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Abstract Bats are efficient reservoirs of a number of viruses with zoonotic potential, and are involved directly in the transmission cycle of many zoonoses. In the present study, which is part of a larger project that is documenting the viromes of the bat species found in the Mid-North states of Maranhão and Piauí, we analyzed 16 pooled samples obtained from four species of bat of the genus Artibeus—Artibeus obscurus, Artibeus cinereus, Artibeus lituratus and Artibeus planirostris. We describe and identify a Hepatovirus, denominated Hepatovirus H isolate sotense, which was found in a pool of internal organs (liver and lungs) extracted from a specimen of A. planirostris, a frugivorous bat, collected in the Cerrado biome of Maranhão state. This material was analyzed using new generation sequencing, which produced a contig of 7390 nucleotides and presented a degree of identity with a number of existing Hepatovirus sequences available for bats (amino acid identity of 61.5% with Bat hepatovirus C of Miniopterus cf. manavi, 66.6% with Bat hepatovirus G of Coleura afra, 67.4% with Hepatovirus G2 of Rhinolophus landeri, and 75.3% with Hepatovirus H2 of Rhinolophus landeri). The analysis of the functional domains of this contig confirmed a pattern consistent with the characteristics of the genus Hepatovirus (Picornaviridae). In the phylogenetic tree with several other Hepatovirus species, this genome also grouped in a monophyletic clade with Hepatovirus H (HepV-H1; HepV-H2, and HepV-H3) albeit on an external branch, which suggests that it may be a distinct genotype within this species. This is the first isolate of Hepatovirus H identified in bats from South America, and represents an important discovery, given that most studies of viruses associated with bats in the state of Maranhão have focused on the family Rhabdoviridae.

Abstract Bats play an essential role in maintaining ecosystems. Their unique characteristics increase the likelihood of interactions with various species, making them a potential source for the emergence and spread of infectious diseases. Hantaviruses are continuously expanding their range of hosts. This study presents the identification of a partial genome associated with Hantavirus in samples collected from neotropical bats. We conducted a metagenomic study using samples from Carollia perspicillata in Maranhão, Brazil. Tissue fragments were used for RNA extraction and subsequent sequencing. The resulting data was subjected to bioinformatic analysis. A sequence showing an identity of 72.86% with the L gene in the reference genome was obtained. The phylogenetic analysis revealed the study sequence, denoted as Buritiense, clustering within the Mobatvirus clade. The intragroup analysis showed a broader dispersion and were markedly asymmetric. This observation suggests the possibility that Buritiense could potentially represent a new species within the bat-borne hantaviruses, but further analyses are needed to provide additional insights if bats plays a role as reservoirs and the potential for transmission to human populations.

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.Molossus melini 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.

Brazil stands out for presenting the highest amphibian anuran diversity in the world. However, taxonomic studies that address characteristic of larval stage of anurans are incipient, representing only 62% of known species. We assess the species diversity of tadpoles from eastern Maranhão state, mid-northern region of Brazil based on morphological and molecular identification (i.e., 16S rRNA gene fragment), and we also provide characteristics of the habitats occupied by each species. We carried out 30 field samplings during 13 months in 16 environments along an ecotonal area, over five cities inside the limits of state of Maranhão, between the Maranhão Babaçu Forest and Cerrado ecoregions. We searched for tadpoles in a variety of water bodies, and the tadpoles that reached the developmental stage between 34 to 40 Gosner were morphologically identified. The tadpoles collected herein represent 26 species belonging to five families. The external morphology enabled the identification of 24 species, while the molecular data recognized 22 unique evolutionary units. The most represented family was Hylidae (Hylinae 11 spp., Phyllomedusinae one spp.) followed by Leptodactylidae (Leptodactylinae seven spp., Leiuperinae three spp.), Microhylidae (Gastrophryninae two spp.), and Bufonidae (two spp.). Our results show that oral morphology was the most important character for identifying tadpoles based on morphology, and the specific 16S rRNA primer was suitable for molecular identification. This study pioneers the use of both morphological and molecular data to identify tadpoles in the state of Maranhão. It also provides, for the first-time, habitat characteristic for the species. Our study reveals a high number of anuran species sampled at the larval stage in the region, identifies species that require further taxonomic and systematic attention, and extends the geographic distribution of six species, three of which represent new occurrences for the state. Our results strengthen the hypothesis that the diversity of amphibians from Maranhão is underestimated and highlight the importance of herpetological inventories in poorly sampled areas, decentralizing the knowledge of biodiversity.

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.

Leporinus is one of the most speciose genera of the order Characiformes, with 81 valid species distributed throughout much of Central and South America. The considerable diversity of this genus has generated extensive debate on its classification and internal arrangement. In the present study, we investigated the species diversity of the genus Leporinus in central northern Brazil, and conclude that six valid species—Leporinus maculatus, Leporinus unitaeniatus, Leporinus affinis, Leporinus venerei, Leporinus cf. friderici, and Leporinus piau—are found in the hydrographic basins of the Brazilian states of Maranhão, Piauí, and Tocantins. We analyzed 182 sequences of the Cytochrome Oxidase subunit I gene, of which, 157 were obtained from Leporinus specimens collected from the basins of the Itapecuru, Mearim, Turiaçu, Pericumã, Periá, Preguiças, Parnaíba, and Tocantins rivers. The species delimitation analyses, based on the ABGD, ASAP, mPTP, bPTP, and GMYC methods, revealed the presence of four distinct molecular operational taxonomic units (MOTUs), identified as L. maculatus, L. unitaeniatus, L. affinis, and L. piau (from the Parnaíba River). The bPTP method restricted L. venerei to a single MOTU, and confirmed the occurrence of this species in the rivers of Maranhão for the first time. The separation of L. cf. friderici into two clades and the subsequent formation of different operational taxonomic units was consistent with polyphyly in this species, which indicates the existence of cryptic diversity. The arrangement of L. cf. friderici and L. piau in two different clades supports the conclusion that the L. piau specimens from Maranhão were misidentified, based on their morphological traits, reflecting the taxonomic inconsistencies that exist among morphologically similar species. Overall, then, the species delimitation methods employed in the present study indicated the presence of six MOTUs—L. maculatus, L. unitaenitus, L. affinis, L. cf. friderici, L. venerei, and L. piau. In the case of two other MOTUs identified in the present study, one (L. venerei) is a new record for the state of Maranhão, and we believe that the other represents a population of L. piau from the basin of the Parnaíba River.

A specimen of Eptesicus furinaliswas collected in the municipality of Cândido Mendes, in the state of Maranhão, Brazil. It was a non-lactating adult female, with dark chestnut dorsal coloration, yellow venter, hairless membranes, short and rounded ears, and a pointed tragus. The analysis of the DNA barcode of the COI mitochondrial gene revealed a 99.80 % similarity with the sequence of E. furinalisdeposited in the BOLDSystems platform. The combined analysis of the morphological and molecular data confirmed the occurrence of E. furinalis in the state of Maranhão. This extends the known distri-bution of the species 676.1 km from the nearest recorded locality in the Ceará State.

Cryptonanus agricolai (Moojen, 1943) is a small didelphid widely distributed across of central and northeastern Brazil and in the Brazilian Atlantic Forest. A recent phylogeographic study of this species identified three major mtDNA clades with strong geographic structure. Here, we present new records of C. agricolai in northeastern Brazil, including the first report for Maranhão state. We also identify a novel mtDNA lineage, sister to one of the three major clades, in some specimens from Maranhão, indicating that C. agricolai is morphologically and genetically more variable than previously recognized.

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.

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.

Abstract The study provides the first record of Eumops glaucinus in the Maranhão state, located in the northern region of Brazil. The collected specimen was a non-lactating adult female, with grayish pelage, broad ears, smooth face, a well-developed and squarish tragus, and elongated snout. The combined analysis of the morphological and molecular data (COI, Cyt b, and rRNA 16S genes) confirmed the occurrence of E. glaucinus in the state of Maranhão. This record extends the known species range area by 660 km easternward from the closest locality, Belém, Pará.

Abstract 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.

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.

Abstract Background Cervical cancer is the fourth most common type of cancer affecting women globally. In Brazil, it is the third most frequent type of cancer in women and HPV is present in approximately 90% of cases. Evidence suggests that variants of HPV 16 can interfere biologically and etiologically during the development of cervical cancer. Methods Cervix tumor fragments were collected, their DNA was extracted, and nested PCR was used to detect HPV. Positive samples were sequenced to determine the viral genotype. To characterize the HPV 16 strains, positive samples PCR was used to amplify the LCR and E6 regions of the HPV 16 virus. Results Data from 120 patients with cervical cancer were analyzed. Most women were between 41 and 54 years of age, had schooling until primary school, a family income between 1 and 2 times the minimum wage and were married/in a consensual union. There was no statistically significant association between HPV or socio-demographic variables and risk factors for cervical cancer (P

Abstract The bat genus Sturnira is widely distributed in the Neotropical region, from northwestern Mexico to northern Argentina, and four species occur in Brazil: Sturnira lilium, Sturnira giannae, Sturnira magna, and Sturnira tildae. The present study is the first to record Sturnira tildae in the state of Maranhão, Brazil, based on morphological and molecular diagnoses. The specimen was identified based on its cranial and morphometric traits. The diagnostic traits include discreetly bilobed inner upper incisors with a broad base, lower first and second molars with lingual cusps separated by shallow grooves, and forearm longer than 45 mm. The molecular sequences of Cytochrome C Oxidase Subunit 1 (COI) and 16S rRNA genes confirmed the morphological identification and thus the occurrence of Sturnira tildae in the Amazon biome of Maranhão. This record represents an eastward extension of the known distribution of the species in the Amazonia, to Cândido Mendes, Maranhão, within an area dominated by dense rainforest and influenced by tides.

Niceforo’s big-eared bat, Trinycteris nicefori (Sanborn, 1949), is a monotypic species which has been recorded in a number of Brazilian states, but has a disjunct distribution in this country. This study presents the first record of T. nicefori in the Brazilian state of Maranhão. The specimens were collected in the municipalities of Godofredo Viana and Cândido Mendes, in fragments of the Amazon forest. One male (forearm: 38.00 mm, weight: 6 g) and one female (39.68 mm, 8 g) specimens were collected. The specimens presented chestnut-colored fur, and a chin with a pair of dermal pads arranged in a V-shape, without a central papilla. The COI gene sequences were plotted in the BOLD Systems platform, which confirmed the morphological identification of the species, with a 99.1% similarity in the male, and 99.4% in the female to existing sequences. This record extends the known distribution of T. nicefori in Brazil by approximately 310 km to the most eastern part of the Amazon Biome.

The geographic distribution of Megalechis thoracata is extended to the basin of the Itapecuru River, based on a specimen collected in the Soledade stream, a tributary of the Itapecuru, located in the Inhamum Municipal Environmental Protection Area in Maranhão, Brazil.

Aedes (Stegomyia) aegypti is the vector responsible for the transmission of the viruses that cause zika, yellow and chikungunya fevers, the four dengue fever serotypes (DENV – 1, 2, 3, 4), and hemorrhagic dengue fever in tropical and subtropical regions around the world. The present study investigated the genetic differentiation of the 15 populations of this vector in the Brazilian state of Maranhão, based on the mitochondrial ND4 marker. A total of 177 sequences were obtained for Aedes aegypti, with a fragment of 337 bps, 15 haplotypes, 15 polymorphics sites, haplotype diversity of h = 0.6938, and nucleotide diversity of π = 0.01486. The neutrality tests (D and Fs) were not significant. The AMOVA revealed that most of the variation (58.47%) was found within populations, with FST = 0.41533 (p

In this study, we aimed to evaluate the genetic diversity within and among chicken breeds from the northeast region of Brazil (states of Bahia and Piauí) using microsatellite markers. In addition, we assessed the identity and genetic relationships of chickens from Europe, Africa, and South America, as well as their influence on the formation of the Brazilian breeds. A total of 25 microsatellite markers and a panel containing 886 samples from 20 breeds (including the Brazilian chickens) were used in this study. Different statistical parameters were used to estimate the genetic diversity and relationship among the genetic groups studied. Our study indicates that the Brazilian Creole chickens have high genetic variability. The results show that chickens reared in the states of Bahia and Piauí could have originated from different ancestors. The Brazilian breeds studied have an evolutionary relationship with chickens from Portugal, Nigeria, Chile, and Spain. Our results will contribute directly to the conservation and recognition of Brazilian Creole chicken breeds and provide a solid basis for the demonstration of their genetic identity and genetic conservation of American Creole chicken populations.

A new molecular phylogeny for akodontine rodents from Brazil was proposed. The phylogenetic tree was enriched with the area of occurrence and with information on the karyotype of the samples. Based on this enriched tree, and with a described methodology, hypotheses were proposed on the karyotype and area of occurrence of the ancestors of each Clade. Thus it was possible to discuss hypotheses on chromosome evolution of the group, and on dispersion events from the "area of original differentiation" of akodontines in the Andes. Chromosome evolution started with high diploid numbers (2n=52) and showed a tendency to reduction (until 2n=14 in more recent clades). Independent side-branches of the tree showed 2n reduction and in one case the 2n increased. At least four dispersion events from the Andes down to South-eastern Brazil were proposed. The results should suggest the direction of new studies on comparative karyology.Uma nova filogenia molecular para roedores akodontinos do Brasil é proposta. A árvore filogenética foi enriquecida com a área de ocorrência e com informações sobre o cariótipo das amostras. Baseado nisto, e com a metodologia descrita, foram propostas hipóteses sobre as características do cariótipo e sobre a área de ocorrência dos ancestrais de cada clado. Assim, foi possível discutir hipóteses sobre evolução cromossômica do grupo, e sobre eventos de dispersão a partir da área de diferenciação original dos akodontinos nos Andes. A evolução cromossômica começou com números diplóides altos (2n=52) e mostrou uma tendência a redução (até 2n=14 em clados mais recentes). Ramos independentes da árvore mostraram redução do 2n e num caso aumentou o numero diplóide. Foram propostos pelo menos quatro eventos de dispersão dos Andes até o Brasil Sul-Oriental. Os resultados indicam a direção de novos estudos em cariologia comparada.

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% para posterior identificação. Dos 96 espécimes analisados provenientes do rio Negro 51 (52%) estavam parasitadas; dos 34 da ilha da Marchantaria 16 (47%) e dos 50 dos açudes do Piauí, apenas dois (4%). 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% 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 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.

We sequenced 12S RNA mtDNA for the majority of the extant species of sloths and anteaters and compared our results with previous data obtained by our group using 16S RNA mtDNA in the same specimens and to GenBank sequences of the extinct giant sloth Mylodon. Our results suggest that pigmy-anteaters may be a case of the long-branch attraction phenomenon and also show the large genetic difference between the Amazonian and Atlantic forest three-toed sloths, contrasting with the small differences observed between the two non-Atlantic forest forms of sloths. These results have important implications for the taxonomy of sloths and anteaters and strongly suggest the placement of pigmy anteaters in their own family (Cyclopidae) and raising the taxonomic status of Bradypus torquatus to a genus.

We sequenced part of the 16S rRNA mitochondrial gene in 17 extant taxa of Pilosa (sloths and anteaters) and used these sequences along with GenBank sequences of both extant and extinct sloths to perform phylogenetic analysis based on parsimony, maximum-likelihood and Bayesian methods. By increasing the taxa density for anteaters and sloths we were able to clarify some points of the Pilosa phylogenetic tree. Our mitochondrial 16S results show Bradypodidae as a monophyletic and robustly supported clade in all the analysis. However, the Pleistocene fossil Mylodon darwinii does not group significantly to either Bradypodidae or Megalonychidae which indicates that trichotomy best represents the relationship between the families Mylodontidae, Bradypodidae and Megalonychidae. Divergence times also allowed us to discuss the taxonomic status of Cyclopes and the three species of three-toed sloths, Bradypus tridactylus, Bradypus variegatus and Bradypus torquatus. In the Bradypodidae the split between Bradypus torquatus and the proto-Bradypus tridactylus / B. variegatus was estimated as about 7.7 million years ago (MYA), while in the Myrmecophagidae the first offshoot was Cyclopes at about 31.8 MYA followed by the split between Myrmecophaga and Tamandua at 12.9 MYA. We estimate the split between sloths and anteaters to have occurred at about 37 MYA.

ABSTRACT Aedes (Stegomyia) aegypti is the vector responsible for the transmission of the viruses that cause zika, yellow and chikungunya fevers, the four dengue fever serotypes (DENV - 1, 2, 3, 4), and hemorrhagic dengue fever in tropical and subtropical regions around the world. The present study investigated the genetic differentiation of the 15 populations of this vector in the Brazilian state of Maranhão, based on the mitochondrial ND4 marker. A total of 177 sequences were obtained for Aedes aegypti, with a fragment of 337 bps, 15 haplotypes, 15 polymorphics sites, haplotype diversity of h = 0.6938, and nucleotide diversity of π = 0.01486. The neutrality tests (D and Fs) were not significant. The AMOVA revealed that most of the variation (58.47%) was found within populations, with FST = 0.41533 (p < 0.05). Possible isolation by distance was tested and a significant correlation coefficient (r = 0.3486; p = 0.0040) was found using the Mantel test. The phylogenetic relationships among the 15 haplotypes indicated the existence of two distinct clades. This finding, together with the population parameters, was consistent with a pattern of genetic structuring that underpinned the genetic differentiation of the study populations in Maranhão, and was characterized by the presence of distinct lineages of Aedes aegypti.