RESUMO
A significant number of microorganisms inhabit the intestinal tract or the body surface of insects. While the majority of research on insect microbiome interaction has mainly focused on bacteria, of late multiple studies have been acknowledging the importance of fungi and have started reporting the fungal communities as well. In this study, high-throughput sequencing was used to compare the diversity of intestinal fungi in Delia antiqua (Diptera: Anthomyiidae) at different growth stages, and effect of differential fungi between adjacent life stages on the growth and development of D. antiqua was investigated. The results showed that there were significant differences in the α and ß diversity of gut fungal communities between two adjacent growth stages. Among the dominant fungi, genera Penicillium and Meyerozyma and family Cordycipitaceae had higher abundances. Cordycipitaceae was mainly enriched in the pupal and adult (male and female) stages, Penicillium was mainly enriched in the pupal, 2nd instar and 3rd instar larval stages, and Meyerozyma was enriched in the pupal stage. Only three fungal species were found to differ between two adjacent growth stages. These three fungal species including Fusarium oxysporum, Meyerozyma guilliermondii and Penicillium roqueforti generally inhibited the growth and development of D. antiqua, with only P. roqueforti promoting the growth and development of female insects. This study will provide theoretical support for the search for new pathogenic microorganisms for other fly pests control and the development of new biological control strategies for fly pests.
Assuntos
Dípteros , Fungos , Microbioma Gastrointestinal , Larva , Animais , Dípteros/microbiologia , Dípteros/crescimento & desenvolvimento , Fungos/classificação , Fungos/isolamento & purificação , Fungos/genética , Larva/microbiologia , Larva/crescimento & desenvolvimento , Masculino , Feminino , Pupa/microbiologia , Pupa/crescimento & desenvolvimento , Biodiversidade , Estágios do Ciclo de Vida , MicobiomaRESUMO
INTRODUCTION AND OBJECTIVE: Hippobosca equina (Diptera: Hippoboscidae), is a widespread blood-feeding ectoparasite associated with the forest ecosystem. The insect is characterized by a wide host range and low host specificity, which increases the risk of feeding on animals that constitute a reservoir of transmissible pathogens, including Bartonella spp. MATERIAL AND METHODS: Hippobosca equina adults were collected from humans and companion animals within a continental mesotrophic oak-pine mixed forest in eastern Poland. DNA was isolated by the ammonia method, and isolates obtained from single individuals were tested by PCR method for the presence of 5 vector-borne pathogens. In case of the positive results, the amplicons were sequenced and examined by a BLAST search. RESULTS: The PCR analysis of DNA isolates obtained from 100 H. equina specimens revealed the presence of the RNA polymerase beta-subunit gene (rpoB) of the genus Bartonella, in 1% of the studied insects, i.e. one H. equina female. The rpoB gene haplotype of Bartonella sp. reported in this study, was identical to a Bartonella sp. sequence obtained from deer keds in Lithuania, and very closely related to strains with zoonotic potential. None of the H. equina specimens studied was positive for the presence of B. burgdorferi s.l., Anaplasma phagocytophilum, Babesia spp., and Coxiella burnetii. CONCLUSIONS: The study indicates the need to screen the occurrence of Bartonella spp., both in potential vectors and reservoirs of this pathogen in various habitats.
Assuntos
Bartonella , Dípteros , Animais , Polônia , Bartonella/isolamento & purificação , Bartonella/genética , Bartonella/classificação , Dípteros/microbiologia , Feminino , Infecções por Bartonella/veterinária , Infecções por Bartonella/microbiologia , Infecções por Bartonella/epidemiologia , Humanos , MasculinoRESUMO
Wolbachia are common heritable endosymbionts that influence many aspects of ecology and evolution in various insects, yet Wolbachia-mediated intracellular metabolic responses to temperature stress have been largely overlooked. Here, we introduced the Wolbachia strain wLhui from the invasive Liriomyza huidobrensis (Blanchard) into a Drosophila Schneider 2 cell line (S2) and investigated the metabolite profile of wLhui-infected (S2_wLhui) and uninfected cell lines (S2_wu) under short-term exposure to either high (37°C), moderate (27°C), or low (7 and 17°C) temperatures. We find that Wolbachia infection, temperature stress, and their interactions significantly affect cellular metabolic profiles. Most significantly, when comparing the changes in metabolites between S2_wLhui and S2_wu, glycerophospholipids, amino acids, and fatty acids associated with metabolic pathways, microbial metabolism in diverse environments, and other pathways were significantly accumulated at either low or high temperatures. Our findings suggest Wolbachia-induced cellular physiological responses to short-term temperature stress, which may in turn affect the fitness and adaptive ability of its host as an invasive species.
Assuntos
Metaboloma , Estresse Fisiológico , Temperatura , Wolbachia , Wolbachia/metabolismo , Wolbachia/fisiologia , Wolbachia/genética , Animais , Linhagem Celular , Drosophila/microbiologia , Simbiose , Dípteros/microbiologia , Ácidos Graxos/metabolismoRESUMO
Non-culture based surveys show that the bacterial family Orbaceae is widespread in guts of insects, including wild Drosophila. Relatively few isolates have been described, and none has been described from Drosophila. We present the isolation and characterization of five strains of Orbaceae from wild-caught flies of the genera Drosophila (Diptera: Drosophilidae) and Neogriphoneura (Diptera: Lauxaniidae). Cells are generally rod-shaped, mesophilic, and measure 0.8-2.0 µm long by 0.3-0.5 µm wide. Optimal growth was observed under ambient atmosphere. Reconstruction of phylogenies from the 16S rRNA gene and from single-copy orthologs verify placement of these strains within Orbaceae. Cells exhibited similar fatty acid profiles to those of other Orbaceae. Strain lpD01T shared 74% average nucleotide identity (ANI) with its closest relatives Ca. Schmidhempelia bombi Bimp and Zophobihabitans entericus IPMB12T. Results from multiple genome-wide similarity comparisons indicate lpD01T should be classified as a novel species within a novel genus. The major respiratory quinone for lpD01T is ubiquinone Q-8. lpD02T, lpD03, lpD04T, and BiBT are more closely related to Orbus hercynius CN3T (76, 77, 76, and 77% ANI, respectively) than to other described Orbaceae. Genomic and phylogenetic analyses suggest that lpD03 and lpD04T belong to the same species and that lpD02T, lpD03/lpD04T, and BiBT are each novel species of the genus Orbus. The proposed names of these strains are Utexia brackfieldae gen. nov., sp. nov. (type strain lpD01T =NCIMB 15517T =ATCC TSD-399T), Orbus sturtevantii sp. nov (type strain lpD02T =NCIMB 15518T =ATCC TSD-400T), Orbus wheelerorum sp. nov. (type strain lpD04T =NCIMB 15520T =ATCC TSD-401T), and Orbus mooreae sp. nov (type strain BiBT=NCIMB 15516T =ATCC TSD-402T). The isolation and characterization of these strains expands the repertoire of culturable bacteria naturally associated with insects, including the model organism D. melanogaster.
Assuntos
Técnicas de Tipagem Bacteriana , DNA Bacteriano , Dípteros , Drosophila , Ácidos Graxos , Filogenia , RNA Ribossômico 16S , Análise de Sequência de DNA , Simbiose , Animais , RNA Ribossômico 16S/genética , DNA Bacteriano/genética , Ácidos Graxos/análise , Dípteros/microbiologia , Drosophila/microbiologia , Composição de Bases , Microbioma GastrointestinalRESUMO
An entomopathogenic nematode, Oscheius tipulae, was isolated from a soil sample. The identification of this species was supported by morphological and molecular markers. The nematode isolate exhibited pathogenicity against different target insects including lepidopteran, coleopteran, and dipteran insects. The virulence of this nematode was similar to that of a well-known entomopathogenic nematode, Steinernema carpocapsae, against the same insect targets. A comparative metagenomics analysis of these two nematode species predicted the existence of a combined total of 272 bacterial species in their intestines, of which 51 bacterial species were shared between the two nematode species. In particular, the common gut bacteria included several entomopathogenic bacteria including Xenorhabdus nematophila, which is known as a symbiotic bacterium to S. carpocapsae. The nematode virulence of O. tipulae to insects was enhanced by an addition of dexamethasone but suppressed by an addition of arachidonic acid, suggesting that the immune defenses of the target insects against the nematode infection is mediated by eicosanoids, which would be manipulated by the symbiotic bacteria of the nematode. Unlike S. carpocapsae, O. tipulae showed high virulence against dipteran insects including fruit flies, onion flies, and mosquitoes. O. tipulae showed particularly high control efficacies against the onion maggot, Delia platura, infesting the Welsh onion in the rhizosphere in both pot and field assays.
Assuntos
Dípteros , Animais , Dípteros/microbiologia , Controle Biológico de Vetores , Rabditídios/patogenicidade , Rabditídios/fisiologia , Virulência , Simbiose , Nematoides , Xenorhabdus/genética , Xenorhabdus/patogenicidade , Xenorhabdus/fisiologiaRESUMO
Brewery by-products are recognised as suitable rearing substrates for Hermetia illucens, better known as black soldier fly (BSF) but information about the impact of different ratios of brewer's spent grains (BSG) and brewer's spent yeast (BSY) are still scarce. This study evaluated the effects of BSG-BSY-based diets on BSF larval growth, survival, bioconversion efficiency, nutritional profile, and microbiota and mycobiota. A total of 3 000 6-day-old BSF larvae were allotted to five dietary treatments (six replicate boxes/diet, 100 larvae/box): (i) BSY2.5 (25 g/kg of BSY+975 g/kg of BSG), (ii) BSY5 (50 g/kg of BSY+950 g/kg of BSG), (iii) BSY7.5 (75 g/kg of BSY+925 g/kg of BSG), (iv) BSY10 (100 g/kg of BSY+900 g/kg of BSG), and (v) control (Gainesville diet). Larval weight and substrate pH were recorded every 4 days. At the end of the trial (5% of prepupae), bioconversion efficiency corrected for residue (BER), reduction rate (RR), and waste reduction index (WRI) were calculated, and the larval proximate composition, microbiota and mycobiota characterised. At 10 and 14 days of age, BSY7.5 and BSY10 larvae displayed higher weight than BSY2.5 and BSY5 (P < 0.05), with BSY10 larvae showing the highest weight among the BSG-BSY-based diets at the end of the trial (P < 0.05). The BSY7.5 and BSY10 larvae also displayed a better BER than BSY2.5 and BSY5 (P < 0.01), whereas similar RR, WRI, survival and development time, as well as pH, were, however, observed among the BSG-BSY-based diets (P > 0.05). The BSY10 larvae displayed lower ether extract content than the other BSG-BSY-based diets (P > 0.001). The use of BSG-BSY-based diets did not influence the alpha diversity of larval microbiota and mycobiota (P > 0.05), but a specific microbial signature was identified per each dietary treatment (Porphyromonadaceae [BSY5], Sphingomonas [BSY7.5], Bacillus [BSY10] and Ruminococcus and Myroides [BSG-BSY-based diets]; P < 0.05). Co-occurrence and co-exclusion analysis also showed that Saccharomyces cerevisiae and Pichia excluded and favoured, respectively, the presence of Streptomyces and Fluviicola, while Clavispora lusitaniae was associated with Myroides (P < 0.05). In conclusion, BSG-BSY-based diets are suitable for rearing HI in terms of larval performance, nutritional profile, and microbiota and mycobiota, with 7.5 and 10% of BSY inclusion levels being able to improve larval growth and bioconversion efficiency.
Assuntos
Ração Animal , Dieta , Larva , Microbiota , Animais , Larva/crescimento & desenvolvimento , Larva/microbiologia , Ração Animal/análise , Dieta/veterinária , Microbiota/efeitos dos fármacos , Simuliidae/crescimento & desenvolvimento , Dípteros/crescimento & desenvolvimento , Dípteros/microbiologia , Micobioma , Grão Comestível/química , Fenômenos Fisiológicos da Nutrição AnimalRESUMO
Introduction: Hypervirulent Klebsiella pneumoniae (hvKp) and carbapenem-resistant K. pneumoniae (CR-Kp) are rapidly emerging as opportunistic pathogens that have a global impact leading to a significant increase in mortality rates among clinical patients. Anti-virulence strategies that target bacterial behavior, such as adhesion and biofilm formation, have been proposed as alternatives to biocidal antibiotic treatments to reduce the rapid emergence of bacterial resistance. The main objective of this study was to examine the efficacy of fatty acid-enriched extract (AWME3) derived from the fat of Black Soldier Fly larvae (Hermetia illucens) in fighting against biofilms of multi-drug resistant (MDR) and highly virulent Klebsiella pneumoniae (hvKp) pathogens. Additionally, the study also aimed to investigate the potential mechanisms underlying this effect. Methods: Crystal violet (CV) and ethidium bromide (EtBr) assays show how AWME3 affects the formation of mixed and mature biofilms by the KP ATCC BAA-2473, KPi1627, and KPM9 strains. AWME3 has shown exceptional efficacy in combating the hypermucoviscosity (HMV) virulent factors of KPi1627 and KPM9 strains when tested using the string assay. The rudimentary motility of MDR KPM9 and KP ATCC BAA-2473 strains was detected through swimming, swarming, and twitching assays. The cell wall membrane disturbances induced by AWME3 were detected by light and scanning electron microscopy and further validated by an increase in the bacterial cell wall permeability and Lewis acid-base/van der Waals characteristics of K. pneumoniae strains tested by MATS (microbial adhesion to solvents) method. Results: After being exposed to 0.5 MIC (0.125 mg/ml) of AWME3, a significant reduction in the rudimentary motility of MDR KPM9 and KP ATCC BAA-2473 strains, whereas the treated bacterial strains exhibited motility between 4.23 ± 0.25 and 4.47 ± 0.25 mm, while the non-treated control groups showed significantly higher motility ranging from 8.5 ± 0.5 to 10.5 ± 0.5 mm. Conclusion: In conclusion, this study demonstrates the exceptional capability of the natural AWME3 extract enriched with a unique combination of fatty acids to effectively eliminate the biofilms formed by the highly drug-resistant and highly virulent K. pneumoniae (hvKp) pathogens. Our results highlight the opportunity to control and minimize the rapid emergence of bacterial resistance through the treatment using AWME3 of biofilm-associated infections caused by hvKp and CRKp pathogens.
Assuntos
Antibacterianos , Biofilmes , Dípteros , Farmacorresistência Bacteriana Múltipla , Ácidos Graxos , Klebsiella pneumoniae , Larva , Fatores de Virulência , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Animais , Klebsiella pneumoniae/efeitos dos fármacos , Klebsiella pneumoniae/patogenicidade , Ácidos Graxos/metabolismo , Fatores de Virulência/metabolismo , Dípteros/microbiologia , Larva/microbiologia , Larva/efeitos dos fármacos , Antibacterianos/farmacologia , Testes de Sensibilidade Microbiana , Virulência/efeitos dos fármacos , Infecções por Klebsiella/microbiologia , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismoRESUMO
Rust fungi (Pucciniales) are plant pathogens that can cause devastating yield losses to economically important crops and threaten native plants with extinction. Rusts are usually controlled with fungicides when rust-resistant plant varieties are unavailable. However, natural enemies may offer an alternative to chemicals by acting as biological controls. The larvae of Mycodiplosis Rübsaamen (49 spp.) feed on the spores of rusts and powdery mildew fungi and have been suggested as a potential biocontrol candidate for disease-causing rusts. However, little is known about the phylogenetic relationships, biogeography, and host range of this genus. We screened 5,665 rust specimens from fungarium specimens and field collections and recovered a total of 363 larvae on 315 rust specimens from 17 countries. Three mitochondrial and 2 nuclear loci were amplified and sequenced for the phylogenetic reconstruction of 129 individuals. We recovered 12 clades, of which 12 and 10 were supported with maximum likelihood and Bayesian inference, respectively. Of the 12 clades, 7 comprised species from multiple continents and climatic regions, and 5 comprised species from a single region. Individuals forming clades were collected from 2 to 18 rust species, suggesting that Mycodiplosis species have a broad host range. In total, Mycodiplosis larvae were identified on 44 different rust species collected from 18 plant families. Future studies should focus on expanding field sampling efforts, including data from additional gene regions, and incorporating morphological data to further elucidate species diversity and distribution patterns.
Assuntos
Basidiomycota , Especificidade de Hospedeiro , Larva , Filogenia , Animais , Basidiomycota/fisiologia , Basidiomycota/genética , Larva/microbiologia , Larva/crescimento & desenvolvimento , Larva/fisiologia , Dípteros/microbiologia , Filogeografia , Esporos Fúngicos/fisiologiaRESUMO
Coelopidae (Diptera), known as kelp flies, exhibit an ecological association with beached kelp and other rotting seaweeds. This unique trophic specialization necessitates significant adaptations to overcome the limitations of an algal diet. We aimed to investigate whether the flies' microbiome could be one of these adaptive mechanisms. Our analysis focused on assessing composition and diversity of adult and larval microbiota of the kelp fly Coelopa frigida. Feeding habits of the larvae of this species have been subject of numerous studies, with debates whether they directly consume kelp or primarily feed on associated bacteria. By using a 16S rRNA metabarcoding approach, we found that the larval microbiota displayed considerably less diversity than adults, heavily dominated by only four operational taxonomic units (OTUs). Phylogenetic placement recovered the most dominant OTU of the larval microbiome, which is the source of more than half of all metabarcoding sequence reads, as an undescribed genus of Orbaceae (Gammaproteobacteria). Interestingly, this OTU is barely found among the 15 most abundant taxa of the adult microbiome, where it is responsible for less than 2% of the metabarcoding sequence reads. The other three OTUs dominating the larval microbiome have been assigned as Psychrobacter (Gammaproteobacteria), Wohlfahrtiimonas (Gammaproteobacteria), and Cetobacterium (Fusobacteriota). Moreover, we also uncovered a distinct shift in the functional composition between the larval and adult stages, where our taxonomic profiling suggests a significant decrease in functional diversity in larval samples. Our study offers insights into the microbiome dynamics and functional composition of Coelopa frigida.
Assuntos
Bactérias , Dípteros , Larva , Microbiota , Filogenia , RNA Ribossômico 16S , Animais , Dípteros/microbiologia , Larva/microbiologia , RNA Ribossômico 16S/genética , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Código de Barras de DNA Taxonômico , Kelp/microbiologiaRESUMO
BACKGROUND: Flies are acknowledged as vectors of diseases transmitted through mechanical means and represent a significant risk to human health. The study aimed to determine the prevalence of enteropathogens carried by flies in Pudong New Area to inform strategies for preventing and controlling flies. METHODS: Samples were collected from various locations in the area using cage trapping techniques between April and November 2021, encompassing various habitats such as parks, residential areas, restaurants, and farmers' markets. The main fly species were identified using cryomicrography and taxonomic enumeration, with 20 samples per tube collected from different habitats. Twenty-five enteropathogens were screened using GI_Trial v3 TaqManTM microbial arrays. RESULTS: A total of 3,875 flies were collected from 6,400 placements, resulting in an average fly density of 0.61 flies per cage. M. domestica were the most common species at 39.85%, followed by L. sericata at 16.57% and B. peregrina at 13.14%. Out of 189 samples, 93 tested positive for enteropathogens, with nine different pathogens being found. 12.70% of samples exclusively had parasites, a higher percentage than those with only bacteria or viruses. The study found that M. domestica had fewer enteropathogens than L. sericata and B. peregrina, which primarily harbored B. hominis instead of bacteria and viruses such as E. coli, Astrovirus, and Sapovirus. During spring testing, all three fly species exhibited low rates of detecting enteropathogens. M. domestica were found in residential areas with the highest number of pathogen species, totaling six. In contrast, L. sericata and B. peregrina were identified in farmers' markets with the highest number of pathogen species, totaling six and seven, respectively. CONCLUSIONS: Flies have the potential to serve as vectors for the transmission of enteropathogens, thereby posing a substantial risk to public health.
Assuntos
Insetos Vetores , Animais , Humanos , Insetos Vetores/microbiologia , Bactérias/isolamento & purificação , Bactérias/classificação , Bactérias/genética , China/epidemiologia , Dípteros/microbiologia , Vírus/isolamento & purificação , Vírus/classificação , Vírus/genética , Muscidae/microbiologiaRESUMO
Hyperparasitism is defined as the interaction where one parasite is infected by another parasite. In bat flies (Streblidae and Nycteribiidae), both hyperparasites and microparasites (bacteria, viruses, fungi, and arthropods such as mites) have been documented. Fungi belonging to the order Laboulbeniales are microscopic parasites of a wide diversity of arthropod hosts. Three genera exclusively target bat flies: Arthrorhynchus, which parasitizes species within Nycteribiidae in the Eastern Hemisphere, while genus Gloeandromyces and Nycteromyces parasitize Streblidae in the Western Hemisphere. Among the hyperparasitic arthropods, mites of family Neothrombidiidae, particularly the monospecific genus Monunguis, are known to parasitize bat flies. Here we present the first records of the hyperparasites Monunguis streblida and Gloeandromyces pageanus f. polymorphus parasitizing Streblidae bat flies in Colombia and a summary of these hyperparasitic interactions in the Neotropics. We detected fungi and mites parasitizing bat flies that were collected in the Magdalena River Basin, Colombia, in field expeditions in 2018, 2022, and 2023. We identified 17 bat flies and two species of hyperparasites, specifically M. streblida and the fungi Gloeandromyces. Our search for reports of these interactions in the Neotropics revealed that seven species of Trichobius (Streblidae) are parasitized by M. streblida, whereas Paratrichobius longicrus (Streblidae) is parasitized by Gloeandromyces pageanus f. polymorphus. These interactions have been reported in 11 countries, but our records are the first of M. streblida and Laboulbeniales fungi parasitizing bat flies in Colombia. So far, a total of 14 species of fungi and one species of mite have been associated with 19 species of bat flies, which in turn, are linked to 15 species of Neotropical bats.
Assuntos
Quirópteros , Dípteros , Animais , Dípteros/microbiologia , Dípteros/parasitologia , Quirópteros/parasitologia , Colômbia , Ácaros/microbiologia , Ácaros/fisiologia , Interações Hospedeiro-ParasitaRESUMO
Hematophagous Stomoxys (stable) fly populations in dairy barns are sustained by a constant availability of cattle hosts and manure, which serve as major reservoirs of both zoonotic and opportunistic bacterial pathogens. However, the composition of the Stomoxys fly microbiota, the mechanisms by which flies acquire their microbiome, and the ability of potentially pathogenic bacteria to colonize and persist in fly hosts remain to be investigated. Here, we longitudinally collected fly and manure samples from two connected dairy facilities. High throughput 16S rRNA gene amplicon sequencing was then used to characterize and compare bacterial communities present on or within flies and in manure collected from the same facility, while culture-dependent methods were used to verify the viability of clinically relevant bacteria. Bacterial alpha diversity was overall higher in manure samples as compared to fly samples, with manure-associated bacterial communities being dominated by members of the Bacteroidales, Eubacteriales, and Oscillospirales. In contrast, flies harbored relatively low-complexity communities dominated by members of the Enterobacterales, Staphylococcales, and Lactobacillales. Clinically relevant bacterial strains, including Escherichia spp. and other taxa associated with mastitic cows housed in the same facilities, were detected in paired fly and manure samples but exhibited dramatically elevated abundances in fly samples as compared to manure samples. Viable colonies of Escherichia, Klebsiella, and Staphylococcus spp. were also readily isolated from fly samples, confirming that flies harbor culturable mastitis-associated bacteria. This study identifies biting flies as bona fide carriers of opportunistically pathogenic bacterial taxa on dairy farms. IMPORTANCE: Disease prevention on dairy farms has significant implications for cattle health, food security, and zoonosis. Of particular importance is the control of bovine mastitis, which can be caused by diverse bacteria, including Klebsiella, Escherichia coli, Streptococcus, and Staphylococcus spp. Despite being one of the most significant and costly cattle diseases worldwide, the epidemiology of bovine mastitis is not well understood. This study provides parallel culture-independent and culture-dependent evidence to support the carriage of opportunistically pathogenic bacteria by Stomoxys flies on dairy farms. We further show that the fly microbiota is enriched in clinically relevant taxa-the vast majority of which can be traced to the manure habitats in which flies breed. Altogether, our results identify biting flies as underrecognized carriers of bacterial taxa associated with environmental bovine mastitis and other opportunistic infections in vertebrates and offer important insights into mechanisms of microbial acquisition by these and other medically important insects.
Assuntos
Bactérias , Esterco , Mastite Bovina , Microbiota , RNA Ribossômico 16S , Animais , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Bovinos , Feminino , Esterco/microbiologia , RNA Ribossômico 16S/genética , Mastite Bovina/microbiologia , Microbiota/genética , Dípteros/microbiologia , Muscidae/microbiologiaRESUMO
In a world with a population exceeding 8 billion people and continuing to grow, pollution from food and plastic waste is causing long-term issues in ecosystems. Potential solutions may be found by exploiting insect-based bioconversion. In this context, we investigated the impact of polyvinyl chloride microparticles (PVC-MPs) on the development of Hermetia illucens (black soldier fly; BSF) and its midgut bacterial and fungal microbiota. The impact of PVC-MPs was evaluated feeding BSF larvae with a PVC-MPs-supplemented diet. The larvae exposed to different PVC-MPs concentrations (2.5%, 5%, 10% and 20% w/w) developed into adults with no significant increase in pupal mortality. Faster development and smaller pupae were observed when 20% PVC-MPs was provided. The BSF larvae ingest PVC-MPs, resulting in a reduction in MPs size. Larvae exposed to PVC-MPs did not exhibit differences in gut morphology. Regarding the impact of PVC-MPs on the structure of both bacterial and fungal communities, the overall alpha- and beta-diversity did not exhibit significant changes. However, the presence of PVC-MPs significantly affected the relative abundances of Enterobacteriaceae and Paenibacillaceae among the bacteria and of Dipodascaceae and Plectospharellaceae among the fungi (including yeast and filamentous life forms), suggesting that PVC-MP contamination has a taxa-dependent impact. These results indicate that BSF larvae can tolerate PVC-MPs in their diet, supporting the potential use of these insects in organic waste management, even in the presence of high levels of PVC-MP contamination.
Assuntos
Dípteros , Microbioma Gastrointestinal , Larva , Microplásticos , Animais , Larva/microbiologia , Dípteros/microbiologia , Microbioma Gastrointestinal/efeitos dos fármacos , Cloreto de Polivinila , Fungos/metabolismo , Bactérias/classificação , Bactérias/metabolismo , Dieta , MicobiomaRESUMO
Wolbachia is a maternally inherited intracellular bacterium that is considered to be the most plentiful endosymbiont found in arthropods. It reproductively manipulates its host to increase the chances of being transmitted to the insect progeny; and it is currently used as a means of suppressing disease vector populations or controlling vector-borne diseases. Studies of the dissemination and prevalence of Wolbachia among its arthropod hosts are important for its possible use as a biological control agent. The molecular identification of Wolbachia relies on different primers sets due to Wolbachia strain variation. Here, we screened for the presence of Wolbachia in a broad range of Brachycera fly species (Diptera), collected from different regions of Iran, using nine genetic markers (wsp, ftsZ, fbpA, gatB, CoxA, gltA, GroEL dnaA, and 16s rRNA), for detecting, assessing the sensitivity of primers for detection, and phylogeny of this bacterium. The overall incidence of Wolbachia among 22 species from six families was 27.3%. The most commonly positive fly species were Pollenia sp. and Hydrotaea armipes. However, the bacterium was not found in the most medically important flies or in potential human disease vectors, including Musca domestica, Sarcophaga spp., Calliphora vicinia, Lucilia sericata, and Chrysomya albiceps. The primer sets of 16s rRNA with 53.0% and gatB with 52.0% were the most sensitive primers for detecting Wolbachia. Blast search, phylogenetic, and MLST analysis of the different locus sequences of Wolbachia show that all the six distantly related fly species likely belonging to supergroup A. Our study showed some primer sets generated false negatives in many of the samples, emphasizing the importance of using different loci in detecting Wolbachia. The study provides the groundwork for future studies of a Wolbachia-based program for control of flies.
Assuntos
Dípteros , Filogenia , Wolbachia , Wolbachia/genética , Wolbachia/isolamento & purificação , Animais , Irã (Geográfico) , Dípteros/microbiologia , RNA Ribossômico 16S/genéticaRESUMO
Wolbachia, a group of Gram-negative symbiotic bacteria, infects nematodes and a wide range of arthropods. Diaphorina citri Kuwayama, the vector of Candidatus Liberibacter asiaticus (CLas) that causes citrus greening disease, is naturally infected with Wolbachia (wDi). However, the interaction between wDi and D. citri remains poorly understood. In this study, we performed a pan-genome analysis using 65 wDi genomes to gain a comprehensive understanding of wDi. Based on average nucleotide identity (ANI) analysis, we classified the wDi strains into Asia and North America strains. The ANI analysis, principal coordinates analysis (PCoA), and phylogenetic tree analysis supported that the D. citri in Florida did not originate from China. Furthermore, we found that a significant number of core genes were associated with metabolic pathways. Pathways such as thiamine metabolism, type I secretion system, biotin transport, and phospholipid transport were highly conserved across all analyzed wDi genomes. The variation analysis between Asia and North America wDi showed that there were 39,625 single-nucleotide polymorphisms (SNPs), 2153 indels, 10 inversions, 29 translocations, 65 duplications, 10 SV-based insertions, and 4 SV-based deletions. The SV-based insertions and deletions involved genes encoding transposase, phage tail tube protein, ankyrin repeat (ANK) protein, and group II intron-encoded protein. Pan-genome analysis of wDi contributes to our understanding of the geographical population of wDi, the origin of hosts of D. citri, and the interaction between wDi and its host, thus facilitating the development of strategies to control the insects and huanglongbing (HLB).
Assuntos
Genoma Bacteriano , Filogenia , Simbiose , Wolbachia , Wolbachia/genética , Wolbachia/classificação , Simbiose/genética , Animais , Ásia , América do Norte , Hemípteros/microbiologia , Hemípteros/genética , Dípteros/microbiologia , Dípteros/genética , Polimorfismo de Nucleotídeo ÚnicoRESUMO
Insects are frequently infected with heritable bacterial endosymbionts. Some of them confer resistance to parasitoids. Such defensive symbionts are sensitive to variation in temperature. Drawing predominantly from the literature on aphids and flies, we show that temperature can affect the reliability of maternal transmission and the strength of protection provided by defensive symbionts. Costs of infection with defensive symbionts can also be temperature-dependent and may even turn into benefits under extreme temperatures, for example, when defensive symbionts increase heat tolerance. Alone or in combination, these mechanisms can drive temperature-associated (latitudinal) clines of infection prevalence with defensive symbionts. This has important consequences for host-parasitoid coevolution, as the relative importance of host-encoded vs. symbiont-provided defenses will shift along such clines.
Assuntos
Simbiose , Temperatura , Animais , Insetos/fisiologia , Insetos/microbiologia , Insetos/parasitologia , Interações Hospedeiro-Parasita , Afídeos/fisiologia , Afídeos/microbiologia , Afídeos/parasitologia , Dípteros/fisiologia , Dípteros/microbiologiaRESUMO
Background: Aeromonas hydrophila (A. hydrophila) is a bacterium with zoonotic potential and is multidrug-resistant. It utilizes hemolysin and aerolysin to spread infection. Black soldier flies (BSFs) can be antibacterial because of the fatty acids it contains. Aims: This study aimed to investigate and compare the fatty acid profiles of BSF prepupae grown in fermented and nonfermented media using bioinformatics tools and assess their potential as antibacterial agents against A. hydrophila. Methods: The study used BSF prepupae reared on various organic substrates. BSF prepupae grown in fermented or nonfermented substrate were observed against fatty acid. The fatty acid analysis was performed using GC-MS. Fatty acids were analyzed statistically using the one-way ANOVA test with a 95% confidence level. Fatty acid bioactivity was predicted using the online PASS-two-way drug program. Molecular docking on BSF fatty acid compounds was analyzed with PyMol 2.2 and discovery Studio version 21.1.1. Results: The molecular docking test showed the strongest bond was oleic acid with aerolysin and linoleic acid with hemolysin. BSF prepupae grown on fermented media showed higher crude fat and saturated fatty acids (SFAs) but lower unsaturated fatty acids than nonfermented media. Conclusion: Black soldier fly prepupae, particularly those grown on fermented media, possess antibacterial activity against A. hydrophila through potential fatty acid-mediated inhibition of crucial virulence factors.
Assuntos
Aeromonas hydrophila , Ácidos Graxos , Fermentação , Aeromonas hydrophila/efeitos dos fármacos , Animais , Ácidos Graxos/metabolismo , Biologia Computacional , Antibacterianos/farmacologia , Dípteros/microbiologia , Proteínas Hemolisinas/metabolismo , Larva/microbiologia , Simulação de Acoplamento MolecularRESUMO
The edible fungus industry is one of the pillar industries in the Yunnan-Guizhou Plateau, China. The expansion of the planting scale has led to the release of various mushroom residues, such as mushroom feet, and other wastes, which are not treated adequately, resulting in environmental pollution. This study investigated the ability of black soldier fly (Hermetia illucens L.) larvae (BSFL) to degrade mushroom waste. Moreover, this study analyzed changes in the intestinal bacterial community and gene expression of BSFL after feeding on mushroom waste. Under identical feeding conditions, the remaining amount of mushroom waste in Pleurotus ostreatus treatment group was reduced by 18.66%, whereas that in Flammulina velutipes treatment group was increased by 31.08%. Regarding gut microbial diversity, compared with wheat bran-treated control group, Dysgonomonas, Providencia, Enterococcus, Pseudochrobactrum, Actinomyces, Morganella, Ochrobactrum, Raoultella, and Ignatzschineria were the most abundant bacteria in the midgut of BSFL in F. velutipes treatment group. Furthermore, Dysgonomonas, Campylobacter, Providencia, Ignatzschineria, Actinomyces, Enterococcus, Morganella, Raoultella, and Pseudochrobactrum were the most abundant bacteria in the midgut of BSFL in P. ostreatus treatment group. Compared with wheat bran-treated control group, 501 upregulated and 285 downregulated genes were identified in F. velutipes treatment group, whereas 211 upregulated and 43 downregulated genes were identified in P. ostreatus treatment group. Using Kyoto Encyclopedia of Genes and Genomes and Gene Ontology enrichment analyses, we identified 14 differentially expressed genes (DEGs) related to amino sugar and nucleotide sugar metabolism in F. velutipes treatment group, followed by 12 DEGs related to protein digestion and absorption. Moreover, in P. ostreatus treatment group, two DEGs were detected for fructose and mannose metabolism, and two were noted for fatty acid metabolism. These results indicate that feeding on edible mushroom waste can alter the intestinal microbial community structure of BSFL; moreover, the larval intestine can generate a corresponding feedback. These changes contribute to the degradation of edible mushroom waste by BSFL and provide a reference for treating edible mushroom waste using BSFL.
Assuntos
Agaricales , Microbioma Gastrointestinal , Larva , Pleurotus , Animais , Larva/microbiologia , Pleurotus/metabolismo , Agaricales/metabolismo , Agaricales/genética , Biodegradação Ambiental , Dípteros/microbiologia , Dípteros/metabolismo , Flammulina/metabolismo , Flammulina/genética , Bactérias/metabolismo , Bactérias/genética , Bactérias/classificaçãoRESUMO
BACKGROUND: Hippoboscid flies are bloodsucking arthropods that can transmit pathogenic microorganisms and are therefore potential vectors for pathogens such as Bartonella spp. These Gram-negative bacteria can cause mild-to-severe clinical signs in humans and animals; therefore, monitoring Bartonella spp. prevalence in louse fly populations appears to be a useful prerequisite for zoonotic risk assessment. METHODS: Using convenience sampling, we collected 103 adult louse flies from four ked species (Lipoptena cervi, n = 22; Lipoptena fortisetosa, n = 61; Melophagus ovinus, n = 12; Hippobosca equina, n = 8) and the pupae of M. ovinus (n = 10) in the federal state of Saxony, Germany. All the samples were screened by polymerase chain reaction (PCR) for Bartonella spp. DNA, targeting the citrate synthase gene (gltA). Subsequently, PCRs targeting five more genes (16S, ftsZ, nuoG, ribC and rpoB) were performed for representatives of revealed gltA genotypes, and all the PCR products were sequenced to identify the Bartonella (sub)species accurately. RESULTS AND CONCLUSIONS: The overall detection rates for Bartonella spp. were 100.0%, 59.1%, 24.6% and 75.0% in M. ovinus, L. cervi, L. fortisetosa and H. equina, respectively. All the identified bartonellae belong to the Bartonella schoenbuchensis complex. Our data support the proposed reclassification of the (sub)species status of this group, and thus we conclude that several genotypes of B. schoenbuchensis were detected, including Bartonella schoenbuchensis subsp. melophagi and Bartonella schoenbuchensis subsp. schoenbuchensis, both of which have previously validated zoonotic potential. The extensive PCR analysis revealed the necessity of multiple PCR approach for proper identification of the ruminant-associated bartonellae.
Assuntos
Bartonella , Dípteros , Ftirápteros , Humanos , Animais , Dípteros/genética , Dípteros/microbiologia , Ftirápteros/genética , DNA Bacteriano/genética , Bartonella/genética , Ruminantes/genética , DNA , Alemanha/epidemiologia , Reação em Cadeia da Polimerase/veterináriaRESUMO
Hermetia illucens larvae are recognized for their ability to mitigate or eliminate contaminants by biodegradation. However, the biodegradation characteristics of microplastics and phthalic acid esters plasticizers, as well as the role of larval gut microorganisms, have remained largely unrevealed. Here, the degradation kinetics of plasticizers, and biodegradation characteristics of microplastics were examined. The role of larval gut microorganisms was investigated. For larval development, microplastics slowed larval growth significantly (P < 0.01), but the effect of plasticizer was not significant. The degradation kinetics of plasticizers were enhanced, resulting in an 8.11 to 20.41-fold decrease in degradation half-life and a 3.34 to 3.82-fold increase in final degradation efficiencies, compared to degradation without larval participation. The depolymerization and biodeterioration of microplastics were conspicuously evident, primarily through a weight loss of 17.63 %-25.52 %, variation of chemical composition and structure, bio-oxidation and bioerosion of microplastic surface. The synergistic effect driven by larval gut microorganisms, each with various functions, facilitated the biodegradation. Specifically, Ignatzschineria, Paenalcaligenes, Moheibacter, Morganella, Dysgonomonas, Stenotrophomonas, Bacteroides, Sphingobacterium, etc., appeared to be the key contributors, owing to their xenobiotic biodegradation and metabolism functions. These findings offered a new perspective on the potential for microplastics and plasticizers biodegradation, assisted by larval gut microbiota.