RESUMO
There is evidence that IL-22 and IL-17 participate in the pathogenesis of allergic asthma. To investigate the role of IL-22, we used IL-22 deficient mice (IL-22 KO) sensitized and challenged with ovalbumin (OVA) and compared with wild type (WT) animals exposed to OVA. IL-22 KO animals exposed to OVA showed a decreased number and frequency of eosinophils, IL-5 and IL-13 in the airways, reduced mucus production and pulmonary inflammation. In addition, IL-22 KO animals exhibited a decreased percentage and number of lung CD11c+CD11b+ cells and increased apoptosis of eosinophils. Th17 cell transfer generated from IL-22 KO to animals previously sensitized and challenged with OVA caused a reduction in eosinophil frequency and number in the airways compared to animals transferred with Th17 cells generated from WT mice. Therefore, IL-22 is deleterious with concomitant secretion of IL-17. Our findings show a pro-inflammatory role for IL-22, confirmed in a model of allergen-free and allergen-specific immunotherapy. Moreover, during the comorbidity asthma and pneumonia that induces neutrophil inflammation, IL-22 was not detrimental. Our results show that targeting IL-22 would negatively affect the survival of eosinophils, reduce the expansion or migration of CD11c+CD11b+ cells, and negatively regulate allergic asthma.
Assuntos
Asma , Pneumonia , Camundongos , Animais , Interleucina-17/genética , Asma/patologia , Pulmão/patologia , Eosinófilos , Pneumonia/patologia , Alérgenos , Comorbidade , Ovalbumina , Modelos Animais de Doenças , Líquido da Lavagem Broncoalveolar , Camundongos Endogâmicos BALB CRESUMO
BACKGROUND: Bees are the most important group of pollinators worldwide and their populations are declining. In natural conditions, Apis mellifera depends exclusively on food from the field to meet its physiological demands. In the period of scarcity, available resources are insufficient and artificial supplementation becomes essential for maintaining the levels of vitamins, proteins, carbohydrates, and minerals of colonies. Among these minerals, zinc is essential in all living systems, particularly for the regulation of cell division and protein synthesis, and is a component of more than 200 metalloenzymes. RESULTS: The total RNA extracted from the brain tissue of nurse bees exposed to different sources and concentrations of zinc was sequenced. A total of 1,172 genes in the treatment that received an inorganic source of zinc and 502 genes that received an organic source of zinc were found to be differentially expressed among the control group. Gene ontology enrichment showed that zinc can modulate important biological processes such as nutrient metabolism and the molting process. CONCLUSIONS: Our results indicate that zinc supplementation modulates the expression of many differentially expressed genes and plays an important role in the development of Apis mellifera bees. All the information obtained in this study can contribute to future research in the field of bee nutrigenomics.
Assuntos
Transcriptoma , Zinco , Animais , Abelhas/genética , Encéfalo , Suplementos Nutricionais , Nutrigenômica , Zinco/farmacologiaRESUMO
The reconstruction of relationships within recently radiated groups is challenging even when massive amounts of sequencing data are available. The use of restriction site-associated DNA sequencing (RAD-Seq) to this end is promising. Here, we assessed the performance of RAD-Seq to infer the species-level phylogeny of the rapidly radiating genus Cereus (Cactaceae). To examine how the amount of genomic data affects resolution in this group, we used datasets and implemented different analyses. We sampled 52 individuals of Cereus, representing 18 of the 25 species currently recognized, plus members of the closely allied genera Cipocereus and Praecereus, and other 11 Cactaceae genera as outgroups. Three scenarios of permissiveness to missing data were carried out in iPyRAD, assembling datasets with 30% (333 loci), 45% (1440 loci), and 70% (6141 loci) of missing data. For each dataset, Maximum Likelihood (ML) trees were generated using two supermatrices, i.e., only SNPs and SNPs plus invariant sites. Accuracy and resolution were improved when the dataset with the highest number of loci was used (6141 loci), despite the high percentage of missing data included (70%). Coalescent trees estimated using SVDQuartets and ASTRAL are similar to those obtained by the ML reconstructions. Overall, we reconstruct a well-supported phylogeny of Cereus, which is resolved as monophyletic and composed of four main clades with high support in their internal relationships. Our findings also provide insights into the impact of missing data for phylogeny reconstruction using RAD loci.
Assuntos
Evolução Biológica , Cactaceae/genética , Genoma de Planta , Análise de Sequência de DNA , Sequência de Bases , Bases de Dados Genéticas , Loci Gênicos , Especiação Genética , Funções Verossimilhança , Filogenia , Polimorfismo de Nucleotídeo Único/genética , Análise de Componente PrincipalRESUMO
Standardization of the methods for extraction of DNA from sand flies is essential for obtaining high efficiency during subsequent molecular analyses, such as the new sequencing methods. Information obtained using these methods may contribute substantially to taxonomic, evolutionary, and eco-epidemiological studies. The aim of the present study was to standardize and compare two methods for the extraction of genomic DNA from sand flies for obtaining DNA in sufficient quantities for next-generation sequencing. Sand flies were collected from the municipalities of Campo Grande, Camapuã, Corumbá and Miranda, state of Mato Grosso do Sul, Brazil. Three protocols using a silica column-based commercial kit (ReliaPrep™ Blood gDNA Miniprep System kit, Promega®), and three protocols based on the classical phenol-chloroform extraction method (Uliana et al., 1991), were compared with respect to the yield and quality of the extracted DNA. DNA was quantified using a Qubit 2.0 fluorometer. The presence of sand fly DNA was confirmed by PCR amplification of the IVS6 region (constitutive gene), followed by electrophoresis on a 1.5% agarose gel. A total of 144 male specimens were analyzed, 72 per method. Significant differences were observed between the two methods tested. Protocols 2 and 3 of phenol-chloroform extraction presented significantly better performance than all commercial kit extraction protocols tested. For phenol-chloroform extraction, protocol 3 presented significantly better performance than protocols 1 and 2. The IVS6 region was detected in 70 of 72 (97.22%) samples extracted with phenol, including all samples for protocols 2 and 3. This is the first study on the standardization of methods for the extraction of DNA from sand flies for application to next-generation sequencing, which is a promising tool for entomological and molecular studies of sand flies.
Assuntos
DNA/isolamento & purificação , Técnicas de Genotipagem/métodos , Psychodidae/genética , Análise de Sequência de DNA/métodos , Animais , Distribuição de Qui-Quadrado , Clorofórmio , DNA/química , Endopeptidase K/metabolismo , Técnicas de Genotipagem/normas , Masculino , Fenol , Reação em Cadeia da Polimerase/métodos , Análise de Sequência de DNA/normas , Estatísticas não ParamétricasRESUMO
Hepatozoon sp. are parasites that commonly infect frogs and arthropod vectors. This species has variability in the morphological and morphometric characteristics. Due to these variations, the naming of the species is thus impaired and only by visualizing the sporogonic cycle in vector and by molecular studies this problem can be solved. Recently, the use of molecular genetics has helped the species denomination. In this work, we collected 145 frogs (68 Leptodactylus chaquensis and 77 Leptodactylus podicipinus) in different sampling sites, where were found 18 (26.47%) L. chaquensis and 24 (31.17%) L. podicipinus parasitized; besides of gamonts, schizogonic forms were also seen in animals organs. The positivity difference between the collection sites for both frog species was not significant (p = 0.958). Comparing gamonts found in each species of anuran, we observed differences in morphology. The comparison in the molecular level for L. podicipinus was not possible due to small amount of blood obtained, just L. chaquensis had their parasites DNA sequenced. The amplified and sequenced samples, named HEP1 to HEP10, are presented in the phylogenetic tree as a different branch from other haemogregarines described on other hosts. Therefore, we have seen that, although the morphology and morphometry of the collected parasites at each site showed differences, the sequencing of these samples revealed identical species of Hepatozoon, and different compared to those from GenBank, thereby demonstrating that the species of Hepatozoon in L. chaquensis observed in this study probably represent a new species.
Assuntos
Anuros/parasitologia , Coccidiose/veterinária , Eucoccidiida/isolamento & purificação , Animais , Anuros/classificação , Brasil , Coccidiose/parasitologia , Eucoccidiida/classificação , Eucoccidiida/genética , Eucoccidiida/fisiologia , Especificidade de Hospedeiro , Dados de Sequência Molecular , FilogeniaRESUMO
In visceral leishmaniasis (VL) endemic areas, a minority of infected individuals progress to disease since most of them develop protective immunity. Therefore, we investigated the risk markers of VL within nonimmune sector. Analyzing infected symptomatic and, asymptomatic, and noninfected individuals, VL patients presented with reduced high-density lipoprotein cholesterol (HDL-C), elevated triacylglycerol (TAG), and elevated very-low-density lipoprotein cholesterol (VLDL-C) levels. A polymorphism analysis of the lipoprotein lipase (LPL) gene using HindIII restriction digestion (N = 156 samples) (H+ = the presence and H- = the absence of mutation) revealed an increased adjusted odds ratio (OR) of VL versus noninfected individuals when the H+/H+ was compared with the H-/H- genotype (OR = 21.3; 95% CI = 2.32-3335.3; P = 0.003). The H+/H+ genotype and the H+ allele were associated with elevated VLDL-C and TAG levels (P < 0.05) and reduced HDL-C levels (P < 0.05). An analysis of the L162V polymorphism in the peroxisome proliferator-activated receptor alpha (PPARα) gene (n = 248) revealed an increased adjusted OR when the Leu/Val was compared with the Leu/Leu genotype (OR = 8.77; 95% CI = 1.41-78.70; P = 0.014). High TAG (P = 0.021) and VLDL-C (P = 0.023) levels were associated with susceptibility to VL, whereas low HDL (P = 0.006) levels with resistance to infection. The mutated LPL and the PPARα Leu/Val genotypes may be considered risk markers for the development of VL.
Assuntos
Leishmaniose Visceral/sangue , Lipase Lipoproteica/genética , Lipoproteínas HDL/genética , PPAR alfa/genética , Genótipo , Humanos , Leishmaniose Visceral/genética , Lipoproteínas VLDL/sangue , Polimorfismo Genético/genética , Triglicerídeos/sangueRESUMO
An analysis of the dietary content of haematophagous insects can provide important information about the transmission networks of certain zoonoses. The present study evaluated the potential of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the mitochondrial cytochrome B (cytb) gene to differentiate between vertebrate species that were identified as possible sources of sandfly meals. The complete cytb gene sequences of 11 vertebrate species available in the National Center for Biotechnology Information database were digested with Aci I, Alu I, Hae III and Rsa I restriction enzymes in silico using Restriction Mapper software. The cytb gene fragment (358 bp) was amplified from tissue samples of vertebrate species and the dietary contents of sandflies and digested with restriction enzymes. Vertebrate species presented a restriction fragment profile that differed from that of other species, with the exception of Canis familiaris and Cerdocyon thous. The 358 bp fragment was identified in 76 sandflies. Of these, 10 were evaluated using the restriction enzymes and the food sources were predicted for four: Homo sapiens (1), Bos taurus (1) and Equus caballus (2). Thus, the PCR-RFLP technique could be a potential method for identifying the food sources of arthropods. However, some points must be clarified regarding the applicability of the method, such as the extent of DNA degradation through intestinal digestion, the potential for multiple sources of blood meals and the need for greater knowledge regarding intraspecific variations in mtDNA.
Assuntos
Comportamento Animal/fisiologia , Citocromos b/genética , Psychodidae/fisiologia , Animais , Comportamento Animal/classificação , Gatos , Bovinos , Cães , Comportamento Alimentar/fisiologia , Cavalos , Humanos , Refeições , Mitocôndrias/enzimologia , Gambás , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Psychodidae/classificação , Ratos , SuínosRESUMO
Hepatozoon spp. are commonly found infecting snakes. Since the latter are parasitized by diverse forms and data in the literature show divergence, we studied Hepatozoon spp. diversity on Crotalus durissus terrificus snakes using both molecular and morphological approaches. Naturally infected animals were employed. Blood was collected, blood smears were prepared and an aliquot was stored at -20°C for DNA extraction. Five specimens of C. durissus terrificus were selected, each of them infected with one gamont type. Morphological and morphometric analyses of the found gamonts led to their grouping into three populations. For molecular characterization, seven oligonucleotide pairs that amplify distinct regions of rDNA gene were tested by adopting the PCR technique. Only the oligonucleotide pairs HepF300/Hep900 and HEMO1/HEMO2 were efficient in amplifying and distinguishing different isolates of Hepatozoon spp. from snakes. The better results were obtained when both oligonucleotide pairs were used in association. Based on the molecular and morphologic differences, three new species were proposed: Hepatozoon cuestensis sp. nov.; Hepatozoon cevapii sp. nov. and Hepatozoon massardii sp. nov. This is the first description of new Hepatozoon species from snakes, based on molecular characterization and morphological data, in South America.
Assuntos
Coccídios/classificação , Coccidiose/veterinária , Crotalus/parasitologia , Animais , Brasil/epidemiologia , Coccídios/genética , Coccídios/isolamento & purificação , Coccídios/ultraestrutura , Coccidiose/epidemiologia , Coccidiose/parasitologia , DNA de Protozoário/genética , DNA de Protozoário/isolamento & purificação , DNA Ribossômico/genética , Eritrócitos/parasitologia , Feminino , Processamento de Imagem Assistida por Computador , Masculino , Filogenia , Reação em Cadeia da Polimerase , Prevalência , Alinhamento de SequênciaRESUMO
The increasing population of Aedes aegypti mosquitoes on Madeira Island (Portugal) resulted in the first autochthonous dengue outbreak, which occurred in October 2012. Our study establishes the first genetic evaluation based on the mitochondrial DNA (mtDNA) genes [cytochrome oxidase subunit I (COI) and NADH dehydrogenase subunit 4 (ND4)] and knockdown resistance (kdr) mutations exploring the colonisation history and the genetic diversity of this insular vector population. We included mosquito populations from Brazil and Venezuela in the analysis as putative geographic sources. The Ae. aegypti population from Madeira showed extremely low mtDNA genetic variability, with a single haplotype for COI and ND4. We also detected the presence of two important kdr mutations and the quasi-fixation of one of these mutations (F1534C). These results are consistent with a unique recent founder event that occurred on the island of Ae. aegypti mosquitoes that carry kdr mutations associated with insecticide resistance. Finally, we also report the presence of the F1534C kdr mutation in the Brazil and Venezuela populations. To our knowledge, this is the first time this mutation has been found in South American Ae. aegypti mosquitoes. Given the present risk of Ae. aegypti re-invading continental Europe from Madeira and the recent dengue outbreaks on the island, this information is important to plan surveillance and control measures.
Assuntos
Aedes/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Insetos Vetores/genética , Mutação/genética , NADH Desidrogenase/genética , Distribuição Animal , Animais , Brasil , DNA Mitocondrial/genética , Dengue/epidemiologia , Surtos de Doenças , Haplótipos/genética , Resistência a Inseticidas/genética , Portugal/epidemiologia , VenezuelaRESUMO
Mosquito females of the genus Mansonia (Blanchard) can be a nuisance to humans and animals since they are voraciously hematophagous and feed on the blood of a variety of vertebrates. Despite their relevance, there is a lack of investigation into the blood-feeding patterns of the Mansonia species. Knowledge of the host preference is crucial in establishing the public health importance of a mosquito species and its potential to be involved in the transmission dynamics of pathogens. Species that are primarily anthropophilic can be more effective in spreading vector-borne pathogens to humans. In this study, we used an Illumina Nextera sequencing protocol and the QIIME2 workflow to assess the diversity of DNA sequences extracted in the ingested blood of mosquito species to evaluate the overall and local host choices for three species: Ma. titillans, Ma. Amazonensis, and Ma. humeralis, in rural areas alongside the Madeira River in the vicinities of the Santo Antonio Energia (SAE) reservoir in the municipality of Porto Velho, Rondônia, Western Brazil. By performing our analysis pipeline, we have found that host diversity per collection site showed a significant heterogeneity across the sample sites. In addition, in rural areas, Ma. amazonensis present a high affinity for B. taurus, Ma. humeralis shows an overall preference for C. familiaris and B. taurus, but also H. sapiens and E. caballus in urban areas, and Ma. titillans showed more opportunistic behavior in rural areas, feeding on wild animals and G. gallus, though with an overall preference for H. sapiens.
Assuntos
Culicidae , Humanos , Animais , Feminino , Brasil , Mosquitos Vetores , Comportamento Alimentar , Saúde PúblicaRESUMO
Genetic diversity and population structuring for the species Haemogogus leucocelaenus, a sylvatic vector of yellow fever virus, were found to vary with the degree of agricultural land use and isolation of fragments of Atlantic Forest in municipalities in the state of São Paulo where specimens were collected. Genotyping of 115 mitochondrial SNPs showed that the populations with the highest indices of genetic diversity (polymorphic loci and mean pairwise differences between the sequences) are found in areas with high levels of agricultural land use (northeast of the State). Most populations exhibited statistically significant negative values for the Tajima D and Fu FS neutrality tests, suggesting recent expansion. The results show an association between genetic diversity in this species and the degree of agricultural land use in the sampled sites, as well as signs of population expansion of this species in most areas, particularly those with the highest forest edge densities. A clear association between population structuring and the distance between the sampled fragments (isolation by distance) was observed: samples from a large fragment of Atlantic Forest extending along the coast of the state of São Paulo exhibited greater similarity with each other than with populations in the northwest of the state.
Assuntos
Culicidae , Febre Amarela , Animais , Febre Amarela/genética , Brasil , Mosquitos Vetores/genética , FlorestasRESUMO
Anopheles darlingi is a major malaria vector in the Amazon region and, like other vectors, harbors a community of microorganisms with which it shares a network of interactions. Here, we describe the diversity and bacterial composition from the midguts and salivary glands of lab-reared and field-captured An. darlingi using metagenome sequencing of the 16S rRNA gene. The libraries were built using the amplification of the region V3-V4 16S rRNA gene. The bacterial community from the salivary glands was more diverse and richer than the community from the midguts. However, the salivary glands and midguts only showed dissimilarities in beta diversity between lab-reared mosquitoes. Despite that, intra-variability was observed in the samples. Acinetobacter and Pseudomonas were dominant in the tissues of lab-reared mosquitoes. Sequences of Wolbachia and Asaia were both found in the tissue of lab-reared mosquitoes; however, only Asaia was found in field-captured An. darlingi, but in low abundance. This is the first report on the characterization of microbiota composition from the salivary glands of An. darlingi from lab-reared and field-captured individuals. This study can provide invaluable insights for future investigations regarding mosquito development and interaction between mosquito microbiota and Plasmodium sp.
RESUMO
In high abundance, females of the genus Mansonia (Blanchard) can be a nuisance to humans and animals because they are voraciously hematophagous and feed on the blood of a myriad of vertebrates. The spatial-temporal distribution pattern of Mansonia species is associated with the presence of their host plants, usually Eichhornia crassipes, E. azurea, Ceratopteris pteridoides, Limnobium laevigatum, Pistia stratiotes, and Salvinia sp. Despite their importance, there is a lack of investigation on the dispersion and population genetics of Mansonia species. Such studies are pivotal to evaluating the genetic structuring, which ultimately reflects populational expansion-retraction patterns and dispersal dynamics of the mosquito, particularly in areas with a history of recent introduction and establishment. The knowledge obtained could lead to better understanding of how anthropogenic changes to the environment can modulate the population structure of Mansonia species, which in turn impacts mosquito population density, disturbance to humans and domestic animals, and putative vector-borne disease transmission patterns. In this study, we present an Illumina NGS sequencing protocol to obtain whole-mitogenome sequences of Mansonia spp. to assess the microgeographic genetic diversity and dispersion of field-collected adults. The specimens were collected in rural environments in the vicinities of the Santo Antônio Energia (SAE) hydroelectric reservoir on the Madeira River.
Assuntos
Ascaridídios , Culicidae , Malvaceae , Adulto , Animais , Brasil/epidemiologia , Feminino , Genética Populacional , Humanos , Mitocôndrias , Mosquitos Vetores , RiosRESUMO
BACKGROUND: Anopheles darlingi is the major vector of malaria in South America, and its behavior and distribution has epidemiological importance to biomedical research. In Brazil, An. darlingi is found in the northern area of the Amazon basin, where 99.5% of the disease is reported. METHODS: The study area, known as Ramal do Granada, is a rural settlement inside the Amazon basin in the state of Acre. Population variations and density have been analysed by species behaviour, and molecular analysis has been measured by ND4 mitochondrial gene sequencing. RESULTS: The results show higher density in collections near a recent settlement, suggesting that a high level of colonization decreases the vector presence. The biting activity showed higher activity at twilight and major numbers of mosquitos in the remaining hours of the night in months of high density. From a sample of 110 individual mosquitoes, 18 different haplotypes were presented with a diversity index of 0.895, which is higher than that found in other Anopheles studies. CONCLUSIONS: An. darlingi depends on forested regions for their larval and adult survival. In months with higher population density, the presence of mosquitoes persisted in the second part of the night, increasing the vector capacity of the species. Despite the intra-population variation in the transition to rainy season, the seasonal distribution of haplotypes shows no change in the structure population of An. darlingi.
Assuntos
Anopheles/crescimento & desenvolvimento , Vetores de Doenças , Animais , Anopheles/genética , Comportamento Animal , Brasil , Comportamento Alimentar , Humanos , Proteínas Mitocondriais/genética , NADH Desidrogenase/genética , Dinâmica Populacional , População RuralRESUMO
Trachoma is a keratoconjunctivitis caused by Chlamydia trachomatis, considered an important leading cause of preventable blindness in the world. This study aimed at verifying if flies can be the vectors for trachoma in our municipality. Flies were assessed in the households of children diagnosed with inflammatory trachoma at the municipality of Botucatu, Sao Paulo State, Brazil. Fly traps were placed in the backyard of the houses during 24 h, in each of the four weather seasons, over a period of one year. The collected dipterans were taxonomically classified and the presence of Chlamydia trachomatis in the flies was evidenced by using the Polymerase Chain Reaction (PCR). During the studied period, 2,188 flies were collected, mainly during the summer and the spring. The most common identified fly was Musca domestica. All fly samples were negative for Chlamydia trachomatis but several other different bacteria were identified in these flies. The authors concluded that flies are probably not the vectors for trachoma in the studied area. Further studies should be conducted to evaluate other possible factors responsible for the maintenance of the disease in our environment.
Assuntos
Dípteros , Gonorreia , Tracoma , Animais , Brasil/epidemiologia , Chlamydia trachomatis/genética , Tracoma/epidemiologiaRESUMO
Mosquito susceptibility to Plasmodium spp. infection is of paramount importance for malaria occurrence and sustainable transmission. Therefore, understanding the genetic features underlying the mechanisms of susceptibility traits is pivotal to assessing malaria transmission dynamics in endemic areas. The aim of this study was to investigate the susceptibility of Nyssorhynchus darlingi-the dominant malaria vector in Brazil-to Plasmodium spp. using a reduced representation genome-sequencing protocol. The investigation was performed using a genome-wide association study (GWAS) to identify mosquito genes that are predicted to modulate the susceptibility of natural populations of the mosquito to Plasmodium infection. After applying the sequence alignment protocol, we generated the variant panel and filtered variants; leading to the detection of 202,837 SNPs in all specimens analyzed. The resulting panel was used to perform GWAS by comparing the pool of SNP variants present in Ny. darlingi infected with Plasmodium spp. with the pool obtained in field-collected mosquitoes with no evidence of infection by the parasite (all mosquitoes were tested separately using RT-PCR). The GWAS results for infection status showed two statistically significant variants adjacent to important genes that can be associated with susceptibility to Plasmodium infection: Cytochrome P450 (cyp450) and chitinase. This study provides relevant knowledge on malaria transmission dynamics by using a genomic approach to identify mosquito genes associated with susceptibility to Plasmodium infection in Ny. darlingi in western Amazonian Brazil.
Assuntos
Anopheles , Malária/genética , Plasmodium/patogenicidade , Animais , Anopheles/genética , Anopheles/parasitologia , Brasil , Suscetibilidade a Doenças , Vetores de Doenças , Feminino , Genética Populacional , Estudo de Associação Genômica Ampla/veterinária , Biblioteca Genômica , Interações Hospedeiro-Parasita/genética , Malária/parasitologia , Malária/transmissão , Mosquitos Vetores/genética , Mosquitos Vetores/parasitologia , Polimorfismo de Nucleotídeo ÚnicoRESUMO
The mosquito microbiota is composed of several lineages of microorganisms whose ecological roles and evolutionary histories have yet to be investigated in depth. Among these microorganisms, Asaia bacteria play a prominent role, given their abundance in the gut, reproductive organs, and salivary glands of different mosquito species, while their presence has also been reported in several other insects. Notably, Asaia has great potential as a tool for the control of mosquito-borne diseases. Here, we present a wide phylogenomic analysis of Asaia strains isolated from different species of mosquito vectors and from different populations of the Mediterranean fruit fly (medfly), Ceratitis capitata, an insect pest of worldwide economic importance. We show that phylogenetically distant lineages of Asaia experienced independent genome reductions, despite following a common pattern, characterized by the early loss of genes involved in genome stability. This result highlights the role of specific metabolic pathways in the symbiotic relationship between Asaia and the insect host. Finally, we discovered that all but one of the Asaia strains included in the study possess the pyrethroid hydrolase gene. Phylogenetic analysis revealed that this gene is ancestral in Asaia, strongly suggesting that it played a role in the establishment of the symbiotic association between these bacteria and the mosquito hosts. We propose that this gene from the symbiont contributed to initial pyrethroid resistance in insects harboring Asaia, also considering the widespread production of pyrethrins by several plants.IMPORTANCE We have studied genome reduction within several strains of the insect symbiont Asaia isolated from different species/strains of mosquito and medfly. Phylogenetically distant strains of Asaia, despite following a common pattern involving the loss of genes related to genome stability, have undergone independent genome reductions, highlighting the peculiar role of specific metabolic pathways in the symbiotic relationship between Asaia and its host. We also show that the pyrethroid hydrolase gene is present in all the Asaia strains isolated except for the South American malaria vector Anopheles darlingi, for which resistance to pyrethroids has never been reported, suggesting a possible involvement of Asaia in determining resistance to insecticides.
Assuntos
Acetobacteraceae/genética , Proteínas de Bactérias/metabolismo , Ceratitis capitata/microbiologia , Culicidae/microbiologia , Genoma Bacteriano , Filogenia , Simbiose , Acetobacteraceae/classificação , Acetobacteraceae/isolamento & purificação , Acetobacteraceae/fisiologia , Animais , Proteínas de Bactérias/genética , Ceratitis capitata/efeitos dos fármacos , Ceratitis capitata/fisiologia , Culicidae/efeitos dos fármacos , Culicidae/fisiologia , Evolução Molecular , Tamanho do Genoma , Resistência a Inseticidas , Inseticidas/farmacologia , Masculino , Piretrinas/farmacologiaRESUMO
The microbiota of the gut-lung axis affects local and far-reaching immune responses and might also trigger chronic and inflammatory diseases. We hypothesized that gut dysbiosis induced by obesity, which coexists in countries with a high tuberculosis burden, aggravates the host susceptibility and the pulmonary damage tolerance. To assess our hypothesis, we used a model of high-fat diet (HFD)-induced obesity, followed by infection of C57BL/6 mice with Mycobacterium tuberculosis. We showed that obesity increased the susceptibility, the pulmonary inflammation and IFN-γ levels in M. tuberculosis-infected mice. During the comorbidity obesity and tuberculosis, there is an increase of Bacteroidetes and Firmicutes in the lungs, and an increase of Firmicutes and butyrate in the feces. Depletion of gut microbiota by antibiotic treatment in the obese infected mice reduced the frequencies of CD4+IFN-γ+IL-17- cells and IFN-γ levels in the lungs, associated with an increase of Lactobacillus. Our findings reinforce the role of the gut-lung axis in chronic infections and suggest that the gut microbiota modulation may be a potential host-directed therapy as an adjuvant to treat TB in the context of IFN-γ-mediated immunopathology.
Assuntos
Disbiose/etiologia , Disbiose/microbiologia , Interferon gama/biossíntese , Obesidade/complicações , Obesidade/microbiologia , Pneumonia/microbiologia , Tuberculose/complicações , Imunidade Adaptativa , Animais , Carga Bacteriana , Suscetibilidade a Doenças , Disbiose/imunologia , Transplante de Microbiota Fecal , Fezes/microbiologia , Feminino , Leucócitos/metabolismo , Pulmão/imunologia , Pulmão/microbiologia , Pulmão/patologia , Camundongos Endogâmicos C57BL , Microbiota , Obesidade/imunologia , Pneumonia/imunologia , Tuberculose/imunologiaRESUMO
BACKGROUND: The nutritional requirements of honeybees (Apis mellifera) for their complete development need to be supplied through food sources available in the environment, since honeybees are insects that depend directly on blossoming food sources. However, at certain times a food-supply reduction can promote nutritional stress, thus necessitating food supplementation for maintenance or production stimulus of the colonies. Thus, the determination of optimal energy supplementation can assist in the maintenance and production of colonies. METHODS: Twenty Apis mellifera beehives were used (with five beehives per treatment): CTL, control (without feeding); SJ, sugarcane juice; SS, sugar syrup; and IS, inverted sucrose. We evaluated the food consumption, population development, and physiological state (expression of vitellogenin and hexamerin 70a genes) of each colony. RESULTS: The results showed that the supplementation of colonies with sugar syrup resulted in an intermediate consumption level (894.6 ± 291 mL) and better development (384.9 ± 237.3 and 158.3 ± 171.6 cm2, sealed and open brood, respectively). Furthermore, this diet ensured that the colonies were in a good physiological state, as bees fed this diet presented the highest relative expression levels of vitellogenin and hexamerin 70a among all the diets tested. CONCLUSIONS: Therefore, sugar syrup is concluded to be the best artificial energetic food for use in the supplementation of honeybee colonies.
RESUMO
Multiple sclerosis is an autoimmune disease that affects the myelinated central nervous system (CNS) neurons and triggers physical and cognitive disabilities. Conventional therapy is based on disease-modifying drugs that control disease severity but can also be deleterious. Complementary medicines have been adopted and evidence indicates that yeast supplements can improve symptoms mainly by modulating the immune response. In this investigation, we evaluated the therapeutic potential of Saccharomyces cerevisiae and its selenized derivative (Selemax) in experimental autoimmune encephalomyelitis (EAE). Female C57BL/6 mice submitted to EAE induction were orally supplemented with these yeasts by gavage from day 0 to day 14 after EAE induction. Both supplements determined significant reduction in clinical signs concomitantly with diminished Th1 immune response in CNS, increased proportion of Foxp3+ lymphocytes in inguinal and mesenteric lymph nodes and increased microbiota diversity. However, Selemax was more effective clinically and immunologically; it reduced disease prevalence more sharply, increased the proportion of CD103+ dendritic cells expressing high levels of PD-L1 in mesenteric lymph nodes and reduced the intestinal inflammatory process more strongly than S. cerevisiae. These results suggest a clear gut-brain axis modulation by selenized S. cerevisiae and suggest their inclusion in clinical trials.