ABSTRACT The family Rivulidae is the fourth most diverse clade of Neotropical fishes. Together with some genera of the related African family Nothobranchiidae, many rivulids exhibit a characteristic annual life cycle, with diapausing eggs and delayed embryonic development, which allows them to survive in the challenging seasonal ponds that they inhabit. Rivulidae also includes two species known as the only the self-fertilizing vertebrates and some species with internal fertilization. The first goal of this article is to review the systematics of the family considering phylogenetic relationships and synapomorphies of subfamilial clades, thus unifying information that is dispersed throughout the literature. From this revision, it is clear that phylogenetic relationships within Rivulidae are poorly resolved, especially in one of the large clades that compose it, the subfamily Rivulinae, where conflicting hypotheses of relationships of non-annual and annual genera are evident. The second goal of this work is to present an updated phylogenetic hypothesis (based on mitochondrial, nuclear, and morphological information) for one of the most speciose genus of Rivulidae, Austrolebias. Our results confirm the monophyly of the genus and of some subgeneric clades already diagnosed, but propose new relationships among them and their species composition, particularly in the subgenus Acrolebias.

RESUMEN La familia Rivulidae es el cuarto clado más diverso dentro de los peces Neotropicales. Junto con algunos géneros de la familia Nothobranchiidae, muchos rivulidos presentan un característico ciclo de vida anual, con huevos resistentes a la desecación y embriones con diapausas que les permiten sobrevivir en los ambientes estacionales donde habitan. Los Rivulidae presentan también dos especies consideradas como los únicos vertebrados hermafroditas suficientes y algunas especies con inseminación interna. El primer objetivo de este artículo es actualizar la sistemática de la familia considerando las relaciones filogenéticas y las sinapomorfías de los clados que la componen, reuniendo información que se encuentra dispersa en la literatura. De esta revisión surge que las relaciones filogenéticas dentro de Rivulidae están todavía sin resolver, especialmente en uno de los grandes clados que la componen, la subfamilia Rivulinae, donde relaciones conflictivas entre géneros anuales y no anuales son evidentes. El segundo objetivo de este trabajo es presentar una hipótesis filogenética, basada en datos morfológicos, mitocondriales y nucleares, de uno de los géneros más diversos de la familia, el género Austrolebias. Nuestros resultados confirman la monofilia del género y de algunos clados subgenéricos previamente definidos, y propone nuevas relaciones entre ellos, particularmente de las especies del subgénero Acrolebias.

The embryological, structural and functional unit of the dentine-pulp complex shares the odontoblast, located in the border of the dentine pulp, with basal nuclei and organelles. The odontoblast process emerges from its apical pole. It is formed by microtubules, microfilaments and vesicles covered by membranes penetrating the dentinal tubules, isolated from the inter-tubular matrix, along the extent of the dentine. The objective of this study was to evaluate the efficacy of three staining techniques: hematoxylin-eosin, periodic acid-Schiff and Schmorl, by staining the process, from beginning to end, and compare the results with the erosion technique. Thirty human teeth were employed in the trial; after their extraction the pulp was fixated, the pieces demineralized in nitric acid at 8%, the collagen filaments eliminated with Type II Collage-nase, the tissue was stained, and the measurements were made. The portions with no pulp were prepared with the erosion technique. Results: Comparing the best results obtained by staining with the values obtained with the erosion technique, the former showed lower values. Conclusion: Staining techniques show lower density of the staining processes compared with the dentinal tubules in the erosion technique.

The dentin-pulp complex shares the odontoblast, whose body is located in the external part of the dental pulp. The odontoblast process emerges from the apical pole and penetrates the dentinal canaliculus through the dentine producing multiple collaterals along the way. Descriibing the process was an arduous task for researchers, who combined histological techniques to preserve it in the interior of the canaliculi using transmission electron microscopy or scanning to visualize it. In a previous work we observed both tissues joined by the odontoblasts and the stained processes. The objective now was to micrometrically measure the length of the processes and canaliculi to verify if they are similar in the crown and in the root. Thirty healthy teeth were used, extracted for orthodontic reasons, from individuals of both sexes whose ages ranged from 6 to 18. The dental pulp was uncovered and attached, and they were divided in equal parts. The halves without pulp were prepared with the erosion technique. The halves that kept the pulp were demineralized with type II collagenase and stained with the Schmorl technique. With an optical microscope and a calibrated grid with a micrometric objective the width of the histological fields was first measured and then the length of the dentinal processes and canaliculi. The results were analyzed with chi2 test, Student t and Fischer's exact test with a 5% significance level. We observed that the width of the stained histological fields had a 92 % retraction compared to the same field in the erosion and that the length of the canaliculi was always greater than the length of the processes, although four coincidences in length were found in the crown and one in the root. These results suggest that the length of the process is still a controversial topic. Professional precaution in odontological treatments is the safest prevention against injuries.

El complejo dentinopulpar comparte el odontoblasto cuyo cuerpo está ubicado en la parte externa de la pulpa dentaria. De su polo apical se desprende el proceso odontoblástico que se introduce en el canalículo dentinal atravesando la dentina originando en su recorrido múltiples colaterales hasta su finalización. Describir el proceso fue ardua tarea de investigadores que combinaron técnicas histológicas para preservarlo en el interior de los canalículos utilizando microscopio electrónico de transmisión o scanning para visualizarlo. En un trabajo previo observamos ambos tejidos unidos por los odontoblastos y los procesos coloreados. El objetivo actual fue medir micrométricamente la longitud de procesos y canalículos para verificar si ambas son similares tanto en corona como en raíz. Se utilizaron treinta dientes sanos, extraídos por razones ortodóncicas de ambos sexos, cuyas edades oscilaron entre 6 y 18 años. Se descubrió la pulpa dental, se fijó y se dividieron en partes iguales. Las mitades sin pulpa se prepararon con técnica por desgaste. Las mitades que conservaron la pulpa se desmineralizaron aplicándoseles Colagenasa tipo II y coloreándolos con la técnica de Schmorl. Con microscopio óptico y una cuadrícula calibrada con micrómetro objetivo se midió primero el ancho de los campos histológicos y luego la longitud de procesos y canalículos dentinales. Los resultados fueron analizados con pruebas de chi2, t de Student y test exacto de Fischer con un nivel de significación del 5%. Observamos que el ancho de los campos histológicos coloreados tuvo una retracción del 92% respecto al mismo campo en el desgaste y que la longitud de canalículos siempre fue mayor que la de procesos, si bien se hallaron cuatro coincidencias de longitudes en corona y una en raíz. Estos resultados sugieren que la longitud del proceso aún sigue siendo tema controversial. La precausión profesional en los tratamientos odontológicos es el único medio para evitar dañarlo.

Pimelodus albicans Valenciennes, 1840 (popularmente conhecida como moncholo ou bagre branco) é uma espécie endêmica da família Pimelodidae na bacia do rio da Prata. Estudos filogeográficos baseados nas seqüências do citocromo b mitocondrial foram realizados para testar a existência de uma única linhagem evolutiva in P. albicans e para discriminar unidades populacionais relacionadas ao comportamento migratório desse táxon na bacia do rio da Prata. Um total de 34 amostras de P. albicans provenientes de diferentes lugares de coleta no estuário do rio da Prata e rio Arrecifes na bacia do rio Paraná foram analisados. Entre as 614 pares de bases do citocromo b no conjunto de dados, 203 deles variaram e 120 foram sítios filogeneticamente informativos para P. albicans. No presente estudo foi encontrado um total de vinte haplótipos, diversidade de nucleotídeos (p) = 0,032 e diversidade de haplótipos = 0,941. O teste de Tajima mostrou valores significativos D= -1,88 (p<0,05) rejeitando a hipótese de mutação neutra para os dados de P. albicans. Todas as análises filogenéticas mostraram que o clado P. albicans apresenta quatro grupos monofiléticos com um forte suporte estatístico e uma elevada probabilidade posterior Bayesiana. A rede de haplótipos para P. albicans mostrou uma forte estrutura desses quatro grupos. Uma grande estruturação genética foi observada em P. albicans nas análises de AMOVA mostrando que as amostras do rio Arrecifes constituem uma linhagem isolada. A alta divergência (10%) encontrada entre as populações do rio da Prata e rio Arrecifes sugere que P. albicans pode constituir um complexo de espécies crípticas. Foi verificada também a ocorrência de estrutura genética na bacia do rio da Prata. A localidade 760 apareceu parcialmente isolada sugerindo que P. albicans migra para procriar em os afluentes do rio da Prata gerando diferentes cardumes de haplótipos que poderiam se misturar no estuário do rio da Prata. A alta diversidade nucleotídica encontrada na localidade 765 e a existência de fluxo gênico entre os demais sítios de coleta são concordantes com o cenário hipotético de existência de populações intercruzantes no meio do estuário do rio da Prata.

Pimelodus albicans Valenciennes, 1840 (common name "moncholo" or "bagre blanco") is an endemic species of the family Pimelodidae in the río de la Plata basin. Phylogenetic approach based on cytochrome b sequences was performed to test the existence of a unique evolutionary lineage in P. albicans and to discriminate populations units or subpopulations related to a migration behavior of this taxon in the río de la Plata basin. This study included 34 samples of P. albicans of different collecting sites in the río de la Plata estuary and in the río Arrecifes belonging to the río Paraná basin. Among 614 base pairs in the cytochrome b sequence data set, 203 were variable and 120 were phylogenetically informative sites in P. albicans. A total of twenty haplotypes, nucleotide diversity (p) = 0.032 and haplotype diversity = 0.941 were found. Tajima's test showed significant value D= -1.88 (p<0.05) rejecting the neutral mutation hypothesis for the P. albicans data set. All phylogenetic approaches showed that P. albicans included four monophyletic assemblages that were supported by high bootstrap and Bayesian posterior probability values. Minimum spanning network corroborated these groups for P. albicans haplotypes. High genetic structure was found in P. albicans by means of AMOVA analysis showing that the río Arrecifes samples constitute an isolated lineage. Moreover, the high value of genetic divergence (10%) between the río de la Plata and the río Arrecifes populations could suggest that P. albicans may be conformed by a sibling species complex. On the other hand, a degree of genetic structuring was detected among different sites of the río de la Plata. A partial isolation of the 760 site may suggest that P. albicans could migrates to different tributaries for reproduction, generating different schools of haplotypes which could mix in the río de la Plata estuary. The high nucleotide diversity found in the 765 site and the existence of gene flow with the remaining collecting sites would be concordant with the outlined hypothetic scenarios of the mixing populations in the middle of the río de la Plata estuary.