Cytochrome Oxidase I Research Articles

Description of a new species of Hobbsinella (Crustacea, Bathynellacea, Bathynellidae) from Colorado (USA) based on morphological and molecular characters

A new species of Bathynellidae is described from Colorado (USA). Hobbsinella gunnisonensis Camacho & Taylor sp. nov. displays a unique combination of morphological characters including seven-segmented antenna lacking medial seta on exopod, antennule slightly longer than antenna, three-segmented mandibular palp, four articles on endopod of thoracopods I to VII and five spines on sympod and three spines on endopod of the uropods. Partial sequences of cytochrome oxidase I (COI) and 18S have been obtained from several specimens of the new species. The mitocondrial and nuclear DNA data complement the traditional morphological taxonomic description support the validity of the new species. Molecular data for the Bathynellidae demonstrate the presence of two highly divergent genetic units, with the new species placed in the genus Hobbsinella. With the description of Hobbsinella gunnisonensis Camacho & Taylor sp. nov. and its molecular characterization, we discovered an interesting distribution of the genus, which occurs in both sides of the Continental Divide (Texas and Colorado) and different habitats.

Efficacy of Molecular Identification Using DNA Barcoding Technique of Freshwater Edible Molluscs from Nagaland, India

Freshwater molluscs are widely utilized for food and medicine in many regions of the world. One such area in India is the northeast, where tribal and economically underdeveloped communities consume freshwater molluscs. Because they are an inexpensive source of protein and offer food security, livelihoods, and medicine, these molluscs are in great demand. In the present work, molecular identification and documentation of the edible gastropods of Nagaland is done. Throughout the twentieth century, identification was based on morphology and anatomy. However, the identification based on morphological structure is challenging due to the existence of cryptic species that shows similar morphology which can lead to erroneous identification of species. Freshwater molluscs shells are highly varied and exhibit convergence evolution and hence may not always accurately indicate systematic boundaries. Thus, DNA bar-coding is a potential method for taxonomic identification process that can mitigate this problem. It uses genetic markers such as cytochrome oxidase I (COI) gene, 16S rDNA and 18S rDNA. In the present study COI mitochondrial gene is used. A fragment of COI gene is sequenced and their DNA is compared to the existing sequences in the genomic library. The sequence is then submitted in NCBI to obtain accession number. A total of 7 species of edible freshwater molluscs belonging to two class bivalvia and gastropoda was collected from local fishery ponds, confined water bodies, rivers, streams, and were identified. The identified species with their accession number is as follows, Lamellidens marginalis (ON209524), Parreysia corrugata (OM074301), Indonaia subclatharata (OM075118), Indonaia caerulea (OM076974), Bellamaya bengalensis (OM056888), Paludomus sp (OM078498), Brotia costula (OM056887). The study provides substantial support for COI's effectiveness as a prime marker for DNA barcoding of Indian edible freshwater gastropods and bivalves.

Phylogenetic and biogeographic history of brook lampreys (Lampetra: Petromyzontidae) in the river basins of the Adriatic Sea based on DNA barcode data.

The Adriatic brook lamprey, Lampetra zanandreai Vladykov 1955, was described from northeastern Italy. Its distribution is thought to include left tributaries of the River Po and the river basins of the Adriatic Sea from the River Po to the River Isonzo/Soča in Italy, Switzerland and Slovenia. It also shows a geographically isolated distribution in the Potenza River on the Adriatic slope in Central Italy. Lampetra from the Neretva River system in Croatia and Bosnia and Herzegovina and the Morača River system in Montenegro that were previously identified as L. zanandreai were recently described as a new species Lampetra soljani Tutman, Freyhof, Dulčić, Glamuzina & Geiger 2017 based on morphological data and a genetic distance between the two species of roughly 2.5% in the DNA barcoding gene cytochrome oxidase I (COI). Since DNA barcodes for L. zanandreai are only available for one population from the upper Po River in northwestern Italy, we generated additional COI nucleotide sequence data of this species from Switzerland, northeastern and central Italy comprising near topotypic material and obtained GenBank sequences of the species from Slovenia to better assess the evolutionary history of the two brook lamprey species in the river basins of the Adriatic Sea. Our data show a low sequence divergence of <1% between L. zanandreai from Switzerland, northeastern and central Italy and Slovenia and the Balkan species L. soljani. However, members of the population previously identified as 'L. zanandreai' from northwest Italy are genetically highly divergent from those of L. zanandreai and likely belong to an undescribed species, L. sp. 'upper Po'. The presence of a unique and highly divergent brook lamprey lineage in the upper Po River suggests that L. zanandreai and Lampetra sp. 'upper Po' may have evolved in separate paleo drainages during the formation of the modern Po Valley subsequent to marine inundations in the Pliocene.

Microsatellite and mitochondrial DNA based evidence reveals a single horseshoe crab (Limulus polyphemus) population on the northern Yucatan Peninsula, Mexico

In Mexico, the American horseshoe crab, Limulus polyphemus, is regarded as in danger of extinction, with a distribution range limited to the Yucatan Peninsula. Population genetics knowledge of this species in Mexico is incipient. We conducted a base-line genetic survey of eight localities on the northern coast of the Yucatan Peninsula to assess patterns of genetic structure, genetic variation, and gene flow among these localities. We analyzed DNA samples from 190 L. polyphemus specimens using 14 nuclear-encoded microsatellites and a region of the mitochondrial Cytochrome Oxidase I (COI) gene. Total allelic richness was 5.432, similar to previous figures for populations at the northern and southern extremes of the species’ global distribution. Observed heterozygosities (Ho) were 0.540–0.590, while expected heterozygosities (He) were 0.479–0.532, and there was no evidence of population bottlenecks. We found 129 mtDNA haplotypes (121 of which were found in single individuals) and 275 polymorphic sites, and the mean haplotype diversity among sites was high (0.852 ± 0.027 SD). AMOVAs based on both microsatellite and mtDNA data revealed that L. polyphemus from the northern Yucatan Peninsula show low genetic structure (Fst < 0.05, p > 0.05) and high gene flow (Nm = 10.967) favored by the absence of geographic barriers between coastal lagoons, suggesting that they constitute a single population. This was further confirmed by discriminant analyses of principal components using microsatellite data. This information is useful in establishing coordinated conservation efforts among federal and state protected areas.

Alaskacladius gen. nov., (Diptera: Chironomidae), a unique new orthoclad from Alaska.

We describe a new genus Alaskacladius gen. nov., based on the adult stages collected from Alaska, USA, and British Columbia, Canada. Molecular and morphological assessment of adult specimens supports the presence of a new genus. Alaskacladius gen. nov., is related to the genera Doithrix Sther & Sublette, 1983; Georthocladius Strenzke, 1941; Parachaetocladius Wlker, 1959; and Pseudorthocladius Goetghebuer, 1932. Based on the molecular analysis result and intergeneric K2P distance obtained from Cytochrome Oxidase I (COI) genes, Alaskacladius is closest and forms a sister group with Doithrix.

Genetic variation and population structure of brachiopods, Lingula anatina Lamarck, 1801 in the Northern Aceh shore, Indonesia

Abstract. Octavina C, Ramadhaniaty M, Daulay RE, Dewiyanti I, Ulfah M. 2023. Genetic variation and population structure of brachiopods, Lingula anatina Lamarck, 1801 in the Northern Aceh shore. Biodiversitas 24: 3951-3959. Lingula anatina is one of the brachiopods recognized as one of the most primitive groups with a fossil record from the early Cambrian period. Although L. anatina has been targeted as an object of biological studies, the information about the genetic variation within species is still limited. The purpose of this research is to identify the genetic variation and population structure of the species L. anatina found in the Aceh northern shore based on the mtDNA Cytochrome Oxidase I (COI) gene sequence. In this study, L. anatina specimens collected from five populations on Aceh's northern shore, namely Krueng Raya, Syiah Kuala, Ulee Lheue, and Ujong Pancu, with complete mitochondrial genome sequences were analyzed and compared with genebank sequences. The total nucleotide base of L. anatina is 657 bp, with 18 haplotypes. Based on the results of a comparison of the distribution of L. anatina in Asia shows that the Japanese population has the highest population structure value compared to the other five populations. The Fst value obtained from this study is 0.7, so it is classified as high and structured. Phylogenetic analysis showed that the correlation between geographic location and geographic distance affects the genetic distance between populations.

New Faunal Data for Black Flies (Diptera: Simuliidae), with the Evidence of COI Sequences, from Mediterranean Region of Türkiye

The present study provides faunal data for Turkish Black flies (Simuliidae, Diptera) from the Central and Western Mediterranean Region in Türkiye. Among the specimens collected from 221 different running water sites, 17 species in 3 genera and 5 subgenera were identified. Two species, Simulium (Nevermannia) ibleum (Rivosecchi 1966) and Simulium (Nevermannia) brevidens (Rubtsov 1956), were recorded for the first time from Türkiye, and 5 additional species were reported from the study area for the first time. The morphological identifications were tested by phylogenetic analyzes using mitochondrial cytochrome oxidase I (COI) barcode sequences. The COI analysis results overlapped with the morphotaxonomic identification results for eight of the 15 species. The first genetic data of 4 species (for World) and 5 species (from Türkiye) were stored in GenBank (NCBI).

Türkiye’de İstalacı bir tür olan Nysius cymoides (Spinola, 1837) (Hemiptera: Lygaeidae)’in Moleküler Tanımlaması

The false chinch bug, Nysius cymoides (Spinola, 1837) (Hemiptera: Lygaeidae), is an important pest with a wide host range. It was first discovered in Italy but now causing agricultural damage worldwide, including Turkey. Specimens of Nysius cymoides were collected from surveys of the sunflower and corn fields in Malkara-Keşan province, in Edirne in July 2020. Because of the population increase of the pest and their damages to sunflowers and corn fields, the pest identification have become an important issue. We investigated molecular characterization and identification of the pest by using mitochondrial cytochrome oxidase I (COI) barcode region. Adult and late immature stage of the specimens were examined under a stereozoom microscope and morphologically identified as well. The partial COI gene (659 bp) was amplified by using LCO1490/ HCO 2198 and PCR products were sequenced directly for molecular characterization. The sequence was registered GenBank database with accession number OL989232. Molecular analyzes were performed using MEGA X software. As a result, the nucleotide frequencies were A = 33.81%, T/U = 35.79%, C = 16.03%, and G = 14.37%. The distance-based species limitation method applied by using ASAP and ABGD, identified 5 species in the partition with the lowest ASAP score (1.5). The barcode gap distance of this partition with the best ASAP score was determined as 1.7%. The threshold distance was determined as 1.62%. The highest nucleotide identity of the studied specimen (OL989232) was detected as 99.8% with the specimen of Nysius cymoides from France (KJ541649.1) in GenBank. The objective of this study was the first attempt to identify Nysius cymoides specimens collected in Edirne based on DNA barcode regions.

Genetic diversity and population structure of Capitulum mitella (Linnaeus, 1767) in Fujian (China) revealed by mtDNA COI sequences.

Background: Capitulum mitella is a widely distributed and ecologically important stalked barnacle that settles extensively on rocky shores. This species contributes to the structural complexity of intertidal habitats and plays a critical role in the marine ecosystem. This study aimed to reveal the genetic diversity and population structure of C. mitella by analyzing the mitochondrial cytochrome oxidase I (COI) gene. Methods: A 683bp fragment of the COI gene was sequenced from 390 individuals sampled from six localities in Fujian, China. Results: A total of 84 distinct haplotypes were identified through the analysis of 82 polymorphic sites, resulting in an average haplotype diversity (h) of 0.660 and nucleotide diversity (π) of 0.00182. Analysis of molecular variance (AMOVA) and pairwise F ST statistics showed no significant population structure. Neutrality tests and mismatch distributions provided evidence of recent population expansion for the species. Conclusions: We suggest that the species' high dispersal ability, and ocean currents coupled with limited physical barriers in the region, contribute to its current phylogeographic structure. These findings enhance our comprehension of the genetic diversity and population structure of C. mitella, providing valuable insights for future conservation efforts.

DNA Barcodes for thrips species and development of multiplex real-time PCR assay for Frankliniella occidentalis Pergande, Frankliniella panamensis Hood, Thrips palmi Karny and Thrips tabaci Lindeman (Thysanoptera: Thripidae)

ABSTRACT Thrips (Order Thysanoptera) species are agriculturally important pests and vectors of plant disease. These tiny insects are commonly found in all life stages on imported commodities at the New Zealand border. Morphological identification of thrips is able to be performed on adults, but the identification keys for immature stages are inadequate and so DNA barcoding is regularly used for their identification. Here, we have generated cytochrome oxidase I (COI) DNA barcode data for 29 thrips species from 124 individuals, focusing on Frankliniella occidentalis, a dominant species intercepted at New Zealand border, followed by F. panamensis, Thrips palmi and T. tabaci. In addition, a multiplex real-time PCR assay has been developed to target the four thrips species. This assay is intended to facilitate the identification of quarantine interceptions with greater accuracy and faster diagnostic turnaround times, regardless of the developmental stages of the thrips. The developed assay demonstrated a high level of specificity for all the four target species and could detect as few as 10 copies/µL of the target DNA. Linear responses and high correlation coefficients between the amount of DNA and Cq values for each species were also achieved. The method was successfully tested on single egg, larva and adult samples and proved to be applicable for all life stages of the four species. Overall, this study has shown that the developed assay is an effective biosecurity tool for rapid and reliable identification of the target thrips species.

Optimizing a rapid LAMP assay for discrimination of Drosophila suzukii (Diptera: Drosophilidae) from common drosophilids captured in monitoring traps from the Midwest, United States.

Spotted-wing Drosophila, Drosophila suzukii, is an economically important pest of small fruits worldwide. Currently, the timing of management strategies relies on detection of adult flies captured in baited monitoring traps; however, identifying D. suzukii in trap catch based on morphology can be challenging for growers. DNA-based diagnostic methods such as loop-mediated isothermal amplification (LAMP) have the potential to improve D. suzukii detection. This study evaluated a LAMP assay as a diagnostic tool to discriminate between D. suzukii and closely related drosophilid species found commonly in monitoring traps in the Midwestern United States. Targeting the mitochondrial cytochrome oxidase I (COI) gene, we found the LAMP assay accurately detected D. suzukii with as little as 0.1 ng/μl of DNA at 63 °C for 50 min. Under these optimal incubation conditions, D. suzukii could be discriminated from D. affinis and D. simulans consistently, when specimens collected from liquid monitoring traps were tested independently. Compared to other DNA-based diagnostic tools for D. suzukii, LAMP offers unique benefits: DNA extraction is not required, testing occurs at one temperature in less than 1 h, and positive results are visible as a colorimetric change from pink to yellow. The LAMP assay for D. suzukii can reduce reliance on morphological identification, enhance the adoption of monitoring tools, and improve accuracy of detection. Further optimization can be conducted to evaluate the accuracy and sensitivity of results when a mixture of DNA from both D. suzukii and congener flies are tested in a single LAMP reaction.

Molecular characterization of Fasciola hepatica in endemic regions of Colombia.

Fasciola hepatica is a zoonotic trematode that affects a wide range of hosts, including cattle, sheep, and goats. The economic impact of the parasite on the cattle industry is significant, with high losses reported worldwide. While its impact on human health was previously underestimated, recent years have seen a rise in fascioliasis cases, leading to increased interest among researchers globally. To characterize the genetic diversity and intraspecific variation of this parasite in South America, specifically in Colombia, we collected 105 adult parasites from cattle bile ducts in seven Colombian departments (Antioquia, Boyacá, Santander, Cauca, Cundinamarca, Nariño, Norte de Santander, and Santander) to assess the parasite's phenotypic analyses, genetic diversity, and population structure. A computer image analysis system (CIAS) was applied based on standardized morphological measurements. Liver-fluke size was studied by principal component analysis (PCA). DNA sequences were obtained for nuclear markers such as the 28S, β-tubulin 3, ITS1, ITS2, and the mitochondrial marker Cytochrome Oxidase I (COI). Multiple statistical tests were performed, and the parasite's population structure was analyzed. Maximum Likelihood (ML) phylogenetic reconstructions were carried out using the sequences obtained herein and sequences available in GenBank. Morphological results revealed that all the obtained individuals matched F. hepatica's morphology. There was no evidence of high genetic diversity, and the absence of genetic structure at the country-level was notable, possibly caused by a demographic expansion of this trematode in Colombia or the low resolution of the molecular markers employed. Future studies are still needed to unveil the genetic population structure of F. hepatica across the country.

A temporal and spatial study of genetic structure in four species of bladed Bangiales (Rhodophyta) from the southeastern Pacific coast of Chile.

The coastline is a heterogeneous and highly dynamic environment influenced by abiotic and biotic variables affecting the temporal stability of genetic diversity and structure of marine organisms. The aim of this study was to determine how much the genetic structure of four species of marine Bangiales vary in time and space. Partial sequences of the cytochrome oxidase I (COI) gene obtained from two Pyropia (Py. sp. CHJ and Py. orbicularis) and two Porphyra (P. mumfordii and P. sp. FIH) species were used to compare the effect of the 40° S/41° S biogeographic break (spatial-regional scale) and the one of the Valdivia River discharges (spatial-local scale) and determine their temporal stability. Four seasonal samplings were taken during 1 year at five sites, one site located in Melinka (Magallanes province) and four sites along the coast of Valdivia (Intermediate area), on both sides of the river mouth. Results showed a strong genetic spatial structure at regional scale (ΦST > 0.4) in Py. sp. CHJ, Py. orbicularis, and P. mumfordii, congruent with the 41° S/42° S biogeographic break. A potential barrier to gene flow, related to the Valdivia River discharge, was detected only in P. mumfordii. In P. sp. FIH, spatial genetic structure was not detected at any scale. The genetic structure of all four species is stable throughout the year. The potential effect of main currents and river discharge in limiting the transport of Bangiales spores are discussed. We propose that both a restricted propagule dispersal and the formation potential for persistent banks of microscopic stages could lead to a temporally stable spatial partitioning of genetic variation in bladed Bangiales.

Accidental discovery of a Tetraodontidae (&lt;i&gt;Sphoeroides marmoratus&lt;/i&gt;) within a cuttlefish (&lt;i&gt;Sepia officinalis&lt;/i&gt;) bought in a fish shop in Italy: risk assessment associated with the presence of Tetrodotoxin

The discovery of a pufferfish specimen (Tetraodontidae) inside a frozen cuttlefish, purchased by a fishmonger, and caught in the central-east Atlantic (FAO 34) is reported. The consumer, who reported this case to FishLab (Department of Veterinary Sciences, University of Pisa) for investigation, was a student of Veterinary Medicine at the University of Pisa. He recognized the Tetraodontidae because he attended practical lessons on fish morphological identification during the course of food inspection and was aware of the risks to human health linked to the Tetrodotoxin (TTX). In this study, the pufferfish was identified morphologically, using the FAO morphological keys, and molecularly, analyzing two markers, the cytochrome oxidase I (COI) and the cytochrome b genes, by DNA barcoding. The pufferfish was identified morphologically as Sphoeroides spp., and molecularly as Sphoeroides marmoratus using the COI gene (99-100% identity values). Literature reports that S. marmoratus from the eastern Atlantic contains high concentrations of TTX in the gonads and the digestive tract. However, the possible passage of TTX from fish to other organisms linked to contact or ingestion has never been reported. This represents the first case of a potentially toxic pufferfish entering the market inside another organism. The fact that a student observed this occurrence highlights the key role of citizen science in the management of emerging risks.

DNA Barcoding of Cardinalfish (Apogonidae) in Gilimanuk Bay, Bali, Indonesia

Highlight Research Eight species of cardinalfish in Gilimanuk Bay were delineated using the barcoding method. This study provides the first nucleotide sequence for Ostorhinchus hartzfeldii in GenBank online database. The phylogenetic tree showed that each sample resided in a distinct cluster, indicating that the barcoding method efficiently differentiated at the species level. The mean genetic distance between genera within the family was 31.8-fold higher than the mean genetic distance within species. Abstract The Apogonidae is estimated to consist of nearly 300 fish species, most of which inhabit coral reef areas. The lack of distinctive body markings and overlapping species distribution makes species assignment challenging. Therefore, this study aimed to delineate species and establish barcoding reference databases of Apogonidae in Gilimanuk Bay (Bali, Indonesia) using the Cytochrome Oxidase I (COI) gene of the mitochondrial DNA. A total of 22 fish tissue samples were extracted with 10% Chelex solution. BLAST analysis was performed and genetic differentiation between species was calculated. The phylogenetic tree was constructed using the Maximum Likelihood method and tree visualization was generated using iTOL V5. The morphology and genetic identification results based on the mitochondrial COI gene revealed eight species of seven genera, and one species was new to GenBank online database. This study was the first-ever addition of COI sequence for Ostorhinchus hartzfeldii into the GenBank database. The average K2P genetic distance within species and K2P distance between genera within the family were 0.60% and 19.10%, respectively. The mean genetic distance between genera within the family was 31.8-fold higher than the mean genetic distance within species. The phylogenetic tree showed that each sample resided in a distinct cluster, which indicates that DNA barcoding is a reliable and effective approach for species delimitation in Apogonidae fishes.

Identification of Acrossocheilus fasciatus (Teleostei: Cyprinidae) from Three Different Phylogeographic Sources using Morphology and COI DNA Barcoding

Background: Accurate species delimitation is essential for phylogenetic, phylogeographic, ecological, conservation and biogeographic investigations. Acrossocheilus species classification has long been contentious, owing to their uncertain evolutionary relationships, high levels of morphological homoplasy and a lack of knowledge of the taxonomic validity of conventionally utilized morphological characters. Methods: A final data set consisting of 102 mitochondrial cytochrome oxidase I (COI) gene sequences was analysed for K2P intraspecific and interspecific divergence in the current study, using reconstruction methods (maximum likelihood) and various models (ABGD, PTP, GMYC) to divide the sequences according to the pattern of genetic variation. We used landmark-based geometric morphometrics to identify specimens. Result: Two taxa were identified according to the phylogenetic tree (maximum likelihood method) and three different models (ABGD, PTP, GMYC). The molecular taxonomy of Acrossocheilus agreed with the morphological identification of specimens from the same collection locality. Therefore, the COI gene and geometric morphometrics were effective in species identification of A. fasciatus.

MtCO1 Analysis of the Butterfly Genus &lt;i&gt;Papilio&lt;/i&gt;, from Indonesia

The genus Papilio is a butterfly that has many subspecies with similar phenotypes. So until now, researchers are still studying the genus Papilio based on morphology and genetics. Partial sequences of mitochondrial cytochrome oxidase-I (mtCOI) genes from individuals from 13 species of Papilionidae from North Sulawesi: Indonesia, Papua New Guinea, Malaysia: Negeri Sembilan, India: Assam, South Korea, China, Taiwan: Nantou, and Thailand: Fang, Chiang Mai have been compared in this study. Nucleotide divergences showed that the mean genetic distance between species was 0.023 (2.3%) to 0.723 (72.32%). Phylogenetic analysis revealed that groups of Papilionidae from North Sulawesi: Indonesia formed their own group; Species from Papua New Guinea are closely related to other Papilionidae species from other regions. There are 50 distinct sites or single nucleotide polymorphism (SNP) in the mtCO1 sequence of eleven species. These mtCO1 sequences can reveal the genetic differentiation of closely related species in the genus Papilio.

DNA Barcoding Approach: Assessing Diversity of Economically Important Anchovy on the Northern Coast of Java, Indonesia

Abstract Anchovy is one of the economically important fish in the western part of the northern coast of Central Java. The production of anchovies extends along the coast from Brebes Regency to Batang Regency. Therefore, this study aims to determine the diversity of anchovies on the northern coast of Central Java (Brebes, Tegal, Pemalang, Batang, and Kendal), using a molecular approach. This study was conducted from April to September 2019 in the Laboratory of Tropical Marine Biotechnology, Faculty of Fisheries and Marine Science, Universitas Diponegoro. The descriptive exploratory method was applied in this study with a random sampling technique, while the cytochrome oxidase I (COI) gene was used for molecular identification. An anchovy sample molecularly identified from each region was found by sampling, including BB 3, PML 2, BTG 1, BTG 2, BTG 4, TGL 2, TGL 4, KDL 1, and KDL 4. The base-pair length of 10 anchovy samples from the COI gene amplification result was 600-700 bp. The sequencing and alignment results of the BLAST analysis showed that the ten anchovy samples were included in the type of Encrasicholina heteroloba, Stolephorus commersoni, Stolephorus waitei and Atherinomorus sp., which ranged from 98 to 99% similarity in location. Therefore, it is concluded that there were at least four different species of anchovy on the northern coast of Java. However, further research is suggested to determine the genetic variation of each species for better fisheries management.

Karyotypic and Molecular Analysis of Pterygoplichthys pardalis (Castelnau 1855) from the Lower Amazon River.

Pterygoplichthys pardalis is an armored catfish native to South America and an important resource for the ornamental fish industry. Recently, several exotic populations have been introduced into rivers on five continents. Despite its commercial and environmental importance, P. pardalis is poorly studied from a genetic perspective. In this study, we analyzed the karyotype of P. pardalis from the Amazon River and molecular variations in the mitochondrial gene Cytochrome oxidase I (COI) between native and exotic populations. The karyotype presented diploid number 2n = 52 and NF = 100 without cytogenetic variation between males and females. Nucleolus organizer regions (Ag-NOR) in the distal region of the long arm of pair 12 coincided with the 18S hybridization signal, whereas 5S was syntenic to this chromosome but localized in the short arm. The constitutive heterochromatin was restricted in the distal regions of pairs 4, 12, 25, and 26. Telomeric probes showed only distal hybridization signals. The karyotype of P. pardalis diverged from that of its congeners, and COI molecular variation revealed four haplotypes. The Philippine population revealed the greatest diversity with three haplotypes, while haplotype H1 was the most abundant and observed in both native and exotic populations. This new genetic data contributes to species management and provides useful information from an aquaculture perspective.

A molecular phylogeny of Noctuini (Lepidoptera: Noctuidae: Noctuinae)

Abstract The tribe Noctuini is comprised over 520 species; many are economically important species that impact human agriculture. Despite their diversity and relevance, relationships of Noctuini have been difficult to resolve. There have been extensive morphological revisions of some of these taxa (e.g., Agrotis Ochsenheimer, Euxoa Hübner); however, there are no studies that focus exclusively on the phylogenetic relationships of the Noctuini. Currently, Noctuini are separated into 2 subtribes, Agrotina and Noctuina. In this study, we tested previous classification schemes with 2 analyses. The total evidence analysis utilized 3 loci for 3 outgroup taxa and 54 noctuine species. Using museum specimens, we collected data from cytochrome oxidase I (COI), Dopa Decarboxylase (DDC), and Elongation factor 1-α (EF1-α) to generate a dataset of 1,378 base pairs (bp) for a total evidence approach. We also used previously published COI sequences for 626 species, resulting in a matrix of 690 bp, termed “the barcode analysis.” We used both Maximum Likelihood and Bayesian approaches for the total evidence analysis, and ML for the barcode analysis. In both sets of results, we found equivocal support for monophyletic Agrotina and Noctuina. We found strong support for smaller, well-described genera (e.g., Spaelotis), but poor support for large genera such as Euxoa, Feltia Walker, and Agrotis. It is likely that a combination of limited taxon/gene sampling, limited gene choice, and the rapid evolution of these species resulted in a lack of phylogenetic resolution in the total evidence topologies. Based on these results, we recommend increased taxon sampling and inclusion of loci that target these rapidly evolving lineages to achieve a better understanding of the phylogeny of Noctuini and its genera.