RESUMO
High-throughput sequencing studies have shown that diet or antimicrobial treatments impact animal gut microbiota equilibrium. However, properties related to the gut microbial ecosystem stability, such as resilience, resistance, or functional redundancy, must be better understood. To shed light on these ecological processes, we combined advanced statistical methods with 16â¯S rRNA gene sequencing, functional prediction, and fitness analyses in the gut microbiota of the cockroach Blattella germanica subject to three periodic pulses of the antibiotic (AB) kanamycin (n=512). We first confirmed that AB did not significantly affect cockroaches' biological fitness, and gut microbiota changes were not caused by insect physiology alterations. The sex variable was examined for the first time in this species, and no statistical differences in the gut microbiota diversity or composition were found. The comparison of the gut microbiota dynamics in control and treated populations revealed that (1) AB treatment decreases diversity and completely disrupts the co-occurrence networks between bacteria, significantly altering the gut community structure. (2) Although AB also affected the genetic composition, functional redundancy would explain a smaller effect on the functional potential than on the taxonomic composition. (3) As predicted by Taylor's law, AB generally affected the most abundant taxa to a lesser extent than the less abundant taxa. (4) Taxa follow different trends in response to ABs, highlighting "resistant taxa," which could be critical for community restoration. (5) The gut microbiota recovered faster after the three AB pulses, suggesting that gut microbiota adapts to repeated treatments.
Assuntos
Antibacterianos , Bactérias , Blattellidae , Microbioma Gastrointestinal , Canamicina , RNA Ribossômico 16S , Animais , Microbioma Gastrointestinal/efeitos dos fármacos , Antibacterianos/farmacologia , RNA Ribossômico 16S/genética , Blattellidae/microbiologia , Bactérias/classificação , Bactérias/genética , Bactérias/efeitos dos fármacos , Bactérias/isolamento & purificação , Canamicina/farmacologia , Masculino , Feminino , Biodiversidade , Baratas/microbiologia , Baratas/efeitos dos fármacos , Filogenia , Adaptação Fisiológica , Sequenciamento de Nucleotídeos em Larga EscalaRESUMO
The abuse of chemical insecticides has led to strong resistance in cockroaches, and biopesticides with active ingredients based on insect pathogens have good development prospects; however, their slow effect has limited their practical application, and improving their effectiveness has become an urgent problem. In this study, the interaction between Serratia marcescens and Metarhizium anisopliae enhanced their virulence against Blattella germanica and exhibited a synergistic effect. The combination of S. marcescens and M. anisopliae caused more severe tissue damage and accelerated the proliferation of the insect pathogen. The results of high-throughput sequencing demonstrated that the gut microbiota was dysbiotic, the abundance of the opportunistic pathogen Weissella cibaria increased, and entry into the hemocoel accelerated the death of the German cockroaches. In addition, the combination of these two agents strongly downregulated the expression of Imd and Akirin in the IMD pathway and ultimately inhibited the expression of antimicrobial peptides (AMPs). S. marcescens released prodigiosin to disrupted the gut homeostasis and structure, M. anisopliae released destruxin to damaged crucial organs, opportunistic pathogen Weissella cibaria overproliferated, broke the gut epithelium and entered the hemocoel, leading to the death of pests. These findings will allow us to optimize the use of insect pathogens for the management of pests and produce more effective biopesticides.
Assuntos
Baratas , Microbioma Gastrointestinal , Metarhizium , Serratia marcescens , Animais , Serratia marcescens/patogenicidade , Serratia marcescens/fisiologia , Metarhizium/patogenicidade , Metarhizium/fisiologia , Microbioma Gastrointestinal/efeitos dos fármacos , Baratas/microbiologia , Prodigiosina/farmacologia , Micotoxinas/metabolismo , Blattellidae/microbiologia , Controle Biológico de Vetores/métodos , Virulência , DepsipeptídeosRESUMO
Cockroaches are significant pests worldwide, being important in medical, veterinary, and public health fields. Control of cockroaches is difficult because they have robust reproductive ability and high adaptability and are resistant to many insecticides. Wolbachia is an endosymbiont bacterium that infects the reproductive organs of approximately 70% of insect species and has become a promising biological agent for controlling insect pests. However, limited data on the presence or strain typing of Wolbachia in cockroaches are available. PCR amplification and sequencing of the wsp and gltA genes were used to study the presence, prevalence and molecular typing of Wolbachia in two main cockroach species, Blattella germanica (German cockroach) and Periplaneta americana (American cockroach), from different geographical locations of Iran. The Wolbachia endosymbiont was found only in 20.6% of German cockroaches while it was absent in American cockroach samples. Blast search and phylogenetic analysis revealed that the Wolbachia strain found in the German cockroach belongs to Wolbachia supergroup F. Further studies should investigate the symbiotic role of Wolbachia in cockroaches and determine whether lack of Wolbachia infection may increase this insect's ability to tolerate or acquire various pathogens. Results of our study provide a foundation for continued work on interactions between cockroaches, bacterial endosymbionts, and pathogens.
Assuntos
Blattellidae , Baratas , Periplaneta , Wolbachia , Animais , Periplaneta/microbiologia , Blattellidae/genética , Blattellidae/microbiologia , Wolbachia/genética , Filogenia , Baratas/microbiologia , AlérgenosRESUMO
Omnivorous synanthropic cockroaches, such as the German cockroach (Blattella germanica), are reservoirs and vectors of enteric bacterial pathogens. A lifestyle conducive to frequent encounters with high loads of diverse bacteria may have led to the evolution of unique innate immune systems in these insects. The innate immune response of insects relies largely on generalized mechanisms to sense and eliminate foreign microbes. However, analyses of the genomes of common synanthropic cockroaches previously revealed a repertoire of pathogen associated molecular pattern (PAMP) receptors and antimicrobial peptides (AMPs) that is significantly expanded relative to most holometabolous insect models and vectors, supporting the intriguing possibility that cockroaches may encode enhanced recognition within their immune system and may possess an enhanced capacity to fine tune innate immune responses. Investigating how cockroaches respond to infection with enterobacteria provides the opportunity to expand our fundamental knowledge of the regulation of insect innate immunity in a context that is biologically and medically relevant. German cockroaches can harbor both Salmonella enterica serovar Typhimurium and Escherichia coli in their gut without experiencing pathogenesis. The former colonizes the gut and replicates while the latter persists only transiently. We hypothesized that differences in the innate immune response may contribute to or result from the difference in infection dynamics between the two enterobacteria. To test this hypothesis, we used qRT-PCR to analyze expression of five genes encoding representative AMPs (Attacins, Blattellicin, Defensins) in the gut of German cockroaches 1 and 24â h after ingestion of live or heat-killed enterobacteria. We found that robust AMP expression was induced in response to ingestion of a live wild-type strain of S. Typhimurium, but not in response to live E. coli, heat-killed S. Typhimurium, or a live mutant strain of S. Typhimurium lacking type III secretion systems. These results indicate that the cockroach immune system does not respond to stimulation with high levels of ingested bacterial PAMPs such as peptidoglycan. Rather, AMP expression in the gut appears to be induced by active bacterial colonization involving type III secretion. We speculate that this form of regulation may have evolved to prevent over activation of the immune system from frequent ingestion of innocuous, non-colonizing, or non-viable bacteria. While additional work is needed to delineate the molecular mechanisms underlying our observations, our findings provide significant novel insight into the immunological adaptation of cockroaches to life in septic environments as well as the factors that regulate bacterial pathogen transmission by these insects.
Assuntos
Blattellidae , Infecções por Enterobacteriaceae , Animais , Peptídeos Antimicrobianos , Bactérias , Blattellidae/genética , Blattellidae/microbiologia , Escherichia coliRESUMO
BACKGROUND: The widespread use of chemical insecticides has resulted in the development of resistance in German cockroaches worldwide, and biopesticides based on entomopathogenic fungi as active ingredients have become a promising alternative strategy. Resistance can change many of the physiological and biochemical characteristics of insect pests, such as cuticle thickness, detoxification enzyme activity, and even intestinal flora composition. Thus, potential interactions between pathogenic fungi and insecticide resistance may lead to unpredictable changes in pest susceptibility to fungi. RESULTS: Beta-cypermethrin-resistant German cockroaches were more susceptible to infection with the fungus Metarhizium anisopliae regardless of age and sex. Histopathological results showed that the infection of resistant strains (R) by M. anisopliae was visibly faster than that of susceptible strains (S). The gut microbiota of the S strain indicated a stronger ability to inhibit fungi in vitro. The abundance of Parabacteroides, Lachnoclostridium, and Tyzzerella_3 decreased significantly in the R strain, and most demonstrated the ability to regulate glucose and lipid metabolism, and antifungal infections. The expression levels of Akirin, BgTPS, and BgPo genes in the R strain were significantly lower than those in the S strain, while BgChi and CYP4G19 gene expression were significantly higher. The mortality of cockroaches infected with M. anisopliae decreased to varying degrees after RNA interference, reflecting the role of these genes in antifungal infection. CONCLUSIONS: Results confirmed that insecticide resistance may enhance cockroach susceptibility to fungi by altering intestinal flora and gene expression. Fungal biopesticides have high utilization value in pest control and insecticide resistance management strategies. © 2021 Society of Chemical Industry.
Assuntos
Blattellidae , Metarhizium , Piretrinas , Animais , Blattellidae/genética , Blattellidae/microbiologia , Resistência a Inseticidas/genética , Metarhizium/genética , Piretrinas/farmacologiaRESUMO
Diarrheal diseases are among the most common illnesses in the world and the bacterium Salmonella enterica serovar Typhimurium is a leading cause of morbidity and mortality from diarrhea globally. The German cockroach (Blattella germanica) frequently harbors and has been linked to human outbreaks of Salmonella, but the mechanisms of vector-borne transmission are not fully clear. Transmission of S. Typhimurium by cockroaches has been previously described as mechanical. Mechanical transmission is a wholly passive process that involves physical transfer of a pathogen from one location or host to another but lacks bacterial replication in the vector and active bacterial processes that promote vector colonization or transmission. Towards the goal of obtaining novel insight into the mechanisms of S. Typhimurium transmission by cockroaches, here we orally provisioned wild type and mutant strains to adult B. germanica and examined several aspects of colonization and shedding. Our results provide evidence of three previously unappreciated phenomena with significant implications. First, we demonstrate that S. Typhimurium undergoes replication at multiple phases during colonization of the cockroach gut. Second, we show the formation of biofilm-like aggregates by S. Typhimurium in the cockroach foregut. Lastly, we identify two mutant strains of S. Typhimurium that are deficient in colonization and shedding relative to isogenic controls, implicating type III secretion and the formation of fimbriae as two processes that are necessary for interaction with the cockroach vector. Together, our data indicate that transmission of S. Typhimurium by cockroaches is not solely mechanical but may resemble biological transmission by other insect vectors that intake human pathogenic bacteria from infected hosts and are subsequently colonized, enabling active dissemination. Thus, these findings suggest that cockroaches and their control may be more important for infection prevention than is currently appreciated. Additional studies to better understand the cycle and biological mechanisms of vector-borne transmission are warranted.
Assuntos
Blattellidae , Salmonella typhimurium , Animais , Bactérias , Blattellidae/microbiologia , Humanos , Salmonella typhimurium/genéticaRESUMO
AIMS: The study aimed to explore the diversity of culturable microbiota colonizing the alimentary tract and outer surfaces of German cockroaches (Blattella germanica) captured in a health care facility. METHODS AND RESULTS: Microbial identification was conducted using Matrix Assisted Laser Desorption-Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) Biotyper and 16S rRNA sequencing. A total of 181 bacteria strains were isolated from 25 cockroach specimens and the MALDI-TOF MS-based assay yielded direct identification of 96·5% (175 out of 181) of the strains at the species level. The proteomic fingerprinting mainly revealed strains belonged to Gram-negative Enterobacteria (103) with six different genera that were characterized including Citrobacter, Klebsiella, Kluyevera, Leclercia, Morganella and Serratia. In addition, Pseudomonas sp. strains ranked in second with 29·8% (54 strains) followed by Staphylococcus sp. (6·62%) and Enterococcus sp. (1·65%). A large number of these bacteria (n = 90, 49·72%) was found in cockroaches captured in the maternity ward, whereas 45 strains (24·8%) were recovered in the paediatric ward. Altogether, 24 bacterial species were identified from both the external surface and digestive tract of the insect, of which Serratia marcescens presented the major group (n = 80, 44·19%) followed by Pseudomonas aeruginosa (n = 53, 29·28%) and Klebsiella oxytoca (n = 9, 4·94%). CONCLUSION: The findings showed a high prevalence of bacterial pathogens harboured in the body and alimentary tract of B. germanica captured in a health care facility. SIGNIFICANCE AND IMPACT OF THE STUDY: This investigation shows the possible role of German cockroaches as a source for bacterial pathogens, increasing the likelihood of direct transmission of healthcare associated infections, and thereby representing a public health risk. In addition, the present study revealed a high discriminatory power of the mass spectra investigation and a competent bacterial typing tool that extends phenotypic and genotypic approaches, which allows new possibilities for fast and accurate diagnosis in medical entomology.
Assuntos
Bactérias/isolamento & purificação , Blattellidae/microbiologia , Hospitais , Insetos Vetores/microbiologia , Animais , Bactérias/classificação , Bactérias/genética , Bactérias/metabolismo , Infecção Hospitalar/epidemiologia , Infecção Hospitalar/transmissão , Humanos , Proteômica , RNA Ribossômico 16S/genética , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
The German cockroach, Blatella germanica (L.), is a suspected vector of several enteric bacterial pathogens, including Escherichia coli, among livestock and humans. However, little is known about the factors that influence bacterial transmission by cockroaches. Here, we orally infected B. germanica with various laboratory and field strains of E. coli and examined bacterial titers over time to shed new light on the factors that influence the dynamics of infection. Our results reveal that a laboratory strain of E. coli is largely cleared within 48 h while one field isolate can persist in a majority of cockroaches (80-100%) for longer than 3 d with minimal impact on cockroach longevity. We also find that the ability to clear some strains of E. coli is greater in cockroach nymphs than adults. Notably, no differential expression of the antimicrobial gene lysozyme was observed between nymphs and adults or in infected groups. However, clearance of E. coli was significantly reduced in gnotobiotic cockroaches that were reared in the absence of environmental bacteria, suggesting a protective role for the microbiota against exogenous bacterial pathogens. Together, these results demonstrate that the interactions between cockroaches and enteric bacterial pathogens are highly dynamic and influenced by a combination of microbial, host, and environmental parameters. Such factors may affect the disease transmission capacity of cockroaches in nature and should be further considered in both lab and field studies.
Assuntos
Blattellidae/microbiologia , Escherichia coli/isolamento & purificação , Microbioma Gastrointestinal , Animais , Blattellidae/crescimento & desenvolvimento , Masculino , Ninfa/crescimento & desenvolvimento , Ninfa/microbiologiaRESUMO
A bacterial strain (BGf-2) with anti-Beauveria bassiana activity was obtained from the feces of Blattella germanica (L.) and identified as Pseudomonas aeruginosa based on biochemical tests and 16S rRNA sequence analysis. An antifungal protein (A0A0H2ZK06) was purified with Sephadex G-100 column and DEAE-sepharose Fast Flowanion exchange from sterile BGf-2 fermentation liquid. Based on MALDI-TOF MS analysis and protein model building, A0A0H2ZK06 showed homology with Pyrrolidone carboxyl peptidases (pcps). Fermentation liquid and antifungal proteins not only reduced the B. bassiana conidial germination rate but also inhibited hyphal growth. A per os test showed that the mortality of cockroaches decreased after treatment with BGf-2 suspension compared with control. We hypothesized that gut microbes with antifungal activity might play an important role in protect cockroaches from pathogenic fungi.
Assuntos
Proteínas de Bactérias/isolamento & purificação , Beauveria/efeitos dos fármacos , Blattellidae/microbiologia , Pseudomonas aeruginosa/química , Piroglutamil-Peptidase I/isolamento & purificação , Animais , Antifúngicos/farmacologia , Fezes/microbiologia , Controle Biológico de VetoresRESUMO
Cockroaches are widely perceived to evolve resistance to insecticides. Over-expression of a resistance-conferring gene can be costly and may require energy and resource reallocation for metabolic and developmental processes. To evaluate whether changes in the composition of gut microbiota in Blattella germanica affected its resistance evolution to beta-cypermethrin and to determine the role of gut microbiota in host growth and development, we studied the relationship between insecticide resistance and the diversity and genetic content of gut microbiota in cockroaches. Results suggest beta-cypermethrin-resistant cockroaches (R strain) exhibited a delayed development period and reduced adult longevity compared with susceptible cockroaches (S strain). Based on 16S rRNA gene sequencing and community metagenomics, we found that the relative abundance of Lactobacillus and Acetobacteraceae were significantly lower in the R strain compared with the S strain in the foregut and midgut of both strains. Functional annotation of Kyoto Encyclopedia of Genes and Genomes (KEGG) modules of midgut genes in the two strains revealed that 10.6% were involved in metabolism, while the relative abundance in the R strain was 7.4%. Unigenes were also translated into amino acid sequences and assigned to protein families based on hits to the Carbohydrate-Active enzymes (CAZy) database. This process identified the glycoside hydrolases, glycosyl transferases and carbohydrate-binding modules of the S strain as all being significantly higher in diversity than those in the R strain. Overall, we conclude that fitness-related costs increased in the resistant strain of cockroaches compared with the susceptible strain, and the variation in insect gut microbiota, especially those related to growth and development, was an important influencing factor.
Assuntos
Blattellidae/efeitos dos fármacos , Inseticidas/farmacologia , Microbiota/fisiologia , Piretrinas/farmacologia , Animais , Bactérias/classificação , Bactérias/genética , Blattellidae/genética , Blattellidae/crescimento & desenvolvimento , Blattellidae/microbiologia , Biologia Computacional , DNA Bacteriano/química , DNA Bacteriano/isolamento & purificação , Feminino , Genótipo , Resistência a Inseticidas/genética , Masculino , Metagenoma/genética , Microbiota/efeitos dos fármacos , Ninfa/crescimento & desenvolvimento , RNA Ribossômico 16S/químicaRESUMO
The German cockroach, Blatella germanica, is a common pest in urban environments and is among the most resilient insects in the world. The remarkable ability of the German cockroach to develop resistance when exposed to toxic insecticides is a prime example of adaptive evolution and makes control of this insect an ongoing struggle. Like many other organisms, the German cockroach is host to a diverse community of symbiotic microbes that play important roles in its physiology. In some insect species, there is a strong correlation between the commensal microbial community and insecticide resistance. In particular, several bacteria have been implicated in the detoxification of xenobiotics, including synthetic insecticides. While multiple mechanisms that mediate insecticide resistance in cockroaches have been discovered, significant knowledge gaps still exist in this area of research. Here, we examine the effects of altering the microbiota on resistance to a common insecticide using antibiotic treatments. We describe an indoxacarb-resistant laboratory strain in which treatment with antibiotic increases susceptibility to orally administered insecticide. We further reveal that this strains harbors a gut microbial community that differs significantly from that of susceptible cockroaches in which insecticide resistance is unaffected by antibiotic. More importantly, we demonstrate that transfer of gut microbes from the resistant to the susceptible strain via fecal transplant increases its resistance. Lastly, our data show that antibiotic treatment adversely affects several reproductive life-history traits that may contribute to the dynamics of resistance at the population level. Together these results suggest that the microbiota contributes to both physiological and evolutionary aspects of insecticide resistance and that targeting this community may be an effective strategy to control the German cockroach.
Assuntos
Antibacterianos/farmacologia , Blattellidae/microbiologia , Inseticidas/metabolismo , Microbiota/fisiologia , Oxazinas/metabolismo , Animais , Blattellidae/efeitos dos fármacos , Blattellidae/fisiologia , Fezes/microbiologia , Feminino , Resistência a Inseticidas/efeitos dos fármacos , Inseticidas/farmacologia , Masculino , Microbiota/efeitos dos fármacos , Oxazinas/farmacologia , SimbioseRESUMO
The successful adaptation of cockroaches is, in part, dependent of the activity of their obligatory endosymbionts, Blattabacterium spp., which are involved in uric acid degradation, nitrogen assimilation and nutrient provisioning. Their strategic localization, within bacteriocytes in the proximities of uric acid storage cells (urocytes), highlights their importance in the recycling of nitrogen from urea and ammonia, end-products not secreted by their host insects. In this study, we present the complete genome sequence of two new Blattabacterium spp. from Periplaneta fuliginosa (BPfu) and P. japonica (BPja), and detailed comparison with other Blattabacterium strains from different cockroach species. The genomes of BPfu and BPja show a high degree of stability as showed with for other Blattabacterium representatives, only presenting a 19-kb fragment inversion between BPja and BPfu. In fact, the phylogenomics showed BPja as an ancestor species of BPfu, BPLAN (P. americana) and BBor (Blatta orientalis), in congruence with their host cockroach phylogeny. Their functional profile is similar and closest to the omnivorous strain BBge (Blattella germanica). Interesting, BPja possesses the complete set of enzymes involved sulfate assimilatory pathway only found in BBge and BMda (Mastotermes darwiniensis). The newly sequenced genomes of BPja and BPfu emphasise the remarkable stability of Blattabacterium genomes supported by their long-term coevolution and obligatory lifestyle in their host insect.
Assuntos
Bacteroidetes/genética , Blattellidae/microbiologia , Genoma Bacteriano , Periplaneta/microbiologia , Amônia/química , Animais , DNA Bacteriano/genética , Masculino , Nitrogênio/química , Filogenia , Simbiose , Ureia/química , Ácido Úrico/químicaRESUMO
Eukaryotes have established symbiotic relationship with microorganisms, which enables them to accomplish functions that they cannot perform alone. In the German cockroach, Blattella germanica, the obligate endosymbiont Blattabacterium coexists with a rich gut microbiota. The transmission of Blattabacterium is vertical, but little is known about how the gut microbiota colonizes newborn individuals. In this study, we treated B. germanica populations with rifampicin, a broad-spectrum antibiotic, during two generations and analyzed gut bacterial composition and the Blattabacterium load in control and rifampicin-treated populations. Rifampicin exerted a drastic effect on gut microbiota composition, which recovered in the second generation in the case where the antibiotic was not added to the diet. Furthermore, we observed that bacterial species present in the diet, and particularly in the feces, contribute significantly to establishing the gut microbiota. Finally, the Blattabacterium population remained unaffected by the antibiotic treatment of adults during the first generation but was strongly reduced in the second generation, suggesting that this intracellular symbiont is sensitive to rifampicin only during the infection of the mature oocytes, when it is in an extracellular stage.
Assuntos
Antibacterianos/farmacologia , Blattellidae/microbiologia , Flavobacteriaceae/efeitos dos fármacos , Flavobacteriaceae/isolamento & purificação , Microbioma Gastrointestinal/efeitos dos fármacos , Rifampina/farmacologia , Adulto , Animais , Técnicas de Tipagem Bacteriana , Fezes/microbiologia , Humanos , Masculino , Filogenia , SimbioseRESUMO
BACKGROUND: Metarhizium anisopliae and Beauveria bassiana mainly infect insects through the cuticle; gut infection occasionally occurs. Micro-organisms existing in the gut may play a crucial role in the evolution and ecology of host defenses against fungal pathogens. To evaluate whether the gut bacteria participate in antifungal activity, and to determine their role in host protection, the interactions between gut bacteria and M. anisopliae and the diversity of gut microbiota in cockroaches were studied. RESULTS: An oral feeding test showed that the mortality of conventional cockroaches was significantly lower than that of germ-free cockroaches; both gut homogenates and aqueous fecal extracts showed antifungal activity, but the samples from germ-free cockroaches did not. Twenty-two bacterial strains with antifungal activity and siderophore-producing ability were isolated from the gut and feces of cockroaches. Using high-throughput sequencing techniques, a total of 23 different phyla and 212 genera were detected. The composition of the microbiota of the hindgut was vastly different from those of the foregut and midgut; higher diversity and abundance of Bacteroides and Pseudomonas were found in the hindgut. CONCLUSION: The gut microbiota of German cockroaches may play a critical role in protecting cockroaches from fungal invasion and colonization. Removing certain bacteria from the B. germanica microbiota may facilitate microbial control using fungal pathogens. © 2017 Society of Chemical Industry.
Assuntos
Blattellidae/microbiologia , Microbioma Gastrointestinal , Metarhizium/fisiologia , Controle Biológico de Vetores/métodos , Animais , Vida Livre de GermesRESUMO
A wild, forest-dwelling cockroach from the subfamily Ectobiidae (order Blattodea) in a nature reserve in Cavalcante, in the state of Goiás, Brazil, was found to be infected by a new, genetically distinct species in the Metarhizium flavoviride species complex that we describe here as Metarhizium blattodeae. The status of this fungus as a new species is supported by both multigenic sequence comparisons and protein profiles generated by MALDI-TOF (matrix-assisted laser desorption/ionization time-of-flight) mass spectrometry. This is one of the first reports of a naturally occurring fungal pathogen affecting any sylvatic (forest-dwelling) cockroach from any part of the world. M. blattodeae caused up to 96 % mortality of Periplaneta americana nymphs (a serious peridomestic cockroach species) after 10 d.
Assuntos
Blattellidae/microbiologia , Metarhizium/classificação , Metarhizium/isolamento & purificação , Micoses/microbiologia , Animais , Brasil , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Proteínas Fúngicas/análise , Genes de RNAr , Metarhizium/química , Metarhizium/genética , Microscopia , Fator 1 de Elongação de Peptídeos/genética , Filogenia , RNA Polimerase II/genética , RNA Fúngico/genética , RNA Ribossômico 5,8S/genética , Análise de Sequência de DNA , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
The German cockroach, Blattella germanica (L.) has been recognized as a serious health problem throughout the world. Control failures due to insecticide resistance and chemical contamination of environment have led some researchers focus on the other alternative strategy controls. Microbial insecticides such as those containing entomo pathogenic fungi could be of high significance. Lecanicillium muscarium and Beauveria bassiana grow naturally in soils throughout the world and act as a parasite on various arthropod species, causing white muscardine disease. Thus, these two species could be considered as entomopathogenic fungi. The current study conducted to evaluate the toxicity of Beauveria bassiana and Lecanicillium muscarium against German cockroach, Blattella germanica. Conidial formulations of L. muscarium (PTCC 5184) and B. bassiana (PTCC5197) were prepared in aqueous suspensions with Tween 20. Bioassays were performed using two methods including submersion of cockroaches in conidial suspension and baiting. Data were analyzed by Probit program and LC50 and LC90 were estimated. The obtained results indicated that both fungi species were toxic against German cockroach however; Beauveria bassiana was significantly 4.8 fold more toxic than L. muscarium against German cockroach using submersion method.
Assuntos
Blattellidae/microbiologia , Blattellidae/fisiologia , Hypocreales/crescimento & desenvolvimento , Animais , Bioensaio , Feminino , Controle de Insetos/métodos , Masculino , Controle Biológico de Vetores/métodos , Análise de SobrevidaRESUMO
Systemic fungal infections are becoming an increasingly important problem. Exploring the development of mechanisms of pathogenesis, immune response of the human organism and the search for new potential antifungal agents requires in vivo testing. Mice, rats and rabbits are indispensable model organisms for this type of study. Unfortunately, such a kinds of studies carried out on a large scale are associated with high costs as well as with logistical and ethical problems. This paper reports proposal for the use of insects as model organisms to study the development of systemic fungal infections and analysis of biological activities of antifungal agents.
Assuntos
Antifúngicos/farmacologia , Modelos Animais de Doenças , Insetos/microbiologia , Micoses/tratamento farmacológico , Micoses/microbiologia , Animais , Blattellidae/microbiologia , Culex/microbiologia , Drosophila melanogaster/microbiologia , Micoses/imunologiaRESUMO
A study was performed to estimate the prevalence of the external bacterial flora of two domestic cockroaches (Blattella germanica and Blatta orientalis) collected from households in Tebessa (northeast Algeria). Three major bacterial groups were cultured (total aerobic, enterobacteria, and staphylococci) from 14 specimens of cockroaches, and antibiotic susceptibility was tested for both Staphylococcus and Pseudomonas isolates. Culturing showed that the total bacterial load of cockroaches from different households were comparable (P<0.001) and enterobacteria were the predominant colonizers of the insect surface, with a bacterial load of (2.1 × 105 CFU/insect), whereas the staphylococci group was the minority. Twenty-eight bacterial species were isolated, and susceptibility patterns showed that most of the staphylococci isolates were highly susceptible to chloramphenicol, gentamycin, pristinamycin, ofloxacin, clindamycin, and vancomycin; however, Pseudomonas strains exhibited resistance to amoxicillin/clavulanic acid, imipenem, and the second-generation antibiotic cephalosporin cefuroxime.
Assuntos
Blattellidae/microbiologia , Farmacorresistência Bacteriana , Pseudomonas/efeitos dos fármacos , Staphylococcus/efeitos dos fármacos , Argélia , Animais , Feminino , Habitação , Masculino , MicrobiotaRESUMO
BACKGROUND: Cockroaches are among the most common pests in public dwellings and health facilities. Their presence can raise safety concerns, especially as they maybe carriers of pathogenic organisms. METHODS: This study was carried out to isolate and identify the bacterial flora from German cockroaches (Blattella germanica). Cockroaches collected by hand catches from two public hospital environments in Tebessa city (northeast Algeria) were screened for microbial load from their external surfaces and alimentary tract using standard bacterial protocols. RESULTS: A total of 174 bacterial isolates were isolated from 39 German cockroach specimens. The most common and abundant bacterial species belonged to the Pseudomonas group (23.5%) and Serratia (13.2%). Pathogens like Staphylococcus aureus were also isolated, as well as opportunistic pathogens like Klebsiella species and food spoilage bacteria such as Enterobacter and Citrobacter species were isolated from both external surface and digestive tract of the insect. Generalized linear models (GLM) were performed to analyze the variation of abundances and occurrences of bacterial isolates harboured by B. germanica. The GLMs revealed that the main factors affecting variation of bacterial diversity and abundance were sex and hospital (P < 0.001). CONCLUSION: The findings of this study suggest that German cockroach acts as reservoir and potential vector of some bacterial pathogens.
Assuntos
Bactérias/classificação , Bactérias/isolamento & purificação , Carga Bacteriana , Blattellidae/microbiologia , Argélia , Animais , Hospitais Públicos , Humanos , Insetos VetoresRESUMO
Although blattid cockroaches and termites share a common ancestor, their diets are distinctly different. While termites consume a highly specialized diet of lignocellulose, cockroaches are omnivorous and opportunistic feeders. The role of the termite gut microbiota has been studied intensively, but little is known about the cockroach gut microbiota and its function in digestion and nutrition, particularly the adaptation to different diets. Our analyses of the bacterial gut microbiota of the blattid cockroach Shelfordella lateralis combining terminal restriction fragment length polymorphism of their 16S rRNA genes with physiological parameters (microbial metabolites, hydrogen and methane emission) indicated substantial variation between individuals but failed to identify any diet-related response. Subsequent deep-sequencing of the 16S rRNA genes of the colonic gut microbiota of S. lateralis fed either a high- or a low-fiber diet confirmed the absence of bacterial taxa that responded to diet. Instead, we found a small number of abundant phylotypes that were consistently present in all samples and made up half of the community in both diet groups. They varied strongly in abundance between individual samples at the genus but not at the family level. The remaining phylotypes were inconsistently present among replicate batches. Our findings suggest that S. lateralis harbors a highly dynamic core gut microbiota that is maintained even after fundamental dietary shifts, and that any dietary effects on the gut community are likely to be masked by strong individual variations.