Antimicrobial-Resistant Bacteria from Free-Living Green Turtles (Chelonia mydas).

Bioindicator species are used to assess the damage and magnitude of possible impacts of anthropic origin on the environment, such as the reckless consumption of antimicrobials. Chelonia mydas has several characteristics that make it a suitable bioindicator of marine pollution and of the presence of pathogens that cause diseases in humans. This study aimed to investigate the green sea turtle as a reservoir of resistant bacteria, mainly because C. mydas is the most frequent sea turtle species in Brazilian coastal regions and, consequently, under the intense impact of anthropic factors. Free-living green sea turtles ranging from 42.8 to 92 cm (average = 60.7 cm) were captured from Itaipú Beach, Brazil. Cloaca samples (characterizing the gastrointestinal tract) and neck samples (representing the transient microbiota) were collected. Bacterial species were identified, and their was resistance associated with the antimicrobials cephalothin, ciprofloxacin, gentamicin, tetracycline, and vancomycin. Citrobacter braaki, Klebsiella oxytoca, K. variicola and Proteus mirabilis were found resistant to cephalothin and Morganella morganii and Enterococcus faecalis tetracycline-resistant isolates in cloaca samples. In neck samples, species resistant to tetracycline were Salmonella sp., Serratia marcescens, S. ureylitica and Proteus mirabilis. This data reinforces that the green turtle is a bioindicator of antimicrobial resistance (AMR).

The microbiome of the sponge Aplysina caissara in two sites with different levels of anthropogenic impact.

Despite the important roles that marine sponges play in ecosystem functioning and structuring, little is known about how the sponge holobiont responds to local anthropogenic impacts. Here we assess the influence of an impacted environment (Praia Preta) on the microbial community associated with the endemic sponge Aplysina caissara in comparison to a less-impacted area (Praia do Guaecá) from the coast of São Paulo state (Brazil, southwestern Atlantic coast). We hypothesized that the local anthropogenic impacts will change the microbiome of A. caissara and that the community assembly will be driven by a different process (i.e. deterministic versus stochastic) under distinct levels of impact. The microbiome at the amplicon sequence variants level was found to be statistically distinct between sponges from the different sites, and this was also seen for the microbial communities of the surrounding seawater and sediments. Microbial communities of A. caissara from both sites were found to be assembled by deterministic processes, even though the sites presented distinct anthropogenic impacts, showing a pivotal role of the sponge host in selecting its own microbiome. Overall, this study revealed that local anthropogenic impacts altered the microbiome of A. caissara; however, assembly processes are largely determined by the sponge host.

Microplastics in coastal and oceanic surface waters and their role as carriers of pollutants of emerging concern in marine organisms.

Microplastics (Mps) pose a significant environmental challenge with global implications. To examine the effect of Mps on coastal and oceanic surface waters, as well as in marine organisms, 167 original research papers published between January 2013 and September 2022 were analyzed. The study revealed an unequal distribution of research efforts across the world. Fragments and fibers were the most frequently detected particles in ocean surface waters and marine biota, which mainly consisted of colored and transparent microparticles. Sampling of Mps was primarily done using collecting nets with a mesh size of 330 µm. Most articles used a stereomicroscope and Fourier-Transform Infrared spectroscopy for identification and composition determination, respectively. Polyethylene and polypropylene were the most frequent polymers found, both in coastal waters and in marine organisms. The major impact observed on marine organisms was a reduction in growth rate, an increase in mortality, and reduced food consumption. The hydrophobic nature of plastics encourages the formation of biofilms called the "plastisphere," which can carry pollutants that are often toxic and can enter the food chain. To better define management measures, it is necessary to standardize investigations that assess Mp pollution, considering not only the geomorphological and oceanographic features of each region but also the urban and industrial occupation of the studied marine environments.

Diversity and structure of bacterial and archaeal communities associated with the vulnerable sponge Halichondria cebimarensis.

Sponges are essential components of the marine benthos and well known for their complex and abundant associated microbial communities. There are five endemic species of the genus Halichondria on the Brazilian coast and H. cebimarensis is one of the least studied. This sponge has a very limited geographic distribution and is classified as vulnerable. In order to understand the bacterial and archaeal communities associated with this sponge, samples of H. cebimarensis were collected from the southwestern Atlantic coast (Brazil, São Paulo state). Choanosome samples were separated and processed to be (i) inoculated in three different culture media; (ii) investigated by transmission electron microscopy; (iii) submitted to 16S rRNA metabarcoding. Forty isolates were obtained and 12 were identified as belonging to the Bacilli class. The culture-dependent approaches allowed us to access unique members of the microbial community. Our analyses revealed that this animal belongs to the Low Microbial Abundance group of sponges. Culture-independent approaches showed that the H. cebimarensis microbiome is dominated by the heterotrophic Gammaproteobacteria AqS2 ("Ca. Amphirhobacter heronislandensis"). This is the first study to reveal details of the microbiome of this vulnerable sponge and is an important step in understanding how this sponge functions, its biotechnological potential and a contribution to conservation efforts.

Prokaryotic, Fungal, and Unicellular Eukaryotic Core Communities Across Three Sympatric Marine Sponges From the Southwestern Atlantic Coast Are Dominated Largely by Deterministic Assemblage Processes.

Marine sponges are known to harbor a diverse and complex microbiota; however, a vast majority of surveys have been investigating the prokaryotic communities in the north hemisphere and Australia. In addition, the mechanisms of microbial community assembly are poorly understood in this pivotal player of the ecosystem. Thus, this survey addressed the holobiome of the sponge species in the São Paulo region (Brazil) for the first time and investigated the contribution of neutral and niche processes of prokaryotic, fungal, and unicellular eukaryotic assemblage in three sympatric species Aplysina caissara, Aplysina fulva, and Tedania ignis along with environmental samples. The compositions of the holobiome associated with the sponges and detected in environmental samples were strikingly different. Remarkably, between 47 and 88% of the assigned operational taxonomic units (OTUs) were specifically associated with sponge species. Moreover, around 77, 69, and 53% of the unclassified OTUs from prokaryotic, fungal, and unicellular eukaryotic communities, respectively, showed less than 97% similarity with well-known databases, suggesting that sponges from the southwestern Atlantic coast are an important source of microbial novelty. These values are even higher, around 80 and 61% of the unclassified OTUs, when excluding low abundance samples from fungal and unicellular eukaryotic datasets, respectively. Host species were the major driver shaping the sponge-associated microbial community. Deterministic processes were primarily responsible for the assembly of microbial communities in all sponge species, while neutral processes of prokaryotic and fungal community assembly were also detected in the sympatric A. caissara and T. ignis replicates, respectively. Most of the species-rich sponge-associated lineages from this region are also found in the Northern seas and many of them might play essential roles in the symbioses, such as biosynthesis of secondary metabolites that exhibit antimicrobial and antiviral activities, as well as provide protection against host predation. Overall, in this study the microbiota was assembled by interactions with the host sponge in a deterministic-based manner; closely related sponge species shared a strong phylogenetic signal in their associated prokaryotic and fungal community traits and Brazilian sponges were a reservoir of novel microbial species.

How a collaborative integrated taxonomic effort has trained new spongiologists and improved knowledge of Martinique Island (French Antilles, eastern Caribbean Sea) marine biodiversity.

Although sponges are important components of benthic ecosystems of the Caribbean Sea, their diversity remained poorly investigated in the Lesser Antilles. By organizing a training course in Martinique, we wanted both to promote taxonomy and to provide a first inventory of the sponge diversity on this island. The course was like a naturalist expedition, with a field laboratory and a classroom nearby. Early-career scientists and environmental managers were trained in sponge taxonomy. We gathered unpublished data and conducted an inventory at 13 coastal sites. We explored only shallow water habitats (0-30 m), such as mangroves, reefs or rocky bottoms and underwater caves. According to this study, the sponge fauna of Martinique is currently represented by a minimum of 191 species, 134 of which we could assign species names. One third of the remaining non-identified sponge species we consider to be new to science. Martinique appears very remarkable because of its littoral marine fauna harboring sponge aggregations with high biomass and species diversity dominating over coral species. In mangroves, sponges cover about 10% of the surface of subtidal roots. Several submarine caves are true reservoirs of hidden and insufficiently described sponge diversity. Thanks to this new collaborative effort, the Eastern Caribbean has gained a significant increase of knowledge, with sponge diversity of this area potentially representing 40% of the total in the Caribbean Sea. We thus demonstrated the importance of developing exploratory and educational research in areas historically devoid of biodiversity inventories and systematics studies. Finally, we believe in the necessity to consider not only the number of species but their distribution in space to evaluate their putative contribution to ecosystem services and our willingness to preserve them.

Characterization of fibropapillomatosis in green turtles Chelonia mydas (Cheloniidae) captured in a foraging area in southeastern Brazil.

Fibropapillomatosis (FP) is a multifactorial disease that affects all species of marine turtles, including green turtles Chelonia mydas (Linnaeus, 1758). It is characterised by the development of internal or external tumours that, depending on their locations and sizes, may intensely impact the health condition of sea turtles. The goal of this study was to characterise the disease in C. mydas found in a foraging area in southeastern Brazil, evaluate the prevalence in this region, and correlate presence and absence, size, body distribution, number of tumours, and disease severity with biometric variables of the captured green turtles. Between 2008 and 2014, the prevalence rate of FP was 43.09%, out of 246 green turtles. The size of the animals with FP was relatively greater than animals without tumours, and the prevalence of FP increased with animal size, peaking in the 60-80 cm size class. From 2013 to 2014, gross evaluation of fibropapillomas was performed. The number of tumours per turtle ranged from 1 to 158. The size of tumours ranged from <1 cm (Size A) to >10 cm (Size D); Size A tumours and turtles slightly affected by the disease (Score 1) predominated. Tumour progression (72.1%) and regression (32.8%) were seen in some recaptured individuals (n = 61). Moreover, 24.6% of these turtles showed both progressions and regressions of tumours.

Lissodendoryx (Ectyodoryx) Lundbeck, 1909 (Coelosphaeridae, Poecilosclerida, Demospongiae) from Southern Chile: new species and a discussion of morphologic characters in the subgenus.

Our ongoing studies of the Chilean sponge fauna revealed four new species of Lissodendoryx (Ectyodoryx) that are described here, including three from the fjord's region (< 30 m depth, L.(E.) ballena sp. nov., L. (E.) corrugata sp. nov.,      L. (E.) coloanensis sp. nov.), and one from the deep waters off Diego Ramírez Archipelago (ca. 2000 m, L. (E.) diegoramirezensis sp. nov.). In addition, the type of L. (E.) anacantha was revised and found to bear much larger acanthostyles than originally reported. Our results revealed skeletal architectures quite distinct from that reported from the type species of L. (Ectyodoryx), and not predicted in the current diagnosis of Lissodendoryx, requiring an amended diagnosis. Ectosomal megascleres also set the majority of the new species apart from the type species of L. (Ectyodoryx), which does not have terminally microspined (sub)tylotes. This character is widespread in Lissodendoryx and might be pointing to phylogenetic affinities across the current subgeneric arrangement. Further morphologic characters and alternative phylogenetic scenarios are discussed, including hymedesmiid and myxillid affinities of the species described here.

Conjugated polyenes as chemical probes of life signature: use of Raman spectroscopy to differentiate polyenic pigments.

Polyenes, which are represented by carotenes, carotenoids and conjugated polyenals, are some of the most important targets for astrobiology, because they can provide strong evidence of the presence of organic compounds in the most extreme environments, such as on Mars. Raman spectroscopy has been used as the main analytical tool in the identification of such compounds, for the greatest variety of living species, from microorganisms to animals and plants. However, using only the position of the characteristic Raman bands can lead to errors in tentatively identifying chemicals. In this work, we present a series of observations that can provide a more complete and robust way to analyse the Raman spectrum of a polyenal, in which the position, the intensity, the use of various laser lines for excitation, and the combination of more than one pigment can be considered in the complete analysis.

Mycalina: another crack in the Poecilosclerida framework.

This is the first phylogenetic analysis integrating both morphological and molecular data of the sponge suborder Mycalina (Poecilosclerida), which was erected in 1994. A cladistic analysis of morphology supported the monophyly of Cladorhizidae (including Euchelipluma), Guitarridae (excluding Euchelipluma), Isodictyidae, Latrunculiidae, and Podospongiidae but rejected monophyly for Desmacellidae, Esperiopsidae, Hamacanthidae, and Mycalidae. Analyses of partial 16S and partial 28S rRNA datasets combined, as well as that of a complete 18S rDNA dataset, suggest that Mycalina is not monophyletic; Biemnidae is only distantly related to other poecilosclerids; Merlia and Desmacella branch near the base of a diverse Poecilosclerida clade; Mycalidae is monophyletic (excluding Mycale [Anomomycale] titubans in 18S); and Esperiopsidae and Isodictyidae form a clade. Analyses of the two molecular datasets differed on the monophyly of Podospongiidae and about the relationship of Podospongiidae to Isodictyidae + Esperiopsidae.

Evidence of regression of fibropapillomas in juvenile green turtles Chelonia mydas caught in Niterói, southeast Brazil.

Fibropapillomatosis is a disease characterized by cutaneous tumors affecting all marine turtle species, but mostly Chelonia mydas. The disease was first reported in 1938, and since then, the number of sightings has been increasing over the years. This disease can cause many complications in the affected animal and can lead to death, and is thus included in the many threats to marine turtle populations. It is still not known for certain what causes this disease, although many studies indicate a herpesvirus as the main etiologic agent. The incidence of fibropapillomatosis is rarely reported in adults, leading to speculations that there may be a cure for the disease or that the animals die before reaching adulthood. In this paper, 2 cases of fibropapillomatosis regression are reported from juvenile C. mydas caught between July 2008 and July 2010 in the coastal zone of Itaipu, Niterói, Rio de Janeiro, Brazil. These individuals were identified photographically upon recapture. One individual had a total regression (disappearance) of external papilloma within 164 d between first capture and recapture, and the other individual had a partial regression (decrease in size) observed within 13 to 188 d of recapture. The mechanism that triggers the regression is still unknown but is likely to be an immune system response or removal of the tumor promoter. There are few reported cases of regression in the world, and constant monitoring through mark-recapture is necessary to assess whether the marine turtles affected by this disease have real chances of survival.

Reconstruction of family-level phylogenetic relationships within Demospongiae (Porifera) using nuclear encoded housekeeping genes.

BACKGROUND: Demosponges are challenging for phylogenetic systematics because of their plastic and relatively simple morphologies and many deep divergences between major clades. To improve understanding of the phylogenetic relationships within Demospongiae, we sequenced and analyzed seven nuclear housekeeping genes involved in a variety of cellular functions from a diverse group of sponges. METHODOLOGY/PRINCIPAL FINDINGS: We generated data from each of the four sponge classes (i.e., Calcarea, Demospongiae, Hexactinellida, and Homoscleromorpha), but focused on family-level relationships within demosponges. With data for 21 newly sampled families, our Maximum Likelihood and Bayesian-based approaches recovered previously phylogenetically defined taxa: Keratosa(p), Myxospongiae(p), Spongillida(p), Haploscleromorpha(p) (the marine haplosclerids) and Democlavia(p). We found conflicting results concerning the relationships of Keratosa(p) and Myxospongiae(p) to the remaining demosponges, but our results strongly supported a clade of Haploscleromorpha(p)+Spongillida(p)+Democlavia(p). In contrast to hypotheses based on mitochondrial genome and ribosomal data, nuclear housekeeping gene data suggested that freshwater sponges (Spongillida(p)) are sister to Haploscleromorpha(p) rather than part of Democlavia(p). Within Keratosa(p), we found equivocal results as to the monophyly of Dictyoceratida. Within Myxospongiae(p), Chondrosida and Verongida were monophyletic. A well-supported clade within Democlavia(p), Tetractinellida(p), composed of all sampled members of Astrophorina and Spirophorina (including the only lithistid in our analysis), was consistently revealed as the sister group to all other members of Democlavia(p). Within Tetractinellida(p), we did not recover monophyletic Astrophorina or Spirophorina. Our results also reaffirmed the monophyly of order Poecilosclerida (excluding Desmacellidae and Raspailiidae), and polyphyly of Hadromerida and Halichondrida. CONCLUSIONS/SIGNIFICANCE: These results, using an independent nuclear gene set, confirmed many hypotheses based on ribosomal and/or mitochondrial genes, and they also identified clades with low statistical support or clades that conflicted with traditional morphological classification. Our results will serve as a basis for future exploration of these outstanding questions using more taxon- and gene-rich datasets.

Twelve new Demospongiae (Porifera) from Chilean fjords, with remarks upon sponge-derived biogeographic compartments in the SE Pacific.

This article reports on 12 new species originating from the Chilean fjords region, namely Clathria (Microciona) mytilifila sp. nov., Haliclona (Reniera) caduca sp. nov., Latrunculia (L.) ciruela sp. nov., Latrunculia (L.) copihuensis sp. nov., Latrunculia (L.) verenae sp. nov., Latrunculia (L.) yepayek sp. nov., Myxilla (Burtonanchora) araucana sp. nov., Neopodospongia tupecomareni sp. nov., Oceanapia guaiteca sp. nov., Oceanapia spinisphaera sp. nov., Suberites cranium sp. nov. and Tethya melinka sp. nov. The material studied was collected between 5 and 30 m depth at latitudes comprised between 42º and 50ºS, and is part of a large collection of Chilean sponges gathered by an international team in a series of expeditions. Identification keys are provided for SE Pacific Suberites and Latrunculia, and the known species of Myxilla (Burtonanchora) and Neopodospongia. A trans-Pacific link to the New Zealand fauna was retrieved for the latter genus. Distribution ranges apparent from the materials studied here are judged too preliminary to allow any inference on biotic boundaries in the SE Pacific. A revision of earlier assertions about these biogeographic units and their boundaries concluded that very little support remains other than for existence of a Magellanic fauna. This is in part a consequence of revising the taxonomy of sponge species originally deemed to underpin these areas. Specifically, the former proposal of a Central to Southern Chile biogeographic unit (33-56ºS) has been markedly undone. 

Nuclear markers reveal a complex introgression pattern among marine turtle species on the Brazilian coast.

Surprisingly, a high frequency of interspecific sea turtle hybrids has been previously recorded in a nesting site along a short stretch of the Brazilian coast. Mitochondrial DNA data indicated that as much as 43% of the females identified as Eretmochelys imbricata are hybrids in this area (Bahia State of Brazil). It is a remarkable find, because most of the nesting sites surveyed worldwide, including some in northern Brazil, presents no hybrids, and rare Caribbean sites present no more than 2% of hybrids. Thus, a detailed understanding of the hybridization process is needed to evaluate natural or anthropogenic causes of this regional phenomenon in Brazil, which could be an important factor affecting the conservation of this population. We analysed a set of 12 nuclear markers to investigate the pattern of hybridization involving three species of sea turtles: hawksbill (E. imbricata), loggerhead (Caretta caretta) and olive ridley (Lepidochelys olivacea). Our data indicate that most of the individuals in the crossings L. olivacea × E. imbricata and L. olivacea × C. caretta are F1 hybrids, whereas C. caretta × E. imbricata crossings present F1 and backcrosses with both parental species. In addition, the C. caretta × E. imbricata hybridization seems to be gender and species biased, and we also found one individual with evidence of multispecies hybridization among C. caretta × E. imbricata × Chelonia mydas. The overall results also indicate that hybridization in this area is a recent phenomenon, spanning at least two generations or ~40 years.

Taxonomic and functional microbial signatures of the endemic marine sponge Arenosclera brasiliensis.

The endemic marine sponge Arenosclera brasiliensis (Porifera, Demospongiae, Haplosclerida) is a known source of secondary metabolites such as arenosclerins A-C. In the present study, we established the composition of the A. brasiliensis microbiome and the metabolic pathways associated with this community. We used 454 shotgun pyrosequencing to generate approximately 640,000 high-quality sponge-derived sequences (∼150 Mb). Clustering analysis including sponge, seawater and twenty-three other metagenomes derived from marine animal microbiomes shows that A. brasiliensis contains a specific microbiome. Fourteen bacterial phyla (including Proteobacteria, Cyanobacteria, Actinobacteria, Bacteroidetes, Firmicutes and Cloroflexi) were consistently found in the A. brasiliensis metagenomes. The A. brasiliensis microbiome is enriched for Betaproteobacteria (e.g., Burkholderia) and Gammaproteobacteria (e.g., Pseudomonas and Alteromonas) compared with the surrounding planktonic microbial communities. Functional analysis based on Rapid Annotation using Subsystem Technology (RAST) indicated that the A. brasiliensis microbiome is enriched for sequences associated with membrane transport and one-carbon metabolism. In addition, there was an overrepresentation of sequences associated with aerobic and anaerobic metabolism as well as the synthesis and degradation of secondary metabolites. This study represents the first analysis of sponge-associated microbial communities via shotgun pyrosequencing, a strategy commonly applied in similar analyses in other marine invertebrate hosts, such as corals and algae. We demonstrate that A. brasiliensis has a unique microbiome that is distinct from that of the surrounding planktonic microbes and from other marine organisms, indicating a species-specific microbiome.

Morphology and molecules on opposite sides of the diversity gradient: four cryptic species of the Cliona celata (Porifera, Demospongiae) complex in South America revealed by mitochondrial and nuclear markers.

A great number of marine organisms lack proper morphologic characters for identification and species description. This could promote a wide distributional pattern for a species morphotype, potentially generating many morphologically similar albeit evolutionarily independent worldwide lineages. This work aimed to estimate the genetic variation of South America populations of the Cliona celata species complex. We used COI mtDNA and ITS rDNA as molecular markers and tylostyle length and width as morphological characters to try to distinguish among species. Four distinct clades were found within the South American C. celata complex using both genetic markers. The genetic distances comparisons revealed that scores among those clades were comparable to distances between each clade and series of previously described clionaid species, some of which belong to different genera. Our results also suggest that one of the clades has a broad discontinuous distribution in the Atlantic Ocean, while another presents high gene flow between the southern Atlantic and Pacific coasts of South America. Conversely, spicule morphology was not able to distinguish each clade, due to the high degree of overlap among them. Therefore, we considered that each recovered clade correspond, in fact, to different species that cannot be differentiated via morphological characters, which are often used to describe species within the C. celata species complex.

Isolation and cultivation of fungal strains from in vitro cell cultures of two marine sponges (Porifera: Halichondrida and Haplosclerida)

Despite the large number of reports describing sponge-microbe associations, limited knowledge is available about associated fungi and their relationships with the hosts. In this work, specific fungal strains were obtained directly from in vitro sponge cell cultures (primmorphs) and single sponge cells (cytospins) and compared with those obtained from whole tissue preparations. A total of 27 fungal strains were isolated from the marine sponges Hymeniacidon heliophila and Haliclona melana. Fifteen strains, nine from H. heliophila and six from H. melana, were obtained from whole tissue and were considered as possible mesohyl associated or transient fungi. Twelve strains were isolated from in vitro sponge cell cultures (primmorphs) and were, therefore, considered as cell associated. From these, five different strains were obtained from H. heliophila isolated cells, while five were identified from cytospins and two from primmorphs of H. melana. The fungal strains obtained from cell cultures from both sponge species were different, and none of them were detected in the whole tissue preparations of the same species. Nine H. heliophila and seven H. melana strains shows low similarity with the sequences available in public databases and belong to potentially new species. This is the first report of fungi isolated directly from sponge cells, which allowed the observation and selection of specific strains that probably would not be obtained by usual culture dependent techniques.

Sex determination in Turdus amaurochalinus (Passeriformes: Muscicapidae): morphometrical analysis supported by CHD gene.

Sex determination is important for conservation and population studies, particularly for reproduction programs of threatened species and behavioural ecology. Turdus amaurochalinus, Creamy-bellied Thrush, only exhibits sexual dimorphism during the breeding season, when males are considered to show intense yellow bills, and females and immature males show dark brown bills. The objectives of this study were: 1) to determine the sex of individuals using genetic techniques, and 2) to test the hypothesis that sex dimorphism can be detected by morphometry. This study was carried out at Parque Nacional da Restinga de Jurubatiba, a preserved area located on the North coast of Rio de Janeiro State. The birds were captured using ornithological nets, singly marked with metal rings, weighed, measured and had blood samples collected before being released. The sex of 42 T. amaurochalinus individuals was determined using the CHD gene marker. A total of 20 males and 22 females were identified from June to August, with peak capture frequency in June. Turdus amaurochalinus females and males differed significantly in morphometrical measures. The most important traits to distinguish males from females were wing length (Student t-test = 4.34, df = 40, p = 0.0001) and weight (Student t-test = 2.08, d f = 40, p = 0.044): females were heavier and had significantly shorter wing length than males. Females and males were correctly classified in 86% and 75% of cases, respectively, using Discriminant Analysis. The molecular analysis was the most secure method for sex determination in the studied species.

Sex determination in Turdus amaurochalinus (Passeriformes: Muscicapidae): morphometrical analysis supported by CHD gene

Sex determination is important for conservation and population studies, particularly for reproduction programs of threatened species and behavioural ecology. Turdus amaurochalinus, Creamy-bellied Thrush, only exhibits sexual dimorphism during the breeding season, when males are considered to show intense yellow bills, and females and immature males show dark brown bills. The objectives of this study were: 1) to determine the sex of individuals using genetic techniques, and 2) to test the hypothesis that sex dimorphism can be detected by morphometry. This study was carried out at Parque Nacional da Restinga de Jurubatiba, a preserved area located on the North coast of Rio de Janeiro State. The birds were captured using ornithological nets, singly marked with metal rings, weighed, measured and had blood samples collected before being released. The sex of 42 T. amaurochalinus individuals was determined using the CHD gene marker. A total of 20 males and 22 females were identified from June to August, with peak capture frequency in June. Turdus amaurochalinus females and males differed significantly in morphometrical measures. The most important traits to distinguish males from females were wing length (Student t-test=4.34, df=40, p=0.0001) and weight (Student t-test=2.08,df=40, p=0.044): females were heavier and had significantly shorter wing length than males. Females and males were correctly classified in 86% and 75% of cases, respectively, using Discriminant Analysis. The molecular analysis was the most secure method for sex determination in the studied species. Rev. Biol. Trop. 59 (2): 789- 794. Epub 2011 June 01.

La determinación del sexo es importante para la conservación y los estudios poblacionales. Turdus amaurochalinus no presenta aparente dimorfismo sexual. El objetivo de este estudio fue determinar el sexo a través de una técnica genética, mediante el uso del marcador del gen CHD y se puso a prueba la hipótesis de que el dimorfismo sexual puede ser detectado por morfometría. Este estudio se llevó a cabo en el Parque Nacional da Restinga de Jurubatiba, una zona protegida situada en la costa norte de Río de Janeiro. Las aves fueron capturadas con redes de niebla, los individuos se marcaron con anillos de metal, se pesaron, medieron y se les tomó una muestra de sangre antes de ser liberados. Un total de 20 machos y 22 hembras fueron identificados en el área de estudio desde junio hasta agosto, con la frecuencia máxima de captura en junio. La prueba de t-student fue usada para evaluar si hembras y machos se diferencian considerablemente en relación a medidas morfométricas. Los rasgos más importantes para distinguir machos de hembras fueron la longitud del ala y el peso: las hembras eran más pesadas y tenían longitud de ala considerablemente más corta que los machos. Hembras y machos fueron correctamente clasificados en un 86% y 75% de casos respectivamente, donde se usó un análisis discriminante. El análisis molecular es el método más seguro para la determinación sexual en la especie estudiada.

Isolation and cultivation of fungal strains from in vitro cell cultures of two marine sponges (Porifera: Halichondrida and Haplosclerida).

Despite the large number of reports describing sponge-microbe associations, limited knowledge is available about associated fungi and their relationships with the hosts. In this work, specific fungal strains were obtained directly from in vitro sponge cell cultures (primmorphs) and single sponge cells (cytospins) and compared with those obtained from whole tissue preparations. A total of 27 fungal strains were isolated from the marine sponges Hymeniacidon heliophila and Haliclona melana. Fifteen strains, nine from H. heliophila and six from H. melana, were obtained from whole tissue and were considered as possible mesohyl associated or transient fungi. Twelve strains were isolated from in vitro sponge cell cultures (primmorphs) and were, therefore, considered as cell associated. From these, five different strains were obtained from H. heliophila isolated cells, while five were identified from cytospins and two from primmorphs of H. melana. The fungal strains obtained from cell cultures from both sponge species were different, and none of them were detected in the whole tissue preparations of the same species. Nine H. heliophila and seven H. melana strains shows low similarity with the sequences available in public databases and belong to potentially new species. This is the first report of fungi isolated directly from sponge cells, which allowed the observation and selection of specific strains that probably would not be obtained by usual culture dependent techniques.