First Record of the Gall-Inducing Insect Ophelimus mediterraneus Borowiec & Burks (Hymenoptera, Chalcidoidea, Eulophidae) in Eucalyptus from America, Including the First Report of Male Specimens.

BackgroundA total of six Neocalanus species inhabit the oceans of the world. Of these, three species plus form variants (N. cristatus, N. plumchrus, N. flemingeri large form, and N. flemingeri small form), which constitute a monophyletic group among Neocalanus copepods, occur in the Northwestern Pacific off Japan. In the present study, we have tried to discriminate the three species plus form variants of Neocalanus copepods based on sequences of four DNA marker regions.Methodology/Principal FindingsDiscrimination was performed based on the DNA sequence information from four genetic markers, including the mitochondrial COI, 12S, nuclear ITS, and 28S gene regions. Sequence dissimilarity was compared using both distance- and character-based approaches. As a result, all three species were confirmed to be distinct based on the four genetic marker regions. On the contrary, distinction of the form variants was only confirmed based on DNA sequence of the mitochondrial COI gene region.Conclusions/SignificanceAlthough discrimination was not successful for the form variants based on the mitochondrial 12S, nuclear ITS, and 28S genes, diagnostic nucleotide sequence characters were observed in their mitochondrial COI gene sequences. Therefore, these form variants are considered to be an important unit of evolution below the species level, and constitute a part of the Neocalanus biodiversity.

Southeast Asian mouth-brooding Betta fighting fish (Teleostei: Perciformes) species and their phylogenetic relationships based on mitochondrial COI and nuclear ITS1 DNA sequences and analyses

Comparative phylogeography of commensal species may show congruent patterns where the species involved share a common history. Temnosewellia is a genus of flatworms, members of which live in commensal relationships with host freshwater crustaceans. By constructing phylogenetic trees based on mitochondrial COI and 28S nuclear ribosomal gene sequences, this study investigated how evolutionary history has shaped patterns of intraspecific molecular variation in two such freshwater commensals. This study concentrates on the flatworm Temnosewellia albata and its critically endangered crayfish host Euastacus robertsi, which have a narrow climatically-restricted distribution on three mountaintops. The genetic data expands upon previous studies of Euastacus that suggested several vicariance events have led to the population subdivision of Euastacus robertsi. Further, our study compared historical phylogeographic patterning of these species. Our results showed that phylogeographic patterns shared among these commensals were largely congruent, featuring a shared history of limited dispersal between the mountaintops. Several hypotheses were proposed to explain the phylogeographic points of differences between the species. This study contributes significantly to understanding evolutionary relationships of commensal freshwater taxa.

The geographic population patterns of Lingula anatina across the Indo-West Pacific region are analyzed based on mitochondrial COI and nuclear EF-1α gene sequences. Compared with the remarkable morphological stasis, genetic evidence of extant Lingula species displays deep genetic divergence. Three distinct COI lineages were detected for L. anatina, one of which is from Qinhuangdao (Hebei, China, Bohai Sea), the other two from Beihai (Guangxi, China, Gulf of Tonkin). Individuals from South Japan have a very close relationship with one of the two COI lineages found in Beihai, which is also supported by EF-1α results, suggesting a relatively recent migration between South China Sea and East China Sea. Genetic distances between the three lineages of L. anatina are rather high (8.9%, 8.6%, and 2.7%), and those between L. anatina and L. adamsi is much higher (44.5%), compared to other marine invertebrates. Both tectonic evolution and the repeated Quaternary glaciations have contributed to the complex phylogeographic pattern found in these recent Lingula anatina populations.

Biodiversity of the Betta smaragdina (Teleostei: Perciformes) in the northeast region of Thailand as determined by mitochondrial COI and nuclear ITS1 gene sequences

Two new species of Conopea (Say 1822) are described from the Gulf of Guinea: Conopea saotomensis sp. n.and Conopea fidelis sp. n. These two new species were collected from the historically isolated volcanic islands of São Tomé and Príncipe. The relationship between Conopea saotomensis sp. n., Conopea fidelis sp. n.and two other Atlantic barnacle species, Conopea calceola (Ellis 1758) and Conopea galeata (Linnaeus 1771), is examined. The methods employed are the construction of a molecular phylogeny using mitochondrial COI and nuclear H3 gene sequence data along with morphological comparisons of calcareous and cuticular body parts. It is found that Conopea saotomensis sp. n., Conopea fidelis sp. n.and Conopea calceola are most closely related to each other but the relationship among them is unresolved. Gorgonian hosts are identified. Preliminary observations show species level host specificity for Conopea fidelis sp. n.

Forty three European population samples of mealy aphids from various winter and summer host plants were attributed to respective species of Hyalopterus by means of their partial sequences of mitochondrial COI gene. Used Hyalopterus samples emerged as monophyletic relative to outgroup and formed three major clades representing three host specific mealy aphid species in the Neighbor joining, Maximum parsimony, Maximum likelihood and Bayesian inference trees. Hyalopterus pruni and Hyalopterus persikonus emerged as a sister species, whilst Hyalopterus amygdali was located basally. Samples representing different clades in the molecular trees were used for canonical discrimination analysis based on twenty two morphological characters. Length of the median dorsal head hair enabled a 97.3 % separation of Hyalopterus amygdali from the remaining two species. No single character enabled satisfactory discrimination between apterous viviparous females of Hyalopterus pruni and Hyalopterus persikonus. A modified key for the morphological identification of Hyalopterus species is suggested and their taxonomic status discussed.

Mitochondrial COI and 16S rRNA gene partial sequences were generated for Pagurus brachiomastus from two locations and P. proximus from single location of the south of Russian Far East. Genetic variation and differentiation of P. brachiomastus Russian and South Korean (GenBank data) populations were estimated and compared on the basis of sequence data on two mitochondrial genes separately. Additionally, P. simulans from South Korea was analyzed. Results show P. brachiomastus from two locations of the south of Russian Far East and South Korea and Korean P. simulans differs at intraspecific level based on molecular markers. Median-Joining analysis indicate presence of two main groups of haplotypes, representing P. proximus and P. brachiomastus + P. simulans, respectively. These data support that P. brachiomastus and P. simulans are conspecific. Mismatch distribution analysis showed population growth for P. brachiomastus at the south of Russian Far East. Sequence pairwise variations of both molecular markers appear differently in two far eastern samples of P. brachiomastus, that can be partially explained by active gene flow through hermit crab migrations between two localities after their isolation.

This study presents the molecular phylogenetic analysis of Korean Aurelia aurita Linnaeus collected from Yeosu in the southern waters of Korea using nuclear ITS1 region and mitochondrial COI gene sequences. The use of oligonucleotide primers F5 (forward) and R5 (reverse) targeted to ITS1 and LCO1490 (forward) and HCO2198 (reverse) targeted to COI amplified 267 bp and 643 bp fragments, respectively. The shortest genetic distance towards the ITS1 region is estimated at 0.023 when comparing Korean A. aurita to Aurelia sp. collected from California, USA. In particular, Korean and American/Swedish A. aurita were located far away in terms of genetic distance, ranging from 0.393 to 0.395. On the other hand, the genetic distance between Korean and English/Turkish/Swedish/ American A. aurita regarding the mitochondrial DNA COI gene ranged from 0.201 to 0.205. However, a sister-ship with Korean and American A. aurita showed an extremely high bootstrap value (100%). The predicted secondary RNA structure of the mitochondrial DNA COI gene showed many different folding structures with a similar energy between Korean and American A. aurita. These results suggest that ITS1 and the mitochondrial DNA COI gene could be used as genetic markers for identification of the biogeographic populations.

Megaselia scalaris (Diptera: Phoridae), commonly known as scuttle fly, is widely distributed all over the world. It is easily cultured in the laboratory condition making it a potential model organism. Besides, it has forensic importance. However, no report from Bangladesh could be retrieved about this fly. So, in the present study, identification of this species was attempted using both morphological and molecular approaches. Characteristics of male hypopygium and legs played key roles in morphological identification. To strengthen identification, mitochondrial COI and 16S rRNA gene fragments were amplified and sequenced. Blast search at NCBI provided highest hits to available COI and 16S rRNA sequences of M. scalaris. A neighbor joining phylogenetic tree was built using sequences of respective COI gene region to show its relationship among other closely related dipteran flies.
 Dhaka Univ. J. Biol. Sci. 25(2): 149-159, 2016 (July)

BackgroundThe Tephritidae family, commonly referred to as true fruit flies, comprises of a substantial group within order Diptera. Numerous species within this family are major agricultural pests, with a tendency to infest a wide array of fruits and vegetables in tropical and sub- tropical regions, leading to considerable damage and consequent reductions in the market value of the crops.Methods and resultsThe current study was aimed to propose a promising solution to the menace posed by fruit flies by offering rapid, accurate and reliable species identification by using character-based DNA barcode methodology. The Tephritid specimens were collected from Cucurbitaceous plants of southern parts of West Bengal, India, and a total of eight species from Tephritidae family were obtained belonging to three genera, namely Bactrocera (Macquart, 1835), Dacus (Fabricius, 1805) and Zeugodacus (Hendel, 1927). Their morphological features were meticulously studied based on available literature, along with genetic analysis based on mitochondrial COI and ND1 gene sequences. A total of 30 uniquely variable sites at nucleotide position 42,48,51,60,66,72, 105,111,144,198,207,243, 273,297,307,318,345,357, 375,378,381,387,399,400, 402,436,444,450,453 and 460 in COI gene were discerned among Tephritid species in the present study.ConclusionsThe character-based DNA barcode holds the potential to differentiate closely related species of fruit flies and morphologically look-a-like ones. The novel method will be very significant in terms of rapid, precise and reliable species identification and might be extremely essential for early detection during pest outbreaks by facilitating timely intervention strategies to mitigate crop damage.

Dry fish is considered as a delicacy in the menu of many people of Bangladesh. The most economically important marine and freshwater dried fishes are ribbon fish, bombay duck, olive barb, pomfret, shrimp, etc. In this study, fresh ribbon fish sample was collected from Cox's Bazar and morphologically identified as ribbon fish. The collected fish was confirmed through a molecular technique using mitochondrial COI gene (cytochrome c oxidase subunit I) to avoid morphological ambiguity, which is first reported data from Bangladesh. Partial sequence of this COI gene was amplified using a genus specific universal primer set. The target nucleotide sequence of COI gene was determined successfully with 616 bp length and the ribbon fish was identified as Eupleurogrammus sp. Within the amplified region GC and AT content were 45.5% and 54.5%, respectively. 16.88% interspecific polymorphism was observed when compared with other species under genus Eupleurogrammus collected from Gen Bank database. This sequence will be used as molecular bar code for authentic identification of Eupleurogrammus sp. at genus level. Bangladesh J. Zool. 50 (2): 231-238, 2022

The leaf beetle, <TEX>$Chrysolina$</TEX> <TEX>$aurichalcea$</TEX> (Coleoptera: Chysomelidae), is a pest damaging plants of Compositae. In order to understand the genetic diversity and geographic variation we sequenced a portion of mitochondrial COI gene (658 bp) and complete nuclear internal transcribed spacer 2 (ITS2) of the species collected from seven Korean localities. A total of 17 haplotypes (CACOI01~CACOI17), with the maximum sequence divergence of 3.04% (20 bp) were obtained from COI gene sequence, whereas 16 sequence types (ITS2CA01~ITS2CA16), with the maximum sequence divergence of 2.013% (9 bp) were obtained from ITS2, indicating substantially larger sequence divergence in COI gene sequence. Phylogenetically, the COI gene provided two haplotype groups with a high nodal support (<TEX>${\geq}87%$</TEX>), whereas ITS2 provided only one sequence type group with a high nodal support (<TEX>${\geq}92%$</TEX>). The result of COI gene sequence may suggest the presence of historical biogeographic barriers that bolstered genetic subdivision in the species. Different grouping pattern between COI gene and ITS2 sequences were interpreted in terms of recent dispersal, reflected in the ITS2 sequence. Finding of unique haplotypes and sequence types only from Beakryeng-Islet population was interpreted as an intact remnant of ancient polymorphism. As more samples are analyzed using further hyper-variable marker, further fruitful inference on the geographic contour of the species might be available.

During April 2018, a sudden appearance of some unknown flies was observed in some villages under Sibsagar district of Assam, Northeast India. The flies attacked in groups and had more attraction towards human than livestock. The present article describes the entomological and molecular identification of the fly at species level in Sibsagar district of Assam, northeast India. Preliminary examination revealed the fly to be an insect of order Diptera, Family Tabanidae and Genus Chrysops as they posses short proboscis, ocelli, third antennal joint with five divisions and wings demarcated with dark median cross-band. This was supported by molecular data where the partial nucleotide sequences of Mitochondrial COI gene revealed maximum identity (90.6%-92.3%) with genus Chrysops. The mitochondrial COI sequence data of Chrysops flavocinctus has been made available in NCBI Gen Bank. Gen Bank Accession No. MH998226.

Low character variation among onychophoran species has been an obstacle for taxonomic and phylogenetic studies in the past, however we have identified a number of new and informative characters using morphological, molecular, and chromosomal techniques. Our analyses involved a detailed examination of Epiperipatus biolleyi from Costa Rica, Eoperipatus sp. from Thailand, and a new onychophoran species and genus from Costa Rica, Principapillatus hitoyensis gen. et sp. nov.. Scanning electron microscopy on embryos and specimens of varying age revealed novel morphological characters and character states, including the distribution of different receptor types along the antennae, the arrangement and form of papillae on the head, body and legs, the presence and shape of interpedal structures and fields of modified scales on the ventral body surface, the arrangement of lips around the mouth, the number, position and structure of crural tubercles and anal gland openings, and the presence and shape of embryonic foot projections. Karyotypic analyses revealed differences in the number and size of chromosomes among the species studied. The results of our phylogenetic analyses using mitochondrial COI and 12S rRNA gene sequences are in line with morphological and karyotype data. However, our data show a large number of unexplored, albeit informative, characters in the Peripatidae. We suggest that analysing these characters in additional species would help unravel species diversity and phylogeny in the Onychophora, and that inconsistencies among most diagnostic features used for the peripatid genera in the literature could be addressed by identifying a suite of characters common to all peripatids.