Search

Three new N-alkylated amino acid derivatives, penichrysoamides A-C (1-3), along with a new citric acid derivative, penichrysoacid A (4), a new chromanone lactone penichrysoacid B (5), and a new amide derivative, penichrysoamide D (6), as well as seven known benzamide derivatives (7-13), were isolated from the endophytic fungus Penicillium chrysogenum derived from the insect Periplaneta americana. The structures of these compounds, including their absolute configurations, were elucidated using spectroscopic and computational techniques. Biological evaluation revealed that compounds 8-13 exhibited significant COX-2 inhibitory activity, with IC 50 values ranging from 275 nM to 1350 nM., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Insect-associated actinomycetes represent a potentially rich source for discovering bioactive metabolites. However, the diversity, antibacterial and phytotoxic activities of symbiotic actinomycetes associated with Periplaneta fuliginosa have not yet been conducted., Results: A total of 86 strains of actinomycetes were isolated from the cornicles and intestines of both nymphs and adults of P. fuliginosa . Diversity analysis revealed that the isolated strains were preliminarily identified as 17 species from two genera, and the dominant genus was Streptomyces . A total of 36 crude extracts (60%) obtained from the supernatant of the 60 fermented strains exhibited a potent antibacterial activity against at least one tested pathogenic bacterium. Among these active strains, 27 crude extracts (75%) exhibited phytotoxic activity against the radicle of Echinochloa crusgalli . Furthermore, seven known compounds, including methoxynicotine ( 1 ), (3Z,6Z)-3-(4-methoxybenzylidene)-6-(2-methylpropyl) piperazine-2,5-dione ( 2 ), XR334 ( 3 ), 1-hydroxy-4-methoxy-2-naphthoic acid ( 4 ), nocapyrone A ( 5 ), β-daucosterol ( 6 ), and β-sitosterol ( 7 ) were isolated from an active rare actinomycete Nocardiopsis sp. ZLC-87 which was isolated from the gut of adult P. fuliginosa . Among them, compound 4 exhibited moderate antibacterial activity against Micrococcus tetragenus , Staphylococcus aureus , Escherichia coli , and Pseudomonas syringae pv. actinidiae with the zone of inhibition (ZOI) of 14.5, 12.0, 12.5, and 13.0 mm at a concentration of 30 μg/disc, respectively, which was weaker than those of gentamicin sulfate (ZOI of 29.5, 19.0, 18.5, and 24.5 mm). In addition, the compound 4 had potent phytotoxic activity against the radicle of E. crusgalli and Abutilon theophrasti with the inhibition rate of 65.25% and 92.68% at the concentration of 100 μg/mL., Conclusion: Based on these findings, this study showed that P. fuliginosa -associated actinomycetes held promise for the development of new antibiotic and herbicide resources., Competing Interests: The authors declare that they have no competing interests., (© 2024 Liu et al.)

Three p-terphenyl metabolites (1-3), three indole-diterpenoids (4-6), an herbicide sesquiterpene (7), a flavonoid (8), and five other small molecules containing nitrogen (9-13) were isolated from the medicinal insect (Periplaneta americana)-derived endophytic Aspergillus taichungensis SMU01. Their chemical structures were elucidated on the basis of spectroscopic data and quantum chemical computational methods. Biological activity of these isolates in the differentiation of mouse CD4 + T cell subsets was evaluated. Importantly, metabolites 2 targeting JAK-STAT signaling pathway could hold potential benefits in maintaining peripheral immune homeostasis and alleviating the progression of autoimmune diseases., Competing Interests: Declaration of competing interest The authors declare no competing financial interest., (Copyright © 2023. Published by Elsevier B.V.)

Periplaneta americana , one of the most widely distributed insects all over the world, can survive and reproduce in harsh environment which may be closely related to the critical roles of intestinal microorganisms in its multiple physiological functions. However, the composition and structure of gut microbiota throughout different life stages and its effects on the strong resilient and environmental adaptability of P. americana remain unclear. In this study, the gut microbiota across life stages including ootheca (embryos), nymph and adult of P. americana were investigated by 16S rRNA high-throughput sequencing. Multivariate statistical analysis showed the richness and diversity of bacterial communities were significantly different among ootheca, nymph and adult stage of P. americana . Taxonomic analysis showed Blattabacterium was the dominant genus in bacterial community of ootheca while the nutrient absorption-related genera including Christensenellaceae and Ruminococcaceae showed high relative abundance in nymph samples. Moreover, functional prediction analysis showed the metabolic categories in ootheca might have more influence on the basic life activities of the host than improved production and viability, while it was more associated to the society activities, reproduction and development of host in nymph and adult. It was suggested that the gut microbiota in each life stage might meet the requirements for environmental adaptability and survival of P. americana via transforming the composition and structure with specific metabolic capabilities. Overall, these results provided a novel sight to better understand the strong vitality and adaptability throughout life stages of P. americana .

Background: Klebsiella pneumoniae is one of the main pathogens of clinical isolation and nosocomial infections, as K. pneumoniae show broad-spectrum resistance to β-lactam and carbapenem antibiotics. It is emerging clinical need for a safe and effective drug to anti-K. pneumoniae. At present, Achromobacter mainly focused on its degradation of petroleum hydrocarbons, polycyclic aromatic hydrocarbons, assisting insects to decompose, degrade heavy metals and utilize organic matter, but there were few reports on the antibacterial activity of the secondary metabolites of Achromobacter., Results: In this study, a strain WA5-4-31 from the intestinal tract of Periplaneta americana exhibited strong activity against K. Pneumoniae through preliminary screening. The strain was determined to be Achromobacter sp. through the morphological characteristics, genotyping and phylogenetic tree analysis, which is homologous to Achromobacter ruhlandii by 99%, its accession numbe in GenBank at National Center for Biotechnology Information (NCBI) is MN007235, and its deposit number was GDMCC NO.1.2520. Six compounds (Actinomycin D, Actinomycin X2, Collismycin A, Citrinin, Neoechinulin A and Cytochalasin E) were isolated and determined by activity tracking, chemical separation, nuclear magnetic resonance (NMR) and mass spectrometry (MS) analysis. Among them, Actinomycin D, Actinomycin X2, Collismycin A, Citrinin and Cytochalasin E showed a good effect on anti-K. pneumoniae, with MIC values of 16-64 µg/mL., Conclusions: The study reported Achromobacter, which was from the intestinal tract of Periplaneta americana with the activity against K. Pneumoniae, can produce antibacterial compounds for the first time. It lays the foundation for development of secondary metabolites of insect intestinal microorganisms., (© 2023. The Author(s).)

Cockroaches are known as mechanical vectors of some pathogens that can infect humans. The present study aims to rapidly identify Periplaneta americana fungal pathogens from sewer systems of public hospitals in Esfahan using the polymerase chain reaction (PCR) technique. A total of 55 P. americana cockroaches were randomly collected by direct trapping from sewer systems of seven hospitals and screened for fungal infectious agents using standard morphological methods and the PCR sequencing. From the American cockroach, we isolated 62 yeasts and 31 molds from the surface, hemocoel, and digestive tract of P. americana. Based on DNA sequence comparisons and other taxonomic characteristics, they were identified as more than four species of yeast and four species of mold. Yeast species including Pichia kudriavzevii, Candida glabrata, Pichia kluyveri, and Candida viswanathii, and molds such as Aspergillus niger, Penicillium italicum, Mucor plumbeus, and Rhizopus oryzae were isolated repeatedly from the surface, hemocoel, and digestive tract of P. americana. Our results show that the use of a combination of morphological, molecular techniques, and phylogenetic analysis can lead to the identification of pathogenic fungal agents in American cockroaches and also knowledge of fungal pathogens-arthropod host relationships.

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., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Choubdar et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Due to the lack of an adaptive immune system, insects rely on innate immune mechanisms to fight against pathogenic infections. Two major innate immune pathways, Toll and IMD, orchestrate anti-pathogen responses by regulating the expression of antimicrobial peptide (AMP) genes. Although the antifungal or antibacterial function of AMPs has been well characterized, the antiviral role of AMPs in insects remains largely unclear. Periplaneta americana (P. americana), or the American cockroach, is used in traditional Chinese medicine as an antiviral agent; however, the underlying mechanism of action of P. americana extracts is unclear. Our previous study showed that the P. americana genome encodes multiple antimicrobial peptide genes. Based on these data, we predicted five novel P. americana defensins (PaDefensins) and analyzed their primary structure, secondary structure, and physicochemical properties. The putative antiviral, antifungal, antibacterial, and anticancer activities suggested that PaDefensin5 is a desirable therapeutic candidate against viral diseases. As the first experimental evidence of the antiviral effects of insect defensins, we also showed the antiviral effect of PaDefensin5 in Drosophila Kc cells and Drosophila embryos in vivo . In conclusion, results of both in silico predictions and subsequent antiviral experiments suggested PaDefensin5 a promising antiviral drug., (Copyright © 2021. Published by Elsevier B.V.)

Background: Digestive tissues are essential for diet processing and nutrient accessibility, especially in omnivores, and these functions occur despite and in collaboration with dynamic microbial communities that reside within and upon these tissues. Prolonged host development and reduced digestive tissue sizes have been observed in germ-free animals, and normal host phenotypes were recovered following the re-introduction of typical gut microbiomes via coprophagy., Results: High-resolution histological analyses of Periplaneta americana cockroach digestive tissues revealed that total prevention of microbial colonization of the gut had severe impacts on the growth and development of gut tissues, especially the posterior midgut and anterior hindgut subcompartments that are expected to be colonized and inhabited by the greatest number of bacteria. Juveniles that were briefly exposed to normal gut microbiota exhibited a partial gut morphological recovery, suggesting that a single inoculation was insufficient. These data highlight gut microbiota as integral to normal growth and development of tissues they are in direct contact with and, more broadly, the organism in which they reside., Conclusions: We draw on these data, host life history traits (i.e. multigenerational cohousing, molting, and filial coprophagy and exuvia feeding), and previous studies to suggest a host developmental model in which gut tissues reflect a conflict-collaboration dynamic where 1) nutrient-absorptive anterior midgut tissues are in competition with transient and resident bacteria for easily assimilable dietary nutrients and whose growth is least-affected by the presence of gut bacteria and 2) posterior midgut, anterior hindgut, and to a lesser degree, posterior hindgut tissues are significantly impacted by gut bacterial presence because they are occupied by the greatest number of bacteria and the host is relying upon, and thus collaborating with, them to assist with complex polysaccharide catabolism processing and nutrient provisioning (i.e. short-chain fatty acids)., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Gnotobiotic animals are a powerful tool for the study of controls on microbiome structure and function. Presented here is a protocol for the establishment and maintenance of gnotobiotic American cockroaches (Periplaneta americana). This approach includes built-in sterility checks for ongoing quality control. Gnotobiotic insects are defined here as cockroaches that still contain their vertically transmitted endosymbiont (Blattabacterium) but lack other microbes that normally reside on their surface and in their digestive tract. For this protocol, egg cases (oothecae) are removed from a (nonsterile) stock colony and surface sterilized. Once collected and sterilized, the oothecae are incubated at 30 °C for approximately 4-6 weeks on brain-heart infusion (BHI) agar until they hatch or are removed due to contamination. Hatched nymphs are transferred to an Erlenmeyer flask containing a BHI floor, sterile water, and sterile rat food. To ensure that the nymphs are not housing microbes that are unable to grow on BHI in the given conditions, an additional quality control measure uses restriction fragment-length polymorphism (RFLP) to test for nonendosymbiotic microbes. Gnotobiotic nymphs generated using this approach can be inoculated with simple or complex microbial communities and used as a tool in gut microbiome studies.

We are becoming increasingly aware of the dazzling diversity of fungi-new taxa are being discovered from poorly sampled habitats and integrative approaches point at (near-)cryptic species in many groups. The class Laboulbeniomycetes, which is composed of three orders (Herpomycetales, Laboulbeniales, Pyxidiophorales), is no exception. However, still, in the orders Herpomycetales and Laboulbeniales, species are predominantly described based on morphology alone. Here, we present a new species of Herpomyces from Argentina parasitic on Periplaneta fuliginosa , the smokybrown cockroach. Herpomyces spegazzinii , sp. nov., is characterized based on morphology and molecular data. Phylogenetic inference based on internal transcribed spacer (ITS, consisting of ITS1-5.8S-ITS2) barcode region supports the status of this fungus as a separate species, sister to the recently described H. shelfordellae .

Cockroaches inhabit various habitats, which will influence their microbiome. Although the microbiome can be influenced by the diet and environmental factors, it can also differ between species. Therefore, we conducted 16S rDNAtargeted high-throughput sequencing to evaluate the overall bacterial composition of the microbiomes of 3 cockroach species, Periplaneta americana, P. japonica, and P. fuliginosa, raised in laboratory for several generations under the same conditions. The experiments were conducted using male adult cockroaches. The number of operational taxonomic units (OTUs) was not significantly different among the 3 species. With regard to the Shannon and Pielou indexes, higher microbiome values were noted in P. americana than in P. japonica and P. fuliginosa. Microbiome composition was also evaluated, with endosymbionts accounting for over half of all OTUs in P. japonica and P. fuliginosa. Beta diversity analysis further showed that P. japonica and P. fuliginosa had similar microbiome composition, which differed from that of P. americana. However, we also identified that P. japonica and P. fuliginosa host distinct OTUs. Thus, although microbiome compositions may vary based on multiple conditions, it is possible to identify distinct microbiome compositions among different Periplaneta cockroach species, even when the individuals are reared under the same conditions.

Beneficial gut microbes can facilitate insect growth on diverse diets. The omnivorous American cockroach, Periplaneta americana (Insecta: Blattodea), thrives on a diet rich in plant polysaccharides and harbors a species-rich gut microbiota responsive to host diet. Bacteroidetes are among the most abundant taxa in P. americana and other cockroaches, based on cultivation-independent gut community profiling, and these potentially polysaccharolytic bacteria may contribute to host diet processing. Eleven Bacteroidetes isolates were cultivated from P. americana digestive tracts, and phylogenomic analyses suggest that they were new Bacteroides , Dysgonomonas , Paludibacter , and Parabacteroides species distinct from those previously isolated from other insects, humans, and environmental sources. In addition, complete genomes were generated for each isolate, and polysaccharide utilization loci (PULs) and several non-PUL-associated carbohydrate-active enzyme (CAZyme)-coding genes that putatively target starch, pectin, and/or cellulose were annotated in each of the isolate genomes. Type IX secretion system (T9SS)- and CAZyme-coding genes tagged with the corresponding T9SS recognition and export C-terminal domain were observed in some isolates, suggesting that these CAZymes were deployed via non-PUL outer membrane translocons. Additionally, single-substrate growth and enzymatic assays confirmed genomic predictions that a subset of the Bacteroides and Dysgonomonas isolates could degrade starch, pectin, and/or cellulose and grow in the presence of these substrates as a single sugar source. Plant polysaccharides enrich P. americana diets, and many of these gut isolates are well equipped to exploit host dietary inputs and potentially contribute to gut community and host nutrient accessibility. IMPORTANCE Gut microbes are increasingly being recognized as critical contributors to nutrient accessibility in animals. The globally distributed omnivorous American cockroach ( Periplaneta americana ) harbors many bacterial phyla (e.g., Bacteroidetes ) that are abundant in vertebrates. P. americana thrives on a highly diverse plant-enriched diet, making this insect a rich potential source of uncharacterized polysaccharolytic bacteria. We have cultivated, completely sequenced, and functionally characterized several novel Bacteroidetes species that are endemic to the P. americana gut, and many of these isolates can degrade simple and complex polysaccharides. Cultivation and genomic characterization of these Bacteroidetes isolates further enable deeper insight into how these taxa participate in polysaccharide metabolism and, more broadly, how they affect animal health and development., (Copyright © 2020 American Society for Microbiology.)

Serratia marcescens are considered to be abundant and optimal resources for obtaining prodigiosin, which can be isolated from soil, water, plants and air but rarely from insects. In the present study, a strain of Serratia marcescens named WA12‑1‑18 was isolated from the gut of Periplaneta americana, which was capable of producing high levels of pigment reaching 2.77 g/l via solid fermentation and was identified as prodigiosin by ultraviolet, high performance liquid chromatography (LC), Fourier‑transform infrared spectroscopy, LC‑mass spectroscopy and nuclear magnetic resonance. The apoptotic tumor cells treated with prodigiosin were examined by 4',6‑diamidino‑2‑phenylindole (DAPI) staining assays and transmission electron microscopy. Flow cytometry (FCM) was utilized to measure the apoptotic rate with Annexin V staining and the expression levels of proteins involved in apoptosis, including B‑cell lymphoma 2 (Bcl‑2), Bcl‑2‑associated X (Bax) and caspase‑3 were determined by western blot analysis and reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR). The experimental results revealed that prodigiosin could inhibit the proliferation of HeLa cells and the half‑maximal inhibitory concentration values of prodigiosin in HeLa were 2.1, 1.2 and 0.5 µg/ml over 24, 48 and 72 h, respectively. Furthermore, DAPI staining assays and transmission electron microscopy clearly demonstrated that prodigiosin could induce HeLa cell apoptosis. FCM results revealed that the cell apoptotic rates were 19.7±1.4, 23.7±2.4 and 26.2±2.3% following the treatment with 0.5, 1.0 and 2.0 µg/ml prodigiosin for 48 h, respectively. Western blot analysis and RT‑qPCR revealed that prodigiosin could activate apoptosis‑associated molecules including Bcl‑2, Bax and caspase‑3. Therefore, the results of the present study demonstrated that the prodigiosin could induce apoptosis in HeLa cells, which may be associated with the upregulation of Bax and caspase‑3, the concomitant downregulation of Bcl‑2 levels and also triggering the extrinsic apoptotic signaling pathway.

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., Competing Interests: The authors have declared that no competing interests exist.

Actinomycetes are well-known for producing numerous bioactive secondary metabolites. In this study, primary screening by antifungal activity assay found one actinomycete strain WA23-4-4 isolated from the intestinal tract of Periplaneta americana that exhibited broad spectrum antifungal activity. 16S rDNA gene analysis of strain WA23-4-4 revealed close similarity to Streptomyces nogalater (AB045886) with 86.6% sequence similarity. Strain WA23-4-4 was considered as a novel Streptomyces and the 16s rDNA sequence has been submitted to GenBank (accession no. KX291006). The maximum antifungal activity of WA23-4-4 was achieved when culture conditions were optimized to pH 8.0, with 12% inoculum concentration and 210 ml ISP2 medium, which remained stable between the 5 th and the 9 th day. 3-Acetyl benzoyl amide was isolated by ethyl acetate extraction of WA23-4-4 fermentation broth, and its molecular formula was determined as C 9 H 9 NO 2 based on MS, IR, 1 H, and 13 C NMR analyses. The compound showed significant antifungal activity against Candida albicans ATCC 10231 (MIC: 31.25 μg/ml) and Aspergillus niger ATCC 16404 (MIC: 31.25 μg/ml). However, the compound had higher MIC values against Trichophyton rubrum ATCC 60836 (MIC: 500 μg/ml) and Aspergillus fumigatus ATCC 96918 (MIC: 1,000 μg/ml). SEM analysis showed damage to the cell membrane of Candida albicans ATCC 10231 and to the mycelium of Aspergillus niger ATCC 16404 after being treatment with 3-acetyl benzoyl amide. In conclusion, this is the first time that 3-acetyl benzoyl amide has been identified from an actinomycete and this compound exhibited antifungal activity against Candida albicans ATCC 10231 and Aspergillus niger ATCC 16404.

Introduction: Fungal infections have emerged worldwide. Cockroaches have been proved vectors of medically fungi., Methods: A systematic meta-analysis review about cockroach fungal contamination was investigated. Relevant topics were collected between January 2016 and January 2017. After a preliminary review among 392 collected papers, 156 were selected to become part of the detailed systematic meta-analysis review., Results: Cockroaches contaminated to 38 fungi species belonging to 19 families and 12 orders. About 38, 25 and 13 fungal species were recovered from the American, German and brown-banded cockroaches, respectively with a variety of medical importance. Except the fungi isolated from German and brown-banded cockroaches, 15 species have been isolated only from the American cockroaches. The global world mean and trend of cockroach fungal contamination were 84.1 and 50.6-100%, respectively in the human dwelling environments. There is a significant difference between cockroach fungal contamination in the urban and rural environments (P<0.05) without a significant difference between hospital and household environments (P>0.05). The external and internal cockroach fungal contamination is more dangerous than entire surfaces, while the internal is more dangerous than the external surface. The German and brown-banded cockroach fungal contamination are more dangerous than the American cockroaches in the hospital environments., Conclusion: The study indicates that globally cockroach fungal contamination has been increased recognizing as agents of human infections and associating with high morbidity and mortality in immune-compromised patients. These facts, along with insecticide resistance emergence and increasing globally cockroach infestation, reveal importance of cockroaches and need for their control more than ever., (Copyright © 2017 Elsevier Masson SAS. All rights reserved.)

Aims: To study the marked resistance of Periplaneta americana to entomopathogenic Metarhizium anisopliae., Methods and Results: The low susceptibility of 4th instar nymphs applied topically with conidia seemed to be related to an active removal of conidia by the cockroach and to a disabled or retarded germination and subsequent development of conidia on the cuticle (up to 80% germination in the next 7 days after application). Inhibitions or delays of germination were related to the composition of the epicuticular fatty acids (30·1% w/w oleic, 28·3% w/w linoleic, 24·5% w/w palmitic and 11·7% w/w stearic acid) reported here. Propagules invading the nymphs through the cuticle took at least 3 days to reach the haemocoel, and no propagules were found after day 8 post-treatment. Strain IP 46 infected >50% of nymphs treated with doses ≥2 × 10 4  hyphal bodies (HB) nymph -1 and reduced the survival of nymphs ≤50%. Most nymphs (>70%) survived after injection of 6 × 10 3 and 2 × 10 3  HB nymph -1 ., Conclusions: Findings emphasize a distinct resistance of nymphs of the American cockroach to infections by M. anisopliae., Significance and Impact of the Study: Our findings provide support for the development of biological control of this synanthropic cockroach pest., (© 2016 The Society for Applied Microbiology.)

The omnivorous cockroach Periplaneta americana hosts a diverse hindgut microbiota encompassing hundreds of microbial species. In this study, we used 16S rRNA gene sequencing to examine the effect of diet on the composition of the P. americana hindgut microbial community. Results show that the hindgut microbiota of P. americana exhibit a highly stable core microbial community with low variance in compositions between individuals and minimal community change in response to dietary shifts. This core hindgut microbiome is shared between laboratory-hosted and wild-caught individuals, although wild-caught specimens exhibited a higher diversity of low-abundance microbes that were lost following extended cultivation under laboratory conditions. This taxonomic stability strongly contrasts with observations of the gut microbiota of mammals, which have been shown to be highly responsive to dietary change. A comparison of P. americana hindgut samples with human fecal samples indicated that the cockroach hindgut community exhibited higher alpha diversity but a substantially lower beta diversity than the human gut microbiome. This suggests that cockroaches have evolved unique mechanisms for establishing and maintaining a diverse and stable core microbiome., Importance: The gut microbiome plays an important role in the overall health of its host. A healthy gut microbiota typically assists with defense against pathogens and the digestion and absorption of nutrients from food, while dysbiosis of the gut microbiota has been associated with reduced health. In this study, we examined the composition and stability of the gut microbiota from the omnivorous cockroach Periplaneta americana. We found that P. americana hosts a diverse core gut microbiome that remains stable after drastic long-term changes in diet. While other insects, notably ant and bee species, have evolved mechanisms for maintaining a stable association with specific gut microbiota, these insects typically host low-diversity gut microbiomes and consume specialized diets. In contrast, P. americana hosts a gut microbiota that is highly species rich and consumes a diverse solid diet, suggesting that cockroaches have evolved unique mechanisms for developing and maintaining a stable gut microbiota., (Copyright © 2016, American Society for Microbiology. All Rights Reserved.)

Cockroaches are parasitized by thelastomatid nematodes, which live in an obligate manner in their hindgut and interact with the resident microbial community. In the present study, a composition analysis was performed on the gut microbiome of Periplaneta fuliginosa and P. americana to investigate natural and artificial infection by thelastomatid nematodes. Nine libraries of the 16S rRNA gene V3-V4 region were prepared for pyrosequencing. We examined the complete gut microbiome (fore-, mid-, and hindgut) of lab-reared P. fuliginosa naturally infected with the parasitic nematode Leidynema appendiculatum and those that were nematode-free, and complemented our study by characterizing the hindgut microbial communities of lab-reared P. americana naturally infected with Hammerschmidtiella diesingi and Thelastoma bulhoesi, artificially infected with L. appendiculatum, and those that were nematode-free. Our results revealed that the fore- and midgut of naturally infected and nematode-free P. fuliginosa have close microbial communities, which is in contrast with hindgut communities; the hindgut communities of both cockroaches exhibit higher microbial diversities in the presence of their natural parasites and marked differences were observed in the abundance of the most representative taxa, namely Firmicutes, Proteobacteria, and Bacteroidetes. Our results have provided basic information and encourage further studies on multitrophic interactions in the cockroach gut as well as the thelastomatid nematodes that play a role in this environment.

Cockroaches are surrogate hosts for microbes that cause many human diseases. In spite of their generally destructive nature, cockroaches have recently been found to harbor potentially beneficial and medically useful substances such as drugs and allergens. However, genomic information for the American cockroach (Periplaneta americana) is currently unavailable; therefore, transcriptome and gene expression profiling is needed as an important resource to better understand the fundamental biological mechanisms of this species, which would be particularly useful for the selection of novel antimicrobial peptides. Thus, we performed de novo transcriptome analysis of P. americana that were or were not immunized with Escherichia coli. Using an Illumina HiSeq sequencer, we generated a total of 9.5 Gb of sequences, which were assembled into 85,984 contigs and functionally annotated using Basic Local Alignment Search Tool (BLAST), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) database terms. Finally, using an in silico antimicrobial peptide prediction method, 86 antimicrobial peptide candidates were predicted from the transcriptome, and 21 of these peptides were experimentally validated for their antimicrobial activity against yeast and gram positive and -negative bacteria by a radial diffusion assay. Notably, 11 peptides showed strong antimicrobial activities against these organisms and displayed little or no cytotoxic effects in the hemolysis and cell viability assay. This work provides prerequisite baseline data for the identification and development of novel antimicrobial peptides, which is expected to provide a better understanding of the phenomenon of innate immunity in similar species.

The present paper reports the effects of Metarhizium anisopliae, Isaria fumosoroseus and Hirsutella thompsonaii on Periplaneta americana. I. fumosoroseus and H. thompsonaii were cultured at 28±1°C on potato carrot agar and M. anisopliae was cultured at 28±1°C on potato dextrose agar for 14days. Conidial suspensions of fungi were given to cockroaches through different routes. M. anisopliae shows high virulence against adult cockroaches and mortality ranges from 38.65% to 78.36% after 48h. I. fumosoroseus and H. thompsonii show less virulence compared to M. anisopliae. We also investigated the effect of these three fungi on the activity of lactate dehydrogenase, lipid peroxidation and catalase in different tissues of the insect to gain an understanding of the different target site. The result suggested that the activity of lactate dehydrogenase, catalase and level of malondialdehyde varies in different organs and through different routes of exposure. Based on mortality percentages, all tested fungi had high potentials for biocontrol agents against P. americana. Our study reveals for the first time that I. fumosoroseus and H. thompsonaii fungal infections initiate oxidative stress in the midgut, fat body, whole body and hemolymph of cockroach thereby suggesting them to be the target organs for oxidative damage., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Leptospirosis cases in Colombia are typically linked to peridomestic rodents; however, empirical data suggest that Leptospira-infected patients with no apparent exposure to these reservoirs are common. Cockroaches (Periplaneta spp.) have equal or greater interaction with humans than rodents, yet their potential role as carriers of Leptospira has not been assessed. We determined if pathogenic Leptospira is harbored by Periplaneta spp. in Cali (Colombia) and the variables influencing this relationship. Fifty-nine cockroaches were captured from seven sites and DNA was extracted from the body surface and digestive tract for a multiplex polymerase chain reaction, targeting genes secY and flaB. Logistic regression models and proportion tests showed a higher likelihood for Leptospira to be isolated from body surfaces (P > 0.001) and from individuals inside houses (six times more likely). These findings are the first to demonstrate an association between Periplaneta spp. and Leptospira, suggesting the need to investigate the potential for cockroaches to serve as reservoirs or transport hosts for Leptospira., (© The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

The American cockroach Periplaneta americana, one of the worlds' most important urban insect pests was tested with entomopathogenic fungi. Most promising Metarhizium anisopliae, Metarhizium robertsii and Beauveria bassiana killed nymphs (≥ 81.7% mortality, 25 days after treatment), and these fungi developed on all dead insects. Other fungi tested were less virulent (Metarhizium frigidum and Purpureocillium lilacinum) or avirulent (Isaria cateniobliqua, Isaria farinosa, Simplicillium lanosoniveum, Sporothrix insectorum and Tolypocladium cylindrosporum). Intrageneric and intraspecific variability of fungal activity was detected. Adults were highly susceptible, and oothecae proved to be more resistant than nymphs and adults to infection with M. anisopliae IP 46. Findings of the study underscore the potential of fungi as biocontrol agents against this pest., (© 2013.)

Objective: The present study was to fully evaluate the intestinal bacterial community of Periplaneta americana, an important model to study insects., Methods: We investigated the bacterial community of P. americana gut by culture-independent methods, involving constructing the 16S rRNA gene library and microbial diversity analysis., Results: The phylotypes were affiliated with Proteobacteria (66.4%), Bacteroidetes (17.8%), Firmicutes (14.5%), Fusobacteria (0.6%) and unclassified bacteria (0.6%). Phylogenetic analysis shows that 15% of the sequences clustered with that from a closely related omnivorous cockroach; and 59% clustered with that from more distantly related animals, including omnivorous, herbivorous, and carnivorous animals, which differ greatly in feeding habits. Moreover, 18% of the clones showed high sequence identity with potential pathogens closely related to human diseases, which also reinforces the concept of the cockroach as a carrier of pathogens., Conclusion: Due to their habits of feeding on a variety of foodstuffs, omnivorous cockroaches harbor a large and diverse microbial community in the gut. The host phylogeny and dietary habits might be critical for the intestinal bacterial community composition of cockroaches.

In insects, trophocytes (adipocytes) are major cells of a storage organ, the fat body, from which stored glycogen and lipids are mobilized under starvation. However, cockroaches have 2 additional types of cell in the fat body: mycetocytes harboring an endosymbiont, Blattabacterium cuenoti, and urocytes depositing uric acid in urate vacuoles. These cells have not been investigated in terms of their roles under starvation conditions. To gain insight into the roles of trophocytes, mycetocytes and urocytes in cockroaches, structural changes were first investigated in the cells associated with starvation in the American cockroach, Periplaneta americana, by light and electron microscopy. The area of lipid droplets in trophocytes, the endosymbiont population and mitotic activity in mycetocytes, and the area of urate vacuoles in urocytes were analyzed in association with survival rates of the starved cockroaches. After 2 weeks of starvation, trophocytes lost glycogen rosettes and their area of lipid droplets decreased, but almost all cockroaches survived this period. However, further starvation did not reduce the area, but the survival rates dropped rapidly and all cockroaches died in 7 weeks. Endosymbionts were not affected in terms of population size and mitotic activity, even if the cockroaches were dying. The area of urate vacuoles rapidly decreased in a week of starvation and did not recover upon further starvation. These results indicate that starved cockroaches mobilize glycogen and lipids stored in trophocytes to survive for 2 weeks and then die after the exhaustion of nutrients in these cells. Endosymbionts are not digested for the recycling of nutrients, but uric acid is reused under starvation., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Food resources contaminated with spoilage or pathogenic microorganisms pose severe problems to all higher organisms. Here, we describe a food-hygienic strategy of the emerald cockroach wasp Ampulex compressa. The wasp larvae develop on and inside the American cockroach Periplaneta americana, a host that can harbor various putrefactive microbes, as well as human and insect pathogens. From P. americana, we isolated the Gram-negative bacterium Serratia marcescens, which is a potent entomopathogen that can rapidly kill insect larvae. It is also known as a food contaminant and as an opportunistic human pathogen. Using behavioral observations and chemical analyses, we demonstrated that A. compressa larvae impregnate their cockroach hosts from inside with large amounts of an oral secretion containing a blend of γ-lactones and isocoumarins with (R)-(-)-mellein [(R)-(-)-3,4-diydro-8-hydroxy-3-methylisocoumarin] and micromolide [(4R,9Z)-octadec-9-en-4-olide] as dominant components. We fractionated hexane extracts of the secretion and investigated the antimicrobial properties of the fraction containing the lactones and isocoumarins, as well as of synthetic (R)-(-)-mellein and micromolide, against S. marcescens and a Gram-positive bacterium, Staphylococcus hyicus, in broth microdilution assays. The test fraction inhibited growth of both tested bacteria. The activity of the fraction against S. marcescens was explained by (R)-(-)-mellein alone, and the activity against S. hyicus was explained by the combined action of (R)-(-)-mellein and micromolide. Our data suggest that the specific combination of antimicrobials in the larval secretion provides an effective frontline defense against the unpredictable spectrum of microbes that A. compressa larvae may encounter during their development inside their cockroach hosts.

Lophomonas striata is a multiflagellate parabasalid commensal in the hindgut of the omnivorous cockroaches Blatta orientalis and Periplaneta americana. Its closest relatives were traditionally thought to include similar multiflagellate parabasalids with a single flagellar area that degenerates during mitosis, such as Joenia and Kofoidia. However, molecular phylogenetic analyses have shown that "lophomonads" are not monophyletic. We have determined the SSU rRNA sequence of L. striata and we find that it branches sister to the Trichonymphida with strong support. This is surprising because all other lophomonads sampled to date branch within the Cristamonadida, and the order Trichonymphida (e.g. Trichonympha, Pseudotrichonympha, and Hoplonympha) is both morphologically coherent and monophyletic in SSU rRNA phylogenies. Trichonymphida, unlike the lophomonads, share a bilateral symmetry, in which their multiple flagella occur in two (or sometimes four) regions, and instead of degenerating upon mitosis, half of the flagella are passed to each daughter cell. The single apical flagellar region characteristic of lophomonads is therefore either plesiomorphic or it has arisen multiple times in parabasalids; our phylogenetic analyses and available ultrastructural evidence suggest the latter. Our results also suggest that parabasalid gut symbionts may have been vertically transmitted in cockroaches before the common ancestor of Cryptocercus and termites., (Copyright © 2011 Elsevier GmbH. All rights reserved.)

Although it has been difficult to prove the direct involvement of cockroaches (i.e. insects of the order Blattaria) in the transmission of pathogenic agents to humans, such insects often carry microorganisms that are important in nosocomial infections, and their medical importance in the spread of bacteria cannot be ruled out. In houses and institutions with poor standards of hygiene, heavy infestations with cockroaches, such as the peridomestic American cockroach (Periplaneta americana L.) and the domestic German cockroach (Blattella germanica L.), can occur. In the present study, cockroaches (126 B. germanica and 69 P. americana) were collected from four buildings (three public training hospitals and one house) in central Tehran, Iran. Each insect was processed, under sterile conditions, so that the bacteria on its external surfaces and in its alimentary tract and faecal pellets could be isolated and identified. The oldest and largest of the three hospitals sampled (a 1400-bed unit built 80 years ago) appeared to be the one most heavily infested with cockroaches, and cockroaches from this hospital accounted for most (65.4%) of the isolates of medically important bacteria made during the study. No significant difference was found between the percentages of P. americana and B. germanica carrying medically important bacteria (96.8% v. 93.6%; P>0.05). At least 25 different species of medically important bacteria were isolated and identified, and at least 22 were Gramnegative. The genus of enteric bacteria most frequently isolated from both cockroach species, at all four collection sites, was Klebsiella. The cockroaches from each hospital were much more likely to be found contaminated with medically important bacteria than those from the house. The hospital cockroaches were also more likely to be carrying medically important bacteria internally than externally (84.3% v. 64.1%; P<0.05). The implications of these and other recent results, for the control of cockroaches and nosocomial infections, are discussed.

We examined the diversity of a marker gene for homoacetogens in two cockroach gut microbial communities. Formyltetrahydrofolate synthetase (FTHFS or fhs) libraries prepared from a wood-feeding cockroach, Cryptocercus punctulatus, were dominated by sequences that affiliated with termite gut treponemes. No spirochete-like sequences were recovered from the omnivorous roach Periplaneta americana, which was dominated by Firmicutes-like sequences.

Background: Flies and cockroaches are two insects in close contact with human beings. They are carriers of human pathogenic bacteria on the external areas of their bodies or in their digestive tracts. This study examines Periplaneta americana and Musca domestica collected from the residential areas of six districts in Tangier, Morocco., Methodology: In total, 251 bacteria were isolated from external areas of the participants' bodies and the antimicrobial susceptibility was calculated., Results: The predominant bacterial species included Escherichia coli (17.9%), Klebsiella spp. (14.7%), Providencia spp. (9.6%), Staphylococcus spp. (15.1%) and Enterococcus spp. (11.6%). The study showed no difference between the species of bacterial strains from American cockroaches and houseflies. Carbapenems and aminoglycosides were active against 100% of the Gram-negative bacilli isolated in this study. Staphylococcus spp. strains were susceptible to linezolid, vancomycin, daptomycin, levofloxacin and cotrimoxazole, and no antibiotic resistance was found in Enterococcus spp., Conclusions: In our setting, although both cockroaches and flies collected from residential areas may be vectors of human pathogenic bacteria, the infections caused by them are easily treatable as a result of the high susceptibility of their bacteria to antibiotics routinely used in the community or in hospitals.

Nitrogen acquisition and assimilation is a primary concern of insects feeding on diets largely composed of plant material. Reclaiming nitrogen from waste products provides a rich reserve for this limited resource, provided that recycling mechanisms are in place. Cockroaches, unlike most terrestrial insects, excrete waste nitrogen within their fat bodies as uric acids, postulated to be a supplement when dietary nitrogen is limited. The fat bodies of most cockroaches are inhabited by Blattabacterium, which are vertically transmitted, Gram-negative bacteria that have been hypothesized to participate in uric acid degradation, nitrogen assimilation, and nutrient provisioning. We have sequenced completely the Blattabacterium genome from Periplaneta americana. Genomic analysis confirms that Blattabacterium is a member of the Flavobacteriales (Bacteroidetes), with its closest known relative being the endosymbiont Sulcia muelleri, which is found in many sap-feeding insects. Metabolic reconstruction indicates that it lacks recognizable uricolytic enzymes, but it can recycle nitrogen from urea and ammonia, which are uric acid degradation products, into glutamate, using urease and glutamate dehydrogenase. Subsequently, Blattabacterium can produce all of the essential amino acids, various vitamins, and other required compounds from a limited palette of metabolic substrates. The ancient association with Blattabacterium has allowed cockroaches to subsist successfully on nitrogen-poor diets and to exploit nitrogenous wastes, capabilities that are critical to the ecological range and global distribution of cockroach species.

The presence of cockroaches has health implications, such as nosocomial infection, as the insects move freely from areas within and around hospitals that may harbor pathogenic organisms. The goals of the present study were to determine species of bacteria isolated from cockroaches found in three Tehran hospitals, and to determine distribution of potential vectors by species and sex. The study is descriptive laboratory research. A total of 305 cockroaches from three species were trapped and identified: Periplaneta americana (65.6%), Blatella germanica (12.1%), and Blatta orientalis (22.3%). From these potential vectors, 19 species of bacteria were isolated and identified. After transportation of cockroaches to the laboratory, separation of the whole-homogenized suspension of each species was carried out. Identification of the isolated bacteria was performed according to Burgey's Manual of Determinative Bacteriology, 9th Edition. The most common species of bacteria isolated from cockroaches were Escherichia coli, Streptococcus Group D, Bacillus spp., Klebsiella pneumoniae, and Proteus vulgaris. No statistical significance was found between sex and species of cockroach carrying bacteria (p > 0.05), but significance was found for sex in Citrobacter freundii, Staphylococcus aureus, and Streptococcus non-group A&B (p < 0.05).

Blattabacteria are intracellular endosymbionts of cockroaches and primitive termites that belong to the class Flavobacteria and live only in specialized cells in the abdominal fat body of their hosts. In the present study we determined genome sizes as well as genome copy numbers for the endosymbionts of three cockroach species, Blattella germanica, Periplaneta americana and Blatta orientalis. The sole presence of blattabacteria in the fat body was demonstrated by rRNA-targeting techniques. The genome sizes of the three blattabacteria were determined by pulsed field gel electrophoresis. The resulting total genome sizes for the three symbionts were all approximately 650 +/- 15 kb. Comparison of the genome sizes with those of free-living Bacteroidetes shows extended reduction, as occurs in other obligatory insect endosymbionts. Genome copy numbers were determined based on cell counts and determination of DNA amounts via quantitative PCR. Values between 10.2 and 18.3 and between 323 and 353 were found for the symbionts of P. americana and B. orientalis respectively. Polyploidy in intracellular bacteria may play a significant role in the genome reduction process.

"ZA-type" cages were used to capture cockroaches in 267 sites of 5 cities in Hainan. Species were identified and bacteria were isolated by routine method. 441 cockroaches were collected and identified as five species belonging to two genera, 75.3% being Periplaneta americana. More cockroaches were found in sewerage. Bacteria were detected from 82.4% of cockroaches, including Escherichia coli, Pseudomonas aeruginosa, Salmonella sp, Staphylococcus aureus, Shigella sp, Bacillus proteus and sort of mycetes. Therefore, the dominant species is Periplaneta americana in Hainan, and the high bacteria-carrying behavior of cockroaches indicates its importance in disease transmission.

Methanomicrococcus blatticola is an obligately anaerobic methanogen that derives the energy for growth exclusively from the reduction of methylated compounds to methane with molecular hydrogen as energy source. Competition for methanol (concentration below 10 microM) and H(2) (concentration below 500 Pa), as well as oxidative stress due to the presence of oxygen are likely to occur in the peripheral region of the cockroach hindgut, the species' normal habitat. We investigated the ecophysiological properties of M. blatticola to explain how it can successfully compete for its methanogenic substrates. The organism showed affinities for methanol (K(m)=5 microM; threshold<1 microM) and hydrogen (K(m)=200 Pa; threshold <0.7 Pa) that are superior to other methylotrophic methanogens (Methanosphaera stadtmanae, Methanosarcina barkeri) investigated here. Thermodynamic considerations indicated that 'methanol respiration', i.e. the use of methanol as the terminal electron acceptor, represents an attractive mode of energy generation, especially at low hydrogen concentrations. Methanomicrococcus blatticola exploits the opportunities by specific growth rates (>0.2 h(-1)) and specific growth yields (up to 7 g of dry cells per mole of methane formed) that are particularly high within the realm of mesophilic methanogens. Upon oxygen exposure, part of the metabolic activity may be diverted into oxygen removal, thus establishing appropriate anaerobic conditions for survival and growth.

Cockroaches carry microorganisms to sterilized materials, equipment and non-contaminated food in hospitals and in homes. Hence, they are regarded as major microbial vectors. We isolated and identified microorganisms from living cockroaches captured from a Brazilian health care institution and we determined the antimicrobial susceptibility profile of the isolates. Following collection, cockroaches were immobilized at 4 degrees C for 10 to 20 minutes, immersed in 0.8% saline solution and then homogenized. Next, the suspension was seeded in culture media: brain and heart agar infusion, MacConkey agar, Naito agar, and they were incubated for up to 48 hours at 37 degrees C. Sabouraud agar cultures were incubated at ambient temperature. The microbial colonies were analyzed with a stereomicroscope to count and phenotypically identify the colony-forming units (cfu). The disc-diffusion method was used to determine the profile of susceptibility to antimicrobials. Among the 103 cockroaches analyzed, fungi were found in 97%, enterobacteria in 74.6% and coagulase-negative staphylococci (CNS) in 25.40%. Among the enterobacteria, 96% were resistant to gentamicin, 84% to ampicillin, 75.3% to caphalothin, 66.7% to ampicillin-sulbactam, 50% to aztreonam and 30% to chloramphenicol. Among the CNS, 61% were resistant to oxacillin. We concluded that cockroaches carry organisms associated with nosocomial infections and that are resistant to antimicrobials. This fact contributes to the epidemiological chain, complicating therapeutics, and consequently, medical costs are increased.

The fungal pathogenic flora of the external surface of 103 cockroaches (Periplaneta americana) collected from the intensive care unit of a hospital were investigated. In this study, a high percentage of test cockroaches (93.2%) were found to carry fungi of medical importance. The main fungi isolated were species of Candida, Aspergillus and Penicillium. Information about the carriage of pathogenic fungi by cockroaches in hospital environment is scanty. The results suggest that cockroaches can play a role in dissemination of fungi, which they can carry on their external surface.

Methanomicrococcus blatticola, a methanogenic archaeon isolated from the cockroach Periplaneta americana, is specialised in methane formation by the hydrogen-dependent reduction of methanol, monomethyl-, dimethyl- or trimethylamine. Experiments with resting cells demonstrated that the capability to utilise the methylated one-carbon compounds was growth substrate dependent. Methanol-grown cells were incapable of methylamine conversion, while cells cultured on one of the methylated amines did not metabolise methanol. Unlike trimethylamine, monomethyl- and dimethylamine metabolism appeared to be co-regulated. The central reaction in the energy metabolism of all methanogens studied so far, the reduction of CoM-S-S-CoB, was catalysed with high specific activity by a cell-free system. Activity was associated with the membrane fraction. Phenazine was an efficient artificial substrate in partial reactions, suggesting that the recently discovered methanophenazine might act in the organism as the physiological intermediary electron carrier. Our experiments also showed that M. blatticola apparently lacks the pathway for methyl-coenzyme oxidation to CO2, explaining the strict requirement for hydrogen in methanogenesis and the obligately heterotrophic character of the organism.

Cockroaches may harbor and disseminate microorganisms to the environment. In this study, Periplaneta americana and Blattella germanica were collected from 40 households in Kaohsiung City and Kaohsiung County, Taiwan. Cockroach infestation was found in 50% of the studied households and 226 cockroaches (123 P. americana and 103 B. germanica) collected by trapping. P. americana was more often found in the kitchen (70.7%) whereas B. germanica in the storage room (51.5%) and kitchen (36.9%). There was no significant difference between the percentages of P. americana (99.9%) and B. germanica (98.0%) carrying bacteria. A total of 25 species of bacteria was isolated from P. americana and only 21 from B. germanica. Antibiotic resistance was found in Staphylococcus aureus, Enterococcus species, Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Serratia marcescens, and Proteus species isolated from the cockroaches. These findings suggest a potential role of cockroaches in the transmission of pathogenic bacteria with antibiotic resistance in households.

Objectives: The routes of human infection with Helicobacter pylori remain unclear. In the present study, we examined cockroaches as possible vectors for transmission of H. pylori., Methods: We used a common species of cockroach (Periplaneta fuliginosa). After a 3-day fast, cockroaches were placed on agar plates containing freshly grown H. pylori (Sydney strain) (challenge group) or on sterile agar plates without H. pylori (control group). After 24 h of challenge, cockroaches were moved to disinfected containers, and sterile food and water were provided. The external surfaces (legs and body) and excreta of the cockroaches were sampled for culture, rapid urease test, and polymerase chain reaction (PCR)., Results: H. pylori were culturable from the excreta of the challenge group for 24 h postchallenge. Positive rapid urease test results were obtained up to day 3, and PCR analysis was positive for H. pylori DNA up to day 7 from the excreta. In contrast, H. pylori were not culturable from the external surfaces of the cockroaches. The rapid urease test was positive for only 8 h, and PCR analysis was positive for H. pylori DNA for 1 day from the external surface., Conclusions: Cockroaches usually live in unsanitary environments and may contaminate foods and food containment areas such as pantries. Transmission of H. pylori infection could be achieved via inadvertent ingestion of foods contaminated with cockroach excreta containing viable H. pylori.

This study attempted to isolate mycobacteria from hospital and household cockroaches from 90 hospitals and 40 households in Kaohsiung City and Kaohsiung County, South Taiwan. Among 203 cockroaches (139 Periplaneta americana and 64 Blattella germanica) collected from the hospitals, six Mycobacterium spp. were isolated and identified by polymerase chain reaction-restriction fragment length polymorphism analysis. In 12 cockroaches (P. americana): four Mycobacterium kansaii, three Mycobacterium xenopi, two Mycobacterium gordonae, one Mycobacterium hemophilium, one Mycobacterium fortuitum, and one Mycobacterium avium. However, no mycobacteria were obtained form the hospital B. germanica or 226 household cockroaches (123 P. americana and 103 B. germanica). As cockroach infestation occurs commonly in the hospital environment, they may potentially be implicated as a cause of hospital-acquired infections due to non-tuberculous mycobacteria.

Objective: To isolate and identify microorganisms from cockroaches that were captured in a public hospital and to test the antimicrobial susceptibility of these microorganisms., Methods: Cockroaches were captured in the morning and at night. They were placed in flasks rinsed with 70% alcohol, transferred to sterilized flasks, and then taken to the laboratory. Only cockroaches captured whole and live were utilized for the study. After being immobilized at 0 degree C, each cockroach was placed in a test tube with sterile saline solution (0.8%) and then homogenized. The resulting solution was then placed in the following five culture media: MacConkey agar, nutrient broth, brain-heart infusion agar, Sabouraud agar, and mannitol. The cultures were examined using a stereomicroscope, and colony-forming units were counted. The disk diffusion test was used to determine antimicrobial susceptibility., Results: We found a 56% prevalence of enterobacteria and an 18% prevalence of coagulase-negative staphylococci. Fifteen species of enterobacteria were identified. The most frequent were Klebsiella pneumoniae (17%), Enterobacter aerogenes (14%), Serratia marcescens (13%), Hafnia alvei (12%), Enterobacter gergoviae and Enterobacter cloacae (each 9%), and Serratia spp. (6%). Both the enterobacteria and the coagulase-negative staphylococci showed significant resistance to antimicrobials, including oxacillin., Conclusions: The prevalence of enteropathogenic bacteria and coagulase-negative staphylococci isolated from Periplaneta americana cockroaches in the studied hospital reflects the weakness of the measures adopted both for vector control and for antimicrobial use. The results show the need to implement effective health-institution programs focusing on hygiene and the rational use of antimicrobials.

A small irregular coccoid methanogenic bacterium (PAT) was isolated from the hindgut of the cockroach Periplaneta americana. Fluorescence microscopy and transmission electron microscopy of the hindgut of P. americana suggest that the organism occurs abundantly in the microbiota attached to the hindgut wall. The strain produces methane by the reduction of methanol and methylated amines with molecular hydrogen. Acetate, coenzyme M, yeast extract, tryptic soy broth and vitamins are required for growth. The cells lack a rigid cell wall and lyse immediately in buffers of low ionic strength. Maximum rate of growth (specific growth rate, 0.22 h(-1)) occurs in a rich medium at 39 degrees C, at a pH range of 7.2-7.7 and at a salt concentration below 100 mM NaCl. Sequence analysis of the small-subunit rDNA indicates that strain PAT is related to the family Methanosarcinaceae but does not belong to any previously described genus. Therefore, it is proposed that strain PAT be classified in a new genus, related to the Methanosarcinaceae, as Methanomicrococcus blatticola (type strain PAT = DSM 13328T).

The bacterial flora associated with external body parts, stomach and intestine of male and female Periplaneta americana inhabitating students' halls, teachers' residences and houses of low-paid employees in Jahangirnagar, University Campus were examined and analysed qualitatively and quantitatively. The qualitative study in P. americana revealed the presence of 31 species of bacteria belonging to 16 genera. Most of these bacteria are pathogenic to man and domestic animals. The total viable bacterial counts (TVBC/g) in students' halls, teachers' residences and houses of low-paid employees were 1.27 x 10(8), 6.04 x 10(7) and 1.65 x 10(8) respectively in the male and 1.43 x 10(8), 5.83 x 10(7) and 1.63 x 10(8) respectively in the female cockroaches. The highest prevalence of bacterial flora both in number and types occurred in stomach followed by intestine and the external body parts.

A total of 221 cockroaches (Blatta and Periplaneta spp.), collected in hospitals, houses, animal sheds, grocery stores and restaurants, in various parts of South Kanara District, a south-west coastal region of India, were studied bacteriologically for the presence of various salmonellas. Salmonellas were isolated from 4.1% of these cockroaches. Nine strains of salmonellas were recovered, belonging to five serotypes--Salmonella bovismorbificans, S. oslo, S. typhimurium, S. mbandaka and S. braenderup, the former two being the commonest serotypes. All salmonellas were resistant to one or other of 11 antibacterial drugs used in the susceptibility test. Isolation of salmonellas from cockroaches collected from the livestock premises and human dwellings suggested that they may act as significant reservoirs of salmonella in nature. Recovery of serotypes, phage types and R-types that were commonly isolated from humans and animals of this locality, suggested a transmission role for cockroaches. By harbouring potentially pathogenic, drug-resistant salmonellas, these wandering arthropods may pose dangerous infective hazards to humans and animals.

Production of methane in the hindgut of the cockroach Periplaneta americana was found to vary, depending on the feeding regimen. Methane production was positively correlated with the numbers of the ciliate Nyctotherus ovalis living in the cockroach hindgut. Defaunation of the cockroaches by means of low concentrations of metronidazole (Flagyl) resulted in a quick drop of methane production. Addition of the methanogenic substrates acetate and formate to isolated hindguts stimulated methane production. Inside the ciliate cells, autofluorescing bacteria could be demonstrated which were presumed to be methanogens. Electron microscopy revealed that the bacteria resembled Methanobrevibacter and that they were closely associated with organelles which contained infolded membranes and which were presumably hydrogenosomes.