Molecular Evidence Reveals Taxonomic Uncertainties and Cryptic Diversity in the Neotropical Catfish of the Genus Pimelodus (Siluriformes: Pimelodidae).

Pimelodus is the most speciose genus of the family Pimelodidae, and is amply distributed in the Neotropical region. The species-level taxonomy and phylogenetic relationships within this genus are still poorly resolved, however. These taxonomic problems and the general lack of data have generated major uncertainties with regard to the identification of specimens from different localities. In the present study, we applied a single-locus species delimitation approach to identify the MOTUs found within the genus Pimelodus and provide sound evidence for the evaluation of the species richness of this genus in the different river basins of the Neotropical region. The study was based on the analysis of sequences of the mitochondrial COI gene of 13 nominal species, which resulted in the identification of 24 consensus MOTUs. Only six nominal species were recovered as well-defined molecular entities by both the traditional barcoding analysis and the molecular delimitation methods, while the other seven presented cryptic diversity or persistent taxonomic uncertainties. The lineages identified from the Parnaíba ecoregions, Amazonas Estuary and Coastal Drainages may represent a much greater diversity of Pimelodus species than that recognized currently, although a more detailed study of this diversity will be necessary to provide a more definitive classification of the genus.

Identification of fish specimens of the Tocantins River, Brazil, using DNA barcoding.

The fish fauna of the Tocantins River possesses many endemic species; however, it is little studied in molecular terms and is quite threatened by the construction of several hydroelectric dams. Therefore, the objective of this study was to identify the ichthyofauna of the Tocantins River using DNA barcoding. For this, collections were carried out in five points of this river, which resulted in the capture of 725 individuals from which partial sequences of the cytochrome oxidase subunit I (COI) gene were obtained for genetic analysis. A total of 443 haplotypes were recovered with the mean intraspecific K2P genetic distance of 1.82%. Altogether, 138 species were identified based on morphological criteria, which was a quantity that was much lower than that indicated by the four molecular methods (assemble species by automatic partitioning [ASAP], barcode index number [BIN], generalized mixed Yule coalescent (GMYC), and Bayesian Poisson tree processes [bPTP]) through which 152-157 molecular entities were identified. In all, 41 unique BINs were obtained based on the data generated in the BOLDSystems platform. According to the result indicated by ASAP (species delimitation approach considered the most appropriate in the present study), there was an increase of 17 molecular entities (12.32%), when compared to the number of species identified through their morphological criteria, as it can show cryptic diversity, candidates for new species, and misidentifications. There were 21 incongruities indicated between the different identification approaches for species. Therefore, it is suggested that these taxonomic problems be cautiously evaluated by experts to solve such taxonomic issues.

Molossusmelini Montani et al. 2021 (Chiroptera, Molossidae) in Brazil: new insights for distribuition, morphology and genetics.

Background: The species M.melini has been observed in both the Pampa and Spinal ecoregions of Argentina. Researchers have underscored that distinguishing M.melini from other species within the same genus relies primarily on craniometric and molecular analyses. Morphological measurements alone do not offer a clear differentiation between M.melini and other members of this genus.This study aims to document the presence of M.melini within the Brazilian ecoregion, focusing on its morphological, morphometric and genetic characteristics. By undertaking a comprehensive examination, we seek to contribute valuable insights into the distribution and differentiation of M.melini in this region. New information: Molossusmelini specimens exhibited a forearm length ranging from 39.9 to 40.08 mm. The average intraspecific divergence was 1.2%, with specimens from the Argentine Pampas clustering in the same clade with a 98% bootstrap support and a posterior probability of: Regarding dorsal colouration, the specimens displayed fur with two bands-a Snow White base colour and apex colours ranging from Olive Brown, Broccoli Brown, Wood Brown to Yellowish-Brown. This marks the first record of M.melini in Brazil, expanding its distribution 1,300 km northeastwards into the Curitiba, Paraná, Atlantic Forest Ecoregion. The findings contribute valuable information on the distribution, morphology, morphometrics and genetics of this species.

Sequencing and Analysis of the Mitochondrial Genome of Aedes aegypti (Diptera: Culicidae) from the Brazilian Amazon Region.

Aedes aegypti is a mosquito native to the African continent, which is now widespread in the tropical and subtropical regions of the world. In many regions, it represents a major challenge to public health, given its role in the cycle of transmission of important arboviruses, such as Dengue, Zika, and Chikungunya. Considering the epidemiological importance of Ae. aegypti, the present study sequenced the partial mitochondrial genome of a sample collected in the municipality of Balsas, in the Brazilian state of Maranhão, followed by High Throughput Sequencing and phylogenetic analyses. The mitochondrial sequence obtained here was 15,863 bp long, and contained 37 functional subunits (thirteen PCGs, twenty-two tRNAs and two rRNAs) in addition to a partial final portion rich in A+T. The data obtained here contribute to the enrichment of our knowledge of the taxonomy and evolutionary biology of this prominent disease vector. These findings represent an important advancement in the understanding of the characteristics of the populations of northeastern Brazil and provide valuable insights into the taxonomy and evolutionary biology of this prominent disease vector.

Genetic Differentiation of Aedes aegypti (Diptera: Culicidae) in Areas with High Rates of Infestation in Mid-North Region of Brazil.

Aedes aegypti is the principal vector of the arboviruses-yellow fever, dengue virus, chikungunya, and zika virus. Given the epidemiological importance of this mosquito, its capacity to adapt to different habitats, and its resistance to many types of control measures, systematic research into the genetic variability of the populations of this mosquito is one of the most important steps toward a better understanding of its population structure and vector competence. In this context, the present study verified the presence of distinct genetic lineages of Ae. aegypti in areas with high infestation rates, based on the analysis of microsatellite markers. The samples were collected in nine municipalities with high building infestation rates in the Mid-North region of Brazil. Six microsatellite loci were genotyped in the 138 samples, producing a total of 32 alleles, varying from one to nine alleles per locus in each of the different populations. The AMOVA revealed greater within-population genetic differentiation with high fixation rates. The general analysis of population structure, based on a Bayesian approach, revealed K = 2, with two Ae. aegypti lineages that were highly differentiated genetically. These data on the connectivity of the populations and the genetic isolation of the lineages provide important insights for the development of innovative strategies for the control of the populations of this important disease vector.

HPV 18 variants in women with cervical cancer in Northeast Brazil.

BACKGROUND: Human Papillomavirus (HPV) is the main etiological factor for the development of cervical cancer. HPV 18 is the second most frequent type, accounting for up to 65% of all cases. HPV intratypic variation may influence the potential for progression to invasive cancer. The aim of this study was to evaluate the prevalence of human papillomavirus 18 intratypic variants in cervical cancer samples from women in the state of Maranhão, Brazil. METHODS: The study included 118 women over 18 years of age with a diagnosis of cervical cancer. Tumor fragments were collected and subjected to DNA extraction and Polymerase Chain Reaction (PCR) for HPV detection using the PGMY09/11 and GP+5/6 primers. Positive samples were submitted to automated sequencing for viral genotyping. To determine the HPV 18 lineages, positive samples were submitted to PCR, using specific primers to amplify the LCR and E6 regions of HPV 18 virus. RESULTS: HPV was present in 88 women (73.3%). Of those, 48 (54%) were HPV 16, the most prevalent, followed by 12 (13.6%) HPV 18. Histologically, squamous cell carcinoma was predominant (79.1%). Among the HPV 18 variants identified, 10 (80%) belonged to lineage A, and sublineages A1, A2, A3, and A4. Two (29%) HPV 18 B variant was also detected, with the sublineages B1 and B2. In this study, the C variant was not found. There was no statistically significant association between the HPV 18 lineages found and sociodemographic and lifestyle variables (p > 0.05). CONCLUSIONS: A higher frequency of HPV 16 and 18 were found in women with cervical cancer in the state of Maranhão, Brazil, with a high prevalence of the lineage A among women with HPV 18.

HPV 18 variants in women with cervical cancer in Northeast Brazil

Abstract Background Human Papillomavirus (HPV) is the main etiological factor for the development of cervical cancer. HPV 18 is the second most frequent type, accounting for up to 65% of all cases. HPV intratypic variation may influence the potential for progression to invasive cancer. The aim of this study was to evaluate the prevalence of human papillomavirus 18 intratypic variants in cervical cancer samples from women in the state of Maranhão, Brazil. Methods The study included 118 women over 18 years of age with a diagnosis of cervical cancer. Tumor fragments were collected and subjected to DNA extraction and Polymerase Chain Reaction (PCR) for HPV detection using the PGMY09/11 and GP+5/6 primers. Positive samples were submitted to automated sequencing for viral genotyping. To determine the HPV 18 lineages, positive samples were submitted to PCR, using specific primers to amplify the LCR and E6 regions of HPV 18 virus. Results HPV was present in 88 women (73.3%). Of those, 48 (54%) were HPV 16, the most prevalent, followed by 12 (13.6%) HPV 18. Histologically, squamous cell carcinoma was predominant (79.1%). Among the HPV 18 variants identified, 10 (80%) belonged to lineage A, and sublineages A1, A2, A3, and A4. Two (29%) HPV 18 B variant was also detected, with the sublineages B1 and B2. In this study, the C variant was not found. There was no statistically significant association between the HPV 18 lineages found and sociodemographic and lifestyle variables (p > 0.05). Conclusions A higher frequency of HPV 16 and 18 were found in women with cervical cancer in the state of Maranhão, Brazil, with a high prevalence of the lineage A among women with HPV 18.

Litomosoides brasiliensis (Nematoda: Onchocercidae) infecting chiropterans in the Legal Amazon region, Brazil.

Chiropterans play an important role in the maintenance of the environmental balance, since they are pollinators, seed dispersers and predators. They contribute to transmission and spreading of microorganisms such as helminths, fungi, protozoa, bacteria and virus. The aim of the present study was to investigate natural filariid infection among bats in the Legal Amazon region, Brazil, by means of parasitological and molecular analyses. Blood samples were collected from 82 bats for blood smears and for DNA extraction via the polymerase chain reaction (PCR) assay. Microfilariae were observed in blood smears from Carollia perspicillata (2), Artibeus lituratus (1), Artibeus fimbriatus (2), Dermanura gnoma (2) and Glossophaga soricina (1). Five positive samples were detected through the PCR assay and four of these were also positive in blood smears. From genome sequencing and comparative analysis with sequences deposited in GenBank, one sample showed 99.31% similarity to the species Litomosoides brasiliensis. The present study expands the geographical distribution of L. brasiliensis, to include the state of Maranhão as an area of occurrence of this species and includes D. gnoma and A. fimbriatus as hosts in Brazil.

Bat fauna of the Cerrado savanna of eastern Maranhão, Brazil, with new species occurrences / Quiropterofauna do cerrado leste maranhense, Brasil, com ocorrência de novos registros

Abstract In Brazil, 179 bat species have been recorded to date, representing 68 genera and nine families. Few data are available on the bat fauna of the state of Maranhão, so the present study was based on a survey of the bat fauna of the Inhamum Municipal Environmental Protection Area (APA Inhamum) in the Cerrado savanna of the eastern extreme of the state. This inventory provides new records of bat species for the state of Maranhão and the Cerrado biome. A total of 31 species (four families) were collected, representing 39% of the bat species known to occur in Maranhão and 28.71% of the species described for the Cerrado. The family Phyllostomidae was the most diverse, with 23 species (74.20% of the total), and also the most abundant, with 86.09% of the specimens collected. The Molossidae (four species) was the second most diverse, followed by the Vespertilionidae (three species) and the Emballonuridae (one species). Carollia perspicillata was the most abundant species (28%).The records of Artibeus fimbriatus and Platyrrhinus fusciventris are the first for the Cerrado and were also recorded in Maranhão for the first time. The cumulative species curve did not reach an asymptote. The species Dermanura gnoma, Lasiurus blossevillii, Lasiurus ega, Micronycteris schmidtorum, Molossops temminckii, Platyrrhinus cf. recifinus, Phylloderma stenops and Trachops cirrhosus, were also recorded in Maranhão for the first time demonstrating the value of for mammal inventories and emphasizing the need for further surveys in this poorly-known region.

Resumo Atualmente são registrados para o Brasil, nove famílias, 68 gêneros e 179 espécies de morcegos. No Maranhão, poucas são as informações sobre a fauna de morcegos, portanto objetivou-se realizar um levantamento faunístico e reportar os novos registros de morcegos para o Cerrado leste maranhense na Área de Proteção Ambiental Municipal do Inhamum. Foram coletadas 31 espécies, distribuídas em quatro famílias representando 39% das espécies de morcegos registradas no Maranhão e 28,71% das espécies descritas para o Cerrado. A família Phyllostomidae apresentou a maior riqueza (23 espécies - 74,20%) e abundância (86,09%). A família Molossidae (quatro espécies) foi a segunda mais representativa, seguida de Vespertilionidae (três espécies) e Emballonuridae (uma espécie). Carollia perspicillata foi a espécie mais abundante (28%). As espécies Artibeus fimbriatus e Platyrrhinus fusciventris constituem novos registros para o Cerrado e o primeiro registro para o Maranhão. A curva de acumulação de espécies mostrou-se não estabilizada. As espécies Dermanura gnoma, Lasiurus blossevillii, Lasiurus ega, Micronycteris schmidtorum, Molossops temminckii, Platyrrhinus cf. recifinus, Phylloderma stenops e Trachops cirrhosus constituem novos registros para o Maranhão. As espécies coletadas na APA do Inhamum correspondem a novos registros para o Cerrado do leste maranhense evidenciando-a como uma área de extrema importância para levantamentos mastozoológicos, mostrando que a região necessita de mais estudos.

[Parasites of natural and artificial populations of Cichla spp]. / Parasitas de populações naturais e artificiais de tucunaré (Cichla spp.).

The cichlids, economically important fish species in the region, were collected along the Negro River (Cichla orinocensis, C. monoculus e C. temensis) and in the Marchantaria Island (C. monoculus) Amazon State and in three reservoirs localized in the Piauí State. In order to verify the occurrence of parasites, gills, nostrils and the intestine were removed and observed by stereomicroscope and the mouth by naked eyes. The observed parasites were isolated and conserved in 70% ethanol for posterior identification. From 96 specimens collected in Negro River, 51 specimens (52%) were parasitized. In the Marchantaria Island from 34 specimens collected, 16 (47%) showed the presence of parasites, and from 50 specimens collected in the reservoirs only 2 (4%) were parasitized. In fish collected in the Amazon State, the grill was parasitized by Ergailus coatiarus, Argulus amazonicus, Acusicola tucunarense and the mouth by Braga cichlae, whereas the species collected in the Piauí State did not show the presence of parasites in the grill, but the occurrence of Procamallanus (Spirocamallanus) inopinatus an intestinal nematode parasite.

Parasitas de populações naturais e artificiais de tucunaré (Cichla spp.) / Parasites of natural and artificial populations of Cichla spp.

Os ciclídeos, peixes economicamente importantes na região, foram coletados no médio rio Negro (Cichla orinocensis, C. monoculus e C. temensis) e na iIlha da Marchantaria (C. monoculus), no Amazonas e em três açudes do Piauí (C. kelberi). Para verificar a ocorrência de parasitas as brânquias, narinas e intestino foram removidos e observados em microscópio esteroscópio, a boca a olho nu. Os parasitas encontrados foram retirados e armazenados em álcool 70 por cento para posterior identificação. Dos 96 espécimes analisados provenientes do rio Negro 51 (52 por cento) estavam parasitadas; dos 34 da ilha da Marchantaria 16 (47 por cento) e dos 50 dos açudes do Piauí, apenas dois (4 por cento). Nos peixes coletados no Amazonas, as brânquias estavam parasitadas por Ergailus coatiarus, Argulus amazonicus, Acusicola tucunarense e a boca por Braga cichlae, enquanto que os peixes do Piauí não apresentaram parasitas branquiais, mas registrou-se a ocorrência do nematóide Procamallanus (Spirocamallanus) inopinatus, parasita intestinal.

The cichlids, economically important fish specie in the region, were collected along the Negro River (Cichla orinocensis, C. monoculus e C. temensis) and in the Marchantaria Island (C. monoculus) Amazon State and in three reservoirs localized in the Piauí State. In order to verify the occurrence of parasites, gills, nostrils and the intestine were removed and observed by stereomicroscope and the mouth by naked eyes. The observed parasites were isolated and conserved in 70 percent ethanol for posterior identification. From 96 specimens collected in Negro River, 51 specimens (52 percent) were parasitized. In the Marchantaria Island from 34 specimens collected, 16 (47 percent) showed the presence of parasites, and from 50 specimens collected in the reservoirs only 2 (4 percent) were parasitized. In fishes collected in the Amazon State, the grill was parasitized by Ergailus coatiarus, Argulus amazonicus, Acusicola tucunarense and the mouth by Braga cichlae, whereas the species collected in the Piauí State did not show the presence of parasites in the grill, but the occurrence of Procamallanus (Spirocamallanus) inopinatus an intestinal nematode parasite.

Enzymatic variability in Aedes aegypti (Diptera: Culicidae) populations from Manaus-AM, Brazil

Eighteen enzymatic loci were analysed in Aedes aegypti populations from four neighbourhoods in the city of Manaus. The analyses showed that the Downtown population was the most polymorphic (p = 55.6 percent) with higher observed and expected mean heterozygosities (Ho = 0.152 ± 0.052; He = 0.174 ± 0.052, respectively). The least variability was detected in the Coroado and Cidade Nova populations, both with polymorphism of 44.4 percent. The latter population presented the least observed heterozygosity (Ho = 0.109 ± 0.037). Wright's F statistics showed that the mean value of Fis was higher than that of Fst (Fis = 0.164 > Fst = 0.048), and from analysis of molecular variance (AMOVA) it was found that 95.12 percent of the variability is found within populations indicating a certain intra-population differentiation possibly of the microgeographic structure resulting from some barrier in the random coupling. Although the four populations were similar genetically (D = 0.003 to 0.016), the 4.88 percent differentiation was significant