Cytochrome C Oxidase Subunit I Research Articles

A new species of genus Monoctonus (Hymenoptera, Braconidae, Aphidiinae) from South Korea

The genus Monoctonus Haliday, 1833 is a small group which consists of 24 species worldwide. In South Korea, Chang and Youn (1983) recorded one species, M. similis Starý & Schlinger, 1967, but the evidence for identification of this species is doubtful and further confirmation is required (personal communication with Prof. Jong-Cheol Paik). An additional Monoctonus species is recorded as new to science from South Korea. Descriptions and illustrations of the new species –Monoctonus koreanus sp. nov. – are provided, together with its mitochondrial cytochrome c oxidase subunit I (COI) data and phylogenetic position. A key to the female of the two species present in Korea is provided.

Molecular phylogeny and biogeography of the aquatic dance fly subfamily Clinocerinae (Diptera: Empididae)

AbstractThis study presents the first molecular phylogenetic analysis of the Clinocerinae, challenging the traditionally accepted monophyly of this subfamily. DNA was extracted from fresh and museum specimens representing all biogeographical regions. Maximum likelihood (ML) and Bayesian inference (BI) phylogenetic analyses were performed based on sequences from two mitochondrial genes, cytochrome c oxidase subunit I (COI) and cytochrome β, and three nuclear genes, carbomoylphosphate synthase domain of rudimentary, elongation factor‐1α and isocitrate dehydrogenase. Through molecular data and morphological examination, our results reveal a division within Clinocerinae, distinguishing ‘typical’ or Clinocerinae (s.s.) from several genera, specifically Afroclinocera Sinclair, Asymphyloptera Collin and Proagomyia Collin, possibly lending support for a reclassification of these genera outside Clinocerinae. Bergenstammia Mik is proposed as a junior synonym of Phaeobalia Mik, syn. n., and the following new combinations are recognized: Phaeobalia albanica (Wagner) comb. n., Phaeobalia aurinae (Pusch & Wagner) comb. n., Phaeobalia carniolica (Horvat) comb. n., Phaeobalia frigida (Vaillant) comb. n., Phaeobalia glacialis (Palaczyk & Słowińska) comb. n., Phaeobalia multiseta (Strobl) comb. n., Phaeobalia nudimana (Vaillant) comb. n., Phaeobalia nudipes (Loew) comb. n., Phaeobalia pulla (Vaillant & Wagner) comb. n., Phaeobalia pyrenaica (Vaillant & Vinçon) comb. n., Phaeobalia slovaca (Wagner) comb. n. and Phaeobalia thomasi (Vaillant & Vinçon) comb. n. Re‐evaluation of the genus Roederiodes resulted in the following new combinations: Clinocerella macedonicus (Wagner & Horvat) comb. n. and Clinocerella montenegrinus (Wagner & Horvat) comb. n. The origins of Clinocerinae (s.s.) are traced back to the Holarctic region, Laurasian origin, with a likely complex history of dispersal events into the Southern Hemisphere. Based on current knowledge, the greatest generic and species richness is confined to the Palaearctic Region. These findings provide valuable insights into the evolutionary relationships and distribution patterns of Clinocerinae (s.s.), challenging existing taxonomic classifications and shedding light on their historical biogeography.

Population Genetics of Culex tritaeniorhynchus (Diptera: Culicidae) in Türkiye.

Mosquitoes are important vectors of pathogens that can affect humans and animals. Culex tritaeniorhynchus is an important vector of arboviruses such as Japanese encephalitis virus, West Nile virus among various human and animal communities. These diseases are of major public health concern and can have huge economic and health burdens in prevalent countries. Although populations of this important mosquito species have been detected in the Mediterranean and Aegean regions of Türkiye; little is known about its population structure. Our study is to examine the population genetics and genetic composition of Cx. tritaeniorhynchus mosquitoes collected from several localities using cytochrome oxidase subunit I (COI) and the NADH dehydrogenase subunit 5 genes (ND5). This is the first extensive study of Cx. tritaeniorhynchus in the mainland Türkiye with sampling spanning many of provinces. In this study, DNA extraction, amplification of mitochondrial COI and ND5 genes and population genetic analyses were performed on ten geographic populations of Culex tritaeniorhynchus in the Aegean and Mediterranean region of Türkiye. Between 2019 and 2020, 96 samples were collected from 10 geographic populations in the Aegean and Mediterranean regions; they were molecularly analyzed and 139 sequences (50 sequence for COI and 89 sequence for ND5) were used to determine the population structure and genetic diversity. For ND5 gene region, the samples produced 24 haplotypes derived from 15 variable sites and for COI gene region, 43 haplotypes were derived from 17 variable sites. The haplotype for both gene regions was higher than nucleotide diversity. Haplotype phylogeny revealed two groups present in all populations. AMOVA test results show that the geographical populations were the same for all gene regions. Results suggest that Cx. tritaeniorhynchus is a native population in Türkiye, the species is progressing towards speciation and there is no genetic differentiation between provinces and regions. This study provides useful information on the molecular identifcation and genetic diversity of Cx. tritaeniorhynchus; these results are important to improve mosquito control programs.

Ngāokeoke Aotearoa: The Peripatoides Onychophora of New Zealand

(1) Background: Originally described as a single taxon, Peripatoides novaezealandiae (Hutton, 1876) are distributed across both main islands of New Zealand; the existence of multiple distinct lineages of live-bearing Onychophora across this spatial range has gradually emerged. Morphological conservatism obscured the true endemic diversity, and the inclusion of molecular tools has been instrumental in revealing these cryptic taxa. (2) Methods: Here, we review the diversity of the ovoviviparous Onychophora of New Zealand through a re-analysis of allozyme genotype data, mitochondrial DNA cytochrome oxidase subunit I sequences, geographic information and morphology. (3) Results: New analysis of the multilocus biallelic nuclear data using methods that do not require a priori assumptions of population assignment support at least six lineages of ovoviviparous Peripatoides in northern New Zealand, and mtDNA sequence variation is consistent with these divisions. Expansion of mitochondrial DNA sequence data, including representation of all existing taxa and additional populations extends our knowledge of the scale of sympatry among taxa and shows that three other lineages from southern South Island can be added to the Peripatoides list, and names are proposed here. In total, 10 species of Peripatoides can be recognised with current data.

A new parasitic barnacle (Crustacea, Cirripedia, Rhizocephala, Mycetomorpha) from the abyssal zone in the northwestern Pacific

I describe the parasitic barnacle Mycetomorpha abyssalissp. nov. from the crangonid shrimp Sclerocrangon zenkevitchi collected from 3893–3890 m depth off the eastern coast of Iwate, Japan, northwestern Pacific. This is the first Mycetomorpha rhizocephalan from the abyssal zone and the third species in Mycetomorpha. Mycetomorpha abyssalissp. nov. differs from its congeners M. vancouverensis and M. albatrossi in (1) triangular shield lacking, (2) stalk one-quarter of length from posterior end of externa, (3) mantle opening clearly anterior to stalk, (4) different host genus, and (5) depth range much deeper. I determined partial sequences for the mitochondrial cytochrome c oxidase subunit I (COI) and 16S rRNA genes and nuclear 18S rRNA and 28S rRNA genes from M. abyssalissp. nov. for future DNA barcoding and phylogeny. Kimura 2-parameter distances between M. abyssalissp. nov. and M. vancouverensis were 21.2% (16S), 0.6% (18S), and 1.5% (28S).

Haplotype diversity of Heterodera koreana (Tylenchida: Heteroderidae), affecting bamboo in Korea

AbstractIn a survey of plant-parasitic nematodes in agricultural fields, cyst-forming nematodes were found in soil planted bamboo in Korea. The aim of this study was to identify the cyst nematodes based on morphological and molecular characteristics. As the results, the morphology and morphometrics of cysts and second-stage juveniles (J2s) were consistent with those of previous descriptions of Heterodera koreana. In phylogenetic analyses based on DNA sequences, these cyst nematodes were clustered together with clade of H. koreana in internal transcribed spacer (ITS) region, and large subunit D2-D3 segments (LSU D2-D3). These nematodes were clustered together with clade of H. koreana in cytochrome c oxidase subunit I (COI) gene, but a haplotype was different when compared with previous reported haplotypes (haplotype A-C) in Japan. This study showed these cyst nematodes were identified as H. koreana, and a new haplotype of H. koreana is distributed in Korea. We suggest that the new haplotype of H. koreana name as haplotype D.

The six-spotted spider mite, Eotetranychus sexmaculatus (Riley) (Trombidiformes: Tetranychidae) in Australia, New Zealand, Japan and USA: a revised morphological and molecular-based concept, synonyms, and related species

Based on morphological and molecular evidence, Eotetranychus asiaticus Ehara is confirmed as a junior synonym of E. sexmaculatus (Riley). Eotetranychus queenslandicus Manson differs to E. sexmaculatus in some morphological characters states, and currently there is insufficient evidence to confirm if it is also a junior synonym. We therefore propose that it remains a valid species until fresh material can be collected from the type host (Codiaeum variegatum (L.) A.Juss.; Euphorbiaceae) in the type location (Freshwater, Queensland, Australia) for molecular analysis and further comparisons. Eotetranychus sexmaculatus is confirmed as present in Australia and New Zealand. Our analyses indicate that specimens identified as E. sexmaculatus from avocado (Persea americana Mill.; Lauraceae) in Western Australia and New Zealand, and specimens of E. asiaticus from the type host (Ficus erecta Thunb.; Moraceae) and type location (Tomigusuku, Okinawa, Japan) are conspecific with specimens of E. sexmaculatus recently collected from the type host (Citrus sp.; Rutaceae) and type location (Florida, USA), with 0–3.6% divergence in a fragment (451 bp) of cytochrome oxidase subunit I (COI), and 0–0.3% in the ribosomal internal transcribed spacer II (ITS2) gene. Herein we redefine E. sexmaculatus, providing a revised diagnosis, notes on leg chaetotaxy, and variation in the shape of the aedeagus—which is a keystone feature critical for determining species boundaries of spider mite species that have males. Our findings also reinforce the potential of the eupathidion on the palptarsus suζ (i.e. the spinneret) for species diagnoses.

Morphometrics and Truss-networking distances among three species of croaker (Johnius borneensis, Johnius argentatus and Johnius belangerii) from Bangladesh coast of the Bay of Bengal

Morphometric features are efficiently used for the differentiation of species and long-term management of fish populations. Truss analysis is a frequently used taxonomic method as well as a stock identification tool to dissimilitude the morphologically similar species. Using landmark-based truss networking techniques following 18 morphometric lengths, 10 meristic counts, and 23 truss-network distance, the morphological variation of three croaker species viz. Johnius borneensis, Johnius argentatus, and Johnius belangerii were examined from Bangladesh coast. Data were analyzed and visualized through univariate ANOVA, canonical discriminate function analysis (DFA), and principal component analysis (PCA). Results found that the first DFA was responsible for 80.6% & 81.1%, whereas the second DFA was responsible for 19.4% & 18.9% variations among three species for the morphometric and truss-distances, respectively. In addition, 14 out of 18 morphometric and 12 out of 23 truss-network measurements were significantly different (p < 0.05) among means of three species. 1st PCA found 70.76% & 64.49% explained variations and 2nd PCA displayed 12.8% & 11.5% explained variations respectively for morphometric and truss distances. The findings of the study demonstrates that J. borneensis and J. belangerii are highly morphologically similar and J. argentatus is completely distinct which might be due to their physiological and genetic variations. The present study might be helpful in differentiating species and managing existing stocks of marine fish. Further studies regarding the molecular characterization of the Cytochrome c oxidase subunit I (COI) gene could confirm the genetic variations among these species. Bangladesh J. Zool. 51(3): 233-251, 2023

Untangling the cephalopod market: authentication of seafood products in Greece with DNA-barcoding

Cephalopod products are very popular in Greece and are considered a delicacy. However, heavy processing and marketing practices interfere with visual inspection and impede morphological identification, allowing species substitution. Mislabeling of seafood products remains a worldwide issue despite existing labeling regulations at local and European level. For the detection of fraudulent products, a variety of identification methods have been developed, however DNA barcoding remains the most favored. This study aims to investigate the cephalopod species sold in the country's seafood market and assess their mislabeling rates. Two mitochondrial genes, the cytochrome c oxidase subunit I (COI) and the 16S ribosomal RNA (16S), were selected for the analyses. A total of 156 samples were collected from fishmongers, markets, and restaurants across four cities. Identification was successful in 93.58% of the samples and 59 discrepancies (40.41%) between the label and the identified species were recorded. However, in some cases, substitution might have been unintentional, caused by negligence, lack of detailed information on the label, and the overall low awareness of the legislation by retailers. High mislabeling rates were estimated, especially when compared against the average global substitution rate in seafood products but align with those reported by studies on cephalopod products. This study is the first to investigate mislabeling rates in cephalopod commodities in Greece. With the aim of a transparent seafood trade, our results highlight the need for monitoring of all seafood products available in the country's market.

Molecular identification and genetic diversity of ten Pierid butterflies based on mitochondrial COI gene

The IUCN Bangladesh has documented a total of 31 species belonging to the Pieridae family in Bangladesh. There is currently limited knowledge regarding the genetic diversity and molecular relationships among these butterflies. The present investigation involved the generation of mitochondrial COI (mtCOI) gene sequences for ten species of butterflies belonging to the Pierid family. After that, BLAST analysis showed 96% to 100% similarity, and then the sequences were submitted to NCBI’s GenBank. MEGA10 and BioEdit were used to identify a 584 bp COI gene fragment with 179 variable sites and 128 parsimony-informative sites. The COI sequences' average base compositions were 38.56% T, 17.12% C, 30.18% A, and 14.12% G. While a significant AT bias (68.74%) existed among the Pierid species. The genetic distance between ten Pierid species was assessed using the Kimura 2-Parameter (K2P) algorithm, and the nucleotide differences ranged from 0.00172% to 0.24261%. Afterwards, the Maximum Likelihood (ML) method was used to construct a phylogenetic tree using ten sequences of Pierid species. These species belongs to two subfamilies, Pierinae and Coliadinae in Bangladesh. On the tree, the Pierinae subfamily of Pieridae formed a paraphyletic arrangement whereas the Coliadinae subfamily was shown to be monophyletic. The data analyses support the following relationships between the two subfamilies as follows: [(Appias + Leptosia) + (Pieris + Appias) + (Delias) + (Eurema + Catopsilia) + (Pareronia)]. This supported the theory that Pierinae and Coliadinae are sister taxa. Nonetheless, Pareronia hippia remains outside the main Pierinae group, requiring further study to resolve this issue. Finally, these studies generated ten mtCOI gene sequences that have the potential to serve as valuable references for the accurate identification of Pierid species. In addition, this result could be used in the future to reveal the subfamily relationships within the Pieridae taxonomic classification. Bangladesh J. Zool. 51(3): 289-299, 2023  

Identification of fish specimens of the Tocantins River, Brazil, using DNA barcoding.

The fish fauna of the Tocantins River possesses many endemic species; however, it is little studied in molecular terms and is quite threatened by the construction of several hydroelectric dams. Therefore, the objective of this study was to identify the ichthyofauna of the Tocantins River using DNA barcoding. For this, collections were carried out in five points of this river, which resulted in the capture of 725 individuals from which partial sequences of the cytochrome oxidase subunit I (COI) gene were obtained for genetic analysis. A total of 443 haplotypes were recovered with the mean intraspecific K2P genetic distance of 1.82%. Altogether, 138 species were identified based on morphological criteria, which was a quantity that was much lower than that indicated by the four molecular methods (assemble species by automatic partitioning [ASAP], barcode index number [BIN], generalized mixed Yule coalescent (GMYC), and Bayesian Poisson tree processes [bPTP]) through which 152-157 molecular entities were identified. In all, 41 unique BINs were obtained based on the data generated in the BOLDSystems platform. According to the result indicated by ASAP (species delimitation approach considered the most appropriate in the present study), there was an increase of 17 molecular entities (12.32%), when compared to the number of species identified through their morphological criteria, as it can show cryptic diversity, candidates for new species, and misidentifications. There were 21 incongruities indicated between the different identification approaches for species. Therefore, it is suggested that these taxonomic problems be cautiously evaluated by experts to solve such taxonomic issues.

Haplotype Affiliation of Varroa destructor in Selected Locations from Poland and Ukraine

Abstract The virulence patterns observed in viruses transmitted by Varroa destructor implied a potential association of these honey bee pests in Poland with the Korean haplotype. Nevertheless, it is noteworthy that this hypothesis lacked direct verification. Therefore, the aim of the study was to determine the haplotype affiliation of V. destructor mites infesting bees in Poland and Ukraine. Adult females were collected from drone brood from honey bee colonies maintained in the apiary at the University of Life Sciences in Lublin, Poland, and the apiary in Synevyr National Park in Ukraine. The haplotype affiliation was determined through the sequencing of the cytochrome oxidase subunit I gene (CO I; 929 bp) fragment. Based on the phylogenetic comparison with 84% bootstrap support, the V. destructor females from both locations were confirmed to represent three haplotypes: Korean 1 (AmK1-1), Korean 2 (AmK1-2), and Chinese 4 (AmK1-4). They are the most virulent types of this parasite worldwide and in Poland. Concurrently, the present study confirms that the Synevyr National Park in Ukraine is isolated from alien bee subspecies, but is not free from alien parasites.

Population genetic structure of bottlenose and whitespotted wedgefishes from the Southwest Indian Ocean using a dual marker approach

Wedgefishes (Rhinidae) are threatened by unsustainable fishing globally, and especially in the Southwest Indian Ocean (SWIO), due to their high-value fins in the shark trade. The whitespotted wedgefish Rhynchobatus djiddensis and the bottlenose wedgefish R. australiae are both classified as Critically Endangered on the IUCN Red List of Threatened Species, yet a lack of species-specific knowledge and taxonomic uncertainty still exists within this genus. Genetic approaches aid in taxonomic classification and identifying distinct populations for targeted conservation. Morphological specimen identification of samples (n = 189) collected across the SWIO was confirmed based on the cytochrome oxidase c subunit I (COI) and/or nicotinamide adenine dehydrogenase subunit 2 (ND2) gene regions. The genetic diversity and population structure within and between species and sampling locations were investigated using a dual marker approach: (1) 2 concatenated mitochondrial gene regions, namely COI and the control region (n = 117), and (2) 9 nuclear microsatellite markers (n = 146). The overall genetic diversity was moderate, with an indication that different evolutionary forces are at play on a mitochondrial versus nuclear level. The 2 species were delineated based on both marker types, and for R. djiddensis, the sampling locations of South Africa and Mozambique were genetically homogeneous. For R. australiae, significant differentiation was found between sampling locations, with Madagascar and Tanzania being genetically the most similar. This information provides critical insights into the distribution range and population structure of the whitespotted wedgefish species complex that can support the sustainable management of wedgefishes.

First Report of Phytophthora ramorum Causing Leaf Spots on Cornus capitata (Evergreen Dogwood) in United States.

Cornus capitata Wall. ex Roxb. (evergreen dogwood) is a bushy evergreen tree or shrub native to East Asia grown for its showy creamy bracts in late spring followed by attractive red fruit. In Feb 2023, a sample of foliage with leaf spots and tip dieback from C. capitata 'Mountain Moon' was submitted from a Humboldt Co. nursery as part of a CDFA inspection program for Phytophthora ramorum. The leaf spots were medium to dark brown, irregularly shaped, and ranged from 5 to 8 mm in diameter. They were located primarily along the leaf midrib and covered up to 1/4 of the leaf surface. Six 6-mm-diameter leaf discs taken from the margins of brown lesions and tip dieback were plated on Phytophthora selective CMA-PARP (PARP) media (Jeffers and Martin 1986). After 6 to 10 days, colonies resembling P. ramorum, with coralloid coenocytic hyphae, chlamydospores, ellipsoidal semi-papillate and caducous sporangia, and a relatively slow growth rate were recovered. Abundant sporangia formed on agar singly or in clusters on sympodially branched sporangiophores (n = 50), varying in size from 35 to 60 μm × 20 to 30 μm (mean 45.6 × 24.8 μm) with a length/breadth ratio ranging from 1.3 to 2.3 (mean 1.8). Chlamydospores (n = 50) ranged from 35 to 62 μm in diameter (mean 51.9 μm) on 14-day-old PARP cultures. The internal transcribed spacer region (ITS) using primers ITS5/ITS4 (White et al. 1990; accession no. OR636225) and cytochrome oxidase subunit 1 region (cox1) using primers OomCox1Levup/Fm85mod (Robideau et al. 2011; OR635665) of one isolate (0254-32A) were amplified and sequenced. BLAST analysis showed 100% identity of both regions to P. ramorum ex-type strain (MG865581 and MH136973). Microsatellite loci placed the P. ramorum isolate in the NA2 clonal lineage (Goss et al, 2011). Pathogenicity of P. ramorum isolate 0254-32A was tested using five C. capitata plants (2.5-year-old, 28-cm-tall, 3.78-liter pot). Zoospore inoculum was produced as described in Blomquist et al. (2021). Above ground parts of each plant were sprayed with inoculum (15 ml, 1.3 × 105 zoospores/ml). Inoculated plants were incubated in a dew chamber in the dark at 23°C for 72 h and then placed in a 23±1°C growth chamber with a 12-h photoperiod. Five control plants were treated as above but with sterile water instead of the zoospore suspension. Two days after inoculation, brown spots were visible on leaves on all inoculated plants, initiating from where the drops of inoculum had persisted. After 3 days, brown lesions, from water drop- to majority of entire leaf-sized, were observed on approximately 75% of inoculated leaves. After 6 days, lesions expanded to the edges of leaves, causing leaf curling and defoliation. Lesions stopped expanding after 3 weeks, and by 4 weeks, most infected leaves had abscised, with no new infections observed. Phytophthora ramorum was consistently isolated from foliar lesions of inoculated plants on PARP. It was not isolated from leaf or stem tissues of control plants, which remained asymptomatic during the 4-week experiment period. Phytophthora ramorum was detected on C. capitata in the UK in 2015 (DEFRA 2015). To our knowledge, this is the first report of P. ramorum infecting C. capitata in the United States and the completion of Koch's postulates on any Cornus spp. Incidence on C. capitata in the California nursery was low. However, their proximity to other infected foliar hosts suggests Cornus spp. may present a potential risk for the spread of P. ramorum.

Redescription of Apanteles mimoristae (Hymenoptera, Braconidae), a parasitoid of the native pyralid cactus moth Melitara cf. nephelepasa in central Mexico

Novel trophic associations have sometimes resulted in fortuitous and significant biological control. After the invasion of North America by the South American cactus moth, Cactoblastis cactorum (Berg) (Pyralidae: Phycitinae), it is pertinent to characterize the assemblage of local natural enemies that could utilize this moth in new host-parasitoid associations. Herein we report on Apanteles mimoristae Muesebeck (Braconidae: Microgastrinae), a North American gregarious endoparasitoid wasp attacking the caterpillar of the phycitine cactus moth Melitara cf. nephelepasa (Dyar) (Pyralidae: Phycitinae, also known as zebra worm), also native to North America; both collected in Opuntia ficus-indica (L.) Mill. (Cactaceae) cultivated fields at rural areas of Mexico City. We provide an updated morphological account for A. mimoristae visualized with light microscopy and scanning electron microscope (SEM); a fragment of its cytochrome oxidase subunit I (COI) gene sequence data is reported for the first time. Additionally, we analyze its taxonomical position relative to other Apanteles species from the Americas including those attacking cactus-feeding moths. Our analyses place A. mimoristae (from Mexico) in a clade with A. esthercentenoae Fernández-Triana (from Costa Rica), a parasitoid of both Cromarcha stroudagnesia Solis (Pyralidae) and Palpita venatalis (Schaus) (Crambidae) (non cactus-feeding), and in a sister clade to A. opuntiarum Martínez & Berta (from Argentina) and A. alexanderi Brèthes (from Argentina and Uruguay), parasitoids of the cactus-feeding phycitines Cactoblastis and Tucumania respectively. Finally, we provide an updated key for the identification of Apanteles species recorded parasitizing cactus moth caterpillars in the American continent.

Violins we see, species we don’t… Species delimitation of the spider genus Loxosceles Heineken & Lowe (Araneae: Sicariidae) from North America using morphological and molecular evidence

In modern systematics, different sources of evidence are commonly used for the discovery, identification, and delimitation of species, especially when morphology fails to delineate between species or in underestimated species complexes or cryptic species. In this study, morphological data and two DNA barcoding markers—cytochrome c oxidase subunit I (COI) and internal transcribed spacer 2 (ITS2)—were used to delimit species in the spider genus Loxosceles from North America. The molecular species delimitation analyses were carried out using three different methods under the corrected p-distance Neighbor-Joining (NJ) criteria: 1) Assemble Species by Automatic Partitioning (ASAP), 2) General Mixed Yule Coalescent model (GMYC), and 3) Bayesian Poisson Tree Processes (bPTP). The analyses incorporated 192 terminals corresponding to 43 putative species of Loxosceles, of which 15 are newly recognized herein, as putative new species, based on morphology and congruence between molecular methods with COI. The average intraspecific genetic distance (p-distance) was <2%, whereas the average interspecific genetic distance was 15.6%. The GMYC and bPTP molecular methods recovered 65-79 and 69 species respectively, overestimating the diversity in comparison with morphology, whereas the ASAP method delimited 60 species. The morphology of primary sexual structures (males palps and female seminal receptacles) was congruent with most of the molecular methods mainly with COI, showing that they are robust characters for identification at the species level. For species delimitation COI was more informative than ITS2. The diversity of Loxosceles species is still underestimated for North America, particularly in Mexico which holds the highest diversity of this genus worldwide.  

Confirmation of the valid specific status of Dolichovespulakuami Kim & Yoon, 1996 (Hymenoptera, Vespidae) based on molecular and morphological evidence.

The taxonomic validity of Dolichovespulakuami, especially in relation to D.flora, has been the subject of a long-term debate. Herein, the valid specific status of the former was supported through an integrated analysis of morphological characters and DNA barcodes. The pronotal rugae and male genitalia of the two species are different, and partial mitochondrial genes (cytochrome oxidase subunit I, COI) indicate that they form significantly distinct lineages. The hitherto unknown male of D.kuami is described for the first time, and a brief discussion of the D.maculata species group is provided.

Abstract 2783: The exosomes from cord blood stem cells, containing novel miRNAs, induce apoptosis in melanoma cells and enhance T cell anticancer function

Abstract Regulatory T (Treg) cells expressing FOXP3 play a critical role in suppressing immune responses against anti-tumor immune responses. However, their presence in tumor tissues is often associated with poor prognosis. Cord blood stem cell-derived exosome vesicles (CBSC-EV) constitute a valuable source of lipids, microRNAs, and proteins, that are essential for cell-to-cell communication and provide therapeutic benefits. This study aimed to investigate the contents of CBSC-EV, identifying several known miRNAs (from Let7 and Let5 families) and two unknown miRNAs. Healthy lymphocytes, fibroblasts, and CHL1 amelanotic melanoma cell lines were transfected with these two novel miRNA inhibitors, and the changes before and after treatment compared to CBSC-EV and H2O2 were examined. Following treatment, the cells underwent a 24-hour incubation period, during which the level of cytotoxicity was assessed using CCK8, and the degree of DNA damage incurred was quantified with the Fast Microplate DNA damage assay. To further investigate the effects of treatment, RNA sequencing was conducted on both healthy and cancerous cells. The results demonstrated that CBSC-EV and novel miRNAs induced cytotoxicity in cancer cells, while in healthy cells, the DNA damage was repaired and cell viability was significantly increased (p<0.001). RNA sequencing revealed that following treatment with CBSC-EV and both CBSC-EV+N1 and N2 inhibitors in malignant melanoma cells, the expression of genes involved in the cytokine-mediated signaling pathway, cytokine-cytokine interaction, and cytokine activity were enhanced. In healthy cells treated with CBSC-EV and novel miRNA inhibitor transfection, the MT-CO1 gene was upregulated, contributing to cytochrome-c oxidase activity. This study presents evidence of the potential use of novel miRNAs as genetic material for cancer therapeutics in the future. Additionally, the findings suggest that these miRNAs could be used as an immune system modulator against melanoma. These findings demonstrate that miRNAs can effectively suppress resistance to anticancer cytotoxic therapy, which is a common feature of cancer cells. This suggests that miRNAs can potentially enhance current cancer therapies. Citation Format: Mojgan Mojgan Najafzadeh, Adi Baumgartner, Shohreh Jafarinejad, Zahra Karimi, Mohammad Isreb, Pouria Akhbari, Nader Ghaderi, Farshid Sefat, Saeed Heidari Keshel, Parisa Naeem, Rojan Ghaderi, Jacobo Elis Gomez, Diana Anderson, Andrew Wright. The exosomes from cord blood stem cells, containing novel miRNAs, induce apoptosis in melanoma cells and enhance T cell anticancer function [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2783.

Unraveling the Life Cycle of Nyssopsora cedrelae: A Study of Rust Diseases on Aralia elata and Toona sinensis

Rust disease poses a major threat to global agriculture and forestry. It is caused by types of Pucciniales, which often require alternate hosts for their life cycles. Nyssopsora cedrelae was previously identified as a rust pathogen on Toona sinensis in East and Southeast Asia. Although this species had been reported to be autoecious, completing its life cycle solely on T. sinensis, we hypothesized that it has a heteroecious life cycle, requiring an alternate host, since the spermogonial and aecial stages on Aralia elata, a plant native to East Asia, are frequently observed around the same area where N. cedrelae causes rust disease on T. sinensis. Upon collecting rust samples from both A. elata and T. sinensis, we confirmed that the rust species from both tree species exhibited matching internal transcribed spacer (ITS), large subunit (LSU) rDNA, and cytochrome oxidase subunit III (CO3) mtDNA sequences. Through cross-inoculations, we verified that aeciospores from A. elata produced a uredinial stage on T. sinensis. This study is the first report to clarify A. elata as an alternate host for N. cedrelae, thus providing initial evidence that the Nyssopsora species exhibits a heteroecious life cycle.

Endurance training increases mitochondrial myoglobin and enhances its interaction with complex IV in rat plantaris muscle.

Endurance exercise training is known to increase mitochondrial respiration in skeletal muscle. However, the molecular mechanisms behind this are not fully understood. Myoglobin (Mb) is a member of the globin family, which is highly expressed in skeletal and cardiac muscles. We recently found that Mb localizes inside mitochondria in skeletal muscle and interacts with cytochrome c oxidase subunit IV (COXIV), a subunit of mitochondrial complex IV, which regulates respiration by augmenting complex IV activity. In the present study, we investigated the effect of endurance training on Mb-COXIV interaction within mitochondria in rat skeletal muscle. Eight-week-old male Wistar rats were subjected to 6-week treadmill running training. Forty-eight hours after the last training session, the plantaris muscle was removed under anesthesia and used for biochemical analysis. The endurance training increased mitochondrial content in the skeletal muscle. It also augmented complex IV-dependent oxygen consumption and complex IV activity in isolated mitochondria from skeletal muscle. Furthermore, endurance training increased Mb expression at the whole muscle level. Importantly, mitochondrial Mb content and Mb-COXIV binding were increased by endurance training. These findings suggest that an increase in mitochondrial Mb and the concomitant enhancement of Mb interaction with COXIV may contribute to the endurance training-induced upregulation of mitochondrial respiration by augmenting complex IV activity.