Carmen Rosa García-Dávila

Phylogeography of the piranha genera Serrasalmus and Pygocentrus: implications for the diversification of the Neotropical ichthyofauna.

The phylogenetic relationships within the piranhas were assessed using mitochondrial sequences with the aim of testing several hypotheses proposed to explain the origin of Neotropical diversity (palaeogeography, hydrogeology and museum hypotheses). Sequences of the ribosomal 16S gene (510 bp) and control region (980 bp) were obtained from 15 localities throughout the main South American rivers for 21 of the 28 extant piranha species. The results indicate that the genus Serrasalmus is monophyletic and comprises three major clades. The phylogeographical analyses of these clades allowed the identification of five vicariant events, extensive dispersal and four lineage duplications suggesting the occurrence of sympatric speciation. Biogeographical patterns are consistent with the prediction made by the museum hypothesis that lineages from the Precambrian shields are older than those from the lowlands of the Amazon. The vicariant events inferred here match the distribution of the palaeoarches and several postdispersal speciation events are identified, thereby matching the predictions of the palaeogeography and hydrogeology hypotheses, respectively. Molecular clock calibration of the control region sequences indicates that the main lineages differentiated from their most recent common ancestor at 9 million years ago in the proto Amazon‐Orinoco and the present rate of diversification is the highest reported to date for large carnivorous Characiformes. The present results emphasize that an interaction among geology, sea‐level changes, and hydrography created opportunities for cladogenesis in the piranhas at different temporal and geographical scales.

Are rapids a barrier for floodplain fishes of the Amazon basin? A demographic study of the keystone floodplain species Colossoma macropomum (Teleostei: Characiformes)

We investigated demographic history and population structuring of Colossoma macropomum sampled from 14 localities in the Amazon basin and the Bolivian sub-basin; the two basins are separated by a series of 16 rapids. Although genetically differentiated, IMa analyses suggest non-zero bi-directional migration rates, and inter-basin divergence of approximately 17 thousand years ago. Analyses in BEAST indicated that Bolivian C. macropomum has been demographically stable except for a moderate population increase in the last 12 thousand years, while Amazonian C. macropomum has been experiencing demographic growth over the last 350 thousand years, resulting in approximately one order of magnitude increase in coalescent N(e).

Revision of Hemoproteid Genera and Description and Redescription of Two Species of Chelonian Hemoproteid Parasites

Abstract:  Pigmented hemosporidian parasites that do not exhibit erthyrocytic schizogony, and infect birds, chelonians, and squamates, have been classified in various genera over time. These classifications have reflected vertebrate hosts, insect vectors, and variations in morphology and life history observed in representative species. Side-necked turtles (Podocnemis spp.) from the Peruvian Amazon were screened for hemoparasites and 2 species of hemosporid parasites infecting these hosts were observed. Molecular phylogenetic analysis of these new isolates, along with parasites from lizards, a snake, and a variety of Haemoproteus species from birds from both the Haemoproteus and Parahaemoproteus subgenera, strongly support the separation of the non-avian parasites into a separate genus. The name with precedent for this group is Haemocystidium Castellani and Willey 1909, and we propose that subgeneric classification of Haemocystidium and Simondia be applied to parasites of squamates and chelonians, respectively. We offer a description of Haemocystidium (Simondia) pacayae n. sp. and a redescription of Haemocystidium (Simondia) peltocephali (Lainson and Naiff 1998, n. comb.) Morphologically, the parasites are quite similar, with H. pacayae slightly more elongated than H. peltocephali. The discovery and identification of parasite species is urgent, especially in endangered species and wildlife inhabiting rapidly declining ecosystems such as the Amazon.

Revisão taxonômica dos camarões de água doce (Crustacea: Decapoda: Palaemonidae, Sergestidae) da Amazônia Peruana

Foi realizada uma revisao taxonomica das especies de camaroes de agua doce da Amazonia peruana que abrangeu oito especies da familia Palaemonidae e uma especie da familia Sergestidae. Sao descritas duas novas especies do genero Pseudopalaemon e feitos os primeiros registros de Euryrhynchus amazoniensis Tiefenbacher, 1978, Macrobrachium jelskii Miers, 1877 e Palaemonetes ivonicus Holthuis, 1950 para o Peru. Sao fornecidos chave de identificacao, dados distribucionais e ilustracoes para as especies estudadas.

Morphometric variability in populations of Palaemonetes spp. (Crustacea, Decapoda, Palaemonidae) from the Peruvian and Brazilian Amazon Basin

Morphometric variability among shrimp populations of the genus Palaemonetes Heller, 1869 from seven lakes (Huanayo and Urcococha, in Peru; Amana, Mamiraua, Camaleao, Cristalino e Irucanga, in Brasil) in the Amazon Basin, presumably belonging to Palaemonetes carteri Gordon, 1935 and Palaemonetes ivonicus Holthuis, 1950, were studied. The morphometric studies were carried out from the ratios obtained from the morphometric characters. Multivariated analysis (Principal Components Analysis-PCA, Discriminant Function Analysis and Cluster Analysis) were applied over the ratios. Intra- and interpopulation variations of the rostrum teeth, and the number of spines in the male appendix, were analyzed through descriptive statistics and bivariate analysis (Spearman Rank Correlation test). Results indicated a wide plasticity and overlapping in the studied ratios between populations. The Principal Components Analysis was not able to separate different populations, revealing a large intrapopulation plasticity and strong interpopulation similarity in the studied ratios. Although the Discriminant Functions Analysis was not able to fully discriminate populations, they could be allocated in three subgroups: 1) Cristalino and Irucanga; 2) Huanayo, Urcococha and Camaleao and 3) Mamiraua and Amana. The first two groups were morphometrically separated from each other, whereas the third one presented a strong overlap with the former two. The Cluster Analysis confirmed the first two subgroups separation, and indicated that the first and third groups were closely related. Rostrum teeth and number of spines in the appendix masculina showed a large intrapopulation variation and a strong overlapping among the studied populations, regardless of the species.

Molecular identification of a cryptic species in the Amazonian predatory catfish genus Pseudoplatystoma (Bleeker, 1962) from Peru

AbstractPseudoplatystoma species are highly prized South American Pimelodid migratory catfishes. Until recently, their taxonomy was not clearly established, with discrepancies between morphological and molecular analyses. Here, Pseudoplatystoma species from the Peruvian Amazon were characterized at the molecular level from a sample representing the observed range of their color pattern variations in the study area. Analyses were performed using seven microsatellite loci for 103 specimens and, for part of them (52), using sequences of two regions of their mitochondrial genome [Cytochrome Oxidase subunit I (COI) and Control Region (CR)]. Factorial correspondence analysis and assignment tests based on microsatellite polymorphism showed that the specimens originally identified as P. punctifer belonged to two different gene pools highly differentiated from P. tigrinum. Morphological examination identified two different morphotypes (with and without black stripes), suggesting the existence of two distinct taxa within P. punctifer. This result was corroborated by the ML tree based on CR sequences, where all individuals but four clustered in a similar way as in the FCA and Bayesian assignment tests. For these four individuals, mitochondrial introgression or retention of ancestral polymorphism was likely. In contrast, the ML tree based on COI sequences showed that reciprocal monophyly was not yet achieved for this marker for the two P. punctifer taxa. The existence of three sympatric species of Pseudoplatystoma in the Peruvian Amazon is discussed in relation to their molecular characteristics, color patterns and ecology. Evolutionary scenarios regarding their divergence are hypothesized.

Histological development of the digestive system of the Amazonian pimelodid catfish Pseudoplatystoma punctifer.

The organogenesis of the digestive system was described in the Amazonian pimelodid catfish species Pseudoplatystoma punctifer from hatching (3.5 mm total length, TL) to 41 days post-fertilization (dpf) (58.1 mm TL) reared at 28°C. Newly hatched larvae showed a simple digestive tract, which appeared as a straight undifferentiated and unfolded tube lined by a single layer of columnar epithelial cells (future enterocytes). During the endogenous feeding period, comprised between 20 and 96 h post-fertilization (3.5 to 6.1 mm TL), the larval digestive system experienced a fast transformation with the almost complete development and differentiation of most of digestive organs (buccopahrynx, oesophagus, intestine, liver and exocrine pancreas). Yolk reserves were not completely depleted at the onset of exogenous feeding (4 dpf, 6.1 mm TL), and a period of mixed nutrition was observed up to 6 to 7 dpf (6.8 to 7.3 mm TL) when yolk was definitively exhausted. The stomach was the organ that latest achieved its complete differentiation, characterized by the development of abundant gastric glands in the fundic stomach between 10 and 15 dpf (10.9 to 15.8 mm TL) and the formation of the pyloric sphincter at the junction of the pyloric stomach and the anterior intestine at 15 dpf (15.8 mm TL). The above-mentioned morphological and histological features observed suggested the achievement of a digestive system characteristic of P. punctifer juveniles and adults. The ontogeny of the digestive system in P. punctifer followed the same general pattern as in most Siluriform species so far, although some species-specific differences in the timing of differentiation of several digestive structures were noted, which might be related to different reproductive guilds, egg and larval size or even different larval rearing practices. According to present findings on the histological development of the digestive system in P. punctifer, some recommendations regarding the rearing practices of this species are also provided in order to improve the actual larval rearing techniques of this fast-growing Neotropical catfish species.

Phylogeography and Sex-Biased Dispersal across Riverine Manatee Populations (Trichechus inunguis and Trichechus manatus) in South America

Phylogeographic patterns and sex-biased dispersal were studied in riverine populations of West Indian (Trichechus manatus) and Amazonian manatees (T. inunguis) in South America, using 410bp D-loop (Control Region, Mitochondrial DNA) sequences and 15 nuclear microsatellite loci. This multi-locus approach was key to disentangle complex patterns of gene flow among populations. D-loop analyses revealed population structuring among all Colombian rivers for T. manatus, while microsatellite data suggested no structure. Two main populations of T. inunguis separating the Colombian and Peruvian Amazon were supported by analysis of the D-loop and microsatellite data. Overall, we provide molecular evidence for differences in dispersal patterns between sexes, demonstrating male-biased gene flow dispersal in riverine manatees. These results are in contrast with previously reported levels of population structure shown by microsatellite data in marine manatee populations, revealing low habitat restrictions to gene flow in riverine habitats, and more significant dispersal limitations for males in marine environments.

DNA Metabarcoding of Amazonian Ichthyoplankton Swarms

Tropical rainforests harbor extraordinary biodiversity. The Amazon basin is thought to hold 30% of all river fish species in the world. Information about the ecology, reproduction, and recruitment of most species is still lacking, thus hampering fisheries management and successful conservation strategies. One of the key understudied issues in the study of population dynamics is recruitment. Fish larval ecology in tropical biomes is still in its infancy owing to identification difficulties. Molecular techniques are very promising tools for the identification of larvae at the species level. However, one of their limits is obtaining individual sequences with large samples of larvae. To facilitate this task, we developed a new method based on the massive parallel sequencing capability of next generation sequencing (NGS) coupled with hybridization capture. We focused on the mitochondrial marker cytochrome oxidase I (COI). The results obtained using the new method were compared with individual larval sequencing. We validated the ability of the method to identify Amazonian catfish larvae at the species level and to estimate the relative abundance of species in batches of larvae. Finally, we applied the method and provided evidence for strong temporal variation in reproductive activity of catfish species in the Ucayalí River in the Peruvian Amazon. This new time and cost effective method enables the acquisition of large datasets, paving the way for a finer understanding of reproductive dynamics and recruitment patterns of tropical fish species, with major implications for fisheries management and conservation.

Metabarcoding by capture using a single COI probe (MCSP) to identify and quantify fish species in ichthyoplankton swarms

The ability to determine the composition and relative frequencies of fish species in large ichthyoplankton swarms could have extremely important ecological applications However, this task is currently hampered by methodological limitations. We proposed a new method for Amazonian species based on hybridization capture of the COI gene DNA from a distant species (Danio rerio), absent from our study area (the Amazon basin). The COI sequence of this species is approximately equidistant from all COI of Amazonian species available. By using this sequence as probe we successfully facilitated the simultaneous identification of fish larvae belonging to the order Siluriformes and to the Characiformes represented in our ichthyoplankton samples. Species relative frequencies, estimated by the number of reads, showed almost perfect correlations with true frequencies estimated by a Sanger approach, allowing the development of a quantitative approach. We also proposed a further improvement to a previous protocol, which enables lowering the sequencing effort by 40 times. This new Metabarcoding by Capture using a Single Probe (MCSP) methodology could have important implications for ecology, fisheries management and conservation in fish biodiversity hotspots worldwide. Our approach could easily be extended to other plant and animal taxa.