On the status of Suifunema peregrinatum (Carreno, 2017) n. comb. (Nematoda: Oxyuridomorpha: Thelastomatidae) and first record of the species for Japan.

Blattophila peregrinata Carreno, 2017 (Nematoda: Oxyuridomorpha: Thelastomatidae) is proposed as a junior synonym of Suifunema peregrinatum (Carreno, 2017) n. comb., based on the revision of the type material of S. caudelli Chitwood, 1932, the type and only species of the genus. The revision permitted the emendation of some features of the original descriptions, namely the position of the nerve ring at level of the corpus instead the isthmus and the female genital tract didelphic-prodelphic instead didelphic-amphidelphic. The males of S. peregrinatum n. comb. are the only ones known for the genus. Thus, an amended generic diagnosis is given, including information on the males. Both S. peregrinatum n. comb. and S. caudelli differ by having a more slender body, a longer tail, and the vulva farther from the anterior end in the former. S. peregrinatum n. comb. is recorded for the first time from Pycnoscelus indicus Fabricius, 1775 (Blattodea: Blaberidae) from Japan. The specimens from Japan are morphologically consistent with other populations of the species from the USA and the Galapagos Islands of Ecuador. Only some morphometrics vary slightly, but these are within the range of the species. The phylogenetic position of the genus among the family Thelastomatidae is discussed, including sequences of the D2-D3 LSU rDNA of specimens from both the type population (Ohio, USA) and Japan.

Systemic RNAi of V-ATPase subunit B causes molting defect and developmental abnormalities in Periplaneta fuliginosa.

The vacuolar (H+ )-ATPases (V-ATPases) are ATP-driven proton pumps with multiple functions in many organisms. In this study, we performed structural and functional analysis of vha55 gene that encodes V-ATPase subunit B in the smokybrown cockroach Periplaneta fuliginosa (Blattodea). We observed a high homology score of the deduced amino acid sequences between 10 species in seven orders. RNAi of the vha55 gene in P. fuliginosa caused nymphal/nymphal molting defects with incomplete shedding of old cuticles, growth inhibition, as well as bent and wrinkled cuticles of thoraxes and abdominal segments. Since growth inhibition caused by vha55 RNAi did not interfere in the commencement of cockroach molting, molting timing and body growth might be controlled by independent mechanism. Our study suggested V-ATPases might be a good candidate molecule for evolutionary and developmental studies of insect molting.

Genome analysis of new Blattabacterium spp., obligatory endosymbionts of Periplaneta fuliginosa and P. japonica.

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.

Broad infectivity of Leidynema appendiculatum (Nematoda: Oxyurida: Thelastomatidae) parasite of the smokybrown cockroach Periplaneta fuliginosa (Blattodea: Blattidae).

Host specificity of parasites is important for the understanding of evolutionary strategies of parasitism that would be a basis of predictions of the disease expansion when parasitized hosts invade new environments. The nematode order Oxyurida is an interesting parasite group for studying the evolution of parasitism as it includes parasites of both invertebrates and vertebrates. In our survey, we found that the smokybrown cockroach Periplaneta fuliginosa was primarily infected with only one nematode species Leidynema appendiculatum. In two cases, L. appendiculatum was isolated from two additional cockroach species Pycnoscelus surinamensis, sold in Japan as a reptile food, and Blatta lateralis, captured in the field and cultured in the laboratory. Inoculation of L. appendiculatum into three additional cockroach species P. japonica, Blattella nipponica, and P. surinamensis also resulted in parasitism. Infection prevalence was high, and timing of postembryonic development from hatched nematode larva to mature adult in these hosts was identical with that in P. fuliginosa. While ecological interactions strongly determine the host range, such broad infectivity is still possible in this parasitic nematode.

First record of Hammerschmidtiella diesingi (Hammerschmidt, 1838) (Oxyuridomorpha: Thelastomatidae) parasite of Periplaneta americana (Linnaeus, 1758) (Blattodea: Blattidae) in Japan, morphological and molecular characterization.

Nematodes, belonging to the family Thelastomatidae, are parasites of saprophytic terrestrial arthropods, mainly cockroaches. American cockroach Periplaneta americana (Linnaeus, 1758) has high environmental adaptability, and has been spread worldwide by human activity. There are several reports of thelastomatid parasitic nematodes of P americana in the world. Here, the thelastomatid nematode Hammerschmidtiella diesingi (Hammerschmidt, 1838) (Oxyuridomorpha: Thelastomatidae), isolated from P americana is recorded for the first time in Japan. Through morphometrics, DIC and SEM observations, we confirmed that specimens of the present study agree with previous records of H. diesingi. DNA sequencing of the partial D2/D3 LSU expansion segment of the 28S ribosomal RNA gene revealed that H. diesingi from Japan matches with H. diesingi from other countries. Our contributions are very useful and fundamental for further analysis of the cockroach and parasite relations.

Composition of the Cockroach Gut Microbiome in the Presence of Parasitic Nematodes.

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.

Morphological, molecular and developmental characterization of the thelastomatid nematode Thelastoma bulhoesi (de Magalhães, 1900) (Oxyuridomorpha: Thelastomatidae) parasite of Periplaneta americana (Linnaeus, 1758) (Blattodea: Blattidae) in Japan.

The American cockroach Periplaneta americana (Linnaeus, 1758) (Blattodea: Blattidae) has been spreading worldwide by commerce and has successfully adjusted to living with humans. There are many reports of thelastomatid parasitic nematode isolated from P. americana in many countries including USA, Canada, India, Argentina, Bulgaria, but not in Japan. We have investigated the parasitic nematodes in P. americana lab strain and field-captured individuals in Japan and found that Thelastoma bulhoesi (de Magalhães, 1900) (Oxyuridomorpha: Thelastomatidae) parasitizes with high infection rates. We described morphological, molecular, and developmental characters of the parasitic nematode because such information was missing despite it has been discovered more than one hundred years ago. We described morphometrics with DIC microscopy and fine structure of male and female adult with SEM observation. We also reveal the embryonic and postembryonic development of this nematode. This is the first report of a thelastomatid nematode isolated from American cockroach in Japan, and the data showed here is also very useful and fundamental for further analysis of the cockroach and parasite relations.

First report of the nematode Leidynema appendiculata from Periplaneta fuliginosa.

The smokybrown cockroach Periplaneta fuliginosa has spread all over the world, and is now one of the most undesired invasive alien pests in Japan. Because cockroaches are generally infected by thelastomatid nematodes, they are being distributed around the world with their parasitic nematodes. Nothing is known about parasitic nematode species in P. fuliginosa differences, or similarity of the parasite's population structures between the different countries of the host cockroaches. Here we investigated the P. fuliginosa invasive to Japan and found that 100% of individuals were infected with one nematode species. According to the morphology and the sequence of the D2/D3 expansion segment of the 28S ribosomal RNA gene, we identified the parasite as Leidynema appendiculata. This nematode reproduced by haplodiploidy and its developmental timing under various conditions is quite divergent. Their population in the hindgut of P. fuliginosa was controlled with a few adult females and a male. This is the first report of the thelastomatid nematode isolated from the smokybrown cockroach, and is the basis for our future research examining the origin, distribution route and immigration history of the cockroach and the impact of L. appendiculata on native Japanese cockroach species.