Genomic screening and molecular dynamics simulations of cyanovirin-N homologs from cyanobacteria phylum.

The phylum cyanobacteria are one of the most ancient groups of organisms on the planet and are well recognized due to its wide distribution, ecological role, and biotechnological potential. Cyanobacterial lectins are being extensively explored due to their antiviral activity, mainly because of their capacity of inhibiting HIV strains from infecting human cells by gp120 and gp41 binding. Cianovirin-N from Nostoc ellipsosporum was the first lectin isolated with this property. Since then, various homologs have been discovered and characterized. In this article, we present results of a genomic screening to find cyanovirin-N homologs (CVNH) in all cyanobacteria genomes available in the GenBank, resulting in 155 CVNH proteins with 63 presenting significant identity differences of cyanovirin-N. Homology modeling and molecular dynamics were employed to characterize 18 unexplored models and their functional capacity of binding to Manα(1-2)Man. Results presented here support the hypothesis of multiple ligand recognition for the CVNH family and may help to understand the function of these lectins for the producer cyanobacteria. Additionally, the theoretical results observed here justify carrying out experimental investigations that can expand the therapeutic potential of cyanobacterial lectins.

Current Knowledge on Microviridin from Cyanobacteria.

Cyanobacteria are a rich source of secondary metabolites with a vast biotechnological potential. These compounds have intrigued the scientific community due their uniqueness and diversity, which is guaranteed by a rich enzymatic apparatus. The ribosomally synthesized and post-translationally modified peptides (RiPPs) are among the most promising metabolite groups derived from cyanobacteria. They are interested in numerous biological and ecological processes, many of which are entirely unknown. Microviridins are among the most recognized class of ribosomal peptides formed by cyanobacteria. These oligopeptides are potent inhibitors of protease; thus, they can be used for drug development and the control of mosquitoes. They also play a key ecological role in the defense of cyanobacteria against microcrustaceans. The purpose of this review is to systematically identify the key characteristics of microviridins, including its chemical structure and biosynthesis, as well as its biotechnological and ecological significance.

A new Piper nigrum cysteine proteinase inhibitor, PnCPI, with antifungal activity: molecular cloning, recombinant expression, functional analyses and molecular modeling.

MAIN CONCLUSION: A new Piper nigrum cysteine proteinase inhibitor, PnCPI, belonging to group I of phytocystatins, with inhibitory activity against papain and growth of Fusarium solani f. sp. piperis, was isolated and characterized. Previous studies (de Souza et al. 2011) have identified a partial cDNA sequence of putative cysteine proteinase inhibitor differentially expressed in roots of black pepper (P. nigrum L.) infected by F. solani f. sp. piperis. Here, we aimed to isolate the full-length cDNA and genomic sequences of the P. nigrum cysteine proteinase inhibitor gene, named PnCPI. Sequence analyses showed that the PnCPI gene encodes a deduced protein of 108 amino acid residues with a predicted molecular mass of 12.3 kDa and isoelectric point of 6.51. Besides the LARFAV-like sequence, common to all phytocystatins, PnCPI contains three conserved motifs of the superfamily cystatin: a glycine residue at the N-terminal region, the QxVxG reactive site more centrally positioned, and one tryptophan in the C-terminal region. PnCPI, belonging to group I of phytocystatins, showed high identity with cystatins isolated from several plant species. Sequence analyses also revealed no putative signal peptide at the N-terminal of PnCPI, as well as no introns within the genomic sequence corresponding to the PnCPI coding region. Molecular modeling showed the ability of PnCPI to interact with papain, while its inhibitory activity against this protease was confirmed after heterologous expression in Escherichia coli. The effects of heat treatments on the inhibitory activity of recombinant PnCPI, rPnCPI, were evaluated. In addition, rPnCPI exhibited in vitro activity against F. solani f. sp. piperis, revealing a new cystatin with the potential antifungal application. The identification of PnCPI as a functional cystatin able to inhibit the in vitro growth of F. solani f. sp. piperis indicates other factors contributing to in vivo susceptibility of black pepper to root rot disease.

Kudoa amazonica n. sp. (Myxozoa; Multivalvulida), a parasite of the esophageal musculature of the freshwater catfish, Hypophthalmus marginatus (Siluriformes: Pimelodidae), from a river of the Amazon region.

The class Myxozoa is a group of spore-producing eukaryote organisms that parasitize both freshwater and marine fish. The multivalvulide myxosporidian parasites of the genus Kudoa infect primarily the musculature of the fish host, some species are producing enzymes (proteases) capable of digesting muscle fibers. In the present study, 50 specimens of the freshwater catfish Hypophthalmus marginatus were collected from the Tocantins River in Pará, Brazil, and were analyzed. Overall, 68% of these specimens presented infections by Kudoa parasite in the esophageal musculature. The morphology of these parasite was examined under light microscopy and nucleotide sequences of the SSU rDNA gene were obtained for phylogenetic analyses. The species formed numerous whitish pseudocysts containing square spores with rounded extremities in the apical view, and four polar capsules of equal size. In the phylogenetic analyses, Kudoa amazonica n. sp. was characterized as a sister taxon of another freshwater species, Kudoa orbicularis. The combination of morphological, morphometric, and molecular data obtained in the present study provided a conclusive diagnosis of Kudoa amazonica n. sp., which is clearly distinct from all other Kudoa taxa described to date.

Effect of gluten diet on blood innate immune gene expressions and stool consistency in Spix's Saddleback Tamarin (Leontocebus fuscicollis) raised in captivity.

The callitrichids are non-human primates that feed on insects and plant matter in nature, but in captivity, they are fed mostly an artificial diet containing amounts of gluten, in their toxic forms in items such as wheat, barley and rye. The aim of this research was to estimate the blood ß-defensin and Toll like receptor 5 (TLR5) gene expressions and to analyze the stool consistency (firm, soft, diarrheic) in Leontocebus fuscicollis raised in captivity. Blood samples of animals under gluten-free and gluten diets were collected and their fecal output quality was periodically monitored and classified during the course of the study. Gene expression was evaluated using real-time PCR. The stool consistencies of individuals fed a gluten diet were most frequently soft or diarrheic, while it was mostly normal in individuals fed a gluten-free diet. ß-Defensin expression increased in individuals fed a gluten diet, but decreased after 15 days. Expression normalized between 30 and 45 days on a gluten-free diet. However, expression of the TLR5 gene did not change under a gluten diet. A gluten diet affects stool quality, and brings about an immediate increase in blood ß-defensin expression in the beginning but decreases after 15 days.

Optimization of Light Intensity and NaNO3 Concentration in Amazon Cyanobacteria Cultivation to Produce Biodiesel.

The objective of this study, for the first time, was to optimize Amazonian cyanobacterial culture conditions for improving cell productivity and lipid content, by analyzing the effect of light intensity and nitrogen concentration, for empirically evaluating biodiesel quality parameters. The strains Synechocystis sp. CACIAM05, Microcystis aeruginosa CACIAM08, Pantanalinema rosaneae CACIAM18, and Limnothrix sp. CACIAM25, were previously identified by morphological and molecular analysis (16S rRNA) and were selected based on their production of chlorophyll a and dry cell weight. Then, factorial planning (22) with central points was applied, with light intensity and NaNO3 concentration as independent variables. As response variables, cell productivity and lipid content were determined. Statistical analysis indicated that for all strains, the independent variables were statistically significant for cell productivity. Analysis of the fatty acid composition demonstrated diversity in the composition of the fatty acid profile from the experimental planning assays of each strain. The Biodiesel Analyzer software predicted the biodiesel quality parameters. CACIAM05 and CACIAM25 obtained better parameters with low levels of light intensity and NaNO3 concentration, whereas CACIAM08 and CACIAM18 obtained better parameters with low NaNO3 concentrations and high luminous intensity.

Genomic screening of new putative antiviral lectins from Amazonian cyanobacteria based on a bioinformatics approach.

Lectins are proteins of nonimmune origin, which are capable of recognizing and binding to glycoconjugate moieties. Some of them can block the interaction of viral glycoproteins to the host cell receptors acting as antiviral agents. Although cyanobacterial lectins have presented broad biotechnological potential, little research has been directed to Amazonian Cyanobacterial diversity. In order to identify new antiviral lectins, we performed genomic analysis in seven cyanobacterial strains from Coleção Amazônica de Cianobactérias e Microalgas (CACIAM). We found 75 unique CDS presenting one or more lectin domains. Since almost all were annotated as hypothetical proteins, we used homology modeling and molecular dynamics simulations to evaluate the structural and functional properties of three CDS that were more similar to known antiviral lectins. Nostoc sp. CACIAM 19 as well as Tolypothrix sp. CACIAM 22 strains presented cyanovirin-N homologues whose function was confirmed by binding free energy calculations. Asn, Glu, Thr, Lys, Leu, and Gly, which were described as binding residues for cyanovirin, were also observed on those structures. As for other known cyanovirins, those residues in both our models also made favorable interactions with dimannose. Finally, Alkalinema sp. CACIAM 70d presented one CDS, which was identified as a seven-bladed beta-propeller structure with binding sites predicted for sialic acid and N-acetylglucosamine. Despite its singular structure, our analysis suggested this molecule as a new putative antiviral lectin. Overall, the identification and the characterization of new lectins and their homologues are a promising area in antiviral research, and Amazonian cyanobacteria present biotechnological potential to be explored in this regard.

Anti-dengue virus activity of scytovirin and evaluation of point mutation effects by molecular dynamics and binding free energy calculations.

The absence of a specific treatment against DENV has led to intensive research into developing strategies for curing the infection. One lectin with high antiviral activity is scytovirin, which was isolated from the cyanobacterium Scytonema varium and has proven activity against HIV and Zaire Ebola Virus. To achieve the results presented here, we tested the affinity of full-length scytovirin, SD1 and SD2 separately, and six SD1 mutants for DENV glycoprotein E carbohydrate by Molecular Dynamics (MD) simulations and binding free energy calculations. It was possible to identify the key residues for protein-ligand interaction such as Glu10, Ala11, Pro17, Ans18, Arg30, Thr41, Ser42 and Arg43, which also has importance action against HIV. All binding free energy calculations showed negative values to ΔGbind of protein-DENV carbohydrate complexation. Additionally, these results are similar to the values of scytovirin and HIV gp120 carbohydrate complexation (-32.20 kcal/mol). Furthermore, we found that SD1 individually has more affinity to the carbohydrate and the Asn9, Glu10, Asn18, Arg30 and Arg43 demonstrated an important role in this matter. We also found that mutant G48R has better affinity (-34.10 kcal/mol) for the DENV carbohydrate than the wild type protein (-27.15 kcal/mol).

In silico analysis of the cyanobacterial lectin scytovirin: new insights into binding properties.

Scytovirin is a lectin isolated from the cyanobacterium Scytonema varium that has shown activity against HIV, SARS coronavirus and Zaire Ebola virus. Its 95 amino acids are divided into two structural domains (SD), the first spanning amino acids 1-48 (SD1) and the second 49-95 (SD2). Interestingly, the domains are nearly identical but differ in their affinities for carbohydrates. With the aim of enhancing understanding of the binding properties of scytovirin, we performed molecular dynamics (MD) simulations of scytovirin complexed with Man4. We set up three systems: (i) Man4 bound to both domains (SD1 + SD2) using the full-length protein; (ii) Man4 bound to an incomplete protein, containing only SD1 and (iii) Man4 bound to an incomplete protein containing only SD2. Contrary to other reports, binding free energy results suggest that Man4 can bind simultaneously to SD1 and SD2 binding regions, but SD1 individually has the best values of energy and the best affinity for Man4. Decomposition of the binding free energy showed that the residues that interact with Man4 were different in the three systems, suggesting that the binding mechanism of Man4 varies between full-length protein, SD1 and SD2. The results presented here may help to formulate strategies to use scytovirin and promote mutagenesis studies to improve the antiviral activity of scytovirin.

Henneguya paraensis n. sp. (Myxozoa; Myxosporea), a new gill parasite of the Amazonian fish Cichla temensis (Teleostei: Cichlidae): morphological and molecular aspects.

The present study describes light microscopy, transmission electron microscopy, and molecular analyses of a myxosporid found parasitizing the gill region of the teleost fish Cichla temensis, collected from the Tocantins River, near Cametá, Pará State, Brazil. The prevalence of infection was 60 %. The spore-containing cysts that were located in the gill lamellae were oval and whitish. The spores had a mean length of 42.3 ± 0.65 µm; fusiform body, 12.8 ± 0.42-µm long and 8.6 ± 0.32-µm wide; each of the two valves exhibited a tapering tail of 29.5 ± 0.73 µm length. The spores had two polar capsules, 7.4 ± 0.16-µm long by 2.6 ± 0.08-µm wide, containing a polar filament with 5-7 twists. The spores differ from the species previously described, and phylogenetic analysis based on spore morphology and molecular aspects indicated that the fish parasite Henneguya sp. has a strong trend to form clades mainly based on the environment and host order/family. Thus, we conclude that the species belongs to the family Myxobolidae, genus Henneguya, which comprises a new species: Henneguya paraensis n. sp.

Complete nucleotide sequences of two blaKPC-2-bearing IncN Plasmids isolated from sequence type 442 Klebsiella pneumoniae clinical strains four years apart.

We sequenced the oldest blaKPC-2-bearing plasmid isolated in Brazil and another plasmid also carried by a Klebsiella pneumoniae strain of sequence type 442 (ST442), isolated 52 months later. Both plasmids present an IncN backbone and few acquired regions. Because the 2005 plasmid presented deletions and a truncated gene within Tn4401b compared to the 2009 plasmid, we can thus infer that IncN blaKPC-2-bearing plasmids pFCF1305 and pFCF3SP had a common ancestor circulating in Brazil prior to May 2005.

New species of Myxobolus in potamodromous catfish from the eastern Amazon, Brazil.

The mapará (Hypophthalmus marginatus) is a commercially important fish in the Brazilian Amazon and has been described as a host for numerous myxosporid species. The integrated taxonomy of a new species, Myxobolus mickeyii n. sp., discovered in the urinary bladder of H. marginatus, is undertaken in this study. In 105 specimens of H. marginatus, plasmodia and myxospores were observed in the urinary bladder fluid, the myxospores measuring 20.5 (19.6-21.3) µm in length and 14.0 (13.2-14.9) µm in width. The posterior valves of the spore body were thick, with valvulogenic nuclei, endoplasmic reticulum, and the presence of secretory vesicles. Two elliptical, rounded appendages attached to the valve, containing tubular filaments. The two polar capsules, symmetry, measuring 6.1 (5.9-6.3) µm in length and 4.4 (3.6-6.2) µm in width, with polar tubules of 3 to 5 turns. Phylogenetic analyses of the small subunit ribosomal RNA gene (SSU rDNA) sequencing revealed that M. mickeyii n. sp. is part of a Myxobolidae family clade with freshwater fish of the Siluriformes order, with a genetic distance of 19% to the nearest species. This work contributes to the wide diversity of myxozoans in this host, as other taxa have previously been reported infecting different tissues.

The potential of third-generation biodiesel from Tolypothrix sp. CACIAM22 as a feedstock.

Renewable energy has been recognized as an alternative to fossil fuels as a step to transform the energy produced and consumed worldwide. Cyanobacteria and microalgae are currently being considered as substitutes to the traditional feedstock used to produce biofuels due to their ability to achieve high amounts of lipids under cellular stress conditions. The aim of this study was to investigate the utilization of Tolypothrix sp. CACIAM 22 cyanobacterial biomass as a feedstock for biodiesel production, specifically by examining the effects of supplementing with hydrolysate of Brazil nutshell (HBNS) on biomass generation, lipid production, fatty acid composition, and quality of synthesized biodiesel. The supplementation of HBNS led to a significant increase of 12g.L-1 in wet biomass production. The lipid content reached 41 % of the biomass produced in HBNS supplemented cultures when nitrate source was deprived. The quality evaluation of cyanobacteria-derived biodiesel was performed using Biodiesel Analyzer ver 2.2 software, revealing superior quality compared to biodiesel produced from plant sources. The biodiesel exhibited values of 23 h for oxidative stability, 65 for cetane number, and an iodine index of 31 (g I2. 100 g-1 fat), indicating promising potential as a renewable source. This study is the first to utilize HBNS as an organic supplement for cyanobacteria culture medium and assess its impact on biomass and lipid production in Tolypothrix sp., supporting the hypothesis of utilizing this biomass as a renewable feedstock for biodiesel production as a viable alternative to plant sources based on biomass production, lipid productivity, and biodiesel quality.

Description of Rhabdias breviensis n. sp. (Rhabditoidea: Rhabdiasidae) in two Neotropical frog species.

Nematodes of the genus Rhabdias Stiles & Hassall, 1905 (Rhabditoidea: Rhabdiasidae) have a dioecious free-living stage and a hermaphroditic stage that parasitises the lungs of amphibians and reptiles. Approximately 94 species of Rhabdias have been described. Because the similar morphological characteristics such as the labial structures, the location of the vulva and the shape of the tail of Rhabdias spp. hinder their identification, molecular biology techniques and scanning electron microscopy have been employed to diagnose species of this genus. This study describes Rhabdias breviensis n. sp., parasitic in the lungs of two Neotropical frog species Leptodactylus petersii Steindachner and Leptodactylus macrosternum Miranda-Ribeiro. The description of this species integrates classical taxonomy, scanning electron microscopy and a molecular analysis of the mitochondrial COI gene. The new species differs from all other Rhabdias species parasitic in Neotropical hosts in certain morphometric parameters, the position of the vulva, the host group and the cephalic characters.

Performance of intron 7 of the ß-fibrinogen gene for phylogenetic analysis: An example using gladiator frogs, Boana Gray, 1825 (Anura, Hylidae, Cophomantinae).

Boana, the third largest genus of Hylinae, has cryptic morphological species. The potential applicability of b-fibrinogen intron 7 - FGBI7 is explored to propose a robust phylogeny of Boana. The phylogenetic potential of FGBI7 was evaluated using maximum parsimony, MrBayes, and maximum likelihood analysis. Comparison of polymorphic sites and topologies obtained with concatenated analysis of FGBI7 and other nuclear genes (CXCR4, CXCR4, RHO, SIAH1, TYR, and 28S) allowed evaluation of the phylogenetic signal of FGBI7. Mean evolutionary rates were calculated using the sequences of the mitochondrial genes ND1 and CYTB available for Boana in GenBank. Dating of Boana and some of its groups was performed using the RelTime method with secondary calibration. FGBI7 analysis revealed high values at informative sites for parsimony. The absolute values of the mean evolutionary rate were higher for mitochondrial genes than for FGBI7. Dating of congruent Boana groups for ND1, CYTB, and FGBI7 revealed closer values between mitochondrial genes and slightly different values from those of FGBI7. Divergence times of basal groups tended to be overestimated when mtDNA was used and were more accurate when nDNA was used. Although there is evidence of phylogenetic potential arising from concatenation of specific genes, FGBI7 provides well-resolved independent gene trees. These results lead to a paradigm for linking data in phylogenomics that focuses on the uniqueness of species histories and ignores the multiplicities of individual gene histories.

Systematic and parasite-host relationship by Baruscapillaria appendiculata in Phalacrocorax brasilianus collected from Marajó Island, State of Pará, Brazil.

The genus Baruscapillaria Moravec, 1982 has six valid species recorded in birds Phalacrocoracidae, namely Baruscapillaria appendiculata Freitas, 1933, B. spiculata Freitas, 1933, B. carbonis (Dubinin & Dubinina, 1940), B. jaenschi (Johnston & Mawson, 1945), B. phalacrocoraxi (Borgarenko, 1975) and B. rudolphii Moravec, Scholz and Nasincová, 1994. Helminthological tests carried out on cormorants of the species Phalacrocorax brasilianus (Gmelin), a migratory bird that occurs in the northeast of the State of Pará, Brazil, demonstrate B. appendiculata parasitizing the cloaca of these birds, through light microscopy, scanning electron microscopy and molecular biology. These studies allowed a redescription of males and females of this nematode in these hosts and in this geographical area through integrative taxonomy. The occurrence of lesions in the cloaca caused by this nematode parasite was registered using histological analysis. This is a new geographic report for this nematode.

Morphological and molecular characterization of Contracaecum australe (Nematoda: Anisakidae) parasitizing Phalacrocorax brasilianus (Aves: Phalacrocoracidae) on the north coast of Brazil.

For the first time in Brazil, Contracaecum australe is recorded parasitizing Phalacrocorax brasilianus (Aves, Suliformes, Phalacrocoracidae) from the Marine Extractive Reserve of Soure on Marajó Island, Brazilian Amazon. Its morphology revealed a body with a transversally striated cuticle, smooth or slightly cleft interlabia, lips with auricles, labial papillae, and conspicuous amphids. In males, the presence of the median papilla on the upper lip of the cloaca and spicules that reach almost half of the body of the parasite. These morphological characters, added to the number and distribution of the pre- and postcloacal papillae of the male specimens, and supported by the molecular phylogeny from the analysis of the ITS-1, 5.8S and ITS-2 genes, allowed the identification of these parasites.

First molecular detection of Leishmania (Leishmania) infantum chagasi in a domestic cat (Felis catus) from an urban area in eastern Amazon.

Background: Domestic cats have been implicated as accidental hosts of Leishmania sp. However, in recent years, the recurrent description of new cases in endemic and nonendemic areas draw attention to the potential epidemiological role of cats as reservoir hosts. Although dogs are considered urban reservoirs, cats could act as a secondary natural reservoirs in these areas. Thus, feline leishmaniasis has become an emerging disease in several countries worldwide. Case presentation: This study aimed to describe the first case of feline leishmaniasis in a stray animal that presented lesions compatible with the disease in Belém, Pará, Brazil, an important urban area in eastern Amazon. Serological tests for Leishmania infantum (ELISA and IFA) were nonreactive, whereas histopathological examination indicated infectious dermatitis caused by Leishmania spp. or Toxoplasma gondii. Cytopathological study of lesion aspirate confirmed the presence of Leishmania sp. amastigotes within macrophages. Finally, molecular analyses revealed that the feline infection was caused by Leishmania (Leishmania) infantum chagasi. Conclusion: To the best of the authors' knowledge, this study reports the first case of natural infection by Leishmania (Leishmania) infantum chagasi in a feline from eastern Amazon. These findings suggest domestic cats as potential secondary reservoir hosts of Leishmania spp. in Belém, which reinforces the importance of further epidemiological investigation of feline leishmaniasis, especially in urban areas with human cases.

Virtual screening and repurposing of approved drugs targeting homoserine dehydrogenase from Paracoccidioides brasiliensis.

Paracoccidioidomycosis is a systemic mycosis endemic in Latin America, and one of the etiological agents of the disease is Paracoccidioides brasiliensis. Currently, available treatments present adversities, such as duration, side effects, and drug interactions. In search of new therapy possibilities, this study evaluates drugs approved for use against the homoserine dehydrogenase enzyme, by an in silico approach, which performs an important biosynthesis phase for the fungus and is not present in the human body. The three-dimensional structure of the homoserine dehydrogenase enzyme from Paracoccidioides brasiliensis was obtained by homology modeling. The model was validated, and simulations were performed for virtual screening of molecules of drugs approved from the Drugs-libs database by the MTiOpenScreen web server. Molecular dynamics in three replicas were used for four drugs with better results, and in two more molecules as a control, the HS9 with inhibition against enzyme and HON which shows inhibition against mold structure. Based on the results of molecular dynamics and the comparison of binding free energy, the drug that obtained the best result was Bemcentinib. In comparison with the controls, it presented a highly relevant affinity with - 44.63 kcal/mol, in addition to good structural stability and occupation of the active site. Therefore, Bemcentinib is a promising molecule for the inhibition of PbHSD protein (homoserine dehydrogenase of Paracoccidioides brasiliensis) and a therapeutic option to be investigated.

Kudoa rousseauxii n. sp. (Cnidaria: Multivalvulida) Infects the Skeletal Muscles of the Freshwater Fish Brachyplatystoma rousseauxii in the Amazon River.

PURPOSE: Members of the genus Kudoa Meglitsch, 1947 are known to infect the muscles of commercially important fishes worldwide, including those in the order Siluriformes. This paper describes the occurrence of a new species of Kudoa in the catfish Brachyplatystoma rousseauxii based on morphological study and molecular analysis of the ribosomal RNA gene (SSU rDNA). METHODS: Fifteen specimens of Brachyplatystoma rousseauxii were purchased from fishing zones near Mosqueiro Island, Belém, Pará, Brazil. After necropsy, tissue samples and cysts were analyzed using a stereomicroscope, and fresh slides were viewed under a light microscope to confirm parasitic infection. The tissue fragments were removed and processed for molecular and histological analyses. RESULTS: Microscopic pseudocysts were found in the epaxial region of skeletal muscle fibers in 80% of the analyzed specimens. The myxospores were quadrangular with four shell valves (SV), pyriform polar capsules (PC), and internal symmetry. Phylogenetic analyses indicated that the new species formed a cluster with the species previously described in the Amazon, being close to two freshwater species. CONCLUSIONS: Morphological differences and molecular data of SSU rDNA support that Kudoa rousseauxii n. sp. is a new species that infects B. rousseauxii, a freshwater fish with intense migratory cycles that is widely captured and consumed in the Amazon.