Partial Cytochrome Oxidase Subunit Research Articles

Description of the Asian chili pod gall midge, Asphondylia capsicicola sp. n., with comparative notes on Asphondylia gennadii (Diptera: Cecidomyiidae) that induces the same sort of pod gall on the same host plant species in the Mediterranean region

A new species of the genus Asphondylia (Diptera: Cecidomyiidae) that infests pods of chili, Capsicum annuum L. and Capsicum frutescens L. (Solanaceae), is described as Asphondylia capsicicola sp. n. based on specimens collected from Indonesia and Vietnam. The new species is similar to Asphondylia gennadii (Marchal) (=Asphondylia capsici) that induces chili pod galls in the Mediterranean region, but is distinguishable from it by the morphological features of pupa such as the nonlinear arrangement of the lower frontal horns, and the narrower longitudinal band of transverse wrinkles on the tergite of the mesothorax. Differences between the two species in the DNA sequencing data were 69 bp (16%) to 77 bp (19%) among 413 bp of the partial cytochrome oxidase subunit I region examined, supporting the morphological identification. This is one of the examples in which two congeners induce the same sort of gall on the same host plant organ and species, which means that the two species are not distinguishable based solely on gall shape and host plant information, unlike many other gall midges.

First Report of Phytophthora hydropathica Causing Wilting and Shoot Dieback on Viburnum in Italy

The genus Viburnum comprises over 150 species of shrubs and small trees such as Laurustinus (Viburnum tinus L.), which is one of the most widely used ornamental plants in private and public gardens. Furthermore, it commonly forms stands of natural woodland in the Mediterranean area. In autumn 2012, a survey was conducted to determine the presence of Phytophthora ramorum on Viburnum in commercial nurseries in the Latium region where wilting, dieback, and death of twigs were observed on 30% of the Laurustinus plants. A Phytophthora species was consistently recovered from soil rich in feeder roots from potted Laurustinus plants showing symptoms. Soil samples were baited with rhododendron leaves. Small pieces of leaf tissue cut from the margin of lesions were plated on P5ARPH selective medium (4). Pure cultures, obtained by single-hypha transfers on potato dextrose agar (PDA), were petaloid. Sporangia formation was induced on pepper seeds (3). Sporangia were almost spherical, ovoid or obpyriform, non-papillate and non-caducous, measuring 36.6 to 71.4 × 33.4 to 48.3 μm (average 53.3 × 37.4 μm) with a length/width ratio of 1.4. Chlamydospores were terminal and 25.2 to 37.9 μm in diameter. Isolates were considered heterothallic because they did not produce gametangia in culture or on the host. All isolates examined had 30 to 35°C as optimum temperatures. Based on these morphological characteristics, the isolates were identified as Phytophthora hydropathica (2). Morphological identification was confirmed by internal transcribed spacer (ITS), and mitochondrial partial cytochrome oxidase subunit 2 (CoxII) with BLAST analysis in the NCBI database revealing 99% identity with ITS and 100% identity with CoxII. The sequences of the three isolates AB234, AB235, and AB236 were deposited in European Nucleotide Archive (ENA) with the accession nos. HG934148, HG934149, and HG934150 for ITS and HG934151, HG934152, and HG934153 for CoxII, respectively. Pathogenicity tests were conducted in the greenhouse on a total of six 1-year-old shoots cut from V. tinus plants with two inoculation points each. Mycelial plugs cut from the margins of actively growing 8-day-old cultures on PDA were inserted through the epidermis into the phloem. Controls were treated as described above except that sterile PDA plugs replaced the inoculum. Shoots were incubated in test tubes with sterile water in the dark at 24 ± 2°C. After 2 weeks, lesions were evident at the inoculation points and symptoms were similar to those caused by natural infection. P. hydropathica was consistently re-isolated from the margin of lesions, while controls remained symptomless. In the United States in 2008, P. hydropathica was described as spreading from irrigation water to Rhododendron catawbiense and Kalmia latifolia (2). This pathogen can also attack several other horticultural crops (1), but to our knowledge, this is the first report of P. hydropathica causing wilting and shoot dieback on V. tinus.

Evidence of cryptic speciation in mesostigmatid mites from South Africa.

Laelaps giganteus and Laelaps muricola (Mesostigmata; Laelapidae) are widespread and locally abundant host generalists on small mammals in southern Africa. The large host range and complex life history of these ectoparasites may allude to possible intraspecific cryptic diversity in these taxa. To assess genetic and morphological diversity in L. giganteus and L. muricola, we sampled 228 rodents at eight localities in South Africa. This sample included nine previously recorded host species and on these, L. muricola was only recorded from Mastomys natalensis and Micaelamys namaquensis while L. giganteus was found on Rhabdomys dilectus and Lemniscomys rosalia. Phylogenetic analyses of partial mtDNA cytochrome oxidase subunit I (COI) and nuclear ITS1 data strongly supported the recognition of L. giganteus and L. muricola, a scenario partly supported by the Tropomyosin intron. Strong support for evolutionary distinct lineages within L. giganteus is found: L. giganteus lineage 1 is confined to R. dilectus and L. giganteus lineage 2 is confined to L. rosalia. These host specific monophyletic lineages were also separated by 9.84% mtDNA sequence divergence and 3.44% nuclear DNA sequence divergence. Since quantitative morphometric analyses were not congruent with these findings, these two lineages more than likely represent cryptic species.

Old Aegean palaeoevents driving the diversification of an endemic isopod species (Oniscidea, Trachelipodidae)

The Greek endemic isopod species Trachelipus aegaeus is distributed in Aegean islands and the adjacent coastal parts of the Greek mainland. Major palaeogeographic events of the Aegean archipelago, such as the formation of the mid‐Aegean trench and the Messinian Salinity Crisis, have been often employed as major causal factors of evolutionary events and phylogeographic patterns exhibited by several taxa. Herein, we infer phylogenetic relationships among T. aegaeus populations using partial cytochrome oxidase subunit I (COI) and 16S rRNA sequences. Due to the poor preservation of the specimens, we propose a modified DNA extraction protocol, which returned highly positive results in terms of the quality of the total extracted DNA. We implement a calibrated molecular clock and path sampling analysis, using alternative palaeogeographic events and rates of substitution, to evaluate the biogeographic history of the species and to estimate the chronology of diversification events among its populations. Our results are clearly in favour of the scenario of the MAT triggering vicariance among most T. aegaus populations. Moreover, the large intraspecific genetic divergence (0–19% for COI and 0–20.3% for the 16S rRNA) and the overall phylogeographic patterns depicted herein seem not to have been obscured by more recent palaeogeological events. A role of dispersal, probably human‐aided, is assumed for certain ‘deviant’ cases.

Vinodolia fiumana Radoman, 1973 (Caenogastropoda: Rissooidea): rediscovery and relationships of a species presumed extinct

Vinodolia fiumana was described by Radoman from Glogi spring in 1973, and later not found, thus considered extinct. In the CLECOM list Anagastina, Dalmatinella and Prespiana were classified, as subgenera, within the genus Vinodolia. After visiting the Glogi spring on several occasions, we found four specimens of V. fiumana. Its shell, soft part external morphology, radula, penis and female reproductive organs are described. Partial cytochrome oxidase subunit I mitochondrial and 18S rRNA nuclear gene sequences were used to infer phylogenetic relationships of Vinodolia. Sadleriana was found to be the sister taxon of Vinodolia, but neither Anagastina (closely related with Radomaniola), nor Dalmatinella were closely related with it.

Phylogeographic analysis of Apis cerana populations on Hainan Island and southern mainland China, based on mitochondrial DNA sequences

The phylogeography of the eastern honeybee (Apis cerana) in southern China is poorly understood. In the present study, we investigated phylogenetic relationships among A. cerana populations on Hainan island and southern mainland China. Analyses were based on the partial cytochrome oxidase subunit 1 gene of mitochondrial DNA and A. cerana samples representing 12 geographic populations. We identified 57 novel haplotypes (GenBank accession numbers: KC175488-KC175544) within our sampled populations and inferred two deeply divergent intraspecific lineages when previously published sequences were included, which could be further divided into six sublineages. We also found a significant phylogeographic structure within lineage A. Our results reveal that A. cerana populations on Hainan island and southern mainland China have undergone a long period of differentiation in isolation followed by historical demographic expansion. Although gene flow now occurs between these locations, gene flow has occurred over comparatively a short time period, and is likely to be the result of anthropogenic effects.

Epinephelus moara: a valid species of the family Epinephelidae (Pisces: Perciformes)

This study documents the major external and internal morphological differences between Epinephelus bruneus and Epinephelus moara, and analyses the complete mitogenomes of both species. The partial cytochrome oxidase subunit I (coI) sequence divergence between E. bruneus and E. moara is significantly higher than specimens within the same species (P < 0·05). Analyses of gene flow (Nm = 0·02) and genetic differentiation (ϕst = 0·92995, P > 0·05) reveal reproductive isolation between E. bruneus and E. moara. These results support the hypothesis that E. moara is a valid species. Further molecular comparisons between E. bruneus and E. moara obtained in this study and a specimen identified in GenBank as E. bruneus from South Korea reveal that the latter is identical to E. moara rather than to E. bruneus.

Evidences of increasing risk of dirofilarioses in southern Italy

Given the spread of Aedes albopictus from northern to southern Italy, and the lack of updated data on Dirofilaria infections, this study was carried out to assess the infection risk for dogs and cats in Apulia region. During a 2-year study, 175 A. albopictus female specimens and samples of blood from 427 dogs (309 privately owned dogs and 118 shelter dogs) and 12 cats were collected. All blood samples were subjected to a modified Knott method, to a test for the detection of circulating Dirofilaria immitis antigen, and to a Dirofilaria species-specific real-time PCR for the simultaneous detection of D. immitis and Dirofilaria repens, targeting on partial cytochrome oxidase subunit 1 and internal transcribed spacer-2, respectively. Two abdomen and one thorax pools from A. albopictus were positive for D. immitis, with minimum infection rates of 1.14 and 0.51, respectively, and a probability of a single positive specimen to be infected of P = 0.6% (95% confidence interval (CI) = 0.12-1.73). Out of 439 examined subjects, 22 (5.0%) tested positive for Dirofilaria spp. in at least one diagnostic test. A specific D. immitis infestation rate of 3.5% was found among the privately owned dogs, while shelter dogs tested positive only for D. repens with a prevalence of 3.4%; one cat tested molecularly positive for D. immitis. There was a significantly higher rate of positivity among guard dogs for D. immitis (odds ratio, 6.24, 95% CI, 1.26-25.28; P < 0.05). The increasing risk of D. immitis infection in southern Italy is supported by the noteworthy positivity of A. albopictus populations and the cat. Our data highlight the usefulness to include filarioid infestation in routine diagnosis.

Endemic angiostrongyliasis in the Brazilian Amazon: Natural parasitism of Angiostrongylus cantonensis in Rattus rattus and R. norvegicus, and sympatric giant African land snails, Achatina fulica

Angiostrongylus cantonensis, the rat lungworm, is one etiological agent of eosinophilic meningoencephalitis in humans. This zoonosis is frequently found in Asia and, more recently, in North America, Caribbean Island and northeastern of South America. Until now, research of A. cantonensis in southern, southeastern and northeastern regions of Brazil has been found natural infections only terrestrial and freshwater intermediate snail hosts (Achatina fulica, Sarasinula marginata, Subulina octona, Bradybaena similaris and Pomacea lineate). In this study, we examined the occurrence of helminthes in the synantropic rodents Rattus rattus and Rattus norvegicus in northern Brazil, focusing on the role of these species as vertebrate hosts of A. cantonensis and A. fulica as intermediate host have found natural. Thirty specimens of R. rattus and twelve of R. norvegicus were collected in the Guamá and Jurunas neighborhoods of the city of Belém, in the Brazilian state of Pará, of which almost 10% harbored adult worms in their pulmonary arteries. Sympatric A. fulica were found to be infected by L3 larvae, which experimental infection confirmed to be A. cantonensis. Natural infection of snails and rodents with A. cantonensis was confirmed through morphological and morphometrical analyses of adults and larvae using light microscopy, scanning electron microscopy and molecular sequences of partial Cytochrome Oxidase subunit I. Phylogenetic analyses showed that A. cantonensis isolated from Pará, Brazil is similar to Japan isolate; once these specimens produced a single haplotype with high bootstrap support with Rio de Janeiro isolate. This study confirms that A. cantonensis is now endemic in northern Brazil, and that R. rattus and R. norvegicus act as natural definitive hosts, and A. fulica as the intermediate host of the parasite in this region.

Description of Jolyelmis spangleri a new species from Churí-tepui (Chimantá Massif, Venezuela), with a description of the larva of J. spangleri and J. reitmaieri (Insecta: Coleoptera: Elmidae)

Four species have been recognized in the genus Jolyelmis Spangler &amp; Faitoute; all collected at isolated table mountains,mostly in cool stenothermal hypocrenal streams. The known distribution of the genus is confined to Venezuela, and eachspecies is here reported as endemic for the respective table mountain where it has been found. Samples from Churí-tepuiand Mount Roraima in south-eastern Venezuela contained numerous adults and larvae of Elmidae among which is a newspecies, Jolyelmis spangleri, which is here described. The new species is illustrated and keyed with all other Jolyelmis,and new distributional data for J. reitmaieriČiampor, Jr. &amp; Kodada are included. Molecular analysis of partial cytochromeoxidase subunit I (cox1) confirmed conspecific life stages for two species: J. spangleri and J. reitmaieri. The larva of eachspecies is described and illustrated, and the generic diagnosis for Jolyelmis larvae is provided. Among the known larvaeof South American Elmidae, larvae of the genus Xenelmis Hinton appear to be the most similar. Morphological similarities are discussed. Habitats of adults and larvae are briefly described and illustrated by photographs.

A new horsehair worm, Chordodes formosanus sp. n. (Nematomorpha, Gordiida) from Hierodula mantids of Taiwan and Japan with redescription of a closely related species, Chordodes japonensis

A new species of horsehair worm, Chordodes formosanus sp. n., is described and compared to a closely related species, Chordodes japonensis. Although both species possess the same six cuticular structures of areoles on the surface, the significantly longer filaments on the female crowned areoles can be used as diagnostic characters for the new species. The different taxonomic status of these two species was also confirmed after analyzing the partial cytochrome oxidase subunit I sequence, and the mantid hosts, which are respectively limited to the genus Tenodera for Chordodes japonensis and Hierodula for Chordodes formosanus sp. n. In addition, the immature stages of eggs and larvae of the new species are also described and discussed in detail.

Reconstructing the colonisation and diversification history of the endemic freshwater crab ( Seychellum alluaudi) in the granitic and volcanic Seychelles Archipelago

The endemic, monotypic freshwater crab species Seychellum alluaudi was used as a template to examine the initial colonisation and evolutionary history among the major islands in the Seychelles Archipelago. Five of the “inner” islands in the Seychelles Archipelago including Mahé, Praslin, Silhouette, La Digue and Frégate were sampled. Two partial mtDNA fragments, 16S rRNA and cytochrome oxidase subunit I (COI) was sequenced for 83 specimens of S. alluaudi. Evolutionary relationships between populations were inferred from the combined mtDNA dataset using maximum parsimony, maximum likelihood and Bayesian inferences. Analyses of molecular variance (AMOVA) were used to examine genetic variation among and within clades. A haplotype network was constructed using TCS while BEAST was employed to date the colonisation and divergence of lineages on the islands. Phylogenetic analyses of the combined mtDNA data set of 1103 base pairs retrieved a monophyletic S. alluaudi group comprised three statistically well-supported monophyletic clades. Clade one was exclusive to Silhouette; clade two included samples from Praslin sister to La Digue, while clade three comprised samples from Mahé sister to Frégate. The haplotype network corresponded to the three clades. Within Mahé, substantial phylogeographic substructure was evident. AMOVA results revealed limited genetic variation within localities with most variation occurring among localities. Divergence time estimations predated the Holocene sea level regressions and indicated a Pliocene/Pleistocene divergence between the three clades evident within S. alluaudi. The monophyly of each clade suggests that transoceanic dispersal is rare. The absence of shared haplotypes between the three clades, coupled with marked sequence divergence values suggests the presence of three allospecies within S. alluaudi.

Aphelenchoides paradalianensis n. sp. (Nematoda: Aphelenchoididae) isolated at Guangzhou, China, in packaging wood from South Korea

Aphelenchoides paradalianensis n. sp. collected in Guangzhou, China from packaging wood exported from South Korea, is described and illustrated based on morphology and molecular analyses of the ITS regions, the near-full-length small subunit rDNA gene and partial cytochrome oxidase subunit I of mitochondrial DNA. This new species has a distinctive tail terminus, possessing a short trunk with two finely rounded and smooth processes of unequal length in the female and a small ventral papilla-like mucronate structure or a short trunk with two fine processes in the male, slightly offset lip region with 6 annules, four incisures in the lateral field, short postuterine sac (extending ca. 13–20% of vulva-anus distance), medium-sized spicules (14.1–18.6 μ m), three pairs of caudal papillae. It is closest to A. dalianensis, but these two species can be distinguished from each other by spicule size and shape, male tail tip shape, postuterine sac length, c ratio and ITS sequence. Molecular analysis reveals that this species has unique ITS, 18S and mt-DNA sequences.

Molecular approaches to the identification ofBulinusspecies in south-west Nigeria and observations on natural snail infections with schistosomes

The current study considers the distribution of a small sample of 138 Bulinus snails, across 28 localities within eight Nigerian states. Snails were identified using a combination of molecular methods involving both DNA sequencing of a partial cytochrome oxidase subunit 1 (cox1) fragment and restriction profiles obtained from ribosomal internal transcribed spacer (its) amplicons. The results showed that the majority of Bulinus samples tested belonged to the species Bulinus truncatus while only two were Bulinus globosus. The use of RsaI restriction endonuclease to cleave the ribosomal its of Bulinus, as a method of species identification, was adopted for the majority of samples, this being a quicker and cheaper method better suited to small laboratory environments. Polymerase chain reaction (PCR) amplification of the schistosome Dra1 repeat within each of the collected Bulinus samples was employed to determine the extent and distribution of infected snails within the sample areas. Successful amplification of the Dra1 repeat demonstrated that 29.7% of snails were infected with schistosomes. Sequencing of the partial schistosome its from a small subset of snail samples suggested that some snails were either penetrated by both Schistosoma haematobium and Schistosoma bovis miracidia or hybrid miracidia formed from the two species.

Morphological and molecular characterization of Aphelenchoides fujianensis n. sp. (Nematoda: Aphelenchoididae) from Pinus massoniana in China

Aphelenchoides fujianensis n. sp. is described and illustrated from a dead Pinus massoniana based on morphology and molecular analyses of the near-full-length small subunit rDNA gene and partial cytochrome oxidase subunit I of mitochondrial DNA. This new species belongs to the Group 3 of Aphelenchoides species sensu Shahina with star-shaped tail terminus and is characterised by a relatively long body (653–843 μ m in the male and 803–941 μ m in the female) and four lateral incisures in the lateral field. The male has relatively large spicules (24–30 μ m). The female has elongate postvulval uterine sac (extending ca 32–44% of vulva-anus distance), usually with sperms. Both male and female have star-shaped mucro. It is distinguished from other species by postvulval uterine sac length, a and c ratios, and spicule size and shape. Molecular analysis reveals that this species has unique 18S and mt-DNA sequences, and is closest to Aphelenchoides besseyi in dendrograms inferred using both markers. The identification codes of OEPP/EPPO for A. fujianensis n. sp. are: A1-B2-C1-D1/3-E1-F1/2.

Elucidating the spread of the emerging canid nematode Angiostrongylus vasorum between Palaearctic and Nearctic ecozones

Angiostrongylus vasorum is an emerging parasite that is currently distributed through Western Europe and parts of South America. An isolated population is also present in Newfoundland, Canada. This presents a risk of onward spread into North America, but its origin is unknown. To ascertain the phylogeographic relationships and genetic diversity of A. vasorum within the western Palaearctic and eastern Nearctic ecozones, a total of 143 adult and larval nematode specimens were collected from foxes (Vulpes vulpes) and dogs (Canis lupus familiaris) in Canada, Denmark, France, Germany, Ireland, the Netherlands, Portugal and the United Kingdom, and a coyote (Canis latrans) in Canada. DNA was extracted and the second internal transcribed spacer and two mitochondrial loci were amplified and sequenced. Multiple haplotypes (n=35) based on combined mitochondrial sequences (1078bp) of the partial cytochrome oxidase subunit I (COI), large subunit ribosomal RNA (rrnL) and the complete nicotinamide adenine dinucleotide dehydrogenase 3 (NADH3) sequences, were observed throughout the Palaearctic countries sampled; however, only a single haplotype was observed for the Canadian A. vasorum population. The likely origin of A. vasorum in Newfoundland is therefore inferred to be within the western Palaearctic. There was no evidence of genetic segregation of parasites in dogs, foxes and coyotes, supporting the hypothesis that transmission occurs between wild and domestic canids. The transmission dynamics and population structure of this nematode are further discussed.

Lithodes formosae, a new species of king crab from Taiwan (Crustacea: Decapoda: Lithodidae)

A new species of king crab is described from Taiwan, Lithodes formosae sp. nov. The new species resembles L. longispina Sakai, 1971, from Japan, L. megacantha Macpherson, 1991, from French Polynesia, and L. paulayi Macpherson &amp; Chan, 2008, from Guam, in having long dorsal spines in adults, though it is most similar to L. paulayi. Lithodes formosae is distinguished from the aforementioned species primarily by differing branchial spine arrangement and proportional spine length. Comparison of partial cytochrome oxidase subunit I (COI) gene sequences from L. formosae sp. nov., L. paulayi and L. longispina supports the specific status of the new species. Six species of Lithodidae are known from Taiwan, of which L. formosae sp. nov. is the first to be described as new from Taiwan.

Reef-associated crustacean fauna: biodiversity estimates using semi-quantitative sampling and DNA barcoding

The cryptofauna associated with coral reefs accounts for a major part of the biodiversity in these ecosystems but has been largely overlooked in biodiversity estimates because the organisms are hard to collect and identify. We combine a semi-quantitative sampling design and a DNA barcoding approach to provide metrics for the diversity of reef-associated crustacean. Twenty-two similar-sized dead heads of Pocillopora were sampled at 10 m depth from five central Pacific Ocean localities (four atolls in the Northern Line Islands and in Moorea, French Polynesia). All crustaceans were removed, and partial cytochrome oxidase subunit I was sequenced from 403 individuals, yielding 135 distinct taxa using a species-level criterion of 5% similarity. Most crustacean species were rare; 44% of the OTUs were represented by a single individual, and an additional 33% were represented by several specimens found only in one of the five localities. The Northern Line Islands and Moorea shared only 11 OTUs. Total numbers estimated by species richness statistics (Chao1 and ACE) suggest at least 90 species of crustaceans in Moorea and 150 in the Northern Line Islands for this habitat type. However, rarefaction curves for each region failed to approach an asymptote, and Chao1 and ACE estimators did not stabilize after sampling eight heads in Moorea, so even these diversity figures are underestimates. Nevertheless, even this modest sampling effort from a very limited habitat resulted in surprisingly high species numbers.

Rapid polymerase chain reaction–restriction fragment length polymorphism method for discrimination of the two Atlantic cryptic deep‐sea species of scabbardfish

The present investigation provides an efficient diagnostic method based on polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis to discriminate between two cryptic species of scabbardfish, Aphanopus carbo and A. intermedius, with commercial relevance in several European fish markets. Two DNA fragments from the mtDNA, including control region and partial cytochrome oxidase subunit I genes of about 1100bp and 700bp, respectively, were isolated by PCR amplification. Digestion of the amplicon including the control region with HaeII and the amplicon including the COI gene with Sau3AI restriction enzymes allowed an unequivocal discrimination between the two scabbardfish species. This PCR-RFLP method allowed a clear and rapid discrimination of the trichiurid species studied.

A taxonomic revision of the genus Stempellinella (Diptera: Chironomidae)

The species of the chironomid genus Stempellinella are revised, described, and figured, and keys to known larvae, pupae, and male and female adults are presented. Five species new to science are diagnosed and described. An emended generic diagnosis of Stempellinella is given and morphological differences from its putative sister genus Zavrelia are discussed. The partial cytochrome oxidase subunit I (COI) gene sequences (DNA barcodes) of 10 Stempellinella species are presented and the variation in genetic distances within and between species is compared. The results indicate that DNA barcodes are suitable molecular markers for identification of Stempellinella species. The following life stages and species are diagnosed and described: the larva, pupa, male, and female of S. brevis, S. chambiensis, S. ciliaris, S. edwardsi, S. fimbriata sp. n., S. flavidula, S. leptocelloides, S. saltuum, and S. truncata; the pupa, male, and female of S. reissi and S. tamaseptima; the larva, pupa, and male of Stempellinella boltoni sp. n.; the pupa and male of S. lamellata sp. n.; the male and female of S. coronata; and the male of S. apicula, S. brevilamellae, S. distincta sp. n., and S. sublettorum sp. n. Two pupal types are described but not formally named. The following changes of combination are made: Stempellina chambiensis and Stempellina truncata are transferred to the genus Stempellinella. Zavrelia inopinata is listed as a new junior synonym of Stempellinella ciliaris.