RESUMEN
BACKGROUND: Nitrogen is considered the most limiting nutrient element for herbivorous insects. To alleviate nitrogen limitation, insects have evolved various symbiotically mediated strategies that enable them to colonize nitrogen-poor habitats or exploit nitrogen-poor diets. In frugivorous tephritid larvae developing in fruit pulp under nitrogen stress, it remains largely unknown how nitrogen is obtained and larval development is completed. RESULTS: In this study, we used metagenomics and metatranscriptomics sequencing technologies as well as in vitro verification tests to uncover the mechanism underlying the nitrogen exploitation in the larvae of Bactrocera dorsalis. Our results showed that nitrogenous waste recycling (NWR) could be successfully driven by symbiotic bacteria, including Enterobacterales, Lactobacillales, Orbales, Pseudomonadales, Flavobacteriales, and Bacteroidales. In this process, urea hydrolysis in the larval gut was mainly mediated by Morganella morganii and Klebsiella oxytoca. In addition, core bacteria mediated essential amino acid (arginine excluded) biosynthesis by ammonium assimilation and transamination. CONCLUSIONS: Symbiotic bacteria contribute to nitrogen transformation in the larvae of B. dorsalis in fruit pulp. Our findings suggest that the pattern of NWR is more likely to be applied by B. dorsalis, and M. morganii, K. oxytoca, and other urease-positive strains play vital roles in hydrolysing nitrogenous waste and providing metabolizable nitrogen for B. dorsalis.
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Nitrógeno , Tephritidae , Animales , Bacterias/genética , Bacterias/metabolismo , Drosophila/metabolismo , Larva/metabolismo , Nitrógeno/metabolismo , Simbiosis , Tephritidae/metabolismo , Tephritidae/microbiologíaRESUMEN
BACKGROUND: The gut bacteria of tephritid fruit flies play prominent roles in nutrition, reproduction, maintenance and ecological adaptations of the host. Here, we adopted an approach based on direct observation of symbiotic or axenic flies feeding on dishes seeded with drops of full diet (containing all amino acids) or full diet supplemented with bacteria at similar concentrations to explore the effects of intestinal bacteria on foraging decision and fitness of Bactrocera dorsalis. RESULTS: The results show that intestinal probiotics elicit beneficial foraging decision and enhance the female reproduction fitness and survival of B. dorsalis (symbiotic and axenic), yet preferences for probiotic diets were significantly higher in axenic flies to which they responded faster compared to full diet. Moreover, females fed diet supplemented with Pantoea dispersa and Enterobacter cloacae laid more eggs but had shorter lifespan while female fed Enterococcus faecalis and Klebsiella oxytoca enriched diets lived longer but had lower fecundity compared to the positive control. Conversely, flies fed sugar diet (negative control) were not able to produce eggs, but lived longer than those from the positive control. CONCLUSIONS: These results suggest that intestinal bacteria can drive the foraging decision in a way which promotes the reproduction and survival of B. dorsalis. Our data highlight the potentials of gut bacterial isolates to control the foraging behavior of the fly and empower the sterile insect technique (SIT) program through the mass rearing.
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Bacterias/crecimiento & desarrollo , Tephritidae/microbiología , Tephritidae/fisiología , Animales , Conducta Apetitiva , Bacterias/genética , Femenino , Fertilidad , Aptitud Genética , Longevidad , Masculino , Simbiosis , Tephritidae/genéticaRESUMEN
Oxidative stress is a state in which oxidants are produced in excess in the body's tissues and cells, resulting in a biological imbalance amid the generation of reactive oxygen and nitrogen species (RONS) from redox reactions. In case of insufficient antioxidants to balance, the immune system triggers signaling cascades to mount inflammatory responses. Oxidative stress can have deleterious effects on major macromolecules such as lipids, proteins, and nucleic acids, hence, Oxidative stress and inflammation are among the multiple factors contributing to the etiology of several disorders such as diabetes, cancers, and cardiovascular diseases. Non-coding RNAs (ncRNAs) which were once referred to as dark matter have been found to function as key regulators of gene expression through different mechanisms. They have dynamic roles in the onset and development of inflammatory and oxidative stress-related diseases, therefore, are potential targets for the control of those diseases. One way of controlling those diseases is through the use of natural products, a rich source of antioxidants that have drawn attention with several studies showing their involvement in combating chronic diseases given their enormous gains, low side effects, and toxicity. In this review, we highlighted the natural products that have been reported to target ncRNAs as mediators of their biological effects on oxidative stress and several inflammation-associated disorders. Those natural products include Baicalein, Tanshinone IIA, Geniposide, Carvacrol/Thymol, Triptolide, Oleacein, Curcumin, Resveratrol, Solarmargine, Allicin, aqueous extract or pulp of Açai, Quercetin, and Genistein. We also draw attention to some other compounds including Zanthoxylum bungeanum, Canna genus rhizome, Fuzi-ganjiang herb pair, Aronia melanocarpa, Peppermint, and Gingerol that are effective against oxidative stress and inflammation-related disorders, however, have no known effect on ncRNAs. Lastly, we touched on the many ncRNAs that were found to play a role in oxidative stress and inflammation-related disorders but have not yet been investigated as targets of a natural product. Shedding more light into these two last points of shadow will be of great interest in the valorization of natural compounds in the control and therapy of oxidative stress- and inflammation-associated disorders.
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Nitrogen is usually a restrictive nutrient that affects the growth and development of insects, especially of those living in low nitrogen nutrient niches. In response to the low nitrogen stress, insects have gradually developed symbiont-based stress response strategies-biological nitrogen fixation and nitrogenous waste recycling-to optimize dietary nitrogen intake. Based on the above two patterns, atmospheric nitrogen or nitrogenous waste (e.g., uric acid, urea) is converted into ammonia, which in turn is incorporated into the organism via the glutamine synthetase and glutamate synthase pathways. This review summarized the reaction mechanisms, conventional research methods and the various applications of biological nitrogen fixation and nitrogenous waste recycling strategies. Further, we compared the bio-reaction characteristics and conditions of two strategies, then proposed a model for nitrogen provisioning based on different strategies.
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In order to understand the role of symbionts for their insect hosts, it is customary to treat them with antibiotics or to sterilize eggs (treatments), resulting in aposymbiotic and axenic insects, respectively. Such axenic insects can then be compared with untreated controls. Fruit flies often bear complex communities which are greatly reduced by such treatments. However, the bacterial community is not completely eliminated. Here, we examine the effect of these procedures on the structure of the remaining bacterial communities in Bactrocera dorsalis (Diptera: Tephritidae) and on the insect longevity. The antibiotics (Norfloxacin and Ceftazedime) were administered to 1-day-old adult flies through sugar meal for 7 days, and eggs were surface sterilized and dechorionated to produce axenic lines. The flies were starved of protein before they were offered full diets or diets containing non-essential amino acids only. Antibiotic and egg disinfection treatments resulted in a significant reduction of the vast majority of gut bacterial populations, especially Proteobacteria, Firmicutes and Bacteroidetes. On the other hand, antibiotic allowed the persistence of Actinobacteria, Cyanobacteria and Acidobacteria populations. In untreated control flies, longevity was extended irrespective of diet quality in comparison to treated flies. Conversely, when gut bacteria were largely reduced (aposymbiotic and axenic flies), longevity was reduced in the non-essential amino acids diet treatment versus slightly improved in the presence of a protein diet. We discuss these results in an ecological-life-history perspective.
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Non-coding RNAs (ncRNAs) including microRNAs (miRs) and long non-coding RNAs (lncRNAs) have emerged as key regulators of gene expression in immune cells development and function. Their expression is altered in different physiological and disease conditions, hence making them attractive targets for the understanding of disease etiology and the development of adjunctive control strategies, especially within the current context of mitigated success of control measures deployed to eradicate these diseases. In this review, we summarize our current understanding of the role of ncRNAs in the etiology and control of major human tropical diseases including tuberculosis, HIV/AIDS and malaria, as well as neglected tropical diseases including leishmaniasis, African trypanosomiasis and leprosy. We highlight that several ncRNAs are involved at different stages of development of these diseases, for example miR-26-5p, miR-132-3p, miR-155-5p, miR-29-3p, miR-21-5p, miR-27b-3p, miR-99b-5p, miR-125-5p, miR-146a-5p, miR-223-3p, miR-20b-5p, miR-142-3p, miR-27a-5p, miR-144-5p, miR-889-5p and miR-582-5p in tuberculosis; miR-873, MALAT1, HEAL, LINC01426, LINC00173, NEAT1, NRON, GAS5 and lincRNA-p21 in HIV/AIDS; miR-451a, miR-let-7b and miR-106b in malaria; miR-210, miR-30A-5P, miR-294, miR-721 and lncRNA 7SL RNA in leishmaniasis; and miR-21, miR-181a, miR-146a in leprosy. We further report that several ncRNAs were investigated as diseases biomarkers and a number of them showed good potential for disease diagnosis, including miR-769-5p, miR-320a, miR-22-3p, miR-423-5p, miR-17-5p, miR-20b-5p and lncRNA LOC152742 in tuberculosis; miR-146b-5p, miR-223, miR-150, miR-16, miR-191 and lncRNA NEAT1 in HIV/AIDS; miR-451 and miR-16 in malaria; miR-361-3p, miR-193b, miR-671, lncRNA 7SL in leishmaniasis; miR-101, miR-196b, miR-27b and miR-29c in leprosy. Furthermore, some ncRNAs have emerged as potential therapeutic targets, some of which include lncRNAs NEAT1, NEAT2 and lnr6RNA, 152742 in tuberculosis; MALAT1, HEAL, SAF, lincRNA-p21, NEAT1, GAS5, NRON, LINC00173 in HIV/AIDS; miRNA-146a in malaria. Finally, miR-135 and miR-126 were proposed as potential targets for the development of therapeutic vaccine against leishmaniasis. We also identify and discuss knowledge gaps that warrant for increased research work. These include investigation of the role of ncRNAs in the etiology of African trypanosomiasis and the assessment of the diagnostic potential of ncRNAs for malaria, and African trypanosomiasis. The potential targeting of ncRNAs for adjunctive therapy against tuberculosis, leishmaniasis, African trypanosomiasis and leprosy, as well as their targeting in vaccine development against tuberculosis, HIV/AIDS, malaria, African trypanosomiasis and leprosy are also new avenues to explore.
Asunto(s)
MicroARNs/inmunología , Enfermedades Desatendidas , ARN Largo no Codificante/inmunología , Regulación hacia Arriba/inmunología , Desarrollo de Vacunas , Proliferación Celular , Humanos , Enfermedades Desatendidas/inmunología , Enfermedades Desatendidas/prevención & controlRESUMEN
BACKGROUND: Pair-rule genes were identified and named for their role in segmentation in animal embryos. Paired, a homolog of mammalian PAX3, acts as one of several pair-rule genes and is key in defining the boundaries of future parasegments and segments during insect embryogenesis. RESULTS: We cloned the paired gene from the oriental fruit fly, Bactrocera dorsalis, and then applied CRISPR/Cas9-mediated genome editing to investigate its physiological function in the embryonic stage of this pest. We identified one transcript for a paired homolog in B. dorsalis, which encodes a protein containing a Paired Box domain and a Homeobox domain. Phylogenetic analysis confirmed that the paired gene is highly conserved and the gene was highly expressed at the 12-14 h-old embryonic stage. Knock-out of Bdpaired led to lack of segment boundaries, cuticular deficiency, and embryonic lethality. Sequence analysis of the CRISPR/Cas9 mutants exhibited different insertion and deletions in the Bdpaired locus. In addition, the relative expression of Wingless (Wg) and Abdominal A (Abd-A) genes were significantly down-regulated in the Bdpaired mutant embryos. CONCLUSION: These results indicate that Bdpaired gene is critical for the embryonic development of B. dorsalis, and could be a novel molecular target for genetic-based pest management practices to combat this serious invasive pest. © 2019 Society of Chemical Industry.
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Tephritidae , Animales , Drosophila , Proteínas de Insectos , Mutación , FilogeniaRESUMEN
Bacillus thuringiensis (Bt) produce diverse insecticidal proteins to kill insect pests. Nevertheless, evolution of resistance to Bt toxins hampers the sustainable use of this technology. Previously, we identified down-regulation of a trypsin-like serine protease gene PxTryp_SPc1 in the midgut transcriptome and RNA-Seq data of a laboratory-selected Cry1Ac-resistant Plutella xylostella strain, SZ-R. We show here that reduced PxTryp_SPc1 expression significantly reduced caseinolytic and trypsin protease activities affecting Cry1Ac protoxin activation, thereby conferring higher resistance to Cry1Ac protoxin than activated toxin in SZ-R strain. Herein, the full-length cDNA sequence of PxTryp_SPc1 gene was cloned, and we found that it was mainly expressed in midgut tissue in all larval instars. Subsequently, we confirmed that the PxTryp_SPc1 gene was significantly decreased in SZ-R larval midgut and was further reduced when selected with high dose of Cry1Ac protoxin. Moreover, down-regulation of the PxTryp_SPc1 gene was genetically linked to resistance to Cry1Ac in the SZ-R strain. Finally, RNAi-mediated silencing of PxTryp_SPc1 gene expression decreased larval susceptibility to Cry1Ac protoxin in the susceptible DBM1Ac-S strain, supporting that low expression of PxTryp_SPc1 gene is involved in Cry1Ac resistance in P. xylostella. These findings contribute to understanding the role of midgut proteases in the mechanisms underlying insect resistance to Bt toxins.
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Toxinas de Bacillus thuringiensis/toxicidad , Endotoxinas/toxicidad , Proteínas Hemolisinas/toxicidad , Proteínas de Insectos/genética , Resistencia a los Insecticidas/genética , Insecticidas/toxicidad , Mariposas Nocturnas/genética , Tripsina/genética , Animales , Quimotripsina/metabolismo , Femenino , Tracto Gastrointestinal , Larva/efectos de los fármacos , Larva/genética , Masculino , Mariposas Nocturnas/efectos de los fármacos , Control Biológico de Vectores , Filogenia , Tripsina/metabolismoRESUMEN
Insects possess highly developed olfactory systems which play pivotal roles in its ecological adaptations, host plant location, and oviposition behavior. Bactrocera minax is an oligophagous tephritid insect whose host selection, and oviposition behavior largely depend on the perception of chemical cues. However, there have been very few reports on molecular components related to the olfactory system of B. minax. Therefore, the transcriptome of B. minax were sequenced in this study, with 1 candidate chemosensory protein (CSP), 21 candidate odorant binding proteins (OBPs), 53 candidate odorant receptors (ORs), 29 candidate ionotropic receptors (IRs) and 4 candidate sensory neuron membrane proteins (SNMPs) being identified. After that, we sequenced the candidate olfactory genes and performed phylogenetic analysis. qRT-PCR was used to express and characterize 9 genes in olfactory and non-olfactory tissues. Compared with GFP-injected fly (control), dsOBP21-treated B. minax and dsCSP-treated B. minax had lower electrophysiological response to D-limonene (attractant), suggesting the potential involvement of BminOBP21 and BminCSP genes in olfactory perceptions of the fly. Our study establishes the molecular basis of olfaction, tributary for further functional analyses of chemosensory processes in B. minax.
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Percepción Olfatoria/genética , Olfato/genética , Tephritidae/genética , Animales , Secuencia de Bases/genética , Femenino , Perfilación de la Expresión Génica/métodos , Genes de Insecto/genética , Proteínas de Insectos/genética , Insectos/genética , Masculino , Filogenia , Transcriptoma/genéticaRESUMEN
Crop straw is an abundant renewable resource whose usage is limited due to its high cellulose, hemicellulose, and lignin contents. Here, Trichoderma viride, Saccharomyces cerevisiae, and Musca domestica were used to transform crop straws, and we investigated their impact on housefly rearing performance and optimized their utilization. The weights of cellulose, hemicellulose, and lignin in fermented crop straw diets significantly decreased after bioconversion by M. domestica larvae. The highest bioconversion rate was recorded in corn straw diet (16.19%), followed by wheat straw diet (10.31%) and wheat bran diet (8.97%). Similarly, high larval weight (yield) and pupation rate and fecundity and fertility rate were recorded in fermented crop straw diets composed of corn straw and wheat bran in 1:1 proportions. These results indicated that fermenting crop straw with T. viride and S. cerevisiae represented an efficient strategy that enhanced crop straw bioconversion and improved the rearing capacity of the housefly larvae. The resulting larvae could further be used as proteinaceous feed in poultry and aquaculture industries. Graphical abstract.
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Celulosa/metabolismo , Lignina/metabolismo , Muscidae/química , Saccharomyces cerevisiae/metabolismo , Trichoderma/metabolismo , Triticum/microbiología , Animales , Reactores Biológicos , Celulosa/química , Fermentación , Moscas Domésticas , Larva/metabolismo , Lignina/química , Polisacáridos , Saccharomyces cerevisiae/química , Trichoderma/química , Triticum/química , Triticum/metabolismoRESUMEN
The gut microbiome of insects directly or indirectly affects the metabolism, immune status, sensory perception and feeding behavior of its host. Here, we examine the hypothesis that in the oriental fruit fly (Bactrocera dorsalis, Diptera: Tephritidae), the presence or absence of gut symbionts affects foraging behavior and nutrient ingestion. We offered protein-starved flies, symbiotic or aposymbiotic, a choice between diets containing all amino acids or only the non-essential ones. The different diets were presented in a foraging arena as drops that varied in their size and density, creating an imbalanced foraging environment. Suppressing the microbiome resulted in significant changes of the foraging behavior of both male and female flies. Aposymbiotic flies responded faster to the diets offered in experimental arenas, spent more time feeding, ingested more drops of food, and were constrained to feed on time-consuming patches (containing small drops of food), when these offered the full complement of amino acids. We discuss these results in the context of previous studies on the effect of the gut microbiome on host behavior, and suggest that these be extended to the life history dimension.
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Conducta Animal/fisiología , Conducta Alimentaria/fisiología , Microbioma Gastrointestinal/fisiología , Tephritidae/fisiología , Animales , Femenino , Larva/fisiología , Masculino , Pupa/fisiología , Simbiosis/fisiología , Tephritidae/microbiologíaRESUMEN
Bacteria inhabiting the gut of insects provide many benefits to their hosts, such as aiding in food digestion, reproduction, and immunity, tissue homeostasis, adaptation to environment and resistance to pathogen and pesticides. The cowpea beetle, Callosobruchus maculatus, is a serious cosmopolitan pest of pulses. This beetle has lent itself as a guinea pig for several ecological studies. It harbors a consortium of bacterial communities in its gut, but the evidence for their role in its physiology is fragmentary. In this work, we hypothesized that gut microbiota mediates C. maculatus resistance to dichlorvos (DDVP or O,O-dimethyl O-2,2-dichlorovinylphosphate) and represent the target of Lippia adoensis (Gambian Tea Bush) essential oil (EO). Symbiotic and aposymbiotic beetles were exposed to artificial cowpea seeds earlier treated with DDVP or EO. Adult mortality and changes in gut bacterial community composition and abundance were examined at F1 and F5 generations. The susceptibility of experimental beetles to DDVP was significantly affected by their symbiotic status. The adult mortality decreased across generations in DDVP treatments, and remained significantly higher in aposymbiotic groups. In EO treatments, the mortality was consistent irrespective of symbiotic status and experimental generations. When compared to DDVP and the Control, EO treatments had significantly lower bacterial richness and diversity, as well as lower abundance of Proteobacteria, Firmicutes, and Bacteroidetes. These results support our hypothesis and describe the responses of gut microbial communities to pesticide treatments. This could be of interest for developing new management strategies of this pest.
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Escarabajos/efectos de los fármacos , Diclorvos/farmacología , Microbioma Gastrointestinal/efectos de los fármacos , Insecticidas/farmacología , Lippia/química , Aceites Volátiles/farmacología , Animales , Bacteroidetes/clasificación , Bacteroidetes/efectos de los fármacos , Bacteroidetes/aislamiento & purificación , Bacteroidetes/fisiología , Escarabajos/microbiología , Escarabajos/fisiología , Firmicutes/clasificación , Firmicutes/efectos de los fármacos , Firmicutes/aislamiento & purificación , Firmicutes/fisiología , Microbioma Gastrointestinal/fisiología , Tracto Gastrointestinal/microbiología , Tracto Gastrointestinal/fisiología , Resistencia a los Insecticidas/fisiología , Longevidad/efectos de los fármacos , Aceites Volátiles/aislamiento & purificación , Proteobacteria/clasificación , Proteobacteria/efectos de los fármacos , Proteobacteria/aislamiento & purificación , Proteobacteria/fisiología , Semillas/parasitología , Simbiosis/fisiología , Vigna/parasitologíaRESUMEN
To examine how nutritional quality and resident gut bacteria interplay in improving the fitness of an oligophagous fruit fly, Bactrocera minax, artificial sucrose diets and full diets (sucrose, tryptone and yeast extract) were fed to flies with and without antibiotic supplementation. Furthermore, Klebsiella oxytoca and Citrobacter freundii were supplemented to sucrose-only diets. Flies were maintained in the laboratory and the fitness parameters, male and female longevity, number of copulations and female fecundity, were recorded. Full diet without bacterial depletion significantly increased fecundity and copulation. In the absence of gut bacteria, flies fed with full diets had significantly decreased mean fecundity and copulation rate. Flies that were fed with sucrose diet had a very low copulation rate and produced no eggs. Diet type and the presence of bacteria did not have any effect on the average longevity of male and female flies. Bacterial supplementation in sucrose diets did not improve any of the measured parameters. The results demonstrate that gut bacteria interact with diet to influence mating and reproduction in B. minax. Symbiotic bacteria significantly and positively impact reproduction in B. minax; however, their impact can only be fully realized when the flies are fed with a nutritionally complete diet.
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The intensive use of synthetic pesticides in cowpea storage has led to the development of resistance by Callosobruchus maculatus and subsequent degradation of grain quality. In an attempt to circumvent these constraints, the susceptibility of C. maculatus to 2,2-dichlorovinyldimethyl phosphate (DDVP) and Lippia adoensis essential oil (EO) was investigated and variations in the proportions of nutritional values of treated grains 150 days after storage were assessed. The survival rate was recorded after five generations. The resistance index and biochemical parameters of grains were determined. The results from this study revealed that the survival rate and resistance index significantly increased proportionally with damage in DDVP treatments (r = 0.889; p = 0.018) while in EO treatments, those values remained low without significant variations (p = 0.0764) throughout the generations. DDVP stored grains yielded higher crude protein values, but lower carbohydrates, tannins, phenolics and minerals compared to EO. Eighteen amino acids were detected in EO treated grains and 14 in DDVP which was devoid of albumin and prolamin. Lippia adoensis EO could therefore represent a safe alternative bio-pesticide to cope with insect resistance and enhance the nutritional qualities of stored cowpea seeds.