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Insect β-1,3-glucanases belong to Glycoside Hydrolase Family 16 (GHF16) and are involved in digestion of detritus and plant hemicellulose. In this work, we investigated the role of GHF16 genes in Aedes aegypti larvae, due to their detritivore diet. Aedes aegypti genome has six genes belonging to GHF16 (Aae GH16.1 - Aae GH16.6), containing two to six exons. Sequence analysis suggests that five of these GHF16 sequences (Aae GH16.1, 2, 3, 5, and 6) contain the conserved catalytic residues of this family and correspond to glucanases. All genomes of Nematocera analyzed showed putative gene duplications corresponding to these sequences. Aae GH16.4 has no conserved catalytic residues and is probably a β-1,3-glucan binding protein involved in the activation of innate immune responses. Additionally, Ae. aegypti larvae contain significant β-1,3-glucanase activities in the head, gut and rest of body. These activities have optimum pH about 5-6 and molecular masses between 41 and 150 kDa. All GHF16 genes above showed different levels of expression in the larval head, gut or rest of the body. Knock-down of AeGH16.5 resulted in survival and pupation rates lower than controls (dsGFP and water treated). However, under stress conditions, severe mortalities were observed in AeGH16.1 and AeGH16.6 knocked-down larvae. Enzymatic assays of β-1,3-glucanase in AeGH16.5 silenced larvae exhibited lower activity in the gut and no change in the rest of the body. Chromatographic activity profiles from gut samples after GH16.5 silencing showed suppression of enzymatic activity, suggesting that this gene codes for the digestive larval β-1,3-glucanase of Ae. aegypti. This gene and enzyme are attractive targets for new control strategies, based on the impairment of normal gut physiology.

Insect β-1,3-glucanases belong to Glycoside Hydrolase Family 16 (GHF16) and are involved in digestion of detritus and plant hemicellulose. In this work, we investigated the role of GHF16 genes in Aedes aegypti larvae, due to their detritivore diet. Aedes aegypti genome has six genes belonging to GHF16 (Aae GH16.1 – Aae GH16.6), containing two to six exons. Sequence analysis suggests that five of these GHF16 sequences (Aae GH16.1, 2, 3, 5, and 6) contain the conserved catalytic residues of this family and correspond to glucanases. All genomes of Nematocera analyzed showed putative gene duplications corresponding to these sequences. Aae GH16.4 has no conserved catalytic residues and is probably a β-1,3-glucan binding protein involved in the activation of innate immune responses. Additionally, Ae. aegypti larvae contain significant β-1,3-glucanase activities in the head, gut and rest of body. These activities have optimum pH about 5–6 and molecular masses between 41 and 150 kDa. All GHF16 genes above showed different levels of expression in the larval head, gut or rest of the body. Knock-down of AeGH16.5 resulted in survival and pupation rates lower than controls (dsGFP and water treated). However, under stress conditions, severe mortalities were observed in AeGH16.1 and AeGH16.6 knocked-down larvae. Enzymatic assays of β-1,3-glucanase in AeGH16.5 silenced larvae exhibited lower activity in the gut and no change in the rest of the body. Chromatographic activity profiles from gut samples after GH16.5 silencing showed suppression of enzymatic activity, suggesting that this gene codes for the digestive larval β-1,3-glucanase of Ae. aegypti. This gene and enzyme are attractive targets for new control strategies, based on the impairment of normal gut physiology.

Dengue is the most prevalent global arboviral disease that affects over 300 million people every year. Brazil has the highest number of dengue cases in the world, with the most severe epidemics in the city of Rio de Janeiro (Rio). The effective control of dengue is critically dependent on the knowledge of population genetic structuring in the primary dengue vector, the mosquito Aedes aegypti. We analyzed mitochondrial and nuclear genomewide single nucleotide polymorphism markers generated via Restriction-site Associated DNA sequencing, as well as traditional microsatellite markers in Ae. aegypti from Rio. We found four divergent mitochondrial lineages and a strong spatial structuring of mitochondrial variation, in contrast to the overall nuclear homogeneity across Rio. Despite a low overall differentiation in the nuclear genome, we detected strong spatial structure for variation in over 20 genes that have a significantly altered expression in response to insecticides, xenobiotics, and pathogens, including the novel biocontrol agent Wolbachia. Our results indicate that high genetic diversity, spatially unconstrained admixing likely mediated by male dispersal, along with locally heterogeneous genetic variation that could affect insecticide resistance and mosquito vectorial capacity, set limits to the effectiveness of measures to control dengue fever in Rio.

The cattle tick Rhipicephalus microplus is a hematophagous ectoparasite that causes important economic losses in livestock. Different species of ticks harbor a symbiont bacterium of the genus Coxiella. It was showed that a Coxiella endosymbiont from R. microplus (CERM) is a vertically transmitted mutualist symbiont, comprising 98% of the 16S rRNA sequences in both eggs and larvae. Sequencing of the bacterial genome revealed genes for biosynthetic pathways for several vitamins and key metabolic cofactors that may provide a nutritional complement to the tick host. The CERM was abundant in ovary and Malpighian tubule of fully engorged female. Tetracycline treatment of either the tick or the vertebrate host reduced levels of bacteria in progeny in 74% for eggs and 90% for larvae without major impact neither on the reproductive fitness of the adult female or on embryo development. However, CERM proved to be essential for the tick to reach the adult life stage, as under antibiotic treatment no tick was able to progress beyond the metanymph stage. Data presented here suggest that interference in the symbiotic CERM-R. microplus relationship may be useful to the development of alternative control methods, highlighting the interdependence between ticks and their endosymbionts. Instituto de Biotecnología Fil: Garcia Guizzo, Melina. Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo de Meis; Brasil Fil: Parizi, Luís Fernando. Universidade Federal do Rio Grande do Sul. Centro de Biotecnologia; Brasil Fil: Nunes, Rodrigo Dutra. Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo de Meis; Brasil Fil: Schama, Renata. Laboratório de Biologia Computacional e Sistemas; Brasil Fil: Albano, Rodolpho M. Universidade do Estado do Rio de Janeiro. Departamento de Bioquímica; Brasil Fil: Tirloni, Lucas. Universidade Federal do Rio Grande do Sul. Centro de Biotecnologia; Brasil Fil: Oldiges, Daiane Patrícia. Universidade Federal do Rio Grande do Sul. Centro de Biotecnologia; Brasil Fil: Pilz Vieira, Ricardo. Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo de Meis; Brasil Fil: Cruz Oliveira, Wanderson Henrique. Universidade Federal Rural do Rio de Janeiro. Departamento de Química; Brasil Fil: De Souza Leite, Milane. Universidade Federal Rural do Rio de Janeiro. Departamento de Química; Brasil Fil: Gonzalez, Sergio Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Farber, Marisa Diana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Martins, Orlando. Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo de Meis; Brasil Fil: Da Silva Vaz, Itabajara. Universidade Federal do Rio Grande do Sul. Centro de Biotecnologia; Brasil. Universidade Federal do Rio Grande do Sul. Faculdade de Veterinária; Brasil. Universidade Federal Rural do Rio de Janeiro. Departamento de Química; Brasil Fil: Oliveira, Pedro L.. Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo de Meis; Brasil. Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular; Brasil

Background The genetic diversity of Neotropical fish fauna is underrepresented in public databases. This distortion is evident for the order Siluriformes, in which the suborders Siluroidei and Loricarioidei share equivalent proportion of species, although far less is known about the genetics of the latter clade, endemic to the Neotropical Region. Recently, this information gap was evident in a study about the structural diversity of fish mitochondrial genomes, and hampered a precise chronological resolution of Siluriformes. It has also prevented molecular ecology investigations about these catfishes, their interactions with the environment, responses to anthropogenic changes and potential uses. Results Using high-throughput sequencing, we provide the nearly complete mitochondrial genomes for 26 Loricariidae and one Callichthyidae species. Structural features were highly conserved. A notable exception was identified in the monophyletic clade comprising species of the Hemiancistrus, Hypostomini and Peckoltia-clades, a ~60 nucleotide-long deletion encompassing the seven nucleotides at the 3′ end of the Conserved Sequence Block (CSB) D of the control region. The expression of mitochondrial genes followed the usual punctuation pattern. Heteroplasmic sites were identified in most species. The retrieved phylogeny strongly corroborates the currently accepted tree, although bringing to debate the relationship between Schizolecis guntheri and Pareiorhaphis garbei, and highlighting the low genetic variability within the Peckoltia-clade, an eco-morphologically diverse and taxonomically problematic group. Conclusions Herein we have launched the use of high-throughput mitochondrial genomics in the studies of the Loricarioidei species. The new genomic resources reduce the information gap on the molecular diversity of Neotropical fish fauna, impacting the capacity to investigate a variety of aspects of the molecular ecology and evolution of these fishes. Additionally, the species showing the partial CSB-D are candidate models to study the replication and transcription of vertebrate mitochondrial genome. Electronic supplementary material The online version of this article (doi:10.1186/s12864-017-3709-3) contains supplementary material, which is available to authorized users.

Phymactis papillosais a rocky shore sea anemone that is commonly found in the Pacific Ocean, from the Gulf of California to Tierra del Fuego, and in the Mar del Plata region, Argentina. The genusPhymactisis closely related toBunodosomaand, due to character plasticity, a number of misidentifications have occurred. Therefore, the presence ofP. papillosain Argentina has been doubted but the matter had not been investigated in detail. Here we analyseP. papillosaspecimens from Argentina and compare them, using molecular and morphological markers, to specimens from the species' type locality. In a phylogenetic analysis using 19 allozyme markers and ribosomal internal transcribed spacers sequences of different sea anemone genera, including all West AtlanticBunodosomaspecies, we have found that the specimens from Argentina were genetically divergent fromP. papillosafrom Chile and closely related to West AtlanticBunodosomaspecies. The genetic and morphological analyses indicate that those specimens belong to a new species of the genusBunodosoma, described here asB. zamponiisp. nov.

In this work we describeActinia ebhayiensissp. nov. from South Africa. The species is externally similar to the type species of the genus,Actinia equina, from which it can be distinguished by nematocyst and allozyme data.Actinia ebhayiensishas a smooth, red column, with well-delimited parapet leading to a deep fosse where conspicuous blue acrorhagi can be found. Tentacles and oral disc are crimson red, and the pedal disc has a lighter pink colour. The microbasicp-mastigophores andb-mastigophores of the mesenterial filaments of the new species are significantly smaller than those ofA. equina. Actinia ebhayiensiscan also be clearly distinguished from other species of the genus in nematocyst measurements, genetic data and muscle morphology.

Rhodnius prolixus not only has served as a model organism for the study of insect physiology, but also is a major vector of Chagas disease, an illness that affects approximately seven million people worldwide. We sequenced the genome of R. prolixus, generated assembled sequences covering 95% of the genome (∼702 Mb), including 15,456 putative protein-coding genes, and completed comprehensive genomic analyses of this obligate blood-feeding insect. Although immunedeficiency (IMD)-mediated immune responses were observed, R. prolixus putatively lacks key components of the IMD pathway, suggesting a reorganization of the canonical immune signaling network. Although both Toll and IMD effectors controlled intestinal microbiota, neither affected Trypanosoma cruzi, the causal agent of Chagas disease, implying the existence of evasion or tolerance mechanisms. R. prolixus has experienced an extensive loss of selenoprotein genes, with its repertoire reduced to only two proteins, one of which is a selenocysteine-based glutathione peroxidase, the first found in insects. The genome contained actively transcribed, horizontally transferred genes from Wolbachia sp., which showed evidence of codon use evolution toward the insect use pattern. Comparative protein analyses revealed many lineage-specific expansions and putative gene absences in R. prolixus, including tandem expansions of genes related to chemoreception, feeding, and digestion that possibly contributed to the evolution of a blood-feeding lifestyle. The genome assembly and these associated analyses provide critical information on the physiology and evolution of this important vector species and should be instrumental for the development of innovative disease control methods., La lista completa de autores que integran el documento puede consultarse en el archivo, Este documento tiene una corrección (ver documento relacionado)., Centro Regional de Estudios Genómicos, Instituto de Investigaciones Bioquímicas de La Plata

The sea anemone Actinia equina was considered a highly variable species with a wide geographical distribution, but molecular systematic studies have shown that this wide distribution may be the result of the lumping of cryptic species. In this work enzyme electrophoresis was used to analyse the genetic variability of A. equina from the Atlantic coasts of Europe and Africa, as well as the relationships between those populations and other species of the genus. Samples of A. equina from the United Kingdom and France were compared with supposedly conspecific populations from South Africa and a recently described species from Madeira, Actinia nigropunctata. The South African and Madeiran populations were genetically very divergent from each other (genetic identity, I=0.15), as well as from the A. equina population from the United Kingdom and from the other species studied (I between 0.23 and 0.78), indicating that the African population of “A. equina” belongs to a different species. High genetic similarities were found between west Atlantic A. bermudensis and east Atlantic A. sali, in spite of the great geographical distance between them, and between the European A. equina and A. prasina. The results confirm the presence of the new species A. nigropunctata from Madeira and suggest a new species within the A. equina species group, from South Africa.

The sea anemone Actinia bermudensis [Proc. Acad. Nat. Sci. Philadelphia 41 (1889) 102] has a widespread distribution over the Western Atlantic, where it has been reported from Bermuda and Bahamas to southern Brazil. Nonetheless, reproductive features of this species, which broods asexually produced offspring, suggest that it has low dispersal capabilities. This raises a question about the specific status of the Brazilian population of A. bermudensis. The aim of this paper was to evaluate the taxonomic status of the Brazilian population, by genetically comparing it with the type locality (Bermuda) population. For this, we used allozyme electrophoresis to analyze samples from both populations. Thirteen enzyme systems (16 loci) yielded interpretable results for all populations. A high differentiation was encountered for the Western Atlantic A. bermudensis population (FST=0.434), partially due to the presence of a diagnostic locus (LAP) between Bermuda and Brazilian populations. This result could be taken as an indication that the Brazilian population should be elevated to a specific status. We do expect some genetic differentiation, however, between natural populations separated over wide geographic distances. Finally, since Mantel and regression analyses did not rule out the isolation by distance model, we have conservatively decided to maintain the specific status of A. bermudensis from Brazil.

Background Aedes aegypti, commonly known as “the yellow fever mosquito”, is of great medical concern today primarily as the major vector of dengue, chikungunya and Zika viruses, although yellow fever remains a serious health concern in some regions. The history of Ae. aegypti in Brazil is of particular interest because the country was subjected to a well-documented eradication program during 1940s-1950s. After cessation of the campaign, the mosquito quickly re-established in the early 1970s with several dengue outbreaks reported during the last 30 years. Brazil can be considered the country suffering the most from the yellow fever mosquito, given the high number of dengue, chikungunya and Zika cases reported in the country, after having once been declared “free of Ae. aegypti”. Methodology/Principal findings We used 12 microsatellite markers to infer the genetic structure of Brazilian Ae. aegypti populations, genetic variability, genetic affinities with neighboring geographic areas, and the timing of their arrival and spread. This enabled us to reconstruct their recent history and evaluate whether the reappearance in Brazil was the result of re-invasion from neighboring non-eradicated areas or re-emergence from local refugia surviving the eradication program. Our results indicate a genetic break separating the northern and southern Brazilian Ae. aegypti populations, with further genetic differentiation within each cluster, especially in southern Brazil. Conclusions/Significance Based on our results, re-invasions from non-eradicated regions are the most likely scenario for the reappearance of Ae. aegypti in Brazil. While populations in the northern cluster are likely to have descended from Venezuela populations as early as the 1970s, southern populations seem to have derived more recently from northern Brazilian areas. Possible entry points are also revealed within both southern and northern clusters that could inform strategies to control and monitor this important arbovirus vector., Author summary Aedes aegypti (“yellow fever mosquito”) is of great medical concern because it is the primary vector of important viruses causing dengue fever, chikungunya and Zika, as well as increasingly transmitting yellow fever once again. Due to the Zika outbreak, which started in Brazil in 2015 and rapidly spread with million cases reported in Central/South America and the Caribbean, this mosquito has attained special notoriety. Brazil is by far the most affected country not only by Zika, but also by dengue and chikungunya. Interestingly, in 1950s Brazil had been declared “free of Ae. aegypti” after a well-documented eradication program. However, during the last 45 years Ae. aegypti reappeared in Brazil causing several dengue outbreaks, with millions of reported cases. Here, we used genetic data to study the reappearance of Ae. aegypti in Brazil after the putative eradication event. Our results support re-invasion; mosquitoes from the non-eradicated areas in Venezuela invaded North Brazil, later expanding their distribution southwards. We also identify specific locations in Brazil, as possible entry points. This knowledge can inform strategies to control the spread of the vector and of the diseases it transmits.

Chagas disease or American trypanosomiasis, is a potentially life-threatening illness caused by the protozoan parasite, Trypanosoma cruzi. Once known as an endemic health problem of poor rural populations in Latin American countries, it has now spread worldwide. The parasite is transmitted by triatomine bugs, of which Rhodnius prolixus (Hemiptera, Reduviidae, Triatominae) is one of the vectors and a model organism. This species occurs mainly in Central and South American countries where the disease is endemic. Disease prevention focuses on vector control programs that, in general, rely intensely on insecticide use. However, the massive use of chemical insecticides can lead to resistance. One of the major mechanisms is known as metabolic resistance that is associated with an increase in the expression or activity of detoxification genes. Three of the enzyme families that are involved in this process – carboxylesterases (CCE), glutathione s-transferases (GST) and cytochrome P450s (CYP) – are analyzed in the R. prolixus genome. A similar set of detoxification genes to those of the Hemipteran Acyrthosiphon pisum but smaller than in most dipteran species was found in R. prolixus genome. All major CCE classes (43 genes found) are present but the pheromone/hormone processing class had fewer genes than usual. One main expansion was detected on the detoxification/dietary class. The phosphotriesterase family, recently associated with insecticide resistance, was also represented with one gene. One microsomal GST gene was found and the cytosolic GST gene count (14 genes) is extremely low when compared to the other hemipteran species with sequenced genomes. However, this is similar to Apis mellifera, a species known for its deficit in detoxification genes. In R. prolixus 88 CYP genes were found, with representatives in the four clans (CYP2, CYP3, CYP4 and mitochondrial) usually found in insects. R. prolixus seems to have smaller species-specific expansions of CYP genes than mosquitoes and beetles, among others. The number of R. prolixus CYP genes is similar to the hemipteran Ac. pisum, although with a bigger expansion in CYP3 and CYP4 clans, along with several gene fragments, mostly in CYP4 clan. Eleven founding members of new families were detected, consisting of ten genes in the CYP3 clan and 1 gene in the CYP4 clan. Members of these clans were proposed to have important detoxification roles in insects. The identification of CCE, GST and CYP genes is of utmost importance for directing detoxification studies on triatomines that can help insecticide management strategies in control programs. Fil: Schama, Renata. Instituto Oswaldo Cruz; Brasil Fil: Pedrini, Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata ; Argentina Fil: Juarez, Marta Patricia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata ; Argentina Fil: Nelson, David R.. University of Tennessee; Estados Unidos Fil: Torres, André. Instituto Oswaldo Cruz; Brasil Fil: Valle, Denise. Instituto Oswaldo Cruz; Brasil Fil: Mesquita, Rafael D.. Universidade Federal do Rio de Janeiro; Brasil. Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular; Brasil

Populations of Ocyurus chrysurus were compared genetically and morphometrically along the West Atlantic coast to test the null hypothesis of population homogeneity in the area. Brazilian populations were found to be differentiated in shape (canonical variates analysis; F[48,515] = 10.84, p < 0.0001). Analyses of mitochondrial DNA sequences (663 bp of the control region) did not show any differences between Brazilian populations but could detect differences between Brazilian and Caribbean (Belize) populations. The samples from Pernambuco differed significantly from the other Brazilian populations in allozyme frequencies (11 loci; F ST = 0.167; p < 0.05), but this may have resulted from the small number of samples analysed for that population. Sequence variation of Belize samples departed from neutral expectations (Fu's FS = -8.88; p < 0.001). A mismatch distribution analysis points to an ancient population expansion in that area. We conclude that the genetic data do not allow the rejection of the null hypothesis of panmixia for Brazilian yellowtail snapper populations which should be treated as a single genetic stock, with a latitudinal gradient on their morphology which probably results from phenotypic plasticity. On the other hand, there is a severe restriction to gene flow between O. chrysurus populations from the Caribbean and from the southwestern Atlantic.