Macroevolutionary consequences of karyotypic changes in the neotropical Serrasalmidae fishes (Ostariophysi, Characiformes) diversification.

In the Neotropical region, one of the most diverse families of freshwater fishes is the monophyletic Serrasalmidae. Karyotypically, the family shows high diversity in chromosome numbers (2n = 54 to 64). However, little is discussed about whether the chromosomal changes are associated with cladogenetic events within this family. In the present study, we evaluated the role of chromosomal changes in the evolutionary diversification of Serrasalmidae. Our phylogenetic sampling included 36 species and revealed three main clades. The ancestral chromosome number reconstruction revealed the basic number 2n = 54 and a high frequency of ascending dysploid events in the most derived lineages. Our biogeographic reconstruction suggests an Amazonian origin of the family at 48-38 Mya, with independent colonization of other basins between 15 and 8 Mya. We did not find specific chromosomal changes or increased diversification rates correlated with the colonization of a new environment. On the other hand, an increase in the diversification rate was detected involving the genus Serrasalmus and Pygocentrus in the Miocene, correlated with the stasis of 2n = 60. Our data demonstrate that chromosomal rearrangements might have played an important evolutionary role in major cladogenetic events in Serrasalmidae, revealing them as a possible evolutionary driver in their diversification.

The Impact of Life Philosophy and Major Field of Study on Brazilian Students' Knowledge of Biological Evolution.

Evolutionary theory (ET) is the unifying theory of Life Sciences, but it is largely misunderstood and the target of dispute in many countries, mainly because of conflicts with religious beliefs. Brazil is a country with a culture that is deeply rooted belief in God. In this paper, we report a study in Brazil where we applied a 12-question questionnaire to understand the relationship among the major field of study and the self-declared religious and philosophical context on the knowledge of evolutionary theory by Brazilian freshmen. We answer the following questions: (1) Is there a difference in ET comprehension according to the major field of study? (2) Are religious and philosophical contexts related to the comprehension of ET? (3) Are major field of study and religious/philosophical contexts together related to the comprehension of ET by Brazilian freshmen? A total of 153 freshmen students answered our questionnaire. Students from the Biological Sciences fared better than students from most other major field of study but were equal to students of Humanities. The philosophical perception of life had a major correlation with their knowledge about evolution, with self-declared atheists and agnostics showing better performance than religious students. There were no clear trends in the interaction of major field of study and philosophy of life. We discuss these results in the light of the rising ideological activism in Brazilian society since philosophy of life was the main driver of evolutionary theory perception. We highlight the importance of scientists and teachers in reassuring the role of science and scientific knowledge in modern societies. Supplementary Information: The online version contains supplementary material available at 10.1007/s11191-021-00286-z.

Integrative systematics unveils the controversial identity of Engraulidae fishing stocks in a Neotropical estuary, northeast Brazil

In Brazil, the use and diversity of the common names of fish species, coupled with taxonomic uncertainties, hinder the reliability of fishing statistical data. In this scenario, there are the so-called pilombetas of the São Francisco River, an important fishing resource in region. Despite its importance, the real diversity of species identified in the area remains obscure. In order to properly identify and delimit the species popularly known as pilombetas, an integrative approach involving traditional taxonomy, geometric morphometrics and molecular systematics was applied. Results from geometric morphometrics and molecular analyses were consistent with the results of the traditional morphological analysis, also indicating the delimitation of six taxa belonging to Engraulidae in the lower São Francisco River. In addition, species delimitation methods revealed an intrapopulation genetic divergence of 1.7% for Lycengraulis grossidens. The results revealed that the currently known richness species of Engraulidae in the studied area has been underestimated. Thus, an updated taxonomic key is herein proposed for the Engraulidae species from the lower São Francisco River and estuary. The integrative analysis approach revealed to be effective to address taxonomic questions and help the management of stocks, ensuring the maintenance of local diversity of fishes in the Neotropical region.(AU)

No Brasil, o uso e a diversidade dos nomes comuns para espécies de peixes, associado a incertezas taxonômicas, dificultam a confiabilidade dos dados estatísticos da pesca. Nesse cenário, existem as chamadas pilombetas do rio São Francisco, um importante recurso pesqueiro da região. Apesar de sua importância, a real diversidade de espécies identificadas na área permanece obscura. Para identificar adequadamente as espécies conhecidas como pilombetas, uma abordagem integrativa envolvendo taxonomia tradicional, morfometria geométrica e sistemática molecular foi aplicada. Os resultados das análises moleculares e de morfometria geométrica foram consistentes com os resultados da análise morfológica tradicional, indicando também a delimitação de seis táxons pertencentes a Engraulidae no baixo São Francisco. Além disso, os métodos de delimitação de espécies revelaram divergência genética intrapopulacional de 1,7% em Lycengraulis grossidens. Nossos dados revelaram que a riqueza de espécies atualmente conhecida de Engraulidae na área estudada é subestimada. Assim, uma chave taxonômica atualizada é aqui proposta para as espécies de Engraulidae do baixo rio São Francisco e seu estuário. A abordagem de análise integrativa revelou ser efetiva para tratar de questões taxonômicas e ajudar no gerenciamento de estoques, garantindo a manutenção da diversidade local de peixes na região Neotropical.(AU)

Trends on the Karyotype Acrocentrization Within Carangidae (Perciformes): A New Phylogenetic Evidence About a Traditional Marine Paradigm.

Carangidae is a morphologically diverse family of marine fish, characterized by stable karyotypes, predominantly with 2n = 48, composed of acrocentric chromosomes (A). This stability is shared with other families of the order Perciformes, which resulted in the hypothesis that 48A is a plesiomorphic karyotype of the group. We tested this hypothesis in the Carangidae family using comparative phylogenetic methods, investigating the evolution of karyotype characters (including chromosome number, morphology, and number of chromosome arms per karyotype [fundamental number, FN]). Our analyses revealed that 2n = 48 is most likely the ancestral chromosome number for the family. However, an extremely variable number of FNs, always above 48, was observed in basal clades within the family and sister groups. On the other hand, the reduced FN = 48 was consistently observed only in the most derived clades, indicating a tendency for acrocentrization. The number of acrocentric chromosomes apparently was accompanied by a trend of reduction in the genome size (1C-value), suggesting that these changes might be correlated. Our data contradict the marine fish hypothesis that the 2n = 48 acrocentric karyotype is plesiomorphic, at least for Carangidae, and reveal the importance for the correct interpretation of karyotype in a temporal and phylogenetic context.

Inferring Diversity and Evolution in Fish by Means of Integrative Molecular Cytogenetics.

Fish constitute a paraphyletic and profusely diversified group that has historically puzzled ichthyologists. Hard efforts are necessary to better understand this group, due to its extensive diversity. New species are often identified and it leads to questions about their phylogenetic aspects. Cytogenetics is becoming an important biodiversity-detection tool also used to measure biodiversity evolutionary aspects. Molecular cytogenetics by fluorescence in situ hybridization (FISH) allowed integrating quantitative and qualitative data from DNA sequences and their physical location in chromosomes and genomes. Although there is no intention on presenting a broader review, the current study presents some evidences on the need of integrating molecular cytogenetic data to other evolutionary biology tools to more precisely infer cryptic species detection, population structuring in marine environments, intra- and interspecific karyoevolutionary aspects of freshwater groups, evolutionary dynamics of marine fish chromosomes, and the origin and differentiation of sexual and B chromosomes. The new cytogenetic field, called cytogenomics, is spreading due to its capacity to give resolute answers to countless questions that cannot be answered by traditional methodologies. Indeed, the association between chromosomal markers and DNA sequencing as well as between biological diversity analysis methodologies and phylogenetics triggers the will to search for answers about fish evolutionary, taxonomic, and structural features.

Discriminatory profile of rDNA sites and trend for acrocentric chromosome formation in the genus Trachinotus Lacépède, 1801 (Perciformes, Carangidae).

Chromosomal traits have provided valuable information for phylogeny and taxonomy of several fish groups. Three Atlantic Carangidae species of the genus Trachinotus Lacépède, 1801 (Trachinotus goodei Jordan et Evermann, 1896, Trachinotus carolinus (Linnaeus, 1766)and Trachinotus falcatus (Linnaeus, 1758)) were investigated, having 2n=48 chromosomes but different chromosomal arms (FN number), i.e., 52, 56 and 58, respectively, in view of the different number of two-armed chromosomes found in their karyotypes. Thus, Trachinotus goodei, Trachinotus carolinus and Trachinotus falcatus present a progressive distancefrom the probable basal karyotype proposed for Perciformes (2n=48 acrocentrics, FN=48). At first sight, these findings do not agree with the phylogenetic hypothesis based on mitochondrial sequences, where Trachinotus goodei appear as the most derived species, followed by Trachinotus falcatus and Trachinotus carolinus, respectively. However, the chromosomal mapping of ribosomal DNAs was informative for clarifying this apparent conflict. Indeed, the multiple 5S and 18S rDNA sites found in Trachinotus goodei corroborate the most derived condition for this species. In this sense, the occurrence of the unexpected number of two-armed chromosomes and FN value for this species, as well as for Trachinotus carolinus, must be due to additional rounds of acrocentric formation in these species, modifying the macrostructure of their karyotypes.

Comparative cytogenetics and heterochromatic patterns in two species of the genus Acanthostracion (Ostraciidae: Tetraodontiformes).

Some groups of fish, such as those belonging to the Order Tetraodontiformes, may differ significantly in the amount and location of heterochromatin in the chromosomes. There is a marked variation in DNA content of more than seven-fold among the families of this Order. However, the karyoevolutionary mechanisms responsible for this variation are essentially unknown. The largest genomic contents are present in species of the family Ostraciidae (2.20-2.60pg). The present study cytogenetically characterized two species of the family Ostraciidae, Acanthostracion polygonius and A. quadricornis, using conventional staining, C-bandings, Ag-NOR, CMA(3)/DAPI, AluI, PstI, EcoRI, TaqI and HinfI restriction enzymes (REs) and double FISH with 18S and 5S rDNA probes. The karyotypes of both species showed 2n=52 acrocentric chromosomes (FN=52; chromosome arms) and pronounced conserved structural characteristics. A significant heterochromatic content was observed equilocally distributed in pericentromeric position in all the chromosome pairs. This condition is unusual in relation to the karyotypes of other families of Tetraodontiformes and probability is the cause of the higher DNA content in Ostraciidae. Given the role played by repetitive sequences in the genomic reorganization of this Order, it is suggested that the conspicuous heterochromatic blocks, present in the same chromosomal position and with apparently similar composition, may have arisen or undergo evolutionary changes in concert providing clues about the chromosomal mechanisms which led to extensive variation in genomic content of different Tetraodontiformes families.

Pleistocene karyotypic divergence in Hoplias malabaricus (Bloch, 1794) (Teleostei: Erythrinidae) populations in southeastern Brazil

The lacustrine system of the middle rio Doce basin is considered a paradigm of Pleistocene geomorphology. In these lakes, two Hoplias malabaricus karyomorphs (2n = 42A and 2n = 42B) live in sintopy in Carioca Lake. Cytogenetic analyses were performed on 65 specimens from 8 lakes (including Carioca Lake) to determine the distribution and relative frequency of these karyomorphs and the degree of cytogenetic divergence caused putatively by recent geographic isolation. All fish were 2n = 42B karyomorphs, except for 1 specimen from the Marola Lake, which was 2n = 42A. Among-population variation was especially high for C-banding patterns. Other characters such as X chromosome size and CMA3/DAPI also varied among populations. Our results suggested that the karyotype of H. malabaricus is able to respond rapidly to geographic isolation, and revealed that heterochromatic variation may represent the lowest hierarchical level of chromosomal evolution.

O sistema lacustre da bacia do médio rio Doce é considerado um paradigma da geomorfologia do Pleistoceno. Nestes lagos, dois cariomorfos de Hoplias malabaricus (2n = 42A e 2n = 42B) vivem em sintopia na lagoa Carioca. Análises citogenéticas foram realizadas em 65 amostras de 8 lagos (incluindo lagoa Carioca) para determinar a distribuição e frequência relativa destes cariomorfos e o grau de divergência citogenética aparentemente causada pelo isolamento geográfico recente. Todos os peixes apresentaram o cariomorfo 2n = 42B, com exceção de 1 espécime da lagoa Marola, que foi 2n = 42A. Entre as populações, a variação foi especialmente elevada nos padrões de bandamento C. Outros caracteres como o tamanho do cromossomo X e os padrões de CMA3/DAPI também variaram entre as populações. Nossos resultados sugerem que o genoma de H. malabaricus é capaz de responder rapidamente ao isolamento geográfico, revelando que a variação de heterocromatina pode representar o nível hierárquico mais baixo de evolução cromossômica.

Chromosome mapping of repetitive sequences in Rachycentron canadum (Perciformes: Rachycentridae): implications for karyotypic evolution and perspectives for biotechnological uses.

The cobia, Rachycentron canadum, a species of marine fish, has been increasingly used in aquaculture worldwide. It is the only member of the family Rachycentridae (Perciformes) showing wide geographic distribution and phylogenetic patterns still not fully understood. In this study, the species was cytogenetically analyzed by different methodologies, including Ag-NOR and chromomycin A(3) (CMA(3))/DAPI staining, C-banding, early replication banding (RGB), and in situ fluorescent hybridization with probes for 18S and 5S ribosomal genes and for telomeric sequences (TTAGGG)(n). The results obtained allow a detailed chromosomal characterization of the Atlantic population. The chromosome diversification found in the karyotype of the cobia is apparently related to pericentric inversions, the main mechanism associated to the karyotypic evolution of Perciformes. The differential heterochromatin replication patterns found were in part associated to functional genes. Despite maintaining conservative chromosomal characteristics in relation to the basal pattern established for Perciformes, some chromosome pairs in the analyzed population exhibit markers that may be important for cytotaxonomic, population, and biodiversity studies as well as for monitoring the species in question.

Biogeography and comparative cytogenetics between two populations of Hoplias malabaricus (Bloch, 1794) (Ostariophysi: Erythrinidae) from coastal basins in the State of Bahia, Brazil

The species Hoplias malabaricus is a predator fish found in nearly all cis-Andean basins. From a cytogenetic point of view, this species comprises, at least, seven differentiated karyomorphs. Several localities have been formerly analyzed in Brazil, however, some regions, such as Bahia State, remain underrepresented. Recently, the Brazilian Environment Ministry classified both Itapicuru and Contas river basins (entirely located within Bahia territory) as priority conservation areas, whose biodiversity status lacks enough information. Therefore, the goal of the present work was to characterize, cytogenetically, populations of H. malabaricus from both basins, by using conventional staining, Ag-NOR and C-banding techniques. All specimens presented a diploid number of 2n = 40 with metacentric/submetacentric chromosomes, without differences between sexes, thereby representing the so-called "karyomorph F". The first metacentric pair presented a remarkably larger size in relation to the other pairs. The NORs were multiple, comprising the terminal region on long arms of two chromosomal pairs in both populations. However, the C-banding pattern was somewhat distinguishable between samples. Although sharing heterochromatic blocks at centromeric region of all chromosomes, the population from Itapicuru River basin appeared to have some more conspicuous blocks than those observed in the population from Contas River basin. The similar karyotype observed in both populations suggests a common geological history between them. The present results represent an advance in the knowledge about the cytogenetic pattern of H. malabaricus populations from poorly studied basins.(AU)

A espécie Hoplias malabaricus é um predador que ocorre em praticamente todas as bacias cis-andinas. Sob o ponto de vista citogenético, ela compreende, pelo menos, sete cariomorfos diferenciáveis. Várias localidades já foram previamente analisadas no Brasil, porém, algumas regiões, como o Estado da Bahia, permanecem pouco amostradas. Recentemente, o Ministério de Meio Ambiente classificou as bacias do rio Itapicuru e Contas (inteiramente localizadas na Bahia), como áreas prioritárias de conservação, cuja biodiversidade carece de informações suficientes. Neste sentido, o objetivo do presente trabalho foi caracterizar citogeneticamente populações de H. malabaricus nessas bacias, por meio de técnicas de coloração convencional, Ag-RON e bandamento C. Todos os espécimes e populações analisadas apresentaram número diploide 2n = 40 com cromossomos metacêntricos/submetacêntricos, sem diferenças entre os sexos, representando assim o denominado "cariomorfo F". O primeiro par metacêntrico apresentou tamanho notavelmente maior que os demais pares. As RONs foram múltiplas, ocupando a região terminal do braço longo de dois pares cromossômicos em ambas populações. Entretanto, os padrões de heterocromatina foram relativamente diferenciáveis entre as bacias hidrográficas estudadas. Apesar de compartilharem blocos heterocromáticos na região centromérica de todos os cromossomos, a população da bacia do Itapicuru apresentou alguns blocos mais conspícuos em relação aos da bacia do rio de Contas. O cariótipo similar encontrado em ambas as populações parece indicar uma história geológica em comum. Os dados obtidos representam um avanço no conhecimento dos padrões citogenéticos de populações de H. malabaricus provenientes de bacias pouco estudadas(AU)