C Oxidase Subunit 1 Research Articles

Exploring the diversity of leafrollers (Lepidoptera: Tortricidae) associated with eucalypts in South African forestry nurseries and plantations, with a new report of Strepsicrates sp.

A number of insect pests cause damage to eucalypts in South Africa. Their accurate identification is a key part of managing and controlling these pests. The aims of this study were to identify leafroller insect species in South African eucalypt nurseries and plantations, and to determine whether the species were native insects that had developed new host associations with eucalypts or introduced non-native species. A section of the mitochondrial cytochrome c oxidase subunit 1 (CO1) gene of multiple specimens from nurseries and plantations was sequenced. The sequence data were compared and validated using sequences available in BOLD systems and GenBank databases. Results revealed the presence of an introduced non-native Strepsicrates sp. in both nurseries and plantations. In addition, the native species Choristoneura occidentalis and Eccopsis incultana were confirmed as present in plantations only and the native species Lozotaenia capensana was confirmed as present in one of the nurseries. Eucalypts are reported as host plants of C. occidentalis, E. incultana and Lozotaenia capensana for the first time. The findings contribute valuable insights into the identification and diversity of leaf rollers in eucalypt plantations in South Africa, with potential implications for pest management.

Distinct Island Lineages of Binturong (Arctictis binturong) from Indonesia and Its Conservation Implications

Binturong (Arctictis binturong) is a threatened carnivore that inhabits the forests of South and Southeast Asia. Despite its wide range, binturong is relatively scarce across its habitat distribution and is currently under the threat of poaching and illegal trade. Captive breeding has unfortunately been conducted rather haphazardly with a lack of origin record maintained, implicating potential risks to the management such as inbreeding or genetic swamping. This study thus aims to characterise the phylogenetic relationship of Indonesian binturong within the context of Southeast Asian binturong and further probe the distinctness of lineages originating from Java, Sumatra, Indonesian Borneo, and Bangka using Cytochrome B (CytB) and Cytochrome C Oxidase Subunit 1 (CO1). Genetic distance, phylogram topology, and haplotype analysis of both encoding genes further corroborate the distinctness of Java, Borneo, and Bangka binturong from other binturong from Indochinese regions such as India, Laos, and Myanmar. Search for prospective single nucleotide polymorphism markers to discriminate island lineages consistently found that each Java, Bangka, and Bornean binturong be distinct from each other and other lineages, especially when assessed using haplotype-based clustering. Assigning binturong originated from Sumatra is nonetheless more complicated, suggesting the possibility. Our findings substantiated the much-needed systematic research of Southeast Asian binturong as ex-situ insurance population management grows in Indonesia and other parts of the world to protect the diversity of binturong lineages and their corresponding unique evolutionary history.

When standard DNA barcodes do not work for species identification: intermixed mitochondrial haplotypes in the Jaera albifrons complex (Crustacea: Isopoda)

Here, we characterise the standard “Folmer region” of the mitochondrial cytochrome c oxidase subunit 1 (CO1) marker and a fragment of nuclear 28S marker in four species of the Jaera albifrons complex. Jaera albifrons (Leach, 1814), Jaera ischiosetosa Forsman, 1949, Jaera praehirsuta Forsman, 1949, and Jaera forsmani Bocquet, 1950 were collected from localities on the Norwegian coast and identified with morphological characters. We compared DNA sequences with sequences available in GenBank and BOLDsystems and calculated haplotype networks and interspecific versus intraspecific genetic distances. These analyses revealed low interspecific genetic distance (CO1 0.00–1.57%, 28S 0.00–0.39%) and extensive haplotype sharing between J. albifrons group species and specimens from both sides of the North Atlantic for both CO1 and 28S. Genetic distances between J. albifrons group species and other Jaera species, however, exceeded 29% for both CO1 and 28S, with no haplotype sharing. These assessments, together with taxonomically unconstrained analyses with software ABGD and ASAP, show that these markers are unable to distinguish between the J. albifrons group of morphospecies. The sequences do, however, clearly identify J. albifrons species complex from other Jaera species. Thus, a likely hypothesis is that taxa in this complex represent a single species. Our results corroborate previous finds where discordance between mitochondrial gene clusters, AFLP, and other data highlights the potential conflict between different “species criteria” and the well-established distinction between gene trees and species trees. In operational terms, common protocols for metabarcoding will potentially underestimate sympatric species diversity with cases like the J. albifrons complex, if the members of this complex indeed represent different species.

A new species of Eutyphoeus earthworm (Clitellata: Acanthodrilidae) from Mizoram, North-East India.

We describe here a new species of terrestrial earthworm Eutyphoeus rihnimensis sp. nov. (family Acanthodrilidae) from specimens collected at Khawrihnim, a village in Mizoram, Northeast India. The new species is distinguished from its congener members in E. gigas group in having an outgrowth that branches into four finger-like projections from the male porophores, five pairs of supra-intestinal glands in 8188 segments, typhlosole that begins from segment 25, ventral intestinal caeca 10 that covers segments 33-34, a bidiverticulate spermathecae with a short and stout duct, and very short lateral and median diverticula among other characters. Sequence analysis based on mitochondrial cytochrome c oxidase subunit 1 (CO1) gene further warrants its distinction from known species of Eutyphoeus.

Phylogenetic analyses and description of a new species of black widow spider of the genus Latrodectus Walckenaer (Araneae, Theridiidae) from Mexico; one or more species?

A new species of the spider genus Latrodectus Walckenaer, 1805 from Mexico is described based on an integrative taxonomic approach. Latrodectus occidentalis Valdez-Mondragón sp. nov. is described using the molecular markers cytochrome c oxidase subunit 1 (CO1) and internal transcribed spacer 2 (ITS2), morphology of male and female specimens, and Species Distribution Models (SDM). Four molecular methods for species delimitation were implemented. The new species is characterized by having a unique dorsal coloration pattern on the abdomen. Latrodectus occidentalis sp. nov. is considered a distinct and valid species for four reasons: (1) it can be distinguished by morphological characters (genital and somatic); (2) the average interspecific genetic variation is > 2%; (3) 12 haplotypes were recovered within the species, being separated by the next close haplogroup of L. hesperus Chamberlin & Ivie, 1935 (30 mutations); and (4) congruence was observed among the four molecular methods. The number of recorded species of Latrodectus from Mexico increases to four: Latrodectus mactans (Fabricius, 1775), L. hesperus Chamberlin & Ivie, 1935, L. geometricus C.L. Koch, 1841 (introduced), and L. occidentalis sp. nov. The diversity of the genus Latrodectus from Mexico is surely underestimated, and more sampling is needed from the different biogeographical provinces and ecoregions to fill in these gaps.

A new species of the genus Achalinus (Squamata: Xenodermidae) from Nanning, Guangxi, China.

We describe a new species of the genus Achalinus Peters, 1869 from Daming Mountain, Shanglin County, Nanning City, Guangxi Zhuang Autonomous Region, China, based on a single adult male specimen. It can be distinguished from all the other species in Achalinus by a combination of the following morphological characters: (1) a bright yellow collar around the neck, extending forward to the ventral of the head; (2) tail length comparatively long, TaL/Tol ratio 0.25; (3) DSR 23-23-23, moderately keeled; (4) VS 3+162; (5) SC 74, unpaired; (6) cloacal plate entire; (7) SPL 6, the fourth and fifth in contact with the eye; (8) IFL 6, the first three touching the first pair of chin shields; (9) a single loreal; (10) length of suture between internasal significantly longer than that between prefrontal, LSBI/LSBP ratio 1.34; (11) two pairs of chin shields; (12) longitudinal vertebral line absent. In addition, the uncorrected p-distances between the new species and other known congeners ranged from 6.3% to 25.4% for the cytochrome c oxidase subunit 1 (CO1). With the addition of the new species the total number of described Achalinus species is increased to 23 of which 17 are found in China.

Development of a real-time PCR (qPCR) method for the identification of the invasive paddle crab Charybdis japonica (Crustacea, Portunidae).

Crabs can be transported beyond their native range via anthropogenic-mediated means such as aquarium trade, live seafood trade and shipping. Once introduced into new locations, they can establish persisting populations and become invasive, often leading to negative impacts on the recipient environment and native species. Molecular techniques are increasingly being used as complementary tools in biosecurity surveillance and monitoring plans for invasive species. Molecular tools can be particularly useful for early detection, rapid identification and discrimination of closely related species, including when diagnostic morphological characters are absent or challenging, such as early life stages, or when only part of the animal is available. In this study, we developed a species-specific qPCR assay, which targets the cytochrome c oxidase subunit 1 (CO1) region of the Asian paddle crab Charybdis japonica. In Australia, as well as many parts of the world, this species is considered invasive and routine biosecurity surveillance is conducted to reduce the risk of establishment. Through rigorous testing of tissue from target and non-target species we demonstrate that this assay is sensitive enough to detect as little as two copies per reaction and does not cross amplify with other closely related species. Field samples and environmental samples spiked with C. japonica DNA in high and low concentrations indicate that this assay is also a promising tool for detecting trace amounts of C. japonica eDNA in complex substrates, making it a useful complementary tool in marine biosecurity assessments.

Historical land use patterns provide insight into contemporary insect diversity loss.

Healthy insect populations are vital for maintaining natural ecosystems and essential to global food security. The ongoing dramatic loss of insect species and biomass is thus a global cause for concern, with much focus on this topic in the media. Yet, determining the mechanism behind these declines remains difficult, particularly when attempting to differentiate between anthropogenic drivers of biodiversity loss and long-term natural fluctuations. In a From the Cover manuscript in this issue of Molecular Ecology, Crossley et al. (2022) examined the long-term impact of land use change on freshwater insects by estimating levels of genetic diversity with publicly available data for the cytochrome c oxidase subunit 1 (CO1) mitochondrial locus from >700 aquatic insect species across the United States. Contemporary genetic diversity measures reflect both current and past demography and therefore are related to both past and present habitat change. Crossley et al. found that environments with greater cropland extent over the last 200 years were associated with lower genetic diversity in contemporary aquatic insect populations. This indicates that historical land use is an important factor in contemporary population dynamics. Most critically, in regions of historical cropland cover that have been converted towards other uses (such as urban environments) more recently, aquatic insect populations exhibited higher levels of genetic diversity, indicating a possible rebound in insect populations after cessation of agricultural activity. This study highlights the power of using publicly available data to answer crucial questions regarding the current biospheric emergency.

Molecular population genetic analysis of Eriocheir japonica inhabiting Andeok Valley

Eriocheir japonica inhabit rivers flowing into the East Sea, South Sea, and the coastal waters of Jeju-do in South Korea, Japan, and Sakhalin. The larvae migrate to freshwater, mature, and move to estuaries to spawn. Jeju-do has habitats that prevent the migration of aquatic organisms. Populations with low within-group genetic diversity experience increased extinction risk. Industrialization causes climate change and also creates species extinctions. The goal of this study was to investigate the genetic diversity of E. japonica inhabiting Andeok Valley to elucidate their genetic stability and encourage future conservation plans. E. japonica samples used for the study were from the Andeok Valley in South Korea. One walking leg was used for DNA extraction. DNA was amplified using polymerase chain reaction to sequence mitochondrial cytochrome c oxidase subunit 1 (CO1) and cytochrome b (CytB) genes. Haplotype and sequence diversity analyses were performed. Sequencing results for CO1 and CytB genes, included 658 and 651 base pairs, respectively. The median network analysis showed that the population inhabiting Andeok Valley had a genetically simple phylogenetic structure. This region provides simple habitat conditions, preventing gene transfer. The probability of extinction of E. japonica that inhabits Andeok Valley is markedly high.

Using eDNA sampling for species-specific fish detection in tropical oceanic samples: limitations and recommendations for future use.

Over the past decade, environmental DNA (eDNA) has become a resourceful tool in conservation and biomonitoring. Environmental DNA has been applied in a variety of environments, but the application to studies of marine fish, particularly at tropical latitudes, are limited. Since many commercially important Caribbean fishes are overexploited, these species are optimal candidates to explore the use of this method as a biomonitoring tool. Specifically, for many of these species, the formation of fish spawning aggregations (FSAs) marks a critical life history event where fishes will gather in large numbers for reproduction. These FSAs are ephemeral in nature, lasting only a few days, but are predictable in time and space which makes them susceptible to overfishing. In this study, we test the feasibility of using an eDNA sampling approach (water and sediment collection) to detect the presence of known FSAs off the west coast of Puerto Rico, with cytochrome c oxidase subunit 1 (CO1) and 12S rRNA (12S) primers designed to target specific species. A total of 290 eDNA samples were collected and, of those, 206 eDNA samples were processed. All eDNA samples varied in DNA concentration, both between replicates and collection methods. A total of 12 primer sets were developed and tested using traditional PCR and qPCR. Despite validation of primer accuracy and sample collection during known peak spawning times, the use of traditional PCR and qPCR with both molecular markers failed to produce species-specific amplification. Thus, a trial test was conducted using the CO1 primers in which target fish DNA was 'spiked' at various concentrations into the respective eDNA samples to determine the target species DNA concentration limit of detection. Upon successful amplification of the trial, results indicated that eDNA samples were below the detection threshold of our methods, suggesting that the number of fish present at the spawning aggregations was inadequate for single-species detection methods. In addition, elements such as the unavoidable presence of non-target DNA, oceanic environmental conditions, shedding rates of target fish, among other biotic and abiotic factors could have affected DNA persistence and degradation rates at the sites. We provide recommendations for species-specific fish detection in lower latitudes, and suggestions for studies aiming to monitor or detect fish spawning aggregations using eDNA sampling.

Identity of the numerous bloodstains at the murder scene: molecular identification of fly artifacts and fly species by CO1 analysis.

Crime scenes may contain insect artifacts as well as samples of human origin. While the presence of insects can be important evidence in forensic medicine and forensic entomology, the insect artifacts sometimes interfere with the interpretation of bloodstain pattern analysis (BPA) which can be critical for accurate crime reconstruction. Fly artifacts are especially complicated to distinguish from true bloodstains. Indeed, we encountered a murder scene with numerous bloodstains inconsistent with the cause of death and had trouble interpreting them. The morphological method has been developed to distinguish them, but this method has to rely on the analyst's experience and opinion. This study aims not only to distinguish fly artifacts from true bloodstains but also to identify fly species by detecting fly DNA in small amounts of bloodstains at the scenes. Melt curve analysis of real-time PCR (qPCR) targeting cytochrome c oxidase subunit 1 (CO1) of mitochondrial DNA (mtDNA) was able to detect fly DNA in bloodstains from a murder scene. The fly DNA was sequenced from the qPCR product, and the fly species were identified by BLAST search. Fluorescence-labeled specific primers for four species of necrophagous flies were designed based on the sequences of the CO1 region, and differences in the length of the amplification products were used to identify fly species from trace amounts of fly DNA in the artifacts.

Morphological and molecular characters of Lemuru fish (Sardinella lemuru) from Tanjung Luar Waters, East Lombok

Lemuru fish from the Clupeidae family in various countries have many similarities that are difficult to distinguish between one species and another. This study aims to determine the morphological and molecular characteristics, genetic diversity, and kinship of lemuru fish in Tanjung Luar waters with lemuru fish species in countries. The molecular feature of lemuru fish used in this study was a marker for the cytochrome c oxidase subunit 1 (CO1) gene. The lemuru fish sample used in this study was lemuru caught in the waters of Tanjung Luar by small-scale fishermen. Lemuru fish sampling method using random, totaling 100 individuals. Firstly, the lemuru fish samples were grouped based on morphological characters. Next, segregation and augmentation of the CO1 gene DNA in five lemuru fish samples using a pair of primers. The amplification of DNA about 700 bp in size was sequenced. CO1 gene sequence data of lemuru samples were analyzed using MEGA 11 Software (Molecular Evolutionary Genetics Analysis) Version 11. The phylogenetic relationships obtained showed that the lemuru samples from Tanjung Luar waters, East Lombok, is in the same clade as Sardinella Jussieu from Malaysia and has a very close kinship relationship.

To be or not to be… Integrative taxonomy and species delimitation in the daddy long-legs spiders of the genus Physocyclus (Araneae, Pholcidae) using DNA barcoding and morphology.

Integrative taxonomy is crucial for discovery, recognition, and species delimitation, especially in underestimated species complex or cryptic species, by incorporating different sources of evidence to construct rigorous species hypotheses. The spider genus Physocyclus Simon, 1893 (Pholcidae, Arteminae) is composed of 37 species, mainly from North America. In this study, traditional morphology was compared with three DNA barcoding markers regarding their utility in species delimitation within the genus: 1) Cytochrome c Oxidase subunit 1 (CO1), 2) Internal Transcribed Spacer 2 (ITS2), and 3) Ribosomal large subunit (28S). The molecular species delimitation analyses were carried out using four methods under the corrected p-distances Neighbor-Joining (NJ) criteria: 1) Automatic Barcode Gap Discovery (ABGD), 2) Assemble Species by Automatic Partitioning (ASAP), 3) General Mixed Yule Coalescent model (GMYC), and 4) Bayesian Poisson Tree Processes (bPTP). The analyses incorporated 75 terminals from 22 putative species of Physocyclus. The average intraspecific genetic distance (p-distance) was found to be < 2%, whereas the average interspecific genetic distance was 20.6%. The ABGD, ASAP, and GMYC methods were the most congruent, delimiting 26 or 27 species, while the bPTP method delimited 33 species. The use of traditional morphology for species delimitation was congruent with most molecular methods, with the male palp, male chelicerae, and female genitalia shown to be robust characters that support species-level identification. The barcoding with CO1 and 28S had better resolution for species delimitation in comparison with ITS2. The concatenated matrix and traditional morphology were found to be more robust and informative for species delimitation within Physocyclus.

Protective efficacy of Shenge San on mitochondria in H9c2 cardiomyocytes.

To investigate whether peroxisome proliferator-activated receptor γ-coactivator-1α/nuclear respiratory factor 1 (PGC-1α/NRF1) activity can protect mitochondrial function in the setting of cardiac hypertrophy and improve cardiomyocyte energy metabolism. Cardiac hypertrophy was modeled in H9c2 cells treated with isoproterenol (ISO) to assess the effects of Shenge San (, SGS) on cell viability and mitochondrial membrane potential. We assessed mitochondrial complex mRNA levels and mitochondrial oxidative phosphorylation factor mRNA and protein levels. Compared with the 100 μM ISO group, cell size was significantly decreased in the 0.3 mg/mL SGS and 20 μM ZLN005 (PGC-1α activator) groups ( < 0.01). Compared with the SGS (0.3) +ISO group, we observed lower phosphorylated adenosine monophosphate-activated kinase (AMPK) protein levels in the ISO and ZLN005+SGS+ISO groups ( < 0.01). Compared with the compound C group, SGS significantly increased PGC-1α expression in ISO-induced cardiac hypertrophy cells ( < 0.01), and this was inhibited by compound C pretreatment ( < 0.05). Compared with the ISO group, the mitochondrial red-green fluorescence ratio increased in the 0.3 mg/mL SGS group ( < 0.05). mRNA levels of cytochrome c oxidase subunit 1 (CO1) in the ISO and compound C groups were lower than those in control group ( 0.01), and the mRNA levels of CO1 and ATP8 were significantly lower in the ISO and compound C groups versus control ( 0.01). Compared with the SGS (0.3) +ISO group, ATP synthetase subunit 8 (ATP8) mRNA was significantly decreased in the ISO group ( < 0.01) and compound C+SGS+ISO group ( < 0.05). Compared with the SGS (0.3) +ISO group, NRF1 mRNA levels were significantly decreased ( < 0.05) in the ISO and compound C+SGS+ISO groups. SGS can attenuate ISO-induced cardiomyocyte hypertrophy, restore the decrease in mitochondrial membrane potential, and upregulate PGC-1α/NRF1 levels. Notably, these effects can be blocked by AMPK inhibitor-compound C.

Updates on Identification and Distribution of the River Mullet “Ludong” in Northern Philippines

“Ludong”, also known as the president’s fish, is a river mullet known to be found in Cagayan River, Northern Philippines. Unfortunately, this species has been heavily exploited over the years. As a conservation measure, the Department of Agriculture - Bureau of Fisheries and Aquatic Resources (DA-BFAR) issued the BFAR Administrative Circular (BAC) 247 of 2013, declaring a closed fishing season for “ludong” during its spawning from October 1 to November 15. Despite these efforts, the identity and distribution of “ludong” remain unclear. Here, the species identification and habitat range of “ludong” have been determined and updated by DNA barcoding using mitochondrial cytochrome c oxidase subunit 1 (CO1) as a marker. Twenty-five (25) “ludong” samples were collected from Cagayan River and Abra River traversing Regions 1, 2, and the Cordillera Administrative Region. BLAST, phylogenetic and pairwise analyses showed that all 25 specimens were Cestraeus goldiei. Consequently, the known distribution of C. goldiei has been extended from Cagayan River to Abra River, from Region 2 to Region 1 and CAR. Knowledge of the definitive species identification and distribution of “ludong” will contribute to further understanding of its biology and fisheries and boost its conservation.

A new species of the genus Achalinus (Squamata: Xenodermidae) from Ningshan County, Shaanxi Province, China

A new species of odd-scaled snake in the genus Achalinus is described from Ningshan County, Shaanxi Province, China, based on seven female specimens. Morphologically, the new species can be distinguished from its congeners by the following combination of characters: (1) dorsum iridescent and uniformly dark brown in preservative, longitudinal vertebral line absent; (2) light brown beneath; (3) dotted black streak in the middle of the subcaudals absent; (4) tail length relatively short, TaL/ToL 12-16%, in females; (5) fewer subcaudals, 41-46 in females; (6) dorsal scales 23 rows throughout, strongly keeled; the outer-most rows on both sides are also keeled and slightly enlarged; (7) one loreal; (8) internasal not fused to prefrontal; (9) suture between internasals is similar in size when compared to the suture between prefrontals; (10) preocular and postocular absent; (11) 6 supralabials; (12) 5 infralabials, the first 3 (rarely 2) touching the first pair of chin shields; (13) 3 pairs of chin shields. A phylogenetic analysis using mitochondrial gene, cytochrome c oxidase subunit 1 (CO1), showed that this new species forms a monophyletic group with strong support. In addition, the uncorrected p-distances between the new species and other known congeners ranged from 3.4% to 13.0%. The recognition of the new species increases the number of described Achalinus species to 21.

Population structure and demographic history of the gastropod Thaisella chocolata (Duclos, 1832) from the Southeast Pacific inferred from mitochondrial DNA analyses.

The present‐day population structure of a species reflects the combination of oceanographic currents, life‐history traits, and historical events. However, little is known about the mechanisms that have shaped the gene lineage distribution of marine species inhabiting the Southeast Pacific. Here, we provide a comprehensive phylogeographical study of a species distributed along the Southeast Pacific coastal region by analyzing the endemic gastropod Thaisella chocolata (Duclos, 1832). Sequencing of mitochondrial cytochrome c oxidase subunit 1 (CO1) and 16S rRNA revealed strikingly high haplotypic nucleotide and genetic diversity but a lack of significant population differentiation within the survey area. In addition, a star‐shaped phylogeny and significantly negative Tajima's D and Fu's Fs tests of neutrality suggested historical occurrence of rapid demographic expansion. Mismatch distributions and Bayesian inference analyses also confirmed T. chocolata to have undergone two ancestral demographic expansions. Calculations suggested that these expansions began in the lower and middle Pleistocene epoch, likely due to continental shelf development and climatic conditions. These findings could help establish a genetic baseline for T. chocolata as the first step toward sustainable spatial management of this species, as well as understand this species’ response to future climate change.

Life history strategies of Cotylurus spp. Szidat, 1928 (Trematoda, Strigeidae) in the molecular era - Evolutionary consequences and implications for taxonomy.

Species of Cotylurus Szidat, 1928 (Diplostomoidea: Strigeidae) are highly specialized digeneans that parasitize the gastrointestinal tract and bursa of Fabricius of water and wading birds. They have a three-host life cycle; the role of first intermediate host is played by pulmonate snails, while a wide range of water snails (both pulmonate and prosobranch) and leeches are reported as second intermediate hosts.Unfortunately, species richness, molecular diversity and phylogeny of metacercariae of Cotylurus spp. (tetracotyle) occurring in snails remain poorly understood. Thus, we have performed the parasitological and taxonomical examination of tetracotyles form freshwater snails from Poland, supplemented with adult Strigeidae specimens sampled from water birds. In this study we report our use of recently obtained sequences of two molecular markers (28S nuclear large ribosomal subunit gene (28S rDNA) and the cytochrome c oxidase subunit 1 (CO1) fragment), supplemented by results of a method of species delimitation (GMYC) and haplotype analysis to analyse some aspects of the ecology, taxonomy, and phylogeny of members of the genus Cotylurus.The provided phylogenetic reconstructions discovered unexpectedly high molecular diversity within Cotylurus occurring in snails, with clearly expressed evidence of cryptic diversity and the existence of several novel-species lineages. The obtained results revealed the polyphyletic character of C. syrius Dubois, 1934 (with three separate molecular species-level lineages) and C. cornutus (Rudolphi, 1809) Szidat, 1928 (with four separate molecular species-level lineages). Moreover, we demonstrated the existence of two divergent phylogenetical and ecological lineages within Cotylurus (one using leeches and other snails as second intermediate hosts), differing significantly in their life history strategies.

Sub-lineages of Taenia solium Asian Genotype Recorded in North India.

Porcine cysticercosis is a neglected zoonotic disease of significant veterinary and medical importance owing to its economic impact and public health significance. The present study aimed at genetic characterization of Taenia solium metacestodes in slaughtered pigs of Haryana (North India). A total of 213 (160 and 53 from Chandigarh and Hisar, respectively) slaughtered pigs intended for human consumption were screened for the presence of T. solium metacestodes. The retrieved metacestodes were confirmed molecularly based on the partial amplification of mitochondrial cytochrome c oxidase subunit 1 (CO1) gene. Evolutionary divergence, haplotype and nucleotide diversities and neutrality indices of the retrieved isolates were also assessed. Out of the 213 pigs, 2 (0.94%) revealed the presence of metacestodes involving 1 pig each from Chandigarh (0.62%) and Hisar (1.9%). The sequences obtained after custom sequencing were submitted to GenBank under the accession numbers LC661682-83. The present study haplotype clustered with haplotypes of Asian origin and showed variation from other haplotypes by 1-23 mutational steps. However, the present study isolates also showed nucleotide polymorphisms (A198T, A199G, A201T, G204A, T206A, C210T, T212G, T213A,T216G/A, T217C, T221C, C524T, G994A) at different positions, which indicated the presence of sub-lineages. Low nucleotide diversity (π = 0.020) and negative value of Tajima's D (-1.304) observed for the haplotypes under consideration was indicative of purifying selection and recent population expansion. Our study confirms the circulation of T. solium Asian genotype (with distinct sub-lineages) in study area and recommends strict control measures to contain the zoonotic disease.

Genetic Variability of Polypedilum (Diptera: Chironomidae) from Southwest Ecuador.

Simple SummaryPolypedilum is a genus of aquatic non-biting midges in the family Chironomidae. This genus is widely distributed in neotropical rivers from lowlands to Andean highlands. Nevertheless, making species identification based on morphology is quite complex, even more so in the Neotropics, since systematic studies of this group are scarce. DNA barcoding can help to overcome this problem using a short DNA sequence as a barcode for species delimitation. A fragment of the mitochondrial gene cytochrome c oxidase I (CO1) has been successfully employed as a barcode in the genus Polypedilum. In this study, our aim was to understand the effect of environmental characteristics on Polypedilum diversity and distribution. We examined the CO1 sequence of 68 Polypedilum specimens from rivers with different environmental conditions located in an important biogeographic area of Ecuador. We identified five morphotypes and seven putative species which revealed high genetic variability among them. Polypedilum distribution seems to be affected mainly by two environmental factors, dissolved oxygen, and temperature. Our study is the first evidence of richness within the genus in Ecuador, highlighting the importance of developing taxonomic studies along with ecological assessments to further describe and identify new species.Chironomids show a wide distribution and can occupy several habitats due to their high adaptive capacity in different freshwater environments. The genus Polypedilum is found along a wide elevational and environmental gradient in the neotropics, and its genetic variability could help to elucidate factors determining its distribution and tolerance to the environmental changes of different species or populations. This study examines the genetic variability of Polypedilum in an important biogeographic area that acts as a geographical barrier of biodiversity at the border of the Choco and Tumbes biomes. We identified five Polypedilum morphotypes using classic taxonomic methods. We examined 68 Polypedilum individuals from eight sampling sites in El Oro Province, Ecuador, analyzing the putative molecular species using the cytochrome c oxidase subunit 1 (CO1) mitochondrial gene fragment. Then, we calculated molecular diversity indices, Haplotype diversity (Hd), and θs and θπ estimators. Seven Polypedilum OTUs were determined from which a high molecular diversity was registered. A CCA was conducted to understand the population composition in relation to environmental characteristics. Results indicated that dissolved oxygen and temperature are the main environmental factors affecting Polypedilum distribution across elevational gradients and between basins.