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Cockroaches are well-known pests and quarantined organisms worldwide. Due to morphological diversity and a lack of molecular data, their classification and identification are facing challenges. This study performed classification, identification, and DNA barcoding for cockroaches collected from China. Seventy-six samples were morphologically identified as seven species of two superfamilies that included Blattella germanica, Eublaberus posticus and Blaptica dubia belonging to the superfamily Blaberoidea, and Periplaneta americana, Periplaneta lateralis, Periplaneta fuliginosa and Periplaneta australasiae belonging to the superfamily Blattoidea. Based on sequence alignments of nine ribosomal and mitochondrial genes across the order Blattaria retrieved from GenBank, rDNA ITS2-517 bp and mtDNA 16S-327 bp were screened as candidates for molecular identification. Universal primers were designed for PCR amplification, cloning, and sequencing of the 37 representative samples. Sequence alignments and phylogeny analysis showed that both ITS2 and 16S confirmed samples 1-9, 20-24, and 25-29 as B. germanica, P. americana, and P. lateralis, respectively; only 16S (not ITS2) confirmed samples 10-14, 15-19, 30-34, and 35-37 as E. posticus, Blap. dubia, P. fuliginosa, and P. australasiae, respectively, indicating that 16S was a better target than ITS2 for molecular identification of cockroaches. Conservative motif and divergence analysis further revealed that ITS2 sequences vary significantly among different taxa, whereas 16S sequences are relatively conserved. There is an obvious barcoding gap between maximum intraspecific divergence and minimum interspecific divergence (2.57 % vs. 5.62 %) for ITS2, but not for 16S (6.15 % vs. 2.63 %). Therefore, it was confirmed that ITS2 is an ideal DNA barcode for molecular identification of cockroaches at lower category., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The Canary Islands is a Macaronesian volcanic archipelago with a depauperate community of three species of Kalotermitidae, including Kalotermes dispar. A total of 54 Kalotermes colonies were collected from Gran Canaria, Tenerife, La Gomera, La Palma, and El Hierro islands. Soldiers and imagos were morphologically examined and sequenced for four mitochondrial markers. Although morphological differences could not be detected, phylogenetic analysis of both cox1/tRNA/cox2 and rrnL markers revealed two distinct clades of K. dispar, suggesting cryptic diversity. The diversification within the Canary Kalotermes lineage most likely occurred around 7.5 Mya, while the divergence within the two clades was reconstructed at about 3.6 Mya and 1.9 Mya. Kalotermes approximatus from the southeastern Nearctic constitutes a sister to the Canary Kalotermes, while the Palearctic K. flavicollis, K. italicus, and K. phoenicae form a separate clade. It is hypothesized that a faunal exchange of Kalotermes from the Nearctic to the Canary Islands occurred via transoceanic rafting during the mid-Miocene., (© The Author(s) 2024. Published by Oxford University Press on behalf of Entomological Society of America.)

Nephridiophagids are unicellular eukaryotes that parasitize the Malpighian tubules of numerous insects. Their life cycle comprises multinucleate vegetative plasmodia that divide into oligonucleate and uninucleate cells, and sporogonial plasmodia that form uninucleate spores. Nephridiophagids are poor in morphological characteristics, and although they have been tentatively identified as early-branching fungi based on the SSU rRNA gene sequences of three species, their exact position within the fungal tree of live remained unclear. In this study, we describe two new species of nephridiophagids (Nephridiophaga postici and Nephridiophaga javanicae) from cockroaches. Using long-read sequencing of the nearly complete rDNA operon of numerous further species obtained from cockroaches and earwigs to improve the resolution of the phylogenetic analysis, we found a robust affiliation of nephridiophagids with the Chytridiomycota-a group of zoosporic fungi that comprises parasites of diverse host taxa, such as microphytes, plants, and amphibians. The presence of the same nephridiophagid species in two only distantly related cockroaches indicates that their host specificity is not as strict as generally assumed.

Phylogenetic relationships among subgroups of cockroaches and termites are still matters of debate. Their divergence times and major phenotypic transitions during evolution are also not yet settled. We addressed these points by combining the first nuclear phylogenomic study of termites and cockroaches with a thorough approach to divergence time analysis, identification of endosymbionts, and reconstruction of ancestral morphological traits and behaviour. Analyses of the phylogenetic relationships within Blattodea robustly confirm previously uncertain hypotheses such as the sister-group relationship between Blaberoidea and remaining Blattodea, and Lamproblatta being the closest relative to the social and wood-feeding Cryptocercus and termites. Consequently, we propose new names for various clades in Blattodea: Cryptocercus + termites = Tutricablattae; Lamproblattidae + Tutricablattae = Kittrickea; and Blattoidea + Corydioidea = Solumblattodea. Our inferred divergence times contradict previous studies by showing that most subgroups of Blattodea evolved in the Cretaceous, reducing the gap between molecular estimates of divergence times and the fossil record. On a phenotypic level, the blattodean ground-plan is for egg packages to be laid directly in a hole while other forms of oviposition, including ovovivipary and vivipary, arose later. Finally, other changes in egg care strategy may have allowed for the adaptation of nest building and other novelties.

Objectives: The current study was conducted to identify cockroach species (Blattodea) of northwestern Iran in public places using morphological characteristics and ribosomal DNA internal transcribed spacer 2 (rDNA-ITS2). Sequences were analyzed with Basic Local Alignment Search Tool (BLAST) searches, Neighbor-Joining methods based on and Tamura-Nei phylogenetic analyses. In addition, eight cockroach rDNA-ITS2 sequences from China, India, Iran and the United States obtained from GenBank were compared to those obtained in this study., Results: Specimens collected in Iran were identified as Periplaneta americana (L.), Shelfordella lateralis (Walker), Blatta orientalis (L.) (Blattodea: Blattidae), Blattella germanica (L.), Supella longipalpa (F.) (Blattodea: Ectobiidae), Polyphaga aegyptiaca (L.), and Polyphaga saussurei (Dohrn) (Blattodea: Corydiidae). rDNA-ITS2 nucleotide sequence analysis showed 100% similarity between P. aegyptiaca and P. saussurei species collected from Iran despite morphological differences. However, ITS2 sequence of P. americana submitted from China showed 30.49-31.71% difference to P. americana sequences from Iran and the United States. The results highlight the importance of morphological identification of cockroach species before conducting molecular techniques.

In this study, the complete mitochondrial genome of Cryptocercus meridianus was sequenced. The circular mitochondrial genome is 15,322 bp in size and contains 13 protein-coding genes, two ribosomal RNA genes (12S rRNA and 16S rRNA), 22 transfer RNA genes, and one D-loop region. We compare the mitogenome of C. meridianus with that of C. relictus and C. kyebangensis . The base composition of the whole genome was 45.20%, 9.74%, 16.06%, and 29.00% for A, G, C, and T, respectively; it shows a high AT content (74.2%), similar to the mitogenomes of C. relictus and C. kyebangensis . The protein-coding genes are initiated with typical mitochondrial start codons except for cox1 with TTG. The gene order of the C. meridianus mitogenome differs from the typical insect pattern for the translocation of tRNA-Ser AGN , while the mitogenomes of the other two Cryptocercus species, C. relictus and C. kyebangensis , are consistent with the typical insect pattern. There are two very long non-coding intergenic regions lying on both sides of the rearranged gene tRNA-Ser AGN . The phylogenetic relationships were constructed based on the nucleotide sequence of 13 protein-coding genes and two ribosomal RNA genes. The mitogenome of C. meridianus is the first representative of the order Blattodea that demonstrates rearrangement, and it will contribute to the further study of the phylogeny and evolution of the genus Cryptocercus and related taxa., Competing Interests: The authors declare no conflict of interest.

Complete mitochondrial genomes (mitogenomes) of two cockroach species, Periplaneta australasiae and Neostylopyga rhombifolia, 15,605 bp and 15,711 bp in length, respectively, were determined. As reported for other cockroach mitogenomes, the two mitogenomes possessed typical ancestral insect mitogenome gene composition and arrangement. Only several small intergenic spacers were found: one, which was common in all sequenced cockroach mitogenomes except for the genus Cryptocercus, was between tRNA-Ser (UCN) and ND1 and contained a 7bp highly conserved motif (WACTTAA). Three different types of short tandem repeats in the N. rhombifolia control region (CR) were observed. The homologous alignments of these tandem repeats with other six cockroach mitogenome CRs revealed a low similarity. Three conserved sequence blocks (CSB) were detected in both cockroach mitochondrial CRs. CSB1 was specific for blattinine mitogenomes and was highly conserved with 95% similarity, speculating that this block was a possible molecular synapomorphy for this subfamily. CSB3 located nearby downstream of CSB1 and has more variations within blattinine mitogenomes compared with CSB1. The CSB3 was capable of forming stable stem-loop structure with a small T-stretch in the loop portion. We assessed the influence of four datasets and two inference methods on topology within Orthopteroidea. All genes excluding the third codon positions of PCGs could generate more stable topology, and higher posterior probabilities than bootstrap values were presented at some branch nodes. The phylogenetic analysis with different datasets and analytical methods supported the monophyly of Dictyoptera and supported strongly the proposal that Isoptera should be classified as a family (Termitidae) of the Blattaria. Specifically, Shelfordella lateralis was inserted in the clade Periplaneta. Considering the K2P genetic distance, morphological characters, and the phylogenetic trees, we suggested that S. lateralis should be placed in the genus Periplaneta.

During the period between 1999 and 2006, wood-feeding cockroaches in the Cryptocercus punctulatus Scudder species complex were collected throughout Great Smoky Mountains National Park, USA. The chromosome numbers of insects from 59 sites were determined, and phylogenetic analyses were performed based on mitochondrial COII and nuclear ITS2 DNA. The distribution of the three male karyotypes found in the park (2n = 37, 39, and 45) is mapped and discussed in relation to recent disturbances and glacial history. Clades of the three karyotype groups meet near the ridgeline separating North Carolina from Tennessee in the center of the park, suggesting that these may have originated from separate lower elevation refugia after the last glacial maximum. The timing of divergence and a significant correlation between elevation difference and genetic distance in two of the clades supports this hypothesis. The ecological role of the cockroaches in the park is discussed., (© The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America.)

We identified the antifungal gene termicin in three species of Cryptocercus woodroaches. Cryptocercus represents the closest living cockroach lineage of termites, which suggests that the antifungal role of termicin evolved prior to the divergence of termites from other cockroaches. An analysis of Cryptocercus termicin and two β-1,3-glucanase genes (GNBP1 and GNBP2), which appear to work synergistically with termicin in termites, revealed evidence of selection in these proteins. We identified the signature of past selective sweeps within GNBP2 from Cryptocercus punctulatus and Cryptocercus wrighti. The signature of past selective sweeps was also found within termicin from Cryptocercus punctulatus and Cryptocercus darwini. Our analysis further suggests a phenotypically identical variant of GNBP2 was maintained within Cryptocercus punctulatus, Cryptocercus wrighti, and Cryptocercus darwini while synonymous sites diverged. Cryptocercus termicin and GNBP2 appear to have experienced similar selective pressure to that of their termite orthologues in Reticulitermes. This selective pressure may be a result of ubiquitous entomopathogenic fungal pathogens such as Metarhizium. This study further reveals the similarities between Cryptocercus woodroaches and termites.

Three complete mitochondrial genomes of Blaberidae (Insecta: Blattodea) (Gromphadorhina portentosa, Panchlora nivea, Blaptica dubia) and one complete mt genome of Blattidae (Insecta: Blattodea) (Shelfordella lateralis) were sequenced to further understand the characteristics of cockroach mitogenomes and reconstruct the phylogenetic relationship of Blattodea. The gene order and orientation of these four cockroach genomes were similar to known cockroach mt genomes, and contained 13 protein-coding genes (PCGs), 2 ribosomal RNA (rRNA) genes, 22 transfer RNA (tRNA) genes and one control region. The mt genomes of Blattodea exhibited a characteristics of a high A+T composition (70.7%-74.3%) and dominant usage of the TAA stop codon. The AT content of the whole mt genome, PCGs and total tRNAs in G. portentosa was the lowest in known cockroaches. The presence of a 71-bp intergenic spacer region between trnQ and trnM was a unique feature in B. dubia, but absent in other cockroaches, which can be explained by the duplication/random loss model. Based on the nucleotide and amino acid datasets of the 13 PCGs genes, neighbor-joining (NJ), maximum parsimony (MP), maximum likelihood (ML) and bayesian inference (BI) analyses were used to rebuild the phylogenetic relationship of cockroaches. All phylogenetic analyses consistently placed Isoptera as the sister cluster to Cryptocercidae of Blattodea. Ectobiidae and Blaberidae (Blaberoidea) formed a sister clade to Blattidae. Corydiidae is a sister clade of all the remaining cockroach species with a high value in NJ and MP analyses of nucleotide and amino acid datasets, and ML and BI analyses of the amino acid dataset., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Cryptocercus punctulatus and Parasphaeria boleiriana are two distantly related xylophagous and subsocial cockroaches. Cryptocercus is related to termites. Xylophagous cockroaches and termites are excellent model organisms for studying the symbiotic relationship between the insect and their microbiota. In this study, high-throughput 454 pyrosequencing of 16S rRNA was used to investigate the diversity of metagenomic gut communities of C. punctulatus and P. boleiriana, and thereby to identify possible shifts in symbiont allegiances during cockroaches evolution. Our results revealed that the hindgut prokaryotic communities of both xylophagous cockroaches are dominated by members of four Bacteria phyla: Bacteroidetes, Firmicutes, Proteobacteria, and Actinobacteria. Other identified phyla were Spirochaetes, Planctomycetes, candidatus Saccharibacteria (formerly TM7), and Acidobacteria, each of which represented 1-2% of the total population detected. Community similarity based on phylogenetic relatedness by unweighted UniFrac analyses indicated that the composition of the bacterial community in the two species was significantly different (P < 0.05). Phylogenetic analysis based on the characterized clusters of Bacteroidetes, Spirochaetes, and Deltaproteobacteria showed that many OTUs present in both cockroach species clustered with sequences previously described in termites and other cockroaches, but not with those from other animals or environments. These results suggest that, during their evolution, those cockroaches conserved several bacterial communities from the microbiota of a common ancestor. The ecological stability of those microbial communities may imply the important functional role for the survival of the host of providing nutrients in appropriate quantities and balance.

Understanding the origin and diversification of organisms requires a good phylogenetic estimate of their age and diversification rates. This estimate can be difficult to obtain when samples are limited and fossil records are disputed, as in Dictyoptera. To choose among competing hypotheses of origin for dictyopteran suborders, we root a phylogenetic analysis (~800 taxa, 10 kbp) within a large selection of outgroups and calibrate datings with fossils attributed to lineages with clear synapomorphies. We find the following topology: (mantises, (other cockroaches, (Cryptocercidae, termites)). Our datings suggest that crown-Dictyoptera-and stem-mantises-would date back to the Late Carboniferous (~ 300 Mya), a result compatible with the oldest putative fossil of stem-dictyoptera. Crown-mantises, however, would be much more recent (~ 200 Mya; Triassic/Jurassic boundary). This pattern (i.e., old origin and more recent diversification) suggests a scenario of replacement in carnivory among polyneopterous insects. The most recent common ancestor of (cockroaches + termites) would date back to the Permian (~275 Mya), which contradicts the hypothesis of a Devonian origin of cockroaches. Stem-termites would date back to the Triassic/Jurassic boundary, which refutes a Triassic origin. We suggest directions in extant and extinct species sampling to sharpen this chronological framework and dictyopteran evolutionary studies.

The so-called "Ogasawara cockroaches" were examined by morphological observations and by breeding experiments to elucidate their actual taxonomical status. Fourteen groups (isolate) of "Ogasawara cockroaches" collected from Iwoto-A, Iwoto-B, Hahajima, Chichijima, Nishijima, Nakodojima, Tokunoshima-A, Tokunoshima-B, Okinawato- A, Okinawa-B, Amamiooshima, Miyakojima, Ishigakijima and Hawaii, were bred and passaged in our laboratory. Cockroaches collected from the field were first reared individually and the sexes of their offspring examined. Cockroaches collected from Iwoto, Tokushima and Okinawa, were found to consist of two groups; those whose offspring were all female and the other whose offspring consist of both male and female. Cross-breeding experiments showed that individuals from the group that did not produce any male but only female offspring were parthenogenetic. On the contrary, the group that have bisexual individuals produced both male and female offspring in a ratio of 1:1. Our results show that the so-called "Ogasawara cockroaches" consist of 2 species, namely, Pycnoscelus surinamensis and Pycnoscelus indicus. There are areas in which both species co-habitated together and there are also areas in which either only one of the two species can be found. The group that reproduces only female offspring and only through parthenogenesis was identified as P. surinamensis. The group that reproduces heterosexually and produce male and female offspring was identified as P. indicus. Thus, the so-called "Ogasawara cockroaches" found in Japan actually consist of 2 species, namely, P. surinamensis and P. indicus, which can be differentiated using the solitary breeding method to demonstrate parthenogenesis in the former and the need for sexual reproduction in the latter.

Whole-genome sequencing has emerged as one of the most effective means to elucidate the biological roles and molecular features of obligate intracellular symbionts (endosymbionts). However, the de novo assembly of an endosymbiont genome remains a challenge when host and/or mitochondrial DNA sequences are present in a dataset and hinder the assembly of the genome. By focusing on the traits of genome evolution in endosymbionts, we herein developed and investigated a genome-assembly strategy that consisted of two consecutive procedures: the selection of endosymbiont contigs from an output obtained from a de novo assembly performed using a TBLASTX search against a reference genome, named TBLASTX Contig Selection and Filtering (TCSF), and the iterative reassembling of the genome from reads mapped on the selected contigs, named Iterative Mapping and ReAssembling (IMRA), to merge the contigs. In order to validate this approach, we sequenced two strains of the cockroach endosymbiont Blattabacterium cuenoti and applied this strategy to the datasets. TCSF was determined to be highly accurate and sensitive in contig selection even when the genome of a distantly related free-living bacterium was used as a reference genome. Furthermore, the use of IMRA markedly improved sequence assemblies: the genomic sequence of an endosymbiont was almost completed from a dataset containing only 3% of the sequences of the endosymbiont's genome. The efficiency of our strategy may facilitate further studies on endosymbionts.

Background: Hebardina concinna is a domestic pest and potential vector of pathogens throughout East and Southeast Asia, yet identification of this species has been difficult due to a lack of diagnostic morphological characters, and to uncertainty in the relationship between macroptyrous (long-winged) and brachypterous (small-winged) morphotypes. In insects male genital structures are typically species-specific and are frequently used to identify species. However, male genital structures in H. concinna had not previously been described, in part due to difficulty in identifying conspecifics., Methods/principal Findings: We collected 15 putative H. concinna individuals, from Chinese populations, of both wing morphotypes and both sexes and then generated mitochondrial COI (the standard barcode region) and COII sequences from five of these individuals. These confirmed that both morphotypes of both sexes are the same species. We then dissected male genitalia and compared genital structures from macropterous and brachypterous individuals, which we showed to be identical, and present here for the first time a detailed description of H. concinna male genital structures. We also present a complete re-description of the morphological characters of this species, including both wing morphs., Conclusions/significance: This work describes a practical application of DNA barcoding to confirm that putatively polymorphic insects are conspecific and then to identify species-specific characters that can be used in the field to identify individuals and to obviate the delay and cost of returning samples to a laboratory for DNA sequencing.

This study examines potential arthropod intermediate hosts for the nematode Abbreviata antarctica. Five species of arthropod (tropical native cricket Teleogryllus oceanicus; native roach Drymoplaneta communis; native termite Nasutitermes exitiosus; and 2 introduced species, the East African roach Naupheta cinerea and the Asian cricket Acheta domesticus) were fed feces from a skink, Tiliqua rugosa, containing embryonated eggs of A. antarctica . The insects were dissected at 4 intervals (1 day, 4 days, 8-16 days, and 25-26 days). Viable third-stage larvae were recovered from cysts on the external wall of the midgut and hindgut of 62.5% nymphal and adult T. oceanicus crickets at 25-26 days post-infection and from a single nymphal A. domesticus cricket. No roaches contained eggs or first-stage larvae after 48 hr, and neither eggs nor larvae were found in termites.

A survey of cockroach fauna was carried out on the 3 inhabited islands of the Ogasawara chain island of Japan, namely, Chichijima island, Hahajima island and Iwo island. Seven species, namely, Periplaneta americana (Linnaeus, 1758), Periplaneta australasiae (Fabricius, 1775), Blattella lituricollis (Walker, 1868), Onychostylus vilis (Brunner von Wattenwyl, 1865), Supella longipalpa (Fabricius, 1798), Pycnoscelus surinamensis (Linnaeus, 1758) and Opisthoplatia orientalis (Burmeister, 1838), were collected on Chichijima island. Four species, namely, P. americana, P. australasiae, O. vilis and P. surinamensis were collected on Hahajima island and 6 species, namely, P. americana, P. australasiae, B. lituricollis, O. vilis, P. surinamensis and Neostylopyga rhombifolia were collected on Iwo island. This is the first record of N. rhombifolia and Onychostylus orientalis on the Ogasawara chain islands. Our study increases the recorded taxon of cockroaches on the Ogasawara from 3 families, 5 genera 10 species to 4 families, 7 genera, 12 species. A list of the cockroach species on Ogasawara islands reported to date as well as a key for their identification is also presented. Periplaneta americana and P. australasiae, being the dominant species, together with S. longipalpa, were collected mostly in the indoor environment, indicating their preference for this habitat. Pycnoscelus surinamensis, which is considered as an outdoor insect has been found in semi-household environments such as greenhouse and shed, indicating their new adaptation to the changing environment.

An experimental excystation assay was used to test the potential species isolating effects of excystation signaling among gregarines. Oocysts of a single gregarine species, Blabericola migrator , were tested for activation, excystation, and sporozoite motility by using intestinal extracts from 11 species of cockroaches representing a cohesive phylogeny of 7 genera, 3 subfamilies, and 2 families of Blattodea. Sporozoite activation, excystation, and motility were observed for all excystation assay replications using intestinal fluid from blaberid hosts, but delayed activation or excystation was observed for all assay replications using intestinal fluid from hosts in the family Blattidae. The results illustrate a trend toward a generalized excystation signal among gregarines that is conserved across the host clade at a subfamily or family level but that is unlikely to play a significant role as a species-isolating mechanism among sibling gregarine species.

We have designed and utilized degenerate primers in the phylogenetic analysis of [FeFe] hydrogenase gene diversity in the gut ecosystems of roaches and lower termites. H(2) is an important free intermediate in the breakdown of wood by termite gut microbial communities, reaching concentrations in some species exceeding those measured for any other biological system. The primers designed target with specificity the largest group of enzymatic H domain proteins previously identified in a termite gut metagenome. "Family 3" hydrogenase sequences were amplified from the guts of lower termites, Incisitermes minor, Zootermopsis nevadensis, and Reticulitermes hesperus, and two roaches, Cryptocercus punctulatus and Periplaneta americana. Subsequent analyses revealed that all termite and Cryptocercus sequences were phylogenetically distinct from non-termite-associated hydrogenases available from public databases. The abundance of unique sequence operational taxonomic units (as many as 21 from each species) underscores the previously demonstrated physiological importance of H(2) to the gut ecosystems of these wood-feeding insects. The diversity of sequences observed might be reflective of multiple niches that the enzymes have been evolved to accommodate. Sequences cloned from Cryptocercus and the lower termite samples, all of which are wood feeding insects, clustered closely with one another in phylogenetic analyses to the exclusion of alleles from P. americana, an omnivorous cockroach, also cloned during this study. We present primers targeting a family of termite gut [FeFe] hydrogenases and provide results that are consistent with a pivotal role for hydrogen in the termite gut ecosystem and point toward unique evolutionary adaptations to the gut ecosystem.

The following domiciliary cockroaches were collected from restaurants in five zones of Kuala Lumpur Federal Territory, Malaysia using 1L glass beaker traps baited with ground mouse-pellets: Periplaneta americana (Linnaeus) (n = 820), Periplaneta brunnea Burmeister (n = 46), Blattella germanica (Linnaeus) (n = 12504), Supella longipalpa (Fabricius) (n = 321), Symploce pallens Stephens (n = 29) and Neostylopyga rhombifolia (Stoll) (n = 5). The following bacteria were isolated from 10 cockroach specimens: Enterobacter cloacae, Klebsiella pneumoniae ssp. pneumoniae, Klebsiella pneumoniae ssp. rhinoscleromatis and Serratia liquefaciens from 5 B. germanica; Acinetobacter calcoaceticus var. anitratus, Citrobacter diversus/amalonaticus, Escherichia vulneris and K.p. pneumoniae from 3 P. brunnea; and Citrobacter freundii, Enterobacter agglomerans 4, Escherichia adecarboxylate, E. vulneris, K. p. pneumonia, K. p. rhinoscleromatis and Proteus vulgeris from 2 P. americana.

Assessing the adulteration of food products and the presence of filth and extraneous materials is one of the measures that the U.S. Food and Drug Administration (FDA) utilizes in implementing regulatory actions of public health importance. To date, 22 common pest species (also known as the "Dirty 22" species) have been regarded by this agency as the spreaders of foodborne diseases. We have further categorized the Dirty 22 species into four groups: I has four cockroach species, II has two ant species, III has 12 fly species, and IV has four rodent species. The presence of any Dirty 22 species is also considered an indicator of unsanitary conditions in food processing and storage facilities. In this study, we describe the development of a two-step nested PCR protocol to amplify the small subunit ribosomal gene of group I Dirty 22 species that include four cockroach species: Blattella germanica, Blatta orientalis, Periplaneta americana, and Supella longipalpa, along with the development of a PCR-restriction fragment length polymorphism method for rapid detection and differentiation of these violative species. This method will be utilized when the specimen cannot be identified with conventional microscopic taxonomic methods, especially when only small body parts are separated and recovered from food samples for analysis or when these body parts are in a decomposed state. This new PCR-restriction fragment length polymorphism will provide correct identification of group I Dirty 22 species; this information can then be used in regulation and prevention of foodborne pathogens.

This study assessed the effectiveness of a biorational control approach using 2% hydramethylnon gel bait on German cockroaches, Blattella germanica (L.) in some residential and hospital buildings in South Western Iran. In total, three buildings consisting of 150 apartment units and 101 hospital units were monitored weekly via sticky trap for German cockroach infestations over a period of eight months. These infested units were randomly subjected to intervention and control treatments. Pamphlets and posters were provided and lectures were given to support the educational programmes as a tactic of the biorational system. Survey on cockroach index for intervention units showed 67-94% recovery to achieve clean level of infestation for intervention units of the residential buildings and 83% for the hospital. Mean percentage reductions for treatment groups throughout the 15-week treatment period were 76.8% for the residential buildings and 88.1% for the hospital, showing significant differences compared to the control groups. Linear regression of infestation rates were recorded weekly after treatment and their negative slope for treatment groups substantiated significant reductions for interventions. The results of this study showed that biorational control method, using gel bait, educational programmes and sanitation, is an effective way to manage German cockroach infestation.

Phylogenetic relationships among termites, mantids and the five traditionally recognized cockroach families have been the subject of several studies during the last half-century. One cockroach lineage that has remained notably absent from such studies is the Nocticolidae. This group of small, elusive surface- and cave-dwelling species from the Old World Tropics has been proposed to represent an additional family. Using molecular sequences, we performed an initial phylogenetic examination of Nocticola spp. The hypothesis that they are phylogenetically divergent was confirmed from the analyses of three genes and a combined dataset. To supplement our phylogenetic analyses, we attempted to amplify 16S rRNA from the obligate mutualistic endosymbiont Blattabacterium cuenoti, present in all cockroaches studied to date. Unexpectedly, amplification was unsuccessful in all Nocticola spp. examined. This result was confirmed by microscopic examinations of fat body tissue. These Nocticola spp. are the first cockroaches found to be uninfected by B. cuenoti, which raise questions about when the bacterium first infected cockroaches.

Termites are instantly recognizable mound-builders and house-eaters: their complex social lifestyles have made them incredibly successful throughout the tropics. Although known as 'white ants', they are not ants and their relationships with other insects remain unclear. Our molecular phylogenetic analyses, the most comprehensive yet attempted, show that termites are social cockroaches, no longer meriting being classified as a separate order (Isoptera) from the cockroaches (Blattodea). Instead, we propose that they should be treated as a family (Termitidae) of cockroaches. It is surprising to find that a group of wood-feeding cockroaches has evolved full sociality, as other ecologically dominant fully social insects (e.g. ants, social bees and social wasps) have evolved from solitary predatory wasps.

Tachykinins and tachykinin-related peptides (TKRPs) play major roles in signaling in the nervous system and intestine of both invertebrates and vertebrates. Here we have identified cDNAs encoding precursors of multiple TKRPs from the cockroaches Leucophaea maderae and Periplaneta americana. All nine LemTKRPs that had been chemically isolated in earlier experiments could be identified on the precursor of L. maderae. Four previously unidentified LemTKRPs were found in addition on the precursor. The P. americana cDNA displayed an open reading frame very similar to that of L. maderae with 13 different TKRPs. MALDI-TOF mass spectra from tissues of both species confirms the presence of all the TKRPs encoded on the precursor plus two additional peptides that are cleavage products of the N-terminally extended TKRPs. A tissue-specific distribution of TKRPs was observed in earlier experiments at isolation from brain and midgut of L. maderae. Our data do not suggest a differential gene expression but a different efficacy in processing of LemTKRP-2 and Lem/PeaTKRP-3 in the brain and intestine, respectively. This results in a gut-specific accumulation of these extended peptides, whereas in the brain their cleavage products, LemTKRP-1 and LemTKRP-3(11-19), are most abundant. Mass spectrometric analysis demonstrated the occurrence of the different TKRPs in single glomeruli of the tritocerebrum and in cells of the optical lobe.

Objective: To determine the possible role of cockroaches in the epidemiology of nosocomial infections., Design: Epidemiologic investigation of bacteria and fungi in cockroaches and evaluation of the antibiotic resistance of the bacteria isolated from the insects., Setting: Ninety hospitals in Kaohsiung City and Kaohsiung County in Taiwan., Methods: Cockroaches were collected in clinical and nonclinical areas and microorganisms were isolated from their external surface and alimentary tract. The susceptibilities of Staphylococcus aureus, Enterococcus species, Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Serratia marcescens, and Proteus species to 17 antibiotics were tested., Results: Cockroach infestation was found in 46.7% of the hospitals studied. Two hundred three cockroaches were collected (139 Periplaneta americana and 64 Blattella germanica). Periplaneta americana was found more often in nonclinical areas (64.5%) and B. germanica in clinical areas (78.1%). There was no statistically significant difference between Periplaneta americana (98.6%) and B. germanica (96.9%) regarding overall isolation rate (P > .05). However, 33 species of bacteria and 16 species of fungi were isolated from Periplaneta americana and only 23 and 12, respectively, from B. germanica. Resistance to ampicillin (13.7% to 100%), chloramphenicol (14.3% to 71.4%), tetracycline (14.3% to 73.3%), and trimethoprim-sulfamethoxazole (14.3% to 57.1%) was found in two gram-positive and five gram-negative bacteria., Conclusion: Because cockroach infestation occurred in more than 40% of the hospitals and nearly all of the cockroaches harbored bacteria with multidrug resistance and fungi, cockroaches may play a potential role in the epidemiology of nosocomial infections in those hospitals.

Unlabelled: Recently, cockroaches have been established as the second most Important allergen, producing allergic diseases, especially in low socioeconomic populations. In Thailand, about 44-61% of atopic patients were positive to cockroach extract by a skin-prick test. This study examined cockroach allergen levels in relation to cockroach species and allergic diseases in the houses of cockroach-sensitive patients. Sixty households of allergic patients in the Bangkok metropolitan area were surveyed using open- and closed-ended questionnaires. Cockroaches were collected using commercial cockroach traps, while dust samples were obtained from the bedrooms, kitchens and living rooms of the houses using a vacuum cleaner. The cockroaches were counted and their species Identified. The levels of cockroach allergens were determined by specific monoclonal antibodies using a monoclonal antibody-polyclonal antibody based sandwich ELISA kit. Six cockroach species were Identified: Periplaneta americana (American cockroach, 72.15%), Supella longlpalpa (2.75%, found in only one house), Periplaneta brunnea (0.78%), Periplaneta australaslae (0.78%), Neostylopyga rhombifolla (0.78%), Blattella germanica (German cockroach, 0.39%) and nymphs (22.35%). Allergens of the predominant species, P. americana, were detectable in all homes studied, with the highest levels in the kitchen areas. The range of allergen levels in house dust varied from 0.40-162.00 microg per g of dust. The median and mean allergen levels in kitchen dust were 59.16 microg and 62.80 microg per g of dust, respectively, while the median allergen level in bedroom dust was only 15.90 microg per g of dust. The German cockroach allergen (Bla g 2) was undetectable in any of the houses., In Conclusion: P. americana was the most common cockroach and may be the species causing allergic diseases, especially asthma, in Thailand, which differs from the USA and Europe

Competitive displacement is the outcome of interspecific competition between invading and native species. Alien insect species invade not only cultivated areas but also natural habitats. We prospected the cockroach fauna on two oceanic islands (Mayotte and La Réunion) to investigate the impact of altitude and anthropic disturbance on the distribution of these species. Most invading cockroach species seem to benefit from cultivation of land whereas endemic species occupy more specific and reduced habitats that are threatened by human impact.

Morphologically similar cockroaches in the subfamilies Panesthiinae and Geoscapheinae (Blattaria: Blaberidae) display contrasting feeding habits, behaviour and biogeographical distributions. Panesthiinae, found throughout Asia and Australia, all live in and feed on decaying wood that they burrow into. Geoscapheinae are restricted to Australia and construct and live in burrows in the soil, where they feed on dry leaves taken from the surface. A lack of knowledge about phylogenetic relationships among these cockroaches hinders an understanding of the factors that have shaped the evolution of their diverse lifestyles and biogeography. To address this issue, we sequenced three genes from representatives of nine of the 10 genera in the two subfamilies, and performed phylogenetic analyses. The well-supported topology revealed the Panesthiinae to be paraphyletic with respect to the Geoscapheinae. Soil-burrowing cockroaches appear to have evolved from a lineage of wood burrowers that invaded Australia from the north some time after the merging of the Asian and Australian tectonic plates ca. 20 Myr ago. The main factor promoting the evolution of soil burrowing is likely to have been one of the periods of strong aridity that Australia has experienced since the Miocene period.

Bacteria of the genus Blattabacterium are intracellular symbionts that reside in specialized cells of cockroaches and the termite Mastotermes darwiniensis. They appear to be obligate mutualists, and are transmitted vertically in the eggs. Such characteristics are expected to lead to equivalent phylogenies for host and symbiont, and we tested this hypothesis using recently accumulated data on relationships among termites and cockroaches and their Blattabacterium spp. Host and symbiont topologies were found to be highly similar, and various tests indicated that they were not statistically different. A close relationship between endosymbionts from termites and members of the wood-feeding cockroach genus Cryptocercus was found, supporting the hypothesis that the former evolved from subsocial, wood-dwelling cockroaches. The majority of the Blattabacterium spp. sequences appear to have undergone similar rates of evolution since their divergence from a common ancestor, and an estimate of this rate was determined based on early Cretaceous host fossils. The results support the idea that the stem group of modern cockroaches radiated sometime between the late Jurassic and early Cretaceous-not the Carboniferous, as has been suggested on the basis of roach-like fossils from this epoch.

Molecular phylogenetic relationships among 25 species of the wood-feeding cockroach belonging to the genus Salganea Stål (Panesthiinae; Blaberidae) in Southeast Asia were analyzed based on the DNA sequence of the complete mitochondrial cytochrome oxidase II (COII) gene. Most basal relationships among species of Salganea are poorly resolved by both neighbor-joining and nonweighted parsimony analyses, suggesting the possibility of a hard polytomy due to a rapid and potentially simultaneous radiation early in the history of the genus. For more apical relationships, however, some interesting phylogenetic relationships were recognized. The monophyly of the two species groups, morio and foveolata, the former of which is distributed mainly in the Sunda lands (containing the Malay Peninsula, Sumatra, Java, and Borneo), whereas the latter is Sulawesi endemic, was strongly supported. Based on the inferred phylogenetic patterns and recent palaeogeographic scenario for Southeast Asia, it is suggested that a radiation of Salganea species occurred in Southeast Asia presumably in the early Tertiary, and several barriers against dispersal and gene flow, such as the formation of straits or high mountains, have arisen from the middle Tertiary.

Cryptocercus are subsocial, xylophagous cockroaches that live in temperate forests. Like other cockroaches, Cryptocercus harbour endosymbiotic bacteria in their fat bodies. Two species of Cryptocercus occur in the palaearctic, one each in eastern Russia and south-central China. In the USA, there are five species: one in the north-west and four in the south-east. Little is known about the relationship between the Eurasian and North American Cryptocercus or the causes of the disjunct distribution. Here, a molecular phylogeny for six out of the seven Cryptocercus species and their endosymbionts is inferred in an attempt to understand the evolution and biogeography of the genus. Our analysis showed that the North American Cryptocercus are monophyletic, suggesting that a single colonization event was followed by vicariance. There was complete concordance between the host and endosymbiont phylogenetic trees. Divergence estimates based on endosymbiont DNA sequences suggested that the palaearctic and nearctic Cryptocercus diverged 70-115 million years (Myr) ago and the eastern- and western-USA species diverged 53-88 Myr ago. These divergence estimates were correlated with biogeographical events, and a hypothesis is presented to explain the current distribution of Cryptocercus. Our findings suggest that Cryptocercus has had a long evolutionary history, dating back to the Jurassic.

The wood-feeding genus Cryptocercus is considered the basal lineage among extant cockroaches. Cryptocercus is the sole representative of the family Cryptocercidae and at present three species are recognized within the genus worldwide: Cryptocercus punctulatus in the United States, C. relictus in Eurasia and C. primarius in the Orient. The geological distribution of C. punctulatus in the USA is disjunct, with populations occurring along the Appalachian Mountains and in the Pacific North-west. In samples collected from several locations of the eastern and western USA, we investigated variation in DNA sequence of portions of the two mitochondrial rRNA genes and in chromosome number. The overall sequence divergence among 30 individuals assayed from 17 locations was 17.3 percent. A phylogenetic analysis revealed that in the east, individuals in Virginia had diverged significantly in their haplotype from those in North Carolina, Georgia and Alabama; individuals in the west (Oregon) had diverged in their haplotype from individuals in the east. The diploid chromosome number for 52 male C. punctulatus sampled from 15 locations varied from 37 (18(II) + X) to 47 (23(II) + X). In the eastern samples, the diploid chromosome number ranged from 37 to 45, whereas in Oregon all individuals had 2n = 47. No polymorphism in DNA sequence or chromosome number among individuals collected within a locality was detected. The DNA sequence and chromosome number variation data, together with preliminary studies on mating incompatibility, strongly suggest that C. punctulatus in the USA is comprised of at least two probably three sibling species, with one species occurring in western USA and one or more species in eastern USA.

Cockroaches are among the most ancient winged insects, the earliest fossils dating back to about 400 million years. Several conflicting phylogenies for cockroach families, subfamilies, and genera have been proposed in the past. In addition, the relationship of Cryptocercidae to other cockroach families and the relationship between the cockroach, Cryptocercus punctulatus, and the termite, Mastotermes darwiniensis, have generated debate. In this paper, a phylogeny for cockroaches, mantids, and termites based on DNA sequence of the mitochondrial ribosomal RNA genes is presented. The results indicated that cockroaches are a monophyletic group, whose sister group is Mantoidea. The inferred relationship among cockroach families was in agreement with the presently accepted phylogeny. However, there was only partial congruence at the subfamily and the generic levels. The phylogeny inferred here does not support a close relationship between C. punctulatus and M. darwiniensis. The apparent synapomorphies of these two species are likely a manifestation of convergent evolution because there are similarities in biology and habitat.