O Complexo Cliona celata Grant, 1826 (Porifera, Demospongiae) na América do Sul: uma abordagem molecular e morfológica

Este trabalho visou a averiguação do status taxonômico das esponjas bioerosivas do complexo Cliona celata da América do Sul por meio de técnicas moleculares, utilizando como marcadores a subunidade I da Citocromo c oxidase (cox1) e os Espaçadores Internos Transcritos do RNAr nuclear (ITS1 e ITS2), além de testar outros marcadores. Igualmente, avaliou o grau de variabilidade morfológica encontrado nessas espécies, principalmente por meio da morfometria dos tilóstilos, a fim de estabelecer uma diagnose para elas. Ainda, tentou determinar as relações filogenéticas dessas espécies com as demais esponjas bioerosivas utilizando o gene 28S do RNAr nuclear. Foi possível determinar a existência de cinco clados de esponjas bioerosivas do complexo Cliona celata para a América do Sul, e dois outros clados não-sulamericanos, por meio dos marcadores moleculares utilizados. Embora seja discutida a validade desses clados como espécies distintas, continua impossível, por meio de caracteres morfológicos, distingui-los, e dessa forma, a proposição formal de novas espécies é evitada. Através da reconstrução filogenética do grupo, é possível verificar que as esponjas bioerosivas analisadas se apresentaram como um grupo monofilético, e se separa em três principais clados: Pione, Spirastrellidae, e Clionaidae. Por meio desta, é sugerida a alocação das espécies do complexo C. viridis e C. schimidti dentro de Spirastrella, além de ser necessária a criação de um novo gênero para alocar as espécies do novo complexo identificado aqui, o complexo C. delitrix.

Massa corporal e morfometria de Conopophaga melanops (Aves: Conopophagidae): uma comparação intrasexual e entre áreas continental e insular de Mata Atlântica, na região da Baía da Ilha Grande, RJ

A forma e o tamanho de um determinado organismo devem caracterizar aspectos ecológicos, uma vez que a morfometria é resultado da evolução. Diferenças nos caracteres morfológicos podem ter sido causadas por isolamento geográfico, mesmo em períodos de tempo relativamente curtos. O estudo da morfologia ecológica é uma tentativa de compreender a relação funcional entre variação morfológica e a ecologia dos animais. A variação nos atributos morfométricos de tamanho corpóreo entre os sexos pode ser um resultado da ação da seleção sexual. O presente estudo aborda uma comparação intrasexual e entre área continental e insular da morfologia de Conopophaga melanops (Vieillot, 1818), tendo sido realizado em uma área na Ilha Grande e em outra área na Reserva Ecológica Rio das Pedras (ReRP), RJ. A espécie, endêmica de Mata Atlântica e estritamente florestal, apresenta dimorfismo sexual, contudo indivíduos jovens possuem plumagem similar a de fêmeas. As aves foram capturadas com redes neblina, e doze medidas morfométricas foram obtidas de 51 indivíduos. A confirmação do sexo foi realizada por métodos moleculares baseados no DNA em 69 amostras. O percentual de erro na identificação do sexo em campo, pela plumagem, foi de 9,7%. A confirmação molecular do sexo é uma importante ferramenta que têm potencial de revelar padrões demográficos em estudos comportamentais e reprodutivos desta espécie. Na ReRP o comprimento da asa e a variável distância da cabeça até a ponta do bico apresentaram uma diferença significativa, sendo maior para machos do que para fêmeas. Já na Ilha Grande, as únicas variáveis que apresentaram diferença significativa foram comprimento da cauda (maior em machos) e altura do bico na base (maior em fêmeas). As diferenças de tamanho da asa entre os sexos corroboram com padrões de diversas outras espécies Neotropicais. A diferença morfométrica do bico pode estar associada à ecologia alimentar desta espécie. Tanto fêmeas quanto machos foram maiores na ilha do que no continente com relação ao comprimento total e comprimento da asa, além de comprimento da cauda maior para os machos.

Alimentación de la merluza en la región del talud continental argentino, época invernal (Merlucciidae, Merluccius merluccius hubbsi)

In the present paper the first results on the food and feeding habityas of hake in the region of the Argentinean continental slope are given. The data were obtained from the hake sampling made on board of the German research ship "Walther Herwig" during his first fishing exploratory trip in the Southwest Atlantic and from the investigation on the collected hake's stomachs.

Genetic identification of four Malaysian mackerel species off Coast of Peninsular Malaysia based on molecular marker

Random Amplified Polymorphic DNA (RAPD) markers and cytochrome b (Cyt-b) gene sequences were utilized to fingerprint and construct phylogenetic relationships among four species of mackerel commonly found in the Straits of Malacca namely Rastrelliger kanagurta, R. brachysoma, Decapterus maruadsi and D. russelli. The UPGMA dendogram and genetic distance clearly showed that the individuals clustered into their own genus and species except for the Decapterus. These results were also supported by partial mtDNA cytochrome b gene sequences (279 bp) which found monotypic sequence for all Decapterus studied. Cytochrome b sequence phylogeny generated through Neighbor Joining (NJ) method was congruent with RAPD data. Results showed clear discrimination between both genera with average nucleotide divergence about 25.43%. This marker also demonstrated R. brachysoma and R. kanagurta as distinct species separated with average nucleotide divergence about 2.76%. However, based on BLAST analysis, this study indicated that the fish initially identified as D. maruadsi was actually D. russelli. The results highlighted the importance of genetic analysis for taxonomic validation, in addition to morphological traits.

Evolutionary associations between sand seatrout (Cynoscion arenarius) and silver seatrout (C. nothus) inferred from morphological characters, mitochondrial DNA, and microsatellite markers

The evolutionary associations between closely related fish species, both contemporary and historical, are frequently assessed by using molecular markers, such as microsatellites. Here, the presence and variability of microsatellite loci in two closely related species of marine fishes, sand seatrout (Cynoscion arenarius) and silver seatrout (C. nothus), are explored by using heterologous primers from red drum (Sciaenops ocellatus). Data from these loci are used in conjunction with morphological characters and mitochondrial DNA haplotypes to explore the extent of genetic exchange between species offshore of Galveston Bay, TX. Despite seasonal overlap in distribution, low genetic divergence at microsatellite loci, and similar life history parameters of C. arenarius and C. nothus, all three data sets indicated that hybridization between these species does not occur or occurs only rarely and that historical admixture in Galveston Bay after divergence between these species was unlikely. These results shed light upon the evolutionary history of these fishes and highlight the genetic properties of each species that are influenced by their life history and ecology.

Pacific Hake, Merluccius productus, Autecology: A Timely Review

Pacific hake, Merluccius productus, the most abundant groundfish in the California Current Large Marine Ecosystem (CCLME), is a species of both commercial significance, supporting a large international fishery, and ecological importance, connecting other species as both predator and prey. Coastal Pacific hake migrations are characterized by movements between northern summer feeding areas and southern winter spawning areas, with variations in annual abundance, distribution, and the extent of these movements associated with varying climate-ocean conditions. In general, warm (cool) years with enhanced (reduced) stratification and poleward (equatorward) transport are often related to good (poor) recruitment, increased (decreased) northward distribution, and reduced (enhanced) growth. However, the classic periodic pattern of annual migration and distribution may no longer be fully representative. Based on recent advances in the understanding of climate-ocean variability off the U.S. west coast, we hypothesize that the annual movements of Pacific hake are more responsive to climate-ocean variability than previously thought, and further, that changes observed in Pacific hake distributions may reflect long-term changes in climate-ocean conditions in the CCLME. Therefore, an updated model of these relations is key to effective monitoring and management of this stock, as well as to devising scenarios of future change in the CCLME as a result of climate variations. The current state of knowledge of the relationship between the Pacific hake and its environment is reviewed, highlighting emerging ideas compared to those of the past, and priorities for future research are suggested.

Caracterização molecular de bastonetes Gram positivos irregulares e actinomicetos aeróbios obtidos de espécimes clínicos, de ensaios de esterilidade e de áreas limpas

Os bastonetes Gram positivos irregulares (BGPIs) compõem um grupo de espécies bacterianas com ampla diversidade fenotípica e que podem estar presente no meio ambiente, na microbiota humana e de animais. A identificação acurada de BGPIs em nível de gênero e espécie empregando métodos bioquímicos convencionais é bastante limitada, sendo recomendado, portanto, o uso de técnicas moleculares. No presente estudo, foram identificadas amostras de BGPIs oriundas de espécimes clínicos de humanos, de produtos farmacêuticos e de áreas limpas através da análise de sequencias do gene 16S rRNA e de outros genes conservados (housekeeping genes). Os resultados obtidos pelo sequenciamento dos genes 16S rRNA e rpoB demonstraram C. striatum multi-resistente (MDR) como responsável por surto epidêmico em ambiente hospitalar da cidade do Rio de Janeiro. Quinze cepas de C. striatum foram isoladas em cultura pura a partir de secreção traqueal de pacientes adultos submetidos a procedimentos de entubação endotraqueal. A análise por eletroforese em gel de campo pulsado (PFGE) indicou a presença de quatro perfis moleculares, incluindo dois clones relacionados com cepas MDR (PFGE I e II). Os dados demonstram a predominância de PFGE I entre cepas MDR isoladas de unidades de terapia intensiva e enfermarias cirúrgicas. Uma potencial ligação causal entre a morte e a infecção por C. striatum MDR (PFGE tipos I e II) foi observada em cinco casos. Adicionalmente, acreditamos que este seja o primeiro estudo de identificação de espécies de Nocardia relacionadas com infecções humanas pela análise da sequencia multilocus (MLSA) no Brasil. Diferente dos dados observados na literatura (1970 a 2013) e obtidos pelos testes fenotípicos convencionais, a caracterização molecular de quatro lócus (gyrB-16S-secA1-hsp65) permitiu a identificação das espécies N. nova, N. cyriacigeorgica, N. asiatica e N. exalbida/gamkensis relacionadas com quadros de nocardiose em humanos. Cepas de N. nova isoladas de diferentes materiais clínicos de um único paciente apresentaram padrões de susceptibilidade antimicrobianos idênticos e dois perfis PFGE, indicando a possibilidade de quadros de co-infecção por N. nova em humanos. Em outra etapa da investigação, amostras de BGPIs obtidos de ambientes de salas limpas que não puderam ser identificadas por critérios convencionais foram submetidas a análise da sequência do gene 16S rRNA e caracterizadas 95,83% em nível de gênero e 35,42% em espécies. Para gêneros mais encontrados no estudo, foram analisados os genes rpoB e recA de dezessete cepas de Microbacterium e utilizado o MLSA para a identificação de sete cepas identificadas como Streptomyces. Os ensaios permitiram a identificação de três cepas de Microbacterium e de uma única amostra de Streptomyces ao nível de espécie. A análise da sequencia do gene rpoB também se mostrou eficaz na identificação de espécies de cepas de Corynebacterium. Finalmente, para as cepas ambientais pertencentes à classe Actinobacteria os dados morfológicos, bioquímicos e genotípicos permitiram documentar a cepa 3117BRRJ como representante de uma nova espécie do gênero Nocardioides, para o qual o nome Nocardioides brasiliensis sp. nov. e as cepas 3712BRRJ e 3371BRRJ como representante de um novo gênero e espécie para o qual o nome Guaraldella brasiliensis nov. foi proposto.

Maternal effects on fecundity and egg quality of the Patagonian stock of Argentine Hake (Merluccius hubbsi)

The influences of age, size, and condition of spawning females on fecundity and oocyte quality were analyzed for the Patagonian stock of Argentine Hake (Merluccius hubbsi). Samples of mature females were collected in the spawning area as part of 2 research surveys conducted in January 2010 and 2011, during the peak of the reproductive season. Batch fecundity (BF) ranged between 40,500 (29 cm total length [TL]) and 2,550,000 (95 cm TL) hydrated oocytes, and was positively correlated with TL, gutted weight, age, hepatosomatic index (HSI), and the relative condition factor (Kn). Relative fecundity ranged between 85 and 1040 hydrated oocytes g–1 and showed significant positive relationships with gutted weight, HSI, and Kn; however, coefficients of determination were low for all regressions. Dry weights of samples of 100 hydrated oocytes ranged between 1.8 and 3.95 mg and were positively correlated with all variables analyzed, including batch and relative fecundity. Multiple regression models created with data of the morphophysiological characteristics of females supported maternal influences on fecundity and egg weights. Within the studied size range (29–95 cm TL), larger individuals had better somatic and egg condition, mainly revealed by higher HSI and hydrated oocytes with larger oil droplets (275.71μm [standard error 1.49]). These results were associated with the higher feeding activity of larger females during the spawning season in comparison with the feeding activity of young individuals (<5 years old); the better nutritional state of larger females, assumed to result from more feeding, was conducive to greater production of high-quality eggs.

Examination of the foraging habits of Pacific harbor seal (Phoca vitulina richardsi) to describe their use of the Umpqua River, Oregon, and their predation on salmonids

The increase in harbor seal (Phoca vitulina richardsi) abundance, concurrent with the decrease in salmonid (Oncorhynchus spp.) and other fish stocks, raises concerns about the potential negative impact of seals on fish populations. Although harbor seals are found in rivers and estuaries, their presence is not necessarily indicative of exclusive or predominant feeding in these systems. We examined the diet of harbor seals in the Umpqua River, Oregon, during 1997 and 1998 to indirectly assess whether or not they were feeding in the river. Fish otoliths and other skeletal structures were recovered from 651 scats and used to identify seal prey. The use of all diagnostic prey structures, rather than just otoliths, increased our estimates of the number of taxa, the minimum number of individuals and percent frequency of occurrence (%FO) of prey consumed. The %FO indicated that the most common prey were pleuronectids, Pacific hake (Merluccius productus), Pacific stag-horn sculpin (Leptocottus armatus), osmerids, and shiner surfperch (Cymatogaster aggregata). The majority (76%) of prey were fish that inhabit marine waters exclusively and fish found in marine and estuarine areas (e.g. anadromous spp.) which would indicate that seals forage predominantly at sea and use the estuary for resting and opportunistic feeding. Salmonid remains were encountered in 39 samples (6%); two samples contained identifiable otoliths, which were determined to be from chi-nook salmon (O. tshawytscha). Because of the complex salmonid composition in the Umpqua River, we used molecular genetic techniques on salmonid bones retrieved from scat to discern species that were rare from those that were abundant. Of the 37 scats with salmonid bones but no otoliths, bones were identified genetically as chinook or coho (O. kisutch) salmon, or steelhead trout (O. mykiss) in 90% of the samples.

Antihemostatic molecules from saliva of blood-feeding arthropods

The ability to feed on vertebrate blood has evolved many times in various arthropod clades. Consequently, saliva of blood-feeding arthropods has proven to be a rich source of antihemostatic molecules. A variety of platelet aggregation inhibitors antagonize platelet responses to wound-generated signals, including ADP, thrombin, and collagen. Anticoagulants disrupt elements of both the intrinsic and extrinsic pathways. Vasodilators include nitrophorins (nitric oxide storage and transport heme proteins), a variety of peptides that mimic endogenous vasodilatory neuropeptides, and proteins that catabolize or sequester endogenous vasoconstrictors. Multiple salivary proteins may be directed against each component of hemostasis, resulting in both redundancy and in some cases cooperative interactions between antihemostatic proteins. The complexity and redundancy of saliva ensures an efficient blood meal for the arthropod, but it also provides a diverse array of novel antihemostatic molecules for the pharmacologist.

A snake venom metalloproteinase that inhibited cell proliferation and induced morphological changes of ECV304 cells

TSV-DM, a basic metalloproteinase with a molecular weight of 110 kDa, was purified from Trimeresurus stejnegeri venom. TSV-DM degraded the A alpha chain of fibrinogen more rapidly than the B beta chain in a dose dependent manner. The cDNA of TSV-DM encode

Molecular systematics of pikas (genus Ochotona) inferred from mitochondrial DNA sequences

The phylogenetic relationships among worldwide species of genus Ochotona were investigated by sequencing mitochondrial cytochrome b and ND4 genes. Parsimony and neighbor-joining analyses of the sequence data yielded congruent results that strongly indicated three major clusters: the shrub-steppe group, the northern group, and the mountain group. The subgeneric classification of Ochotona species needs to be revised because each of the two subgenera in the present classification contains species from the mountain group. To solve this taxonomic problem so that each taxon is monophyletic, i.e., represents a natural clade, Ochotona could be divided into three subgenera, one for the shrub-steppe species, a second for the northern species, and a third for the mountain species. The inferred tree suggests that the differentiation of this genus in the Palearctic Region was closely related to the gradual uplifting of the Tibet (Qinghai-Xizang) Plateau, as hypothesized previously, and that vicariance might have played a major role in the differentiation of this genus on the Plateau, On the other hand, the North American species, O. princeps, is most likely a dispersal event, which might have happened during the Pliocene through the opening of the Bering Strait. The phylogenetic relationships within the shrub-steppe group are worth noting in that instead of a monophyletic shrub-dwelling group, shrub dwellers and steppe dwellers are intermingled with each other. Moreover, the sequence divergence within the sister tars of one steppe? dweller and one shrub dweller is very low. These findings support the hypothesis that pikes have entered the steppe environment several times and that morphological similarities within steppe dwellers were due to convergent evolution. (C) 2000 Academic Press.

Phylogeny and biogeography of the Petaurista philippensis complex (Rodentia : Sciuridae), inter- and intraspecific relationships inferred from molecular and morphometric analysis

With modified DNA extraction and Purification protocols, the complete cytochrome b gene sequences (1140 bp) were determined from degraded museum specimens. Molecular analysis and morphological examination of cranial characteristics of the giant flying squirrels of Petaurista philippensis complex (P. grandis, P. hainana, and P. yunanensis) and other Petaurista species yielded new insights into long-standing controversies in the Petaurista systematics. Patterns of genetic variations and morphological differences observed in this study indicate that P. hainana, P. albiventer, and P. yunanensis can be recognized as distinct species, and P. grandis and P. petaurista are conspecific populations. Phylogenetic relationships reconstructed by using parsimony, likelihood, and Bayesian methods reveal that, with P. leucogenys as the basal branch, all Petaurista groups formed two distinct clades. Petaurista philippensis, P. hainana, P. yunanensis, and P. albiventer are clustered in the same clade, while P. grandis shows a close relationship to P. petaurista. Deduced divergence times based on Bayesian analysis and the transversional substitution at the third codon suggest that the retreating of glaciers and upheavals or movements of tectonic plates in the Pliocene-Pleistocene were the major factors responsible for the present geographical distributions of Petaurista groups. (c) 2005 Elsevier Inc. All rights reserved.

Study on some biological and feeding factors of spined loach Cobitis taenia from the Babolrud, Talar and Siahrud Rivers

Spined loach, Cobitis taenia, is a predominant fish in the river systems of the southern Caspian Sea basin. Although there is evidence of the geographical divergence of this taxon, but no information is available on morphological differences within the species populations. This study was designed to evaluate some biological factors including; morphometric and meristic characters, length-weight, age-growth, condition factor, diet, reproduction, variation and differentiation, in the Babolrud, Talar and Siahrud Rivers in south of the Caspian Sea basin. Age, sex ratio, fecundity, ova diameter and gonadosomatic index were estimated. Also, regression analyses was tested the relation between fecundity and fish length, weight, gonad weight, and also age. Totally 858 fish of which 721 were matures, were collected from these rivers by electrofishing. 37 morphometric characters, 9 meristic characters and 78 truss network system characters were estimated. Resulats of DFA analysis based on data of morphometric and meristic showd that these populations are highly (94.5%) varios from each other. In discriminate function analysis, the proportion of individuals correctly classified into their original groups was 61%, 65.4% and 86.5% for upstream and downstream, respectively. Clustering based on Euclidean distances among groups of centroids using an UPGMA and also principal component analysis’ results for morphometric data indicated that these populations from these three rivers were clearly distinct from each other. Regression equations between length and weight in these three populations were significantly different from Folton factor (b=3), that showed the fish has a negative Alometric growth process. Condition factor was estimated between 0.8912 to 1.2736 and 0.8131 to 1.4489 for males and females, respectively. Sex ratio (female: male) in these populations was 1.2816:1. The difference between the number of females and males was significant and females were more than males. The female and male specimens reach maturity by Tl more than 40 and 30 mm and at the age of 2+ and 1+, respectively. The mean of ova diameter was 0.5824±0.2882. The spawning took place from May to late July, at the water temperature from 18.7 to 24.0°C. The GSI values average at the beginning of the reproduction period was about 9%, with ranged from 2 to 26% in ripe mature females. The absolute and relative fecundity were 2109±792 and 579±208 respectively. The absolute fecundity was significantly related to body weight and gonads weight. Based on the pattern of gonado-somatic index, it was concluded that this fish has prolong active reproductive period, which is a type of adaptation by short-lived small fishes to environmental conditions. The macroscopic and histological results showed that the female and male have 5 and 4 stages in their maturation process, respectively. The RLG index was about 0.4732, which showed the fish is a carnivorous species. Significant difference was observed between fishes with different length and diet. The main foods of the fish were Trichoptera, Chironomidae larvae and Ephemeroptera which were their prefered food as well, however it was estimated that the food selection and diet are affected by environmental conditions.

Molecular evidence for diphyly of the Asian catfish family Amblycipitidae (Teleostei: Siluriformes) and exclusion of the South American Aspredinidae from Sisoroidea

Amblycipitidae Day, 1873 is an Asian family of catfishes (Siluriformes) usually considered to contain 28 species placed in three genera: Amblyceps (14 spp.), Liobagrus (12 spp.) and Xiurenbagrus (2 spp.). Morphology-based systematics has supported the monophyly of this family, with some authors placing Amblycipitidae within a larger group including Akysidae, Sisoridae and Aspredinidae, termed the Sisoroidea. Here we investigate the phylogenetic relationships among four species of Amblyceps, six species of Liobagrus and the two species of Xiurenbagrus with respect to other sisoroid taxa as well as other catfish groups using 6100 aligned base pairs of DNA sequence data from the rag1 and rag2 genes of the nuclear genome and from three regions (cyt b, COL ND4 plus tRNA-His and tRNA-Ser) of the mitochondrial genome. Parsimony and Bayesian analyses of the data indicate strong support for a diphyletic Amblycipitidae in which the genus Amblyceps is the sister group to the Sisoridae and a clade formed by genera Liobagrus and Xiurenbagrus is the sister group to Akysidae. These taxa together form a well supported monophyletic group that assembles all Asian sisoroid taxa, but excludes the South American Aspredinidae. Results for aspredinids are consistent with previous molecular studies that indicate these catfishes are not sisoroids, but the sister group to the South American doradoid catfishes (Auchenipteridae + Doradidae). The redefined sisoroid clade plus Bagridae, Horabagridae and (Ailia + Laides) make up a larger monophyletic group informally termed "Big Asia." Likelihood-based SH tests and Bayes Factor comparisons of the rag and the mitochondrial data partitions considered separately and combined reject both the hypothesis of amblycipitid monophyly and the hypothesis of aspredinid inclusion within Sisoroidea. This result for amblycipitids conflicts with a number of well documented morphological synapomorphies that we briefly review. Possible nomenclatural changes for amblycipitid taxa are noted.