Reverse taxonomy for elucidating diversity of insect-associated nematodes: a case study with termites

BACKGROUND: The molecular operational taxonomic unit(MOTU)has recently been applied to microbial and microscopic animal biodiversity surveys. However, in many cases, some of the MOTUs cannot be definitively tied to any of the taxonomic groups in current databases. To surmount these limitations, the concept of "reverse taxonomy" has been proposed, i.e. to primarily list the MOTUs with morphological information, and then identify and/or describe them at genus/species level using subsamples or by re-isolating the target organisms. Nevertheless, the application of "reverse taxonomy" has not been sufficiently evaluated. Therefore, the practical applicability of "reverse taxonomy" is tested using termite-associated nematodes as a model system for phoretic/parasitic organisms which have high habitat specificity and a potential handle (their termite host species) for re-isolation attempts. METHODOLOGY: Forty-eight species (from 298 colonies) of termites collected from the American tropics and subtropics were examined for their nematode associates using the reverse taxonomy method and culturing attempts (morphological identification and further sequencing efforts). The survey yielded 51 sequence types (= MOTUs) belonging to 19 tentatively identified genera. Within these, four were identified based on molecular data with preliminary morphological observation, and an additional seven were identified or characterized from successful culturing, leaving eight genera unidentified. CONCLUSIONS: That 1/3 of the genera were not successfully identified suggests deficiencies in the depth of available sequences in the database and biological characters, i.e. usually isolated as phoretic/parasitic stages which are not available for morphological identification, and too many undiscovered lineages of nematodes. Although there still is the issue of culturability of nematodes, culturing attempts could help to make reverse taxonomy methods more effective. However, expansion of the database, i.e., production of more DNA barcodes tied to biological information by finding and characterizing additional new and known lineages, is necessary for analyzing functional diversity.Natsumi Kanzaki, Robin M. Giblin-Davis, Rudolf H. Scheffrahn, Hisatomo Taki, Alejandro Esquivel, Kerrie A. Davies and E. Allen Herr

Bursaphelenchus suri n. sp. : a second Bursaphelenchus syconial parasite of figs supports adaptive radiation among section Sycomorus figs

The nematode genus Bursaphelenchus is a highly divergent group. This genus mainly consists of mycophagous entomophilic species, but some species have specialized as obligate or facultative plant parasites, facultative insect parasites, or exhibit feeding dimorphism (phenotypic plasticity) leading to mycophagous and predatory forms. In the present study, a new Bursaphelenchus species, B. suri n. sp. was isolated from fresh syconia (figs) of Ficus sur and is described and illustrated based on its typological characters and molecular phylogenetic status. The new species is characterized by its highly derived feeding structures found in obligate plant parasites, lip possessing a labial disc and a long and thick stylet with a long conus and extremely well-developed basal swellings. In addition, slender body of both sexes is characteristic of the species. The new species is phylogenetically and typologically closely related to B. sycophilus, i.e., these two species share the characteristic feeding structures and form a well-supported clade within the B. fungivorus group in the genus. Biologically, these two species are both isolated from fresh figs of the section Sycomorus. However, the new species differs from B. sycophilus by the length of the female post-uterine sac and the shape of the male spicule, i.e., the new species has a long post-uterine sac and spicule condylus without dorsal recurvature. Thus, the new species is the second obligate fig parasite of the genus, and the evolutionary relationship between the B. suri n. sp. and B. sycophilus clade and section Sycomorus figs is hypothesized as an example of adaptive radiation with more species to be discovered.SUPPORTING INFORMATION : S1 Text. Typological description of Bursaphelenchus suri n. sp. in traditional telegraphic style.The National Research Foundation of South Africa and Grant-in-Aid for Scientific Research (B) from the Japan Society for the Promotion of Science.http://www.plosone.orgdm2022BiochemistryGeneticsMicrobiology and Plant Patholog

Molecular diversity and relationships of fig associated nematodes from South Africa

Nematodes of figs and fig wasps have received limited attention in Africa since their discovery in 1973. Sixteen of the 25 species of native South African figs were sampled for nematode associates using molecular barcoding with three loci (SSU, LSU D2-D3 and mtCOI) and fourteen (93%) were positive for at least one nematode species. Thirty-three putative species of nematodes were identified and classified according to the loci that were amplified and successfully sequenced. Fourteen putative nematode species were classified as Aphelenchoididae, of which nine were identified as Ficophagus from four species of Ficus from the section Galoglychia (i.e., five ex F. burkei including one shared with F. natalensis, one ex F. glumosa, one ex F. lutea, and one ex F. stuhlmannii) and one species ex F. sur from the section Sycomorus. In addition, there were four nematode species classified as Schistonchus s.s. from section Galoglychia figs (i.e., one ex F. burkei, two ex F. trichopoda, and one ex F. glumosa). There was also one species of Bursaphelenchus nematode recovered from F. sur from the section Sycomorus. Sixteen putative nematode species were classified as Diplogastridae, of which eight occurred in two clades of what is currently called Parasitodiplogaster with one (P. salicifoliae) being recovered from two Ficus species in the section Urostigma (F. salicifolia and F. ingens) and seven diplogastrids being associated with six species of Ficus from the section Galoglychia (i.e., two ex F. burkei including P. sycophilon, one ex F. stuhlmannii, one ex F. burtt-davyi, one ex F. trichopoda, one ex F. abutilifolia and one ex F. sansibarica). Three Acrostichus spp., a Teratodiplogaster and a Pristionchus species were recovered from F. sur and two Teratodiplogaster spp. and Pristionchus sycomori were recovered from F. sycomorus with both Ficus species belonging to the subgenus and section Sycomorus. The identities of the previously described T. martini and Parasitodiplogaster doliostoma (= Pristionchus sp. 35) are discussed. Lastly, there was a panagrolaimid identified from F. petersii.The National Research Foundation of South Africa and the Japan Society for the Promotion of Science.http://www.plosone.orgpm2022BiochemistryGeneticsMicrobiology and Plant Patholog

\u3ci\u3eFicotylus laselvae\u3c/i\u3e n. sp. (Tylenchomorpha: Anguinidae) associated with \u3ci\u3eFicus colubrinae\u3c/i\u3e in Costa Rica

Ficotylus laselvae n. sp. was recovered from under the bracts of figs (syconia) of Ficus colubrinae from La Selva, Costa Rica, during a survey of nematode rainforest biodiversity and is described herein. This is only the second report of an association between the nematode suborder Tylenchina and the sycones of figs. Previous reports of most nematode associates of the sycones of figs have been from the lumen and involved transmission by female fig wasp pollinators (Agaonidae) during pollination/oviposition (e.g., Schistonchus and Parasitodiplogaster spp.). The association between F. laselvae n. sp. and Ficus colubrinae may involve an invertebrate host, but none was recovered from dissections of the bracts during this study. It is also possible that this is a rainforest understory nematode that feeds ectoparasitically in protected areas on the aerial parts of F. colubrinae. Molecular analysis using near-full-length sequences of the small subunit (SSU) rRNA and D2-D3 expansion segments of the large subunit (LSU) rRNA genes of Ficotylus laselvae n. sp. suggests that it is a member of the suborder Tylenchina (infraorder: Tylenchomorpha; family: Anguinidae) and that the closest sequenced species is F. congestae from the lumen of sycones of Ficus congesta from Queensland, Australia. Although both nematode species are associated with figs, they are morphologically divergent, suggesting that the different micro-niches that they fill provide different selective pressures for evolution of differing morphological characters or they represent different life history morphotypes of a dicyclic genus

Molecular and morphological observations on Parasitodiplogaster sycophilon Poinar, 1979 (Nematoda : Diplogastrina) associated with Ficus burkei in Africa

A Parasitodiplogaster sp. was isolated from syconia of Ficus burkei from Pretoria, South Africa, and determined to be conspecific with P. sycophilon, originally described by Poinar in 1979 from Harare, Zimbabwe, and also from F. burkei. Examination of type material of P. sycophilon revealed inaccuracies in the former description necessitating a redescription which is provided herein. Additionally, the original description lacked molecular data, which is also provided. Originally, the stoma of P. sycophilon was described as reduced without teeth. However, we observed a large dorsal stegostomatal tooth and an almost equally-sized right subventral tooth which was typologically similar to the stoma of P. laeviagata from Florida. In addition, a pore-like phasmid was observed in both males and females just above the tail tip. Most other characters were as formerly described. Based upon molecular inferences from sequences of the D2/D3 expansion segments of the rDNA of the large subunit (LSU), P. sycophilon is not clearly defined relative to the neotropical Parasitodiplogaster species that have been described and sequenced from figs in the section Urostigma, subsection Americana (i.e., P. laevigata, P. popenema, P. citrinema, and P. trigonema), or to P. australis from Australia ex F. virens (section Urostigma, subsection Urostigma), or to P. maxinema from neotropical figs from the section Pharmacosycea, subsection Pharmacosycea. Further work is needed to elucidate the molecular phylogeny of the Parasitodiplogaster lineages that may have co-speciated with the African figs of the section Urostigma, subsection Galoglychia.Grant 77256 from the National Research Foundationhttp://www.brill.comhb201

Pheromone chirality of african palm weevil, Rhynchophorus phoenicis (F.) and palmetto weevil, Rhynthophorus crueniatus (F.) (Coleoptera: Curculionidae)

There are four stereoisomers of both 3-methyl-octan-4-ol, the aggregation pheromone of the African palm weevil,Rhynchophorus phoenicis (F.) and 5-methyl-octan-4-ol, the aggregation pheromone of the palmetto weevil,Rhynchophorus cruentatus (F.). Synthetic stereoisomers of 3-methyl-octan-4-ol and 5-methyl-octan-4-ol were baseline-separated on a Cyclodex-B fused silica column. Use of this column in gas chromatographic-electroantennographic detection (GC-EAD) and GC-mass spectrometric (GC-MS) analyses revealed that only one stereoisomer, (3S,4S)-3-methyl-octan-4-ol and (4S,5S)-5-methyl-octan-4-ol, is produced by maleR. phoenicis and maleR. cruentatus, respectively, and elicits good antennal responses by conspecific male and female weevils. In field trapping experiments, withR. phoenicis in Côte d'Ivoire andR. cruentatus in Florida, (3S,4S)-3-methyl-octan-4-ol and (4S,5S)-5-methyl-octan-4-ol strongly enhanced attraction of fresh palm tissue, whereas other stereoisomers were behaviorally benign. Stereoisomeric 3-methyl-octan-4-ol and 5-methyl-octan-4-ol may be utilized to monitor and/or manage populations of these two palm weevils.UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro de Investigaciones en Productos Naturales (CIPRONA

Mortality and pulmonary complications in patients undergoing surgery with perioperative sars-cov-2 infection: An international cohort study

Background The impact of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) on postoperative recovery needs to be understood to inform clinical decision making during and after the COVID-19 pandemic. This study reports 30-day mortality and pulmonary complication rates in patients with perioperative SARS-CoV-2 infection. Methods This international, multicentre, cohort study at 235 hospitals in 24 countries included all patients undergoing surgery who had SARS-CoV-2 infection confirmed within 7 days before or 30 days after surgery. The primary outcome measure was 30-day postoperative mortality and was assessed in all enrolled patients. The main secondary outcome measure was pulmonary complications, defined as pneumonia, acute respiratory distress syndrome, or unexpected postoperative ventilation. Findings This analysis includes 1128 patients who had surgery between Jan 1 and March 31, 2020, of whom 835 (740%) had emergency surgery and 280 (248%) had elective surgery. SARS-CoV-2 infection was confirmed preoperatively in 294 (261%) patients. 30-day mortality was 238% (268 of 1128). Pulmonary complications occurred in 577 (512%) of 1128 patients; 30-day mortality in these patients was 380% (219 of 577), accounting for 817% (219 of 268) of all deaths. In adjusted analyses, 30-day mortality was associated with male sex (odds ratio 175 [95% CI 128-240], p<00001), age 70 years or older versus younger than 70 years (230 [165-322], p<00001), American Society of Anesthesiologists grades 3-5 versus grades 1-2 (235 [157-353], p<00001), malignant versus benign or obstetric diagnosis (155 [101-239], p=0046), emergency versus elective surgery (167 [106-263], p=0026), and major versus minor surgery (152 [101-231], p=0047). Interpretation Postoperative pulmonary complications occur in half of patients with perioperative SARS-CoV-2 infection and are associated with high mortality. Thresholds for surgery during the COVID-19 pandemic should be higher than during normal practice, particularly in men aged 70 years and older. Consideration should be given for postponing non-urgent procedures and promoting non-operative treatment to delay or avoid the need for surgery. Funding National Institute for Health Research (NIHR), Association of Coloproctology of Great Britain and Ireland, Bowel and Cancer Research, Bowel Disease Research Foundation, Association of Upper Gastrointestinal Surgeons, British Association of Surgical Oncology, British Gynaecological Cancer Society, European Society of Coloproctology, NIHR Academy, Sarcoma UK, Vascular Society for Great Britain and Ireland, and Yorkshire Cancer Research

Mortality and pulmonary complications in patients undergoing surgery with perioperative SARS-CoV-2 infection: an international cohort study

Background: The impact of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) on postoperative recovery needs to be understood to inform clinical decision making during and after the COVID-19 pandemic. This study reports 30-day mortality and pulmonary complication rates in patients with perioperative SARS-CoV-2 infection. Methods: This international, multicentre, cohort study at 235 hospitals in 24 countries included all patients undergoing surgery who had SARS-CoV-2 infection confirmed within 7 days before or 30 days after surgery. The primary outcome measure was 30-day postoperative mortality and was assessed in all enrolled patients. The main secondary outcome measure was pulmonary complications, defined as pneumonia, acute respiratory distress syndrome, or unexpected postoperative ventilation. Findings: This analysis includes 1128 patients who had surgery between Jan 1 and March 31, 2020, of whom 835 (74·0%) had emergency surgery and 280 (24·8%) had elective surgery. SARS-CoV-2 infection was confirmed preoperatively in 294 (26·1%) patients. 30-day mortality was 23·8% (268 of 1128). Pulmonary complications occurred in 577 (51·2%) of 1128 patients; 30-day mortality in these patients was 38·0% (219 of 577), accounting for 81·7% (219 of 268) of all deaths. In adjusted analyses, 30-day mortality was associated with male sex (odds ratio 1·75 [95% CI 1·28–2·40], p\textless0·0001), age 70 years or older versus younger than 70 years (2·30 [1·65–3·22], p\textless0·0001), American Society of Anesthesiologists grades 3–5 versus grades 1–2 (2·35 [1·57–3·53], p\textless0·0001), malignant versus benign or obstetric diagnosis (1·55 [1·01–2·39], p=0·046), emergency versus elective surgery (1·67 [1·06–2·63], p=0·026), and major versus minor surgery (1·52 [1·01–2·31], p=0·047). Interpretation: Postoperative pulmonary complications occur in half of patients with perioperative SARS-CoV-2 infection and are associated with high mortality. Thresholds for surgery during the COVID-19 pandemic should be higher than during normal practice, particularly in men aged 70 years and older. Consideration should be given for postponing non-urgent procedures and promoting non-operative treatment to delay or avoid the need for surgery. Funding: National Institute for Health Research (NIHR), Association of Coloproctology of Great Britain and Ireland, Bowel and Cancer Research, Bowel Disease Research Foundation, Association of Upper Gastrointestinal Surgeons, British Association of Surgical Oncology, British Gynaecological Cancer Society, European Society of Coloproctology, NIHR Academy, Sarcoma UK, Vascular Society for Great Britain and Ireland, and Yorkshire Cancer Research

FECUNDITY AND FERTILITY OF RHYNCHOPHORUS CRUENTATUS (COLEOPTERA: CURCULIONIDAE)

The palmetto weevil (R. cruentatus F.) breeds in a variety of stressed or dying palms (Giblin-Davis & Howard 1988, 1989). These large (1.9 - 3.0 cm long) weevils are associated with the native cabbage palmetto, Sabal palmetto (Walter), in Florida (Woodruff 1967). Semiochemicals emanating from stressed or dying palms and male conspecifics (Weissling et al. 1992, 1993, 1994, Giblin-Davis et al. 1994) are attractive to R. cruentatus adults. Although not fully understood, mating apparently takes place on dying palms and females lay their eggs in the leaf bases or directly into the wounds of the host. Larvae develop primarily in the crown region but can occasionally be found in the stem tissue. Last instar larvae migrate to the fibrous stem periphery or petiolar bases and construct cocoons from fiber (Giblin-Davis & Howard 1988). Research on an improved method to culture R. cruentatus has required the collection of a large number of eggs to produce neonate larvae for evaluation of diets. Using pineapple, Anana comosus (L.), as an ovipositional substrate, Giblin-Davis et al. (1989) reported the mean lifetime fecundity of field-collected females as 26 ± 15 eggs per female. However, pineapple proved to be a difficult media to dissect for removal of eggs. More suitable ovipositional substrates were investigated and we found that apple (Pyrus malus L.) slices were easily dissected and were readily accepted by R. cruentatus females. Using apple slices, we reinvestigated the fecundity of R. cruentatus females and determined fertility