Cytochrome C Oxidase Subunit I Research Articles

Taxonomic review of the genus Johnius (Perciformes: Sciaenidae) from East Malaysian waters, with comments on newly recorded Johnius species

In terms of sheer abundance, taxonomic diversity, and species richness, Malaysia has one of the most diverse Sciaenidae in the Indo-West Pacific. Up to ten valid species of the sciaenid fish genus Johnius (e.g., J. amblycephalus (Bleeker, 1855), J. belangerii (Cuvier, 1830), J. borneensis (Bleeker, 1850), J. carouna (Cuvier, 1830), J. coitor (Hamilton, 1822), J. heterolepis Bleeker, 1873, J. macrorhynus (Lal Mohan, 1976), J. plagiostoma (Bleeker, 1849), J. sasakii Hanafi et al., 2022 and J. weberi Hardenberg, 1936), and with additional newly recorded species as J. trewavasae Sasaki, 1992, have been recorded to occur in East Malaysia (Borneo Island) coastal waters. The newly recorded species is a close congener to J. heterolepis Sasaki, 1992 and can be well distinguished by several characters and molecular evidence. The goal of this study is to review the taxonomic status of Johnius species in East Malaysia, Borneo Island, by using a literature review, morphological inference and molecular characters. Principal Component Analysis (PCA) successfully discriminates based on second anal spine length and gill raker length within sampled specimens. A phylogenetic tree based on cytochrome oxidase subunit I (COI) showed intraspecific levels of genetic differentiation within Johnius of East Malaysia, which forms a monophyletic group. Moreover, the new record of Johnius trewavasae in East Malaysia represented that the distribution of the subtropical genus Johnius extended to the southernmost part of the tropical region.

Phylogeny and Genetic Population Structure of Dominant Copepods in Two Ponds with Contrasting Salinities in the Solar Saltern of Sfax (Tunisia) Based on Mitochondrial (COI and Cytb) and Nuclear (18S) DNA Sequences

Due to the complexity of taxonomic classification based on the classical morphological characters of copepods, phylogenies have been ambiguous. In this study, we investigate the phylogeny of copepods, including four species from three orders, in the saltern of Sfax using the small subunit of nuclear ribosomal RNA genes (18S). In the studied area, copepods seemed to be a polyphyletic group, and the genetic structure of these crustaceans is complex and problematic. We have also used two mitochondrial markers, the cytochrome c oxidase subunit I (mtCOI) gene and the cytochrome b (Cytb) sequence data, in order to investigate the genetic diversity and differentiation in a total of 96 individuals from two sets of Paracartia grani, sampled from two ponds with different salinities (42 PSU and 61 PSU). All of the results presented here suggest a low genetic diversity among P. grani species and a weak genetic structure between the sets. The nucleotide and haplotype diversity of P. grani were extremely low, indicating the homogeneity of the two sets, which could be combined into one set living in different ranges of salinity. This small genetic diversity is possibly due to the confined natural distribution range and strong selective pressure in a saltern environment. These data also suggest that gene flow is the main factor shaping the genetic structure of the studied sets.

Eriophyes pouteriae sp. nov., a New Mite Species Infesting Pouteria sapota

Pouteria sapota, or “mamey sapote”, is a tropical fruit tree native to Central America and Southern Mexico, producing sweet, nutrient and vitamin-rich fruit. Several insect pests are known to infest P. sapota, but none have been associated with plant growth alterations. Eriophyoid mites are well known to cause plant malformations, but mites that cause this type of damage to mamey sapote have not been reported. Trees with abnormal leaf growth, including stunted leaves, outward curling, leaf yellowing, and diminishing overall tree vigor, were found in multiple locations in Southern Florida and one location in Brazil. Numerous plant samples were examined for the presence of minute eriophyoid-like mites, and a new species was found. It was morphologically described, and DNA fragments of the mitochondrial gene cytochrome c oxidase subunit I (COI), the nuclear subunit D2 region in 28S rDNA, and the ITS nuclear regions were PCR-amplified and sequenced. Morphological and molecular descriptions of the new species, named E. pouteriae sp. nov., are provided to aid the identification and future detection of this mite. Even though several species within the genus Eriophyes have been reported on other Sapotaceae species, this is the first eriophyoid mite known to be associated with mamey sapote.

Arenicolidae (Annelida) in Norwegian waters: Species occurrence, bathymetric distribution and identification of juvenile specimens

The two species of Arenicolidae, Arenicola marina and Arenicolides ecaudata, are common along the Norwegian coast. Juveniles and anterior fragments are often encountered when grab sampling, making traditional morphological characteristics insufficient for species identification. This study examined the robustness of the characteristics and development of branchiae in juvenile specimens of Arenicolides ecaudata (n = 109) and Arenicola (n = 44) (A. marina and Arenicola sp.) and additional specimens to assess geographic and bathymetric distribution. Neuropodia were discovered to be efficient in distinguishing the species. Branchiae of A. ecaudata appear when specimens measure between 7–13 mm, while in A. marina they appear in specimens measuring between 3–5 mm. The ring formula in anterior chaetigers is a key characteristic in differentiating between Arenicola species. This characteristic alone is insufficient for reliable species identifications, as study findings corroborated error rates in identification based on the ring formula. The bathymetric range of both species has increased considerably. Arenicola marina was documented at 194 m depth and A. ecaudata at 387 m depth. DNA barcodes with the cytochrome c oxidase subunit I (COI) gene were produced for three specimens of Arenicolides ecaudata, representing the first publicly available COI barcode sequences for this species. Five specimens of Arenicola marina, with similar sequences to specimens from other north-eastern Atlantic localities were also barcoded in BOLDsystems.

First report of Lipoptena axis Maa, 1965, from captive cervids in Thailand, based on morphological and molecular data

Deer louse flies (Lipoptena spp.) are hematophagous ectoparasites of cervids. The genus Lipoptena comprises 32 species, some of which are of veterinary importance as vectors of various pathogens, and are also known to attack human hosts. Recently, deer louse flies have been observed during annual checkups of captive cervids at Khon Kaen Zoo in Khon Kaen, Thailand. However, data on their specific identity and prevalence remain limited. This study aims to identify louse fly samples from captive cervids at Khon Kaen Zoo using morphological and molecular analyses. A total of 60 louse flies were collected from 17 captive cervids and identified based on their morphology. Major morphological characteristics, including mesothoracic bristle patterns, abdominal tergal plate bristles, and terminalia structure indicated that the Khon Kaen louse fly is Lipoptena axis Maa, 1965. Phylogenetic analysis of sequences from a portion of the mitochondrial cytochrome c oxidase subunit I (COI) gene was performed, which confirmed that L. axis of this study belongs to the cervi group, which is distinct from other groups of Lipoptena species. This study represents the first report of L. axis in Thailand. We provide an updated taxonomic key for the identification of Lipoptena species in the cervi group.

Inferring long‐distance movements of insects using combined hydrogen isotope and genetic analyses: A case study of the African edible bush‐cricket

Abstract The degree to which individuals within populations vary in whether, how far and what direction they disperse is central in order to understand many foundational questions in ecological entomology, including factors determining population and invasion dynamics, species' ability to respond to climate change or when designing conservation strategies for threatened species. This study combined stable hydrogen isotope analysis of nymphal and adult wing chitin with population genetic analysis of the cytochrome c oxidase subunit I (COI) gene to understand long‐distance movements of African edible bush cricket (Ruspolia differens) in East Africa. Results revealed that at most 77% of non‐swarming adults and 85% swarming adults originated locally, that is, within a range of a few hundred kilometres. In contrast, non‐local adults captured in Uganda likely originated from Ethiopia, Kenya or South Sudan based on the H isotopic evidence. Genetic analysis revealed 150 COI haplotypes with no significant differentiation among geographic source regions and no evidence of genetic isolation by movement distance. The observed frequent long‐distance movements, together with the historical records, which indicate that the swarm movements can be irregular in their direction, suggest that the movements represent nomadism rather than regular long‐distance migration. Our results thus provide a valuable case study of what characterizes nomadic movements in insects, which have been one of the outstanding open questions in movement ecology of animals.

Identification and analysis of the genetic diversity of Dermacentor everestianus using mitochondrial 16S rRNA and COI sequences in the Xizang Autonomous Region of China

Ticks serve as vectors and reservoirs for a variety of pathogens, including protozoa, bacteria, and viruses, that cause tick-borne diseases in both humans and livestock. Despite their widespread distribution, their genetic diversity and population structure in Xizang have not been thoroughly investigated. In this study, sheep ticks were identified from four counties by microscopy, followed by genetic analysis of Dermacentor everestianus populations in Xizang based on mitochondrial 16S rRNA and cytochrome oxidase subunit 1 (COI) gene sequences. D. everestianus was the predominant species and 16S rRNA gene sequence analysis revealed nine haplotypes with seven nucleotide variations, showing a mean haplotype diversity (Hd) of 0.505 and mean nucleotide diversity (Pi) of 0.00129. Similarly, the COI sequences revealed 28 haplotypes with 26 nucleotide variations, yielding a mean Hd of 0.828 and a mean Pi of 0.00279. Molecular variance analysis (AMOVA) indicated that most genetic variation occurs within populations. In addition, neutral test analyses suggested potential bottleneck events in the D. everestianus populations. The haplotype network had a star-like distribution with the common haplotypes A1, A2, B1, and B5. The phylogenetic tree separated haplotypes of both genes into two branches on the haplotype network indicating no distinct geographical clustering. This study is the first to highlight the prevalence of D. everestianus in Xizang, revealing moderate genetic differentiation among the four populations, possibly due to host mobility. This intensive investigation of genetic variation and population structure contributes valuable insights into the current population genetics of D. everestianus in Xizang, China.

Phylogeography of the Invasive Fruit Fly Species Bactrocera carambolae Drew & Hancock (Diptera: Tephritidae) in South America

The carambola fruit fly, Bactrocera carambolae Drew & Hancock, is native to Southeast Asia, infests about 150 plant species, and is considered a quarantine pest insect in several regions of the world. Bactrocera carambolae has invaded Suriname, French Guyana, and northern Brazil. In Brazil, it was first recorded in 1996 and has been restricted to the states of Amapá and Roraima due to official control efforts of the Ministry of Agriculture and Food Supply (Ministério da Agricultura e Pecuária—MAPA). This is the first study to estimate the genetic structure and diversity of South American populations of B. carambolae. A total of 116 individuals from 11 localities in Brazil and 7 localities in Suriname were analyzed. Additional sequences available at GenBank from Indonesia (Lampung) and Thailand (San Pa Tong and Muang District) were also used in the analysis. We sequenced a fragment of the mitochondrial gene cytochrome oxidase subunit I. A total of 35 haplotypes were found. Haplotypes from Indonesia were closest to the haplotypes from South America, separated only by a few mutational steps. This suggests that Indonesia is the likely source for the introduction of B. carambolae into South America. The Southeast Asian populations appeared as the most ancestral group in the phylogenetic trees. The high similarity and sharing of several haplotypes among populations within South America indicate a lack of genetic structure. The mismatch distribution and neutrality tests suggest that South American populations have undergone a rapid growth and expansion following a single founder event. The low genetic diversity and the population expansion evidenced by the neutrality tests lend support to the hypothesis of a recent introduction of a single lineage of the carambola fruit fly into South America.

DNA barcode reference database and gap analysis for biomonitoring Hong Kong’s marine animals

DNA-based taxonomic identification relies on comprehensive DNA reference libraries, but uneven barcode coverage poses a global challenge. Here we determined the gaps in BOLD and GenBank for marine animals in Hong Kong’s subtropical waters. We compiled 170,529 sequences from four marker genes [cytochrome c oxidase subunit I (COI), 12S ribosomal RNA (rRNA), 16S rRNA, and 18S rRNA] representing 4074 marine animal species spanning 17 phyla. Gap analysis highlighted variations in COI barcode coverage across phyla, ranging from 0 % in Ctenophora, Gnathostomulida, Hemichordata, Phoronida and Platyhelminthes to 90.44 % in Chordata. The average coverage of COI (53.24 % in BOLD and 58.47 % in GenBank) was substantially higher than that of the other three rRNA genes (19.46–32.25 %). In total, 4392 Barcode Index Numbers (BINs) were assigned to 2169 species, of which 41.13 % were taxonomically concordant and 50.71 % were displayed multiple BINs, indicating the presence of cryptic species and the potential for undiscovered diversity. Overall, this study provided a valuable DNA barcode library for marine animals in Hong Kong, revealed that many marine animals lack a barcode, and many species may be undescribed, highlighting the need for enhanced reference libraries. We also outlined a strategy for filling the gaps in the database. Utilizing the local reference database can significantly enhance molecular-based biodiversity assessment and improve the identification of cryptic (morphologically similar) and invasive species, thereby providing more informed support for coastal management and conservation efforts in Hong Kong waters and the surrounding areas of China’s Hong Kong-Zhuhai-Macau Greater Bay Area.

Description of two new species of Lycopale (Diptera: Syrphidae) from the northwestern Tropical Andes hotspot, with the redescription of Lycopale magnifica

Lycopale Hull, 1944 is a small Neotropical flower fly genus (Syrphidae: Eristalinae: Eristalini: Helophilina) with six described species. Recent surveys in the Colombian and Ecuadorian Andean Forest and Paramo ecosystems revealed the discovery of two species new to science: Lycopale mendozai sp. nov. and Lycopale radioheadi sp. nov. Simultaneously, the species Lycopale magnifica (Bigot, 1880) was rediscovered and is redescribed here, nearly a century after its original description, including photographs of its habitus and illustrations of the male genitalia. A new key is proposed, including illustrations of thoracic and abdominal patterns of all known species to distinguish them from the new taxa. Distributional patterns are illustrated and discussed. Mitochondrial cytochrome c oxidase subunit I (COI) sequences are provided for the three species, L. magnifica, L. mendozai sp. nov., and L. radioheadi sp. nov. The discovery of these two new species suggests that the Tropical Andes diversity of flower flies is still underestimated and many more unnamed species remain to be discovered and described from this biodiversity hotspot.

Population connectivity of the European squid Loligo vulgaris along the West Iberian Peninsula coast: comparing mtDNA and SNPs

Understanding genetic diversity and population connectivity in marine organisms is essential for fisheries management. In the present study, we examined the population genetics of the European squid, Loligo vulgaris, along the western Iberian Peninsula at two genetic resolutions using the mitochondrial cytochrome oxidase subunit I gene (COI) and genomic markers obtained via double digest restriction-site associated DNA sequencing (ddRADseq). The results obtained revealed 79 haplotypes out of 160 COI sequences, while the SNP data set included 86,431 loci after filtering for 38 individuals with 86,319 neutral data. Mitochondrial COI analyses revealed high haplotype (0.961) and nucleotide (0.010) diversities, and the haplotype network reveals complex sub-structure in Turkish waters within a panmictic population. Both Tajima’s D and Fu’s Fs tests suggest that the population of L. vulgaris analysed is evolving neutrally. Pairwise Fst for neutral SNPs were low (0 < Fst<0.002) and not significant showing high homogeneity among populations, while pairwise comparations for candidate adaptive SNPs (112 loci) showed Fst values ranging from 0.026 to 0.234. While neutral SNPs showed admixture, the candidate adaptive SNPs showed a moderate significant structure with a latitudinal discrimination. Overall, both genetic approaches showed homogeneity and strong genetic flux identifying a unique population along the Western Iberian Peninsula.

Genetic diversity, population genetic structure and demographic history of the Ribbontail stingray Taeniura lymma (Fabricius, 1775) (elasmobranchii: myliobatiformes: dasyatidae) along the Tanzanian coastline

The Ribbontail stingray Taeniura lymma is an economically important fish and attractive species for the aquarium trade industry. Overfishing, habitat degradation, and pollution, however, pose a threat to this species. This study used partial mitochondrial cytochrome oxidase subunit I (COI) sequences (603 base pairs long) from 96 samples of T. lymma collected at five fish-landing sites (Deep Sea-Tanga, Malindi-Unguja, Kaole-Bagamoyo, Kivukoni-Dar es Salaam, and Bandarini-Mtwara) located along the coast of Tanzania to determine the species’ genetic diversity, population genetic structure, and demographic history. The findings revealed an average nucleotide diversity of 0.24 ± 0.16% and a haplotype diversity of 0.75 ± 0.04. Nucleotide and haplotype diversities were relatively low at Kaole-Bagamoyo compared to the other studied localities. An Analysis of Molecular Variance (AMOVA) indicated limited but statistically significant genetic differences among populations (Overall FST = 0.09, p < 0.01). Pairwise AMOVA revealed genetic difference between the Deep Sea-Tanga population and all other populations studied with exception of Malindi-Unguja. Analyses of mismatch distribution, demographic history, and a haplotype network support a scenario of historical population expansion in the studied species. Immediate effort is required to protect population exhibiting low genetic diversity in this commercially important ray.

From Korea to Croatia: the first record of the polyclad flatworm Notocomplana koreana in the Mediterranean

Polyclads are large-bodied, free-living flatworms that occur in diverse marine habitats. They are known predators of crustaceans, ascidians, snails, bivalves, polychaetes, and other flatworms. The acotylean genus Notocomplana Faubel, 1983 currently comprises 32 species that primarily inhabit the Pacific but are also found in the Atlantic and Indian Oceans. Here, we report the discovery of Notocomplana koreana (Kato, 1937) in the Mediterranean Sea. N. koreana was originally described from Korean and Japanese waters but was recently found to have expanded its range to the Atlantic. In this report, we shed light on the likely route of migration of N. koreana to the Mediterranean and discuss the challenges in identifying such an inconspicuous species. Through an integrative taxonomic approach, which merged morphological trait analysis using live animals and histology with a comparative genetic analysis of partial 28S ribosomal DNA and mitochondrial cytochrome c oxidase subunit I (COI) barcoding markers, we uncovered the characteristics of this introduced species and provide insights into its potential effect on Mediterranean ecosystems.

Enhanced data on Stolephorus grandis and Stolephorus waitei (Teleostei: Clupeiformes: Engraulidae), two species endemic to the Sahul Shelf, western Pacific Ocean.

Eighteen and 23 specimens of two species of the poorly known anchovies Stolephorus grandis Hata & Motomura, 2021b and Stolephorus waitei Jordan & Seale, 1926 (Clupeiformes: Engraulidae), respectively, are examined. This study presents their detailed morphological characters including fresh colorations along with their precise distributional ranges within the Sahul Shelf coastal region. The "barcode" regions of the mitochondrial cytochrome c oxidase subunit I (COI) gene were examined in four specimens of S. waitei and one specimen of S. grandis. Both species diverged by 10.6% and > 10.3% mean (uncorrected) genetic distances from each other and from any other species of Stolephorus, respectively. Stolephorus waitei, S. grandis, and Stolephorus nelsoni Wongratana, 1987 form a monophyletic group of species endemic to the Sahul Shelf region. Additionally, the role and evolution of double lines on the dorsum observed in several species of Stolephorus, including S. grandis, are discussed.

Genetic Variation and Population Structure of Clonorchis sinensis: An In Silico Analysis.

Clonorchis sinensis is an important zoonotic parasite that is mainly prevalent in China, Korea, Vietnam and the Russian Far East. To explore the genetic variation and population structure of C. sinensis, an in silico analysis was conducted based on mitochondrial cytochrome c oxidase subunit 1 (COX1), ribosomal internal transcribed spacer 1 (ITS1) and ribosomal internal transcribed spacer 2 (ITS2) sequences. The sequences obtained from NCBI were truncated for further analyses, including haplotype network, phylogenetic, gene flow, diversity and neutrality analyses. The results showed that there were 20, 11 and 4 haplotypes for COX1, ITS1 and ITS2, respectively. The results of both the haplotype network and phylogenetic analyses indicated that the haplotypes for each type of sequence from the same country were not all clustered together. Haplotype diversity values were all lower than 0.5. Values of nucleotide diversity were higher than 0.005, except for ITS2. Tajima's D and Fu's Fs values were all negative, and p-values showed significant differences, indicating that the population of C. sinensis is growing. Fst values were all lower than 0.05. In conclusion, this study found that there are specific variations of C. sinensis in different countries, and the population of this parasite is growing with less genetic variation. The findings provide a crucial foundation for understanding the molecular epidemiology and population dynamics of C. sinensis.

Abstract 4132149: Elevated Levels of Plasma Mitochondrial DNA Damage-Associated Molecular Patterns (DAMPS) in Humans and Dogs with Heart Failure and Reduced Ejection Fraction

Background: Heart failure (HF) with reduced ejection fraction (HFrEF) is associated with increased inflammatory response that contributes to progressive worsening of the HF state. Potential endogenous triggers of this process include damage-associated molecular patterns (DAMPs). DAMPs originate from within cells following tissue stress or injury and activate pattern recognition receptors of the immune system, triggering an inflammatory response. Mitochondria are the major cell organelles that release DAMPs into the circulation. Chronic HF is associated with multi-organ mitochondrial dysfunction and cell injury and death. Purpose: The present study sought to quantify differences in mitochondrial DNA (mtDNA) DAMPs in the setting of HFrEF in humans and dogs compared to normal humans and dogs. Methods: Plasma samples were obtained from 9 HFrEF patients with left ventricular ejection fraction (LVEF) ≤35% and 9 age-matched healthy subjects, and from 6 dogs with coronary microembolization-induced HF (LVEF ≤35%) and 6 normal dogs. DNA fragments were isolated from 1 ml of plasma using a commercially available kit. Quantitative PCR and specifically designed primers were used to measure the amount of DNA in plasma from COX1 (cytochrome c oxidase subunit 1), ND1 (NADH dehydrogenase subunit 1) and ND6 (NADH dehydrogenase subunit 6) using an Applied Biosystems 7500 Fast-PCR unit. The relative abundances of plasma mtDNA DAMPs were expressed as threshold cycles (C T ). Results: Compared to normal human subjects, mtDNA DAMPs levels of COX1 , ND1 and ND6 in plasma of HFrEF patients was 4-fold, 9-fold and 8-fold greater, respectively. Similarly, compared to normal dogs, mtDNA DAMPs levels of COX1 , ND1 and ND6 in plasma of HFrEF dogs was increased 48-fold, 12-fold and 4-fold, respectively. Conclusions: These findings indicate a marked increase of mtDNA DAMPs in plasma of humans and dogs with HFrEF. The increase of mtDNA DAMPs is consistent with the elevated systemic inflammatory state of HFrEF. The results underscore mitochondrial abnormalities as integral players in the progressive worsening of HFrEF.

Revalidation of Enteromius crocodilensis (Fowler, 1934) from synonymy with the disjunctly distributed Enteromius argenteus (Günther, 1868) based on molecular and morphological evidence.

The rosefin barb, Enteromius argenteus, as currently described, is a freshwater fish with a distribution that is geographically separated and divided into the northern population in the Kwanza River system in Angola, and the southern population in the Inkomati River system in South Africa and Eswatini. Due to this disjunct distribution pattern, it is likely that the two populations represent distinct species. mtDNA sequence data and detailed examination of morphometric characters revealed considerable genetic (2.8%-3.5%) and morphological differentiation of the two populations, suggesting their recognition as two distinct species. Enteromius crocodilensis is revalidated and redescribed for the southern population that is readily distinguished from E. argenteus by nonoverlapping circumpeduncular scale counts (12-13 vs. 14) and color pattern (presence of a conspicuous dark band along the midline vs. absence of characteristic melanin patterns). Phylogenetic analysis based on mtDNA cytochrome oxidase subunit I (COI) data places both E. argenteus and E. crocodilensis in a well-supported clade that includes a number of morphologically similar sawfin barb species from southern and central Africa.

Hidden diversity in anthropophilic sand flies of the Monticola Series (Diptera, Psychodidae)

The Monticola series comprises two anthropophilic and widely distributed species in Brazil: Pintomyia (Pifanomyia) monticola (Costa Lima, 1932) and Pintomyia (Pifanomyia) misionensis (Castro, 1959). They mainly occur in the Atlantic Rainforest, and it is known that Pi. monticola comprises at least two well-structured genetic lineages regarding a fragment of the cytochrome c oxidase subunit I (COI) gene. Here, we aim to elucidate the taxonomic status of this group using integrative taxonomy tools. Collections were performed in nine localities of four Brazilian states, and COI fragments were sequenced and merged with publicly available data. Several single-locus species delimitation algorithms, genetic distance metrics, phylogenetic trees, and haplotype networks were used to uncover cryptic diversity and population structure within Pi. monticola and Pi. misionensis. The resulting genetic clusters were then tested for morphological differences through linear and geometric morphometry of several characters. We analyzed 152 COI sequences, comprising 48 haplotypes. The maximum intraspecific p distances were 8.21% (mean 4.17%) and 9.12% (mean 4.4%) for Pi. monticola and Pi. misionensis, respectively, while interspecific ones ranged from 10.94 to 14.09% (mean 12.33%). Phylogenetic gene trees showed well-supported clades for both species, with clear structuring patterns within them. Species-delimitation algorithms split our dataset into at least three putative species for each taxon. Moreover, population structure analysis showed a strong correlation between Atlantic Forest areas of endemism as sources of molecular variation in Pi. monticola. Morphometric analyses were significant for wing shape variation and some linear measurements (mainly of the head) when comparing specimens of different genetic clusters for both taxa. These results indicate strong genetic structuring of Monticola series species, confirmed by morphometry, indicating two possible cryptic species complexes.

Molecular Identification of Species Belonging to Culex vishnui Subgroup (Diptera: Culicidae), Vectors of Japanese Encephalitis Virus, in Taiwan.

Classification of mosquitoes with overlapping features remains problematic when using traditional morphological identification alone. In this study, we used molecular methods to elucidate the taxonomic status of Culex tritaeniorhynchus, Culex annulus, and Culex pseudovishnui species as vectors of the Japanese encephalitis virus belonging to the Culex vishnui subgroup and gene flow among them. In this study, 76, 59, and 3 samples of Cx. annulus, Cx. tritaeniorhynchus, and Cx. pseudovishnui, respectively, were collected around Taiwan. Phylogenetic analysis and genetic divergence were based on genomic sequence variations in ribosomal DNA and the internal transcribed spacer (rDNA) and cytochrome c oxidase subunit I (COI). Our results revealed that Cx. annulus and Cx. vishnui are genetically similar and share a gene pool among the species from Taiwan and other Asian countries. However, two hidden taxa of Cx. tritaeniorhynchus, which clustered together according to the rDNA sequences, were discovered based on the COI sequences. In addition, Cx. pseudovishnui has different gene pools from those of the strains from other countries, implying that the population from Taiwan is probably either a unique strain or a sibling species. This study provides molecular information on the taxonomic status of the species in the Cx. vishnui subgroup in Taiwan and gene flow between these species, providing valuable information for vector control operations and the delineation of the evolutionary process.

The Mosquitoes (Diptera: Culicidae) of Sonora: Distribution, Ecology, and the First Records of Aedes deserticola Zavortink and Toxorhynchites septentrionalis (Dyar and Knab) in México

The diversity and distribution of mosquitoes from Sonora, Mexico, was documented through entomologic surveys conducted in the four physiographic regions and sub-regions of Sonora: the Sonoran Plain, the Sierra Madre Occidental, the Northern Mountains and Plains, and the Pacific Coastal Plain. Immature stages were collected from aquatic habitats, while adult mosquitoes were collected using Shannon traps, resting in vegetation, and by human landing collections. Overall, 11,316 specimens, which comprised 493 larvae, 224 larval exuviae, 400 pupal exuviae, 33 pupae, 4552 females, 5607 males, and seven male genitalia, were identified. Two subfamilies: Anophelinae and Culicinae, seven tribes, 10 genera, 23 subgenera, and 56 species are reported below. Of these, one tribe, one genus, five subgenera, and 15 species were recorded for the first time in Sonora. Two species, Aedes deserticola Zavortink and Toxorhynchites septentrionalis (Dyar and Knab), are the first recordings of their kind nationwide. Toxorhynchites septentrionalis was also barcoded with the cytochrome oxidase subunit 1 (COI) gene. The presence of Psorophora columbiae (Dyar and Knab) is confirmed in Mexico. Taxonomic notes, new geographic distribution limits of mosquitoes in Sonora, and information regarding their importance as disease vectors are provided. By adding Ae. deserticola and Tx. septentrionalis and confirming Ps. columbiae in Mexico, there are currently 251 mosquito species in the Country.