae and juveniles reveals the difference in the spawning site among Cyprininae fish species/subspecies in Lake Biwa

To understand the differences in the spawning sites among Cyprininae fishes in Lake Biwa, we conducted periodic sampling of larvae and juveniles at three sites (irrigation ditch, St. 1; river, St. 2; and satellite lake, St. 3). On the basis of species/subspecies identification by using RAPD analysis, we examined the species composition of the larvae and juveniles at these three sampling sites. The number of specimens was 616, 68, and 117 at St. 1, St. 2, and St. 3, respectively. Based on morphological and genetic identification, the specimens were found to include nine fish species/subspecies, namely, Carassius auratus grandoculis, Carassius cuvieri, Carassius auratus langsdorfii, Cyprinus carpio, Sarcocheilichthys sp., Silurus asotus, Oryzias latipes, Odontobutis obscura obscura, and Rhinogobius sp. The species composition at the three sites also differed. Among the Cyprininae fishes, C. auratus grandoculis, C. auratus langsdorfii, and Cyprinus carpio were found in abundance at St. 1; C. cuvieri was not collected from St. 1 but was found at the other two sites, particularly St. 3. Among the other fishes, Rhinogobius sp. was collected at St. 1 and St. 3, whereas the other four occurred only at St. 1. These results suggest that the selection of spawning sites by C. cuvieri differs to a certain extent from that of the other Cyprininae fishes, and the irrigation ditch in the lake is an important habitat for the larvae and juveniles of native fish species.     

Agropastoral activities increase fluctuating asymmetry in tadpoles of two neotropical anuran species

Agriculture and pasture activities are the main drivers for habitat reduction, directly affecting amphibian assemblages. In the Cerrado, the progress of agricultural and pasture areas negatively affects aquatic environments and their organisms, once the suppression of marginal vegetation reduces the natural protection against allochtone stressors. One possible way to measure the stress level is the Fluctuating Asymmetry (FA), which is calculated based on the deviations in the development of bilateral morphological traits of the organism. Herein, we evaluated whether environments with a higher degree of agropastoral influence and reduced marginal vegetation can increase FA levels in tadpoles of two common and widely distributed anuran species in the Cerrado (Physalaemus cuvieri and Scinax fuscomarginatus). We sampled and classified water bodies according to the percentage of agropastoral land use and marginal vegetation, and measured four morphological traits of tadpoles to evaluate the degree of FA. We found that in environments with intensive agropastoral land use, tadpoles of P. cuvieri and S. fuscomarginatus had higher FA in nostril‐snout distance (NSD). In environments with reduced marginal vegetation, tadpoles of S. fuscomarginatus had higher FA in eye width (EW), but no effect was detected for tadpoles of P. cuvieri. These morphological traits (i.e. nostrils and eyes) are associated to individual fitness of tadpoles. Thus, these developmental deviations can affect species fitness and population homeostasis over time, contributing to generate stochastic population dynamics and increasing the vulnerability of native species to local extinctions. The use of FA as a tool to measure environmental impact on species with potential to be used as bioindicators can contribute to generate and test hypothesis when integrated with long‐term studies.     

Effect of flood pulses on the trophic ecology of four piscivorous fishes from the eastern Amazon

This study investigated the effect of hydrological periods on the feeding activity and trophic interactions of four piscivorous fishes from the middle Xingu River, Brazil: pike‐characid Boulengerella cuvieri, dogtooth characin Hydrolycus armatus, dogtooth characin Hydrolycus tatauaia and South American silver croaker Plagioscion squamosissimus. Repletion Index (I_R%), Alimentary Index (I_Ai%) and food web properties were calculated for each species. A total of 825 specimens were collected. The I_R showed changes in feeding intensity of B. cuvieri, H. armatus and H. tatauaia among hydrological periods. Flood pulse showed no influence on composition and importance of food items consumed. Trophic connections showed that connectivity ranged from 0.025 to 0.038. The highest number of trophic connections (75) occurred in the high‐water period, when 51 food items were recorded and the lowest number of trophic connections (43) occurred in receding water, with 31 food items. In all food webs, over 45% of food items were consumed by only one species (ultra‐peripheral items), which is common in piscivorous fishes.     

Defining spatial and temporal patterns of phylogeographic structure in Madagascar's iguanid lizards (genus Oplurus)

Understanding the remarkably high species diversity and levels of endemism found among Madagascar’s flora and fauna has been the focus of many studies. One hypothesis that has received much attention proposes that Quaternary climate fluctuations spurred diversification. While spatial patterns of distribution and phylogenetic relationships can provide support for biogeographic predictions, temporal estimates of divergence are required to determine the fit of these geospatial patterns to climatic or biogeographic mechanisms. We use multilocus DNA sequence data to test whether divergence times among Malagasy iguanid lizards of the subfamily Oplurinae are compatible with a hypotheses of Pliocene–Pleistocene diversification. We estimate the oplurine species tree and associated divergence times under a relaxed‐clock model. In addition, we examine the phylogeographic structure and population divergence times within two sister species of Oplurus primarily distributed in the north‐west and south‐west of Madagascar (Oplurus cuvieri and Oplurus cyclurus, respectively). We find that divergence events among oplurine lineages occurred in the Oligocene and Miocene and are thus far older and incompatible with the hypothesis that recent climate fluctuations are related to current species diversity. However, the timing of intraspecific divergences and spatial patterns of population genetic structure within O. cuvieri and O. cyclurus suggest a role for both intrinsic barriers and recent climate fluctuations at population‐level divergences. Integrating information across spatial and temporal scales allows us to identify and better understand the mechanisms generating patterns diversity.     

Relationships between environmental gradients and geographic variation in the intraspecific body size of three species of frogs (Anura)

The relationship between environmental gradients and patterns of geographic variation in body size has been a controversial topic for ectothermic organisms globally. To examine whether the patterns that generally hold in more temperate species also hold for tropical ones, we examined the intraspecific body size variation in three species of Neotropical frogs, Dendropsophus minutus, Hypsiboas faber and Physalaemus cuvieri, along different environmental gradients (e.g. temperature, precipitation and topography). We analysed four competing hypotheses: (i) the water availability hypothesis that predicts a negative relationship between body size and precipitation; (ii) the heat balance hypothesis that predicts a negative relationship between body size and temperature; (iii) the topography hypothesis that predicts a negative relationship between body size and altitude; and (iv) the mixed‐effect hypothesis that predicts that individuals occurring in wet and cold sites would be larger than individuals occurring in dry and warm sites. The spatial pattern of geographic variation in body size among populations of H. faber was associated with the mixed‐effect hypothesis. In localities with low precipitation seasonality and cold conditions, H. faber individuals were larger than in localities with high precipitation seasonality and warm conditions. Variation in the body size of D. minutus was the opposite of that predicted by the heat balance hypothesis. Individuals in localities with high temperatures were larger than in localities with low temperatures. On the other hand, variation in the body size of P. cuvieri was not associated with the variables used in this study. Our results suggest that intraspecific variation in anuran body size is more dependent on species‐specific response than on the region (i.e. temperate or tropical) where they occur.     

Weight‐length relationships of 14 species of icefishes (Salangidae) endemic to East Asia

Length‐weight relationships (LWRs) of 14 species from 17 populations of salangids, which cover most species in the family Salangidae, were analyzed from 12 regions in China and Japan. A total of 952 specimens were used to estimate the relationship parameters. The allometric factor b values varied from 2.56 in Salanx cuvieri to 3.53 in Neosalanx oligodontis. Additionally, the condition factor a value was calculated setting b = 3 in the remaining six species. Nine maximum‐size records and 17 LWRs of salangids are established for the first time, which will fill in some of the empty spaces on rare neotenic species in the Fishbase.     

Molecular and morphological differences between two geographic populations of Salanx ariakensis (Salangidae) from Korea and Japan

Geographic variations of a salangid, Salanx ariakensis, from Korea and Japan, based on partial mtDNA cyt b gene sequences and morphometric data, are presented and compared with Salanx cuvieri from China. Analyses of molecular variance (AMOVA) suggested that the two populations of S. ariakensis differed significantly from each other in pairwise fixation index (F _ST = 0.43), although their reciprocal monophyly was not constructed in the phylogenetic analysis. The phylogenetic analysis, however, supported the reciprocal monophyly of S. ariakensis (Korean population + Japanese population) and S. cuvieri. Differences in genetic structure between the two populations of S. ariakensis were suggested by diversity indices, mismatch distribution shape, and minimum spanning network. In particular, diversity indices indicated that the Korean population was historically larger and more stable than the Japanese population. In addition, both the principal component analysis (PCA) and canonical discriminant analysis (CDA) for morphometric characters, and the Kruskal–Wallis test for some meristic characters, showed that the two populations of S. ariakensis differed from each other, although morphological differences between the two populations of S. ariakensis were smaller than those between S. ariakensis and S. cuvieri. Such morphological differences were consistent with the differences in mtDNA.     

Genetic constitution and phylogenetic relationships of Japanese crucian carps (Carassius)

The genetic constitution and phylogenetic relationships among the proposed species and subspecies of the crucian carp complex in Japan (Carassius cuvieri, C. auratus subspp. 1 and 2, C. a. grandoculis, C. a. buergeri, and C. a. langsdorfii) were investigated based on analyses of the partial nucleotide sequences of the mitochondrial DNA control region and amplified fragment length polymorphisms of nuclear DNA. Our results demonstrate that C. cuvieri and C. auratus are different entities. However, although several distinct lineages were observed for C. auratus, none corresponded to the proposed subspecies. Moreover, the five subspecies of C. auratus were not necessarily separated genetically from each other. Triploid fish, which are currently classified as a single subspecies (C. a. langsdorfii) but are sometimes treated as an independent species, fell into various clades along with diploid individuals of other subspecies that have the same or similar mitochondrial haplotypes. This suggests that gynogenetic triploid crucian carps distributed throughout the Japanese Archipelago have polyphyletic maternal origins. Our results indicate that Japanese crucian carps are a much more complex assemblage than previously believed.     

Chromosome synapsis and recombination during meiotic division in gynogenetic triploid ginbuna,Carassius auratus langsdorfii

Electron microscope examinations of primary oocytes of gynogenetic triploid ginbuna (Carassius auratus langsdorfii) and diploid gengoroubuna (Carassius auratus cuvieri) were carried out. Typical synaptonemal complexes were observed in both subspecies. In addition, clear differences in PGM-A electrophoretic patterns between a female parent ginbuna and her offspring were detected. It was concluded that synapsis and recombination occur between at least some homologous chromosomes in triploid ginbuna.     

Placing cryptic,recently extinct,or hypothesized taxa into an ultrametric phylogeny using continuous character data: A case study with the lizard Anolis roosevelti

In recent years, enormous effort and investment has been put into assembling the tree of life: a phylogenetic history for all species on Earth. Overwhelmingly, this progress toward building an ever increasingly complete phylogeny of living things has been accomplished through sophisticated analysis of molecular data. In the modern genomic age, molecular genetic data have become very easy and inexpensive to obtain for many species. However, some lineages are poorly represented in or absent from tissue collections, or are unavailable for molecular analysis for other reasons such as restrictive biological sample export laws. Other species went extinct recently and are only available in formalin museum preparations or perhaps even as subfossils. In this brief communication we present a new method for placing cryptic, recently extinct, or hypothesized taxa into an ultrametric phylogeny of extant taxa using continuous character data. This method is based on a relatively simple modification of an established maximum likelihood (ML) method for phylogeny inference from continuous traits. We show that the method works well on simulated trees and data. We then apply it to the case of placing the Culebra Island Giant Anole (Anolis roosevelti) into a phylogeny of Caribbean anoles. Anolis roosevelti is a “crown‐giant” ecomorph anole hypothesized to have once been found throughout the Spanish, United States, and British Virgin Islands, but that has not been encountered or collected since the 1930s. Although this species is widely thought to be closely related to the Puerto Rican giant anole, A. cuvieri, our ML method actually places A. roosevelti in a different part of the tree and closely related to a clade of morphologically similar species. We are unable, however, to reject a phylogenetic position for A. roosevelti that places it as sister taxon to A. cuvieri; although close relationship with the remainder of Puerto Rican anole species is strongly rejected by our method.     

Molecular identification of five Indian sciaenids (pisces: perciformes,sciaenidae) using RAPD markers

The randomly amplified polymorphic DNA (RAPD) markers were used to detect interspecific genetic variability and genetic relatedness among five Indian sciaenids namely Otolithes cuvieri, Johnieops sina, Johnieops macrorhynus, Johnieops vogleri and Protonibea diacanthus for the first time. Eight RAPD primers (OPA01, OPA06, OPA07, OPA18, OPP12, OPP14, OPP16 and OPP11) generated 40 species specific diagnostic bands. The highest genetic divergence was detected between J. macrorhynus and P. diacanthus (0.586) where as the lowest one was observed between J. sina and J. vogleri (0.274). Handling editor: C. Strumbauer     

A revision of the Lower Guinean Hepsetus species (Characiformes; Hepsetidae) with the description of Hepsetus kingsleyae sp. nov

An α‐taxonomic revision of the African pike, Hepsetus odoe, from Lower Guinea is provided. The results show that three different species occur in Lower Guinea instead of one. Hepsetus akawo, recently described from West Africa, is present in the northern part of Lower Guinea; Hepsetus lineata, the most widespread species within Lower Guinea, is known from the Sanaga (Cameroon) in the north to the Shiloango (Democratic Republic of the Congo) in the south and Hepsetus kingsleyae sp. nov. is endemic to the Ogowe Basin. The new species H. kingsleyae is described and H. lineata, which is elevated here to the species level, is redescribed. Hepsetus lineata can easily be recognized by its prominent horizontal line pattern on the flanks and differs further from H. akawo and H. kingsleyae in the number of lateral‐line scales and the number of gill rakers. Hepsetus kingsleyae differs from H. lineata and H. akawo by its narrow head, elongated snout and narrow, knife‐shaped body. All three species are also distinguishable from H. odoe and the recently revalidated H. cuvieri. A few exceptional specimens could not be allocated to one of the three species and may represent hybrids because of their mixed diagnostic characters or their intermediate values.     

Molecular identification and phylogenetic relationships of seven Indian Sciaenids (Pisces: Perciformes,Sciaenidae) based on 16S rRNA and cytochrome c oxidase subunit I mitochondrial genes

The partial sequences of 16S rRNA and cytochrome c oxidase subunit I (COI) mitochondrial genes were analyzed for species identification and phylogenetic relationships among the commercially important Indian sciaenids (Otolithes cuvieri, Otolithes ruber, Johnius dussumieri, Johnius elongatus, Johnieops vogleri, Otolithoides biauritus and Protonibea diacanthus). Sequence analysis of both genes revealed that the seven species fell into three distinct groups, which were genetically distant from each other and exhibited identical phylogenetic resolution. Partial sequences of both the genes provided sufficient phylogenetic information to distinguish the seven sciaenids indicating the usefulness of mtDNA-based approach in species identification.     

Variation of the advertisement call of Physalaemus centralis Bokermann, 1962 (Anura: Leptodactylidae) in the Cerrado of central Brazil

Acoustic signals are employed to support the correct designation of species and enables the discrimination between individuals. We analyzed both intra and interindividual variability of the advertisement call of Physalaemus centralis on the basis of specimens recorded in eight localities of the Cerrado in central Brazil. Within-individual variation, most of the call parameters were static properties. Both spectral and temporal call properties were dynamic between individuals. All call proprieties varied more between individuals than within an individual. Our study adds important data for future studies on the taxonomy and phylogeny of the P. cuvieri clade.     

Intraspecific variability of beaver teeth (Castoridae: Rodentia)

The metric variability of the teeth of Castor fiber Linné, 1758 and Castor canadensis Kuhl, 1820 with age are assessed. The ages of the studied specimens of C. fiber range from about 2 months to 16–17 years. Measurements of all teeth are given, and show a large range of size variation over all ages. The greatest size ranges can be observed for incisors, premolars, and third molars. Size variations of the fossil beavers Steneofiber eseri v. Meyer, 1846, Steneofiber castorinus Pomel, 1847, Anchitheriomys suevicus Schlosser, 1884, and Trogontherium cuvieri Fischer de Waldheim, 1809 are compared with those of C. fiber, and are of the same range and magnitude. It seems that it is nearly impossible to age beavers exactly with non‐invasive methods on the basis of tooth morphology, namely by the form and pattern of enamel islands on the chewing surface of the cheek teeth. Therefore, in conclusion, it is suggested that the observed variability in the size of fossil beaver teeth should be interpreted as changes with age in accordance with the overall size range in C. fiber. © 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2009, 155 , 926–936.     

Articular and vertebral lesions in the Pleistocene sloths (Xenarthra,Folivora) from the Brazilian Intertropical Region

We performed a macroscopic study on six species of Pleistocene sloths from the Brazilian Intertropical Region (BIR) including Valgipes bucklandi, Catonyx cuvieri, Mylodonopsis ibseni, Glossotherium sp., Ocnotherium giganteum and Eremotherium laurillardi and compared the results with a previous investigation of Nothrotherium maquinense and Ahytherium aureum. Differential diagnostic analysis revealed six varieties of musculoskeletal diseases: Calcium Pyrosphosphate Deposition Disease (CPPD), Spondyloarthropathy (SpA), Osteoarthritis (OA), Osteochondritis Dessicans (OD), Schmorl’s Node (SN) and Spondylosis Deformans (SD). We also carried out multivariate statistical analyses on all Pleistocene sloths to verify the similarity between taxa based on diagnosed diseases.     

Calcium uptake by juvenile western rock lobster,Panulirus cygnus George,from dietary coralline algae

Foliose coralline algae form a large proportion of the diet of juvenile Panulirus cygnus George at some locations but its rôle in the diet is not clear. In the laboratory, early post-moult rock lobsters fed Corallina cuvieri Lamouroux, labelled with ⁴⁵Ca primarily in the algal skeleton showed significant uptake of calcium compared with lobsters maintained in sea water containing dissolved ⁴⁵Ca. Shortly after ingestion by the rock lobsters the isotopic calcium was distributed through all the soft tissues examined and the exoskeleton. After a 5-day period isotopic calcium levels had fallen in most soft tissues but were not significantly lower in the digestive gland, the mid-gut or the exoskeleton. The data indicate that calcium in the algal skeleton of dietary C. cuvieri is absorbed by the digestive system and may be important in the postmoult re-mineralization of the exoskeleton.     

Parasites of fishes in the recently inundated ephemeral Lake Liambezi,Namibia

Lake Liambezi forms the periodic connection between the upper Zambezi, Kwando and Okavango rivers. A full parasitological assessment was conducted on 86 fish, representing 14 species in six families sampled in August 2011. Parasite diversity was low and dominated by species with complex life cycles involving intermediate hosts. Most prevalent were larval nematodes (Contracaecum sp.) infecting 12 and Trypanasoma sp. infecting nine of the 14 host species. The intra-erythrocytic parasite Babesiosoma mariae was found in the blood of Coptodon rendalli and Oreochromis andersonii with prevalence of 50% and 60%, respectively. The host-specific monogenean Annulotrema hepseti was recorded only from H. cuvieri with a prevalence of 100%. Notable absences were the copepod and branchiuran parasites that have direct lifecycles and usually occur in high prevalence and abundance in the region. Because parasites with direct life cycles can only be transported into the lake on the host fish, their absence suggests limited immigration of infected fishes into the lake. This suggests that internal recruitment dominates over immigration in the fish population dynamics in Lake Liambezi.     

Fertility of triploid backcross progeny, (GengoroubunaCarassius auratus cuvieri ♀ × carpCyprinus carpio ♂)F11 ♀ × carp or gengoroubuna ♂

The fertility of triploid backcross progeny, (gengoroubunaCarassius auratus cuvieri ♀ × carpCyprinus carpio ♂) F₁ ♀ × gengoroubuna or carp ♂ is reported. In females, spawning could be induced by the injection of pituitary extracts. The fertilization rates were from 52.4 to 88.6%, and free swimming rates from 2.6 to 34.4%. Many normal larvae were obtained after crossing with the sperm of gengoroubuna. However, after crossing with sperm of the loachMisgurnus anguillicaudatus, most of the embryos died before hatching and no normal larvae were obtained. Sperm could not be stripped from the progeny of either type of triploid backcross after the injection of pituitary extracts. Under light microscope, each seminal lobule in the testis was mainly occupied by spermatocytes in various stages. Some spermatids were found in part of the lobules, but no spermatozoa were observed.