Wolbachia infection in the newly described Ecuadorian sand flea, Tunga trimamillata

Wolbachia pipientis is an intracellular endosymbiont producing reproductive alterations in its hosts. This bacterium have been reported in many arthropods and nematodes. By PCR amplification and sequencing of the 16S rDNA and ftsZ genes we have identified a Wolbachia strain in the newly described sand-flea, Tunga trimamillata. Prevalence of this endosymbiont in the 26 individuals screened is equal to 35%. Sympatric and allopatric specimens of the related species Tunga penetrans were also analysed, but in contrast to literature data, Wolbachia appears absent in the presently analysed 24 specimens. Field studies evidence a female-biased sex-ratio in T. trimamillata, suggesting that Wolbachia may cause sex-ratio distortion in this species. By means of BLAST search and phylogenetic analysis we found that the Wolbachia strain from T. trimamillata pertains to the arthropod-infecting Wolbachia; this strain is highly differentiated from the Wolbachia strain of T. penetrans described in literature.     

Molecular characterization of Tunga trimamillata and T. penetrans (Insecta, Siphonaptera, Tungidae): taxonomy and genetic variability

A new species of the genus Tungo, T. trimamillata has recently been described on the basis of several morphological traits. To explore the taxonomic status of this flea with respect to T. penetrans, we undertook a molecular analysis of cytochrome oxydase II and 16S rDNA mitochondrial genes and of the internal transcribed spacer 2 nuclear marker on samples of both species. Maximum Parsimony evaluations of the three data set indicate a differentiation compatible with a specific rank between the two fleas with very high levels of divergence. Both mitochondrial and nuclear data are in line with a recent bottleneck in the Malagasy population of T. penetrans, possibly due to the recent colonisation of Africa via human transportation. Further, significantly lower mitochondrial variability in the Ecuadorian populations of T. penetrans with respect to the T. trimamillata ones is also evidenced.     

Blood-feeding of Tunga penetrans males

The jigger Tunga penetrans (Linnaeus, 1758: type-species of the family Tungidae) is the smallest known species of flea (Siphonaptera), causing serious ectoparasitosis of humans and domestic animals. The adult female Tunga lodges in the epidermis of the mammalian host, grows by neosomy, becomes gravid and expels eggs. Relatively little is known about the free-living male Tunga adults. Among impoverished communities of Fortaleza in north-east Brazil, we observed T. penetrans males as well as females penetrating the skin of human hosts. After penetrating the epidermis for a few hours, evidently for capillary feeding from the dermis, males withdrew their mouthparts and crawled away, whereas the females remained completely embedded, hypertrophying to become gravid, eventually dying in situ after oviposition. Caged rats were placed on the sandy soil and examined periodically for Tunga infestation. On five rats we obtained 140 females embedded and we detected 75 males biting, with rat erythrocytes observed in the proventriculus and midgut of all five males dissected and examined microscopically. This confirms that T. penetrans males are hamatophagous ectoparasites of mammals.     

A new species of Tunga perforating the osteoderms of its armadillo host in Argentina and redescription of the male of Tunga terasma

A new species of Tunga (Siphonaptera: Tungidae) collected from armadillos in Argentina is described. The new species is characterized by large and pigmented eyes, the presence of two bristles on antennal segment II, two bristles at the base of the maxilla, and a discoid neosome compressed anteroposteriorly. The gravid female is located in the carapace of the host, perforating the osteoderms. The new species resembles Tunga penetrans and Tunga terasma in general appearance. However, it differs by the greater anteroposterior compression of the neosome, a more angular head, and a manubrium with a pointed proximal end and convex ventral margin (the proximal end of the manubrium is rounded or slightly pointed in T. terasma, and the ventral margin is straight in both T. penetrans and T. terasma). In addition, specimens of T. penetrans have more bristles in antennal segments II and III, and lack bristles in the posterior tibia. This is the first report of a species of Tunga perforating the osteoderms of its host and thereby showing a high degree of specialization. Tunga terasma is recorded for the first time in Argentina; the male is described again and the characteristics of the species amended. This information may be useful in epidemiological studies of diseases caused by species of Tunga.     

Tunga penetrans: molecular identification of Wolbachia endobacteria and their recognition by antibodies against proteins of endobacteria from filarial parasites

In search of Wolbachia in human parasites, Wolbachia were identified in the sand flea Tunga penetrans. PCR and DNA sequencing of the bacterial 16S rDNA, the ftsZ cell division protein, the Wolbachia surface protein (wsp) and the Wolbachia aspartate aminotransferase genes revealed a high similarity to the respective sequences of endosymbionts of filarial nematodes. Using these sequences a phylogenetic tree was generated, that indicates a close relationship between Wolbachia from T. penetrans and from filarial parasites, but possibly as a member of a new supergroup. Ultrastructural studies showed that Wolbachia are abundant in the ovaries of neosomic fleas, whereas other, smaller and morphologically distinct, bacteria were observed in the lumen of the intestine. Wolbachia were labeled by immunohistology and immunogold electron microscopy using polyclonal antibodies against wsp of Drosophila, of the filarial parasite Dirofilaria immitis, or against hsp 60 from Yersinia enterocolitica. These results show that as in filariasis, humans with tungiasis are exposed to Wolbachia. Furthermore, antisera raised against proteins of Wolbachia from arthropods or from filarial parasites can be immunologically cross-reactive.     

Anatomy of Tunga trimamillata Pampiglione et al., 2002 (Insecta, Siphonaptera, Tungidae) and developmental phases of the gravid female

This paper deals with some internal anatomical features observed in histological sections and freshly dissected mounts of Tunga trimamillata, a Siphonaptera recently discovered in Andean regions of Ecuador from several mammals, including man. It was possible to study in males and also non-gravid and gravid females, the location and anatomy of several organs not previously described for this species: the testes, epididymis, ganglia, Malpighian tubules, eyes, rectal ampulla with one of its pads and structures which could be interpreted as midgut diverticula, whose presence has not been recorded in the Siphonaptera. The process of neosomy in the female during pregnancy is illustrated by photographs of the consecutive developmental phases, taken at the stereomicroscope. Furthermore, some details of the exoskeleton, spermatheca during different phases of pregnancy of the gravid female and the presence of a foreign body (parasite?) within the haemocoel have been displayed in specimens cleared with Hoyer's medium.     

Giselinidae fam. nov., a new monophyletic group of cyclopoid copepods (Copepoda, Crustacea) from the Atlantic deep sea

Four new species of Cyclopoida from deep-sea waters are described and placed in two new genera: Giselina gen. n. and Sensogiselina gen. n. The new genera and species belong to a new monophyletic group within the cyclopinid cyclopoids. A new name, Giselinidae, is proposed for this monophylum. The new family is characterised by the combination of the following characters: (1) tergite of leg 1 fused to cephalosome dorsally, but incompletely fused laterally, (2) absence of aesthetascs on ancestral antennulary segments XVI, XXI and XXV, (3) absence of antennary exopodal setae, (4) presence of only three spines on distal exopodal segment of leg 1, (5) absence of inner setae on first endopodal segments of legs 1–4, (6) outer terminal and distal inner elements of distal endopodal segment of leg 4 transformed into spines, (7) distal outer element of leg 5 exopod transformed into a spine, (8) leg 6 with only one seta, and (9) furcal setae I and III located on dorsal margin. Received in revised form: 16 June 2000 Electronic Publication     

A new genus of Platycopioida (Copepoda) from a marine cave on Bermuda

Nanocopia minuta was collected from an inland marine cave on Bermuda. This new genus is reminiscent of Platycopia in its cephalic appendages and in the 5th legs of the male. The 1st leg has a 3-segmented exopod and a 1-segmented endopod. The other legs show reductions with no armature along the inner margin of the exopods and with 2-segmented endopods in the female. Only 2nd and 3rd legs bear two outer spines on the first exopodal segment.Owing to similarities in sexual characters, mouthparts, and modifications of the swimming legs, Nanocopia and Platycopia are considered more closely related to each other than either is to Antrisocopia.Platycopioida now contains three genera of which two are found only in Roadside Cave. The order has retained several primitive characters and seems to have separated early from the gymnoplean stem.     

Phylogeny of scolopendromorph centipedes (Chilopoda): morphological analysis featuring characters from the peristomatic area

Alternative schemes for the higher‐level systematics of the centipede order Scolopendromorpha have been established from characters of trunk segmentation, including the segmental position of spiracles, and the presence or absence of eyes. A comparative survey of the preoral chamber by light and scanning electron microscopy contributes 16 new characters of the epipharynx and hypopharynx, sampled from 26 species that represent 20 genera. These new data together with 49 additional morphological characters permit cladistic analysis of the major scolopendromorph groups. The shortest cladograms resolve blind Scolopendromorpha as a basal grade within which a clade now classified as Plutoniuminae + Scolopocryptopidae (supported by unreversed characters from the preoral chamber) is sister to the remaining scolopendromorphs. A unique row of bullet‐shaped sensilla between the labral and clypeal parts of the epipharynx provides a new autapomorphy of the Scolopendromorpha. Either 21 or 23 trunk segments optimize at the base of the Scolopendromorpha but in either case homoplasy is forced on the cladogram. New characters from the epipharynx give additional support for the monophyly of several traditional groupings, including Cryptopinae, Scolopendridae, Otostigmini, and Scolopendrini, and a basal resolution of Asanadini within Scolopendridae. Of the two competing hypotheses for the position of the enigmatic Mediterranean Plutonium zwierleini—being either sister to the cryptopid Theatops or sister to all other Scolopendromorpha—the former hypothesis is strongly supported; spiracles on all trunk segments in Plutonium are homoplastic with the state in Geophilomorpha. Observations on feeding behaviour are needed to illuminate convergence in characters of the epipharynx and mandible in Edentistoma (Otostigminae) and Campylostigmus (Scolopendrini). © The Willi Hennig Society 2008.     

The animal reservoir of Tunga penetrans in severely affected communities of north-east Brazil

Tungiasis is a zoonotic ectoparasitosis caused by the sand flea Tunga penetrans L. (Siphonaptera: Tungidae). This disease is hyperendemic in poor communities of north-east Brazil, causing considerable morbidity in affected human populations, but the animal reservoirs have not been investigated previously in Brazil. To assess the prevalence and intensity of T. penetrans infection in domestic and peri-domestic animals, as well as in the human population, we surveyed two typical communities of north-east Brazil: an urban slum and a traditional fishing village. In the slum we examined 849 humans, 121 cats, 82 dogs, 2 pigs, 2 rabbits, 1 monkey and 56 rodents, comprising 34 rats (Rattus rattus L.) and 22 mice (Mus domesticus L). In the fishing village we examined 505 humans, 68 dogs, 37 cats, 7 donkeys, 4 cattle, 3 pigs and 1 monkey. Tungiasis was common among dogs and cats of both communities, with respective prevalence rates of 67.1% (95% CI: 55.8-77.1) and 30.9% (95% CI: 20.2-43.3) in dogs, 49.6% (95% CI: 40.4-58.8) and 32.4% (95% CI: 18.0-49.8) in cats. Slum rats were 41.2% (95% CI: 24.6-59.3) infested, but the other animals were not. Human prevalence rates were 54.4% (95% CI: 51.0-57.8) in the slum and 52.1% (95% CI: 47.6-56.5) in the fishing village. High prevalence rates (range 31-67%) of tungiasis in humans, pets and rats (but apparently not other animals) indicate the need for an eco-epidemiological approach to control of this anthropo-zoonotic problem.     

A phylogeny of the families of Scarabaeoidea (Coleoptera)

Abstract. A study, based on examination of thirteen scarabaeoid families, was made of 134 adult and larval characters from the following character suites: 105 adult characters of the antennae, eye, epipharynx, mandible, maxillae, labium, tentorium, trochantin, procoxae, mesocoxae, mesothoracic spiracles, hind wing articulation, hind wing base, hind wing venation, hind wing folding, abdominal sternites, abdominal spiracles, male genitalia, ovarioles and karyotype; twenty larval characters of the antennae, fronto-clypeal suture, stemmata, labial palpi, maxillae, mandibles, legs, stridulatory apparatus, spiracles and ecdysial process; and nine adult and larval biological characters. In order to assess the reliability of different characters in resolving scarabaeoid family relationships, six data sets were subjected to cladistic analysis: the total evidence character set (134 characters), restricted adult character set (thirty-two characters, not including those of the wings), wing character set (seventy-three characters), larval character set (twenty characters), biological character set (nine characters) and re-coded Howden (1982) character set (thirty-nine characters). The complete character set and wing character set both produced phylograms with all nodes resolved; the restricted adult data set, larval data set, Howden (1982) data set and biological data set produced phylograms with diminishing levels of node resolution. The reconstructed phylogeny, from the preferred phylogram of the total evidence character set, shows that the Scarabaeoidea comprises three major lineages; a glaresid, passalid and scarabaeid lineage. The glaresid lineage consists only of the Glaresidae. The passalid lineage comprises two major lines; a glaphyrid line (containing Glaphyridae, Passalidae, Lucanidae, Diphyllostomatidae, Trogidae, Bolboceratidae and Pleocomidae) and a geotrupid line (containing Geotrupidae, Ochodaeidae, Ceratocanthidae and Hybosoridae). The scarabaeid lineage contains those taxa traditionally included within the Scarabaeidae (Aphodiinae, Scarabaeinae, Orphninae, Melolonthinae, Acoma, Chasmatopterinae, Hopliinae, Oncerinae, Rutelinae, Dynastinae, Trichiinae, Cetoniinae and Valginae).     

Postembryonic development of Antygomonas incomitata (Kinorhyncha: Cyclorhagida)

Postembryonic development in the kinorhynch species Antygomonas incomitata was examined using scanning electron microscopy. The morphology of the six juvenile stages, J‐1 to J‐6, varies at numerous details, but they can also be distinguished by a few key characters. Juvenile stage 1 by its composition of only nine trunk segments; J‐2 by the combination of possessing 10 trunk segments, but no cuspidate spines on segment 9; J‐3 by the presence of cuspidate spines on segment 9, but only one pair of cuspidate spines on segment 8; J‐4 by the combination of 10 trunk segments only, but having two pairs of cuspidate spines on segment 8; J‐5 by possessing 11 trunk segments and same spine compositions as adults but is still maintaining postmarginal spiculae; J‐6 specimens closely resemble adults and are most easily identified by their reduced trunk lengths. New segments are formed in a growth zone in the anterior part of the terminal segment. The complete number of segments is reached in J‐5. Development of cuticular head and trunk structures are described through all postembryonic stages and following developmental patterns could be outlined: the mouth cone possesses outer oral styles from J‐1, but in J‐1 to J‐3, the styles alternate in size. Scalids of the introvert are added after each molt, and scalids appear earliest in the anterior rings, whereas scalids in more posterior rings are added in older postembryonic stages. The early J‐1 stage is poor in spines and sensory spots and both structures increase in number after each molt. The complete spine composition is reached in J‐4, whereas new sensory spots appear after all molts, inclusive the final one from J‐6 to adult. Sensory spots in the paraventral positions often appear as Type 3 sensory spots but are through development transformed to Type 2. This transformation happens earliest on the anterior segments. J. Morphol., 2010. © 2010 Wiley‐Liss, Inc.     

Meristoderes gen. nov., a new kinorhynch genus, with the description of two new species and their implications for echinoderid phylogeny (Kinorhyncha: Cyclorhagida, Echinoderidae)

A new kinorhynch genus, Meristoderes gen. nov., and two new species from Spain and the Solomon Islands, respectively, are described. The new genus is distinguished from all other genera by the first segment consisting of a closed cuticular ring, and the second segment having partial tergosternal junctions, and a superficial midventral fold. This is a new cuticular configuration that may shed light into the phylogenetic relationships of echinoderid kinorhynchs. Meristoderes macracanthus gen. et sp. nov. from the Mediterranean coast of Spain is recognised by the presence of middorsal spines on segments 4, 6 and 8, ventrolateral tubules on segment 2, lateroventral tubules on segment 5, lateroventral spines on segments 6-9, lateral accessory tubules on segment 8, one pair of laterodorsal tubules on segment 10. Meristoderes galatheae sp. nov. from the Solomon Islands is recognized by having a middorsal spine on segment 4 only, ventrolateral tubules on segment 2, lateroventral tubules on segment 5, lateroventral spines on segments 6-9, lateral accessory tubules on segment 8 and subdorsal tubules on segment 10. Both species have a pattern of paraventral perforation site clusters on segments 3-9, with conspicuously long bracteate hairs from the posteriormost perforations sites on the segments 3-7 and 3-6, respectively.The new genus Meristoderes gen. nov. is included into the family Echinoderidae Bütschli, 1876 and appears closely related with the genera Cephalorhyncha Adrianov, 1999 and Echinoderes Claparède, 1863. The new information it provides is discussed to clarify the internal phylogeny of Echinoderidae. The terminology for cuticular characters in the overlapping area between consecutive segments is also standardized.     

The segmental origin of the appendages of the head and anterior body segments of a spiroboloid milliped, Narceus annularis

Study of a series of embryos showed that the spiroboloid leg arrangement (1 pair of legs on each of the first 5 segments) is derived from the typical leg arrangement (no legs on segment 1, 1 pair on segments 2 through 4, and 2 pairs on segment 5) by a shifting forward one segment of the first four pairs of legs. A careful re-examination of the literature, especially papers by Robinson ('07), Silvestri ('03, '49), Pflugfelder ('32), and Manton ('61), combined with observations of Narceus embryos led to the conclusion that (1) the anterior body segments are primatively single (2) the gnathochilarium is composed of only one pair of mouthparts, the diplopod head having but two gnathal segments, and (3) the intercalary segment is present in the Diplopoda.     

Troglocyclops janstocki,new genus,new species,a very primitive cyclopid (Copepods: Cyclopoida) from an anchialine cave in the Bahamas

Troglocyclops janstocki is proposed to accommodate cyclopids from Hatchet Bay Cave, Eleuthera Island, Bahamas. The species is the most primitive member of the Halicyclopinae because of the presence of 15-segmented antennules, mandibular palp reduced to 3 setae, one of them quite long and plumose, a bisegmented maxillary endopodite, and 3 segments in the maxilliped endopodite. These copepods possess the first pediger still distinct, being partially enclosed dorsally and laterally by a carapace-like extension of the posterior margin of the dorsal cephalic shield. This latter character represents a plesiomorphic state shared with the primitive cyclopinids and, within the Cyclopidae, only with the Euryteinae. The new taxon is the only known species of Halicyclopinae having two apical spines on the terminal segment of the exopodite of legs 2 to 4, and the intercoxal sclerites of the legs 1 and 2 sexually dimorphic. The phylogenetic importance of each of these characters is discussed.     

The problem of the number of tarsomeres in the regenerated cockroach leg.

There are 5 tarsomeres in the normal cockroach leg, but this number is often reduced in regenerated legs. In order to examine this complicated situation, fore-, mid-, and hindlegs of German cockroaches were amputated at 11 different tarsal levels and at 18 different times during the last instar. When tarsi were amputated at or proximal to the 3rd tarsomere, 4-segmented tarsi regenerated. When legs were amputated distal to the 3rd tarsomere, the regenerated tarsi had 5 segments. Three-segmented tarsi rarely regenerated when legs were amputated proximal to 3rd tarsomere and in the latter half of the instar period. The lengths of all tarsomeres of regenerated tarsi were measured together with those of unoperated contralateral tarsomeres, and the ratios of the former to the latter were calculated. The ratios ranged from 28 to 138% for the various tarsomeres and levels of amputation. From a comparison of the ratios and morphological observations, it was suggested that the 3rd tarsomere of the normal 5-segmented tarsus has disappeared in the regenerated 4-segmented tarsus. Pads and disto-lateral spines of tarsomeres were observed on unoperated and regenerated tarsi. It was of interest that double spines were often found on the 4-segmented tarsi, mostly on the 2nd tarsomere, just proximal to the position of the missing 3rd tarsomere. This observation supported the idea that the 3rd tarsomere has not simply disappeared, but has probably fused with the 2nd tarsomere.     

新疆阳蝇属Helina R.—D.二新种（双翅目：蝇科）

记述采自新疆蝇属Helina R.-D．2新种,托木尔阳蝇Helina tuomuerra,sp,nov。和新疆阳蝇Helina xinjiangensis,sp.nov.模式标本保存于中国科学院动物研究所。