NADH Dehydrogenase Subunit Research Articles

Taenia asiatica: Mitochondrial signatures based analysis of an emerging public health threat in India.

Taenia asiatica is a zoonotic tapeworm, commonly known as Asian Taenia. It is an emerging sister species of T. saginata with pigs as intermediate hosts. The present study aimed at genetic characterization and population structure analysis of T. asiatica metacestodes in slaughtered pigs in Haryana, north India. In total, the vital organs of 253 slaughtered pigs were screened for the presence of T. asiatica metacestodes. The molecular identification and phylogenetics were performed targeting the mitochondrial NADH dehydrogenase subunit 1 (nad1) and cytochrome C oxidase subunit 1 (cox1) genes. The median-joining haplotype network and population structure analyses were performed with the sequences generated herein and GenBank-archived T. asiatica sequences for both mitochondrial signatures. Out of 253 pigs screened, the liver of only one animal showed the presence of T. asiatica metacestodes. The sequences generated herein exhibited 99.60% and 98.85% similarity to the GenBank-archived sequences of T. asiatica corresponding to the nad1 and cox1 genes, respectively. Overall, 2 and 6 haplotypes for the overall data set with low nucleotide (0.00399±0.00237 and 0.00095±0.00042) and low haplotype (0.400±0.237 and 0.131±0.054) diversities were recorded for the nad1 and cox1 genes, respectively. The negative values recorded for the neutrality indices exhibited deviations from neutrality and hence, propounded recent population expansion or purifying selection or selective sweep. The findings of the present study are of significant medical importance considering an emerging global public health threat of the neglected tapeworm T. asiatica.

First Report of Apple Chlorotic Leaf Spot Virus in Pome and Stone Fruit Trees in Azerbaijan.

Azerbaijan is major producer of fruit crops, such as pome and stone fruits, in the Caspian Sea and Caucasus Mountains areas (FAO Stat, 2022). No information is available on the occurrence of apple chlorotic leaf spot virus (ACLSV, genus Trichovirus, family Betaflexiviridae) in the country. Therefore, the main fruit tree production areas in Azerbaijan were surveyed for ACLSV during the 2017-2019 growing seasons by DAS-ELISA using ACLSV reagents (Neogen - Scotland, UK) (Clark and Adams 1977). Of the 510 plant samples tested, 139 were positive for ACLSV. To confirm the presence of ACLSV, total RNA was isolated from leaf samples (0.5 g) exhibiting deformation, yellow veins, ringspots, and/or chlorosis symptoms as previously described (Foissac et al. 2005), and analyzed by RT-PCR using the OneStep RT-PCR kit (Qiagen) and primers designed in the coat protein gene of ACLSV: F (5'-CTACAATTTGAAGGAGGTGGTCAAC-3'), R (5'-GACGCCGGTTCATATTAGTTATGAC-3') and primers for the NADH dehydrogenase subunit 5 fragment as a housekeeping gene (Menzel et al. 2002). Reactions were performed at 5 min at 95 °C, 34 cycles of 10 s at 95 °C, 20 s at 50 °C, 20 s at 72 °C, followed by a final extension of 2 min at 72 °C. DNA amplicons were resolved by electrophoresis on 2% agarose gels and visualized under UV illumination after staining with GelRed (Biotium, Fremont, CA). Results showed the occurrence of ACLSV in 14 apple samples (Malus domestica L.; n=38), four cherry plum sample (P. cerasifera L.; n=17), one almond samples (P. amygdalus L.; n=13), and one plum sample (P. domestica L.: n=71). All the pear (Pyrus communis L.; n=27), quince (Cydonia oblonga L.; n=2), sweet cherry (P. avium L.; n=27), sour cherry (P. cerasus L.; n=1), peach (P. persica L.; n=22), and apricot (P. armeniaca L.; n=8) samples tested negative for ACLSV in RT-PCR. Next, the 287-bp amplicons obtained by RT-PCR from two Azerbaijani ACLSV isolates from the Ganja and Kurdamir region, i.e., Ganja 43 from apple and Kurdamir 80 from plum, were cleaned with Exo SAP-IT and characterized by bidirectional Sanger sequencing at the Cornell Biotechnology Resource Center in Ithaca, NY. Sequence analyses were performed with the DNASTAR Lasergene software package (v17.3.0.57). Results revealed 87.2%-96.7% and 98.3%-100% nucleotide and amino acid identities, respectively, of ACLSV isolate Ganja 43 isolate (GenBank PP894314) with Afghan (KU749440) and Ukrainian (JQ866623) isolates, and 86.6%-95.2% and 97.9%-99.0% nucleotide and amino acid identities, respectively, of ACLSV isolate Kurdamir 80 (GenBank PP898052) with Afghan (KU749440) and Indian (FN666579) isolates. This is the first report of ACLSV in pome and stone fruit trees in Azerbaijan. Since the occurrence of mixed virus infections was not investigated in this study, the viral symptoms displayed by the infected trees cannot be ascribed to ACLSV. The occurrence of ACLSV suggests the need for certification programs in the country to reduce the presence of the virus in the propagation material of pome and stone fruit trees.

Research on the mechanism of eugenol in the treatment of liver cancer based on network pharmacology, molecular docking technology, and in vitro experiments.

Eugenol, a phenolic natural product with diverse pharmacological activities, remains unexplored in liver cancer. Using network pharmacology, we investigated eugenol's therapeutic mechanisms in liver cancer. We obtained eugenol's molecular structure from PubChem and screened its targets using similarity ensemble approach in Swiss Target Predictiondatabases. Overlapping genes with liver cancer-related genes from GeneCards were identified. Protein-protein interaction networks, Gene Ontology annotations, and Kyoto Encyclopedia of Genes and Genomes pathway analyses were conducted. A target-pathway network revealed eugenol's interaction with 122 liver cancer-related genes. Molecular docking confirmed eugenol's high affinity for mitochondrial nicotinamide adenine dinucleotide, reduced form (NADH) dehydrogenase 1 (MT-ND1), AKT1, NDUFB7, and NADH dehydrogenase (complex I) subunit S3 (NDUFS3). Expression levels of these targets in normal liver and liver cancer tissues were examined using GEPIA2 and HPA databases. The CCK-8 assay and colony formation assay demonstrated that eugenol significantly inhibited the proliferation of hepatocellular carcinoma cells. Western blot analysis confirmed that eugenol upregulated MT-ND1 while downregulating the expression of targets such as AKT1, NDUFB7, and NDUFS3. Furthermore, it was found that eugenol could influence the expression of the AKT1 target through the AKT/p70 S6K pathway. This study provides new insights into the potential mechanisms of eugenol in liver cancer and offers novel perspectives for network-based liver cancer research.

Genome-wide analyses provide insights into genetic variation, phylo- and co-phylogenetic relationships, and biogeography of the entomopathogenic nematode genus Heterorhabditis.

Multigene, genus-wide phylogenetic studies have uncovered the limited taxonomic resolution power of commonly used gene markers, particularly of rRNA genes, to discriminate closely related species of the nematode genus Heterorhabditis. In addition, conflicting tree topologies are often obtained using the different gene markers, which limits our understanding of the phylo- and co-phylogenetic relationships and biogeography of the entomopathogenic nematode genus Heterorhabditis. Here we carried out phylogenomic reconstructions using whole nuclear and mitochondrial genomes, and whole ribosomal operon sequences, as well as multiple phylogenetic reconstructions using various single nuclear and mitochondrial genes. Using the inferred phylogenies, we then investigated co-phylogenetic relationships between Heterorhabditis and their Photorhabdus bacterial symbionts and biogeographical patterns. Robust, well-resolved, and highly congruent phylogenetic relationships were reconstructed using both whole nuclear and mitochondrial genomes. Similarly, whole ribosomal operon sequences proved valuable for phylogenomic reconstructions, though they have limited value to discriminate closely related species. In addition, two mitochondrial genes, the cytochrome c oxidase subunit I (cox-1) and the NADH dehydrogenase subunit 4 (nad-4), and two housekeeping genes, the fanconi-associated nuclease 1 (fan-1) and the serine/threonine-protein phosphatase 4 regulatory subunit 1 (ppfr-1), provided the most robust phylogenetic reconstructions compared to other individual genes. According to our findings, whole nuclear and/or mitochondrial genomes are strongly recommended for reconstructing phylogenetic relationships of the genus Heterorhabditis. If whole nuclear and/or mitochondrial genomes are unavailable, a combination of nuclear and mitochondrial genes can be used as an alternative. Under these circumstances, sequences of multiple conspecific isolates in a genus-wide phylogenetic context should be analyzed to avoid artefactual species over-splitting driven by the high intraspecific sequence divergence of mitochondrial genes and to avoid artefactual species lumping driven by the low interspecific sequence divergence of some nuclear genes. On the other hand, we observed that the genera Heterorhabditis and Photorhabdus exhibit diverse biogeographic patterns, ranging from cosmopolitan species to potentially endemic species, and show high phylogenetic congruence, although host switches have also occurred. Our study contributes to a better understanding of the biodiversity and phylo- and co-phylogenetic relationships of an important group of biological control agents and advances our efforts to develop more tools that are compatible with sustainable and eco-friendly agricultural practices.

Genetic Insights and Phylogenetic Relationships of Lycorma delicatula (Hemiptera: Fulgoridae, 1845) Revealed Through Mitochondrial Genome Sequence Analysis

Lycorma delicatula, a phytophagous agricultural pest, is invading various regions across the globe. We employed the mitochondrial genome and mitochondrial marker NADH dehydrogenase subunit 2 (ND2) to elucidate the mitochondrial genome characterization and phylogenetic relationships. The mitochondrial genome revealed a circular mitochondrial structure with a length of 16 138 bp encompassing 37 genes and a GC% of 23.41%. Codon usage preferences exhibited significant variation, with A or U being the most preferred bases in the third codon position. Most tRNAs displayed a typical cloverleaf structure, while a few exhibited base mismatches, primarily in G-U pairs. Phylogenetic analysis using the ND2 genes indicated that the population in northwest China could be tightly near together, with a genetic distance ranging from 0 to 0.01. Speculations arising from the mitochondrial genome analysis suggest that species from northern China were closer relatives to invaders from other regions, Lycorma meliae emerged as a close relative of Lycorma delicatula among the Hemipterans. Collectively, our findings enhance the understanding of phylogenetic relationships and complement the mitochondrial genomic information of Lycorma delicatula, thereby contributing valuable data for subsequent studies on this pest species.

Polymorphism of Russian Populations of Rhopalosiphum padi L. Based on DNA Markers

Using the next-generation sequencing (NGS) technology, the nucleotide polymorphism in a fragment of the ND4 gene encoding NADH dehydrogenase subunit 4 was studied in 14 samples from three populations of the bird cherry-oat aphid (Rhopalosiphum padi L.) and the range of nucleotide polymorphism was determined. The insects were collected in 2021 and 2022 in the North-West of Russia (in the vicinity of St. Petersburg) and in the northern Caucasus (Krasnodar Territory and Dagestan). Mitochondrial DNA haplotypes were identified, which have 97.95–99.80% sequence identity with the reference GenBank accession number KT447631.1. The level of intraspecific polymorphism of a 438 bp ND4 gene fragment in Rh. padi varied from 0.2 to 4.3%. In the two-year experiments, 33 polymorphic sites (17 transitions and 16 transversions) were found in the ND4 sequences, which made it possible to identify 30 mitochondrial DNA haplotypes. The Rh. padi populations collected simultaneously on different host plants or at different times on bird cherry (spring) and cereals (summer) differed in the proportion of the main haplotype, as well as in the composition of unique minor haplotypes. Analysis of the ratio of mitochondrial DNA haplotypes suggests the important role of the host plant genotype in the formation of the structure of Rh. padi populations.

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.

Comparative Mitogenomics Provides Valuable Insights for the Phylogeny and New DNA Barcodes of Ganoderma.

Ganoderma is the most important genus in the family Ganodermataceae; many species have attracted much attention and widely cultivated because of their medicinal values, but so far, not a sequenced mitogenome derived from dikaryon strains has been explicitly recorded. Herein, four novel mitogenomes of commonly cultivated Ganoderma (G. leucocontextum H4, G. lucidum G6, G. sinense MZ96 and G. tsugae SS) were de novo assembled and given detail functional annotations. Collinearity analysis revealed that the four mitogenomes shared 82.93-92.02% similarity with their corresponding reference mitogenomes at the nucleotide level. A total of 15 core protein-coding genes (PCGs), along with rrnL and rrnS (mtLSU and mtSSU) were chosen as potential candidates for constructing their individual phylogenetic trees. These trees were compared with those derived from the concatenated sequences of 15 core PCGs. And finally, we found that the atp9 and nad4L were the most reliable markers for the phylogenetic analysis of Ganoderma and chosen as standard sequences to generate new DNA barcodes. This finding was further verified by comparing it against almost all available Ganoderma mitogenomes in the NCBI, with Trametes versicolor (Polyporaceae) and Rigidoporus microporus (Meripilaceae) as two outgroups. A total of 52 mitogenomes from three families were highly conserved, with identical gene lengths for atp9 (222 bp) and nad4L (267 bp). These genes were capable of distinguish distinctly different various species, which are grouped into separate clades within the phylogenetic trees. The closest related clades (I and II), including at least 30 samples of the three classical taxonomic species (G. lingzhi, G. sichuanense and G. lucidum), differed in only one SNP. The single base mutation rate increased with the evolutionary divergence of the phylogenetic clades, from two to three SNPs in earlier clades (e.g., clade IV containing G. leucocontextum) to five to six SNPs in later clades (e.g., clade X containing G. sinense). Despite these variations between species, the atp9 and nad4L genes of Ganoderma mitogenomes consistently encoded the same ATP synthase F0 subunit c (73 aa) and NADH dehydrogenase subunit 4L (88 aa). These two genes have been identified as reliable markers of new DNA barcodes, offering valuable insights and contributing significantly to understanding the evolutionary relationships and phylogeny of the Ganoderma genus and even the Ganodermataceae family.

Contrasting the role of historic factors in phylogeograpic patterns in the native Johnny darter (Etheostoma nigrum) and invasive round goby (Neogobius melanostomus) in lower michigan.

Round goby (Neogobius melanostomus) is an invasive fish present in all five Great Lakes and is becoming increasingly common in their tributaries. Johnny darter (Etheostoma nigrum) is a native species that often coexists with N. melanostomus. In this work, historic factors are addressed as a source of genomic variation in study populations of these species. To do this, patterns of variation in the mitochondrial gene NADH dehydrogenase subunit 2 (ND2) were characterized for both species throughout Lower Michigan. Populations of N. melanostomus and E. nigrum were sampled from 17 localities representing both eastern and western basins of Lower Michigan to test the hypothesis that populations differ between the eastern and western basins of the Great Lakes. Neogobius melanostomus populations were largely homogenous with no significant differences detected among populations or between the eastern and western basins. Additionally, N. melanostomus exhibited no evidence of overarching historical genetic structure, consistent with the recent invasion and rapid expansion of this species. Etheostoma nigrum exhibited significant differentiation among local populations; however, similarity among mtDNA haplotypes indicated that differences among populations are recent, suggesting that local forces are a more important factor in shaping patterns of variation than historical factors. Contrary to predictions, there were no significant differences detected between the eastern and western basins of the Great Lakes; however, construction of a neighbor-joining tree with F ST estimates revealed clustering of populations by basin with some anomalies. These anomalies may be the result of recent stream capture events facilitating gene flow between the two basins.

Electroacupuncture Slows Experimental Myopia Progression by Improving Retinal Mitochondrial Function: A Study Based on Single-Cell RNA Sequencing.

This study aimed to establish a complete atlas of retinal cells in lens-induced myopia (LIM) and electroacupuncture (EA) intervention by single-cell RNA sequencing (scRNA-seq) and to explore the potential mechanism of EA in improving experimental myopia progression in guinea pigs. scRNA-seq is used to assess changes in individual cellular gene levels in the retina of LIM- and EA-treated guinea pigs. In addition, the role of EA in slowing myopia progression by improving retinal mitochondrial function is further investigated. scRNA-seq identified ten cell clusters in the retina of LIM and EA guinea pigs and mitochondrial respiratory chain-related genes in Cones and Muller-glia cells-Cytochrome oxidase subunit III (COX3), NADH dehydrogenase subunit 4 (ND4), and NADH dehydrogenase subunit 2 (ND2) are closely related to lens-induced myopia. A comprehensive atlas in the retina of LIM and EA guinea pigs at a single-cell level is established, and the positive role of EA in improving retinal mitochondrial function to slow the experimental myopia progression in guinea pigs is revealed.

The seventh species of the newt genus Tylototriton in Thailand: a new species (Urodela, Salamandridae) from Tak Province, northwestern Thailand.

A new species of the crocodile newt genus Tylototriton from Doi Soi Malai located at Mae Tuen Wildlife Sanctuary, Tak Province, northwestern Thailand is described based on molecular and morphological evidence, and named as Tylototritonsoimalai sp. nov. The new species is the seventh recorded species of the genus Tylototriton reported in Thailand. It differs morphologically from its congeners by a combination of the following morphological characteristics: head longer than wide; snout blunt or truncate; sagittal ridge on head narrow, short and distinct; dorsolateral bony ridges on head pronounced and rough; parotoids distinct; vertebral ridge prominent, wide and not segmented; 14-16 distinct, rounded and isolated rib nodules but posterior nodules connected; tips of fore- and hind limbs overlapping when adpressed along the body. The body background color is black, while the color markings are orange. Molecular analysis indicated that Tylototritonsoimalai sp. nov. is a distinct lineage and sister to T.uyenoi with a 4.1% genetic sequence divergence based on the mitochondrial NADH dehydrogenase subunit 2 gene. The new species is currently restricted to the hill evergreen forests of Doi Soi Malai. The implementation of a strategic plan is recommended to protect both the species and its habitat from anthropogenic activities.

Phylogeographic structure of Italian Formica pratensis (Retzius 1783) populations in the framework of the species Eurasian range

AbstractThe phylogeography and demographical history of Italian Formica pratensis populations were examined and compared with the Eurasian-wide dataset available for this species and the other red wood ant species Formica lugubris. Forty-eight workers belonging to eight populations from both Alps and Apennines were analysed sequencing a 1.5-kilobase mitochondrial DNA fragment, including the cytochrome b gene and part of the NADH dehydrogenase subunit 6 gene. A total of 127 sequences were screened, scoring 53 different haplotypes amongst all specimens, with five new haplotypes discovered in the Italian populations. All the Italian haplotypes clustered in a monophyletic clade, underlining a clear phylogeographical separation of this group from the other Eurasian groups and suggesting a glacial separate forest refugia and different post-glacial colonisation patterns. The haplotypes from the Alps and the Apennines showed a high genetic proximity, pointing out an ancient (Pleistocene) wide distribution of this species across all these areas and common ancestral lineages. No shared haplotypes were scored between Northern and Central Apennine populations, but the low inter-population genetic distance indicated similar post-glacial selective processes acting on these groups. The diversity we recorded may be influenced by the actual fragmentation of F. pratensis populations across its entire Eurasian range, and by the limited geographical origin and sample dimension of the dataset analysed. Future studies with a more extensive sampling in the Alps and Eastern Europe are needed to confirm our result.

Selection of mtDNA Marker Genes In The Study Of Bird Genetic Diversity: A Review

Mitochondrial DNA (mtDNA) has emerged as a crucial tool in understanding bird genetic diversity due to its unique characteristics, including maternal inheritance and high mutation rates. This review synthesizes the current understanding of various mtDNA marker genes, focusing on their applications in avian genetic research. The selection of mtDNA marker genes such as cytochrome c oxidase subunit I (COI), NADH dehydrogenase subunit 2 (ND2), cytochrome b (cyt b), and the D-loop region is evaluated for their advantages and limitations. The review highlights the significance of mtDNA markers in conservation genetics and phylogenetics, emphasizing their utility in resolving evolutionary relationships and assessing population structure.

Whole genome sequence analysis of population structure and insecticide resistance markers in Anopheles melas from the Bijagós Archipelago, Guinea-Bissau

BackgroundAnopheles melas is an understudied malaria vector with a potential role in malaria transmission on the Bijagós Archipelago of Guinea-Bissau. This study presents the first whole-genome sequencing and population genetic analysis for this species from the Bijagós. To our knowledge, this also represents the largest population genetic analysis using WGS data from non-pooled An. melas mosquitoes.MethodsWGS was conducted for 30 individual An. melas collected during the peak malaria transmission season in 2019 from six different islands on the Bijagós Archipelago. Bioinformatics tools were used to investigate the population structure and prevalence of insecticide resistance markers in this mosquito population.ResultsInsecticide resistance mutations associated with pyrethroid resistance in Anopheles gambiae s.s. from the Bijagós were absent in the An. melas population, and no signatures of selective sweeps were identified in insecticide resistance-associated genes. Analysis of structural variants identified a large duplication encompassing the cytochrome-P450 gene cyp9k1. Phylogenetic analysis using publicly available mitochondrial genomes indicated that An. melas from the Bijagós split into two phylogenetic groups because of differentiation on the mitochondrial genome attributed to the cytochrome C oxidase subunits COX I and COX II and the NADH dehydrogenase subunits 1, 4, 4L and 5.ConclusionsThis study identified an absence of insecticide-resistant SNPs common to An. gambiae in the An. melas population, but did identify structural variation over insecticide resistance-associated genes. Furthermore, this study presents novel insights into the population structure of this malaria vector using WGS analysis. Additional studies are required to further understand the role of this vector in malaria transmission.Graphical abstract

A new species of karst-dwelling bent-toed gecko of the Cyrtodactylusintermedius group (Squamata, Gekkonidae) from eastern Thailand and the phylogenetic placement of C.intermedius.

A new karst-dwelling bent-toed gecko of the Cyrtodactylusintermedius group is described from Khlong Hat District, Sa Kaeo Province, eastern Thailand, based on an integrative taxonomic analysis of genetic data and morphological characteristics. Phylogenetic analyses using the mitochondrial NADH dehydrogenase subunit 2 (ND2) gene revealed that topotypes of C.intermedius were sister to a clade containing C.kulenensis from Cambodia, an unnamed lineage from Sakaerat Biosphere Reserve in Nakhon Ratchasima Province, Thailand, and the Khlong Hat lineage described here as Cyrtodactyluskhlonghatensis sp. nov. Multivariate analyses of morphometric and meristic characters showed that C.khlonghatensis sp. nov. is morphologically distinct from all other species in the group by having the combination of SVL 76.5-82.8 mm in adult males and 88.5 mm in an adult female; eight supralabial and nine infralabial scales; 30-32 paravertebral tubercles; 20 or 21 longitudinal rows of dorsal tubercles; 43 or 44 ventral scales; seven or eight expanded subdigital lamellae on the 4th toe; 12 unmodified subdigital lamellae on the 4th toe; 19 or 20 total subdigital lamellae on the 4th toe; 31 or 32 total number of enlarged femoral scales; enlarged femoral and precloacal scales continuous; 6-8 pore-bearing precloacal scales in males; three or four rows of enlarged post-precloacal scales; 1-3 postcloacal tubercles; proximal femoral scales less than one-half the size of distal femoral scales; absence of interdigital pocketing between digits of forefeet and hindfeet; and posterior border of the nuchal loop rounded. Uncorrected pairwise genetic divergences (p-distances) between the new species and other species of the intermedius group ranged from 4.73-22.55%. The discovery of this new species exclusively in isolated karst formations from the Thai-Cambodia border suggests that there may be further undiscovered Cyrtodactylus in unexplored karst landscapes along the border of eastern Thailand and western Cambodia.

Decreased in Mitochondrial Complex I Subunit NDUFS2 Is Critical for Oocyte Quality During Postovulatory Aging in Pigs.

The levels of nicotinamide adenine dinucleotide (NADH) dehydrogenase [ubiquinone] iron-sulfur protein 2 (NDUFS2, a subunit of NADH dehydrogenase) decrease in aged tissues, and these reductions may be partly associated with age-related conditions such as Parkinson's disease. Aging leads to many mitochondrial defects, such as biogenesis disruption, dysfunction, defects in the mitochondrial membrane potential, and production of reactive oxygen species, that may be highly related to NDUFS2 expression. The relationship between NDUFS2 and postovulatory oocyte aging in pigs remains unknown. In this study, we investigated changes in NDUFS2 expression during postovulatory aging (POA). Furthermore, NDUFS2 was knocked down via dsRNA microinjection at the MII stage to evaluate the effects on mitochondrial-related processes during POA. The mRNA expression of NDUFS2 decreased significantly after 48-h aging compared with that in fresh oocytes. NDUFS2 knockdown (KD) significantly impaired the maintenance of oocyte morphology and blastocyst development of embryos after POA. The levels of PGC1α (mitochondrial biogenesis-related proteins) decreased significantly after NDUFS2 KD, while the level of GSNOR, a protein denitrosylase, was reduced by NDUFS2 KD after 48 h of aging. These data suggest that NDUFS2 is vital for maintaining the oocyte quality during POA in pigs.

Study of the population genetic structure of Opisthorchis-like eggs in northern Thailand using mitochondrial genes.

Opisthorchis-like eggs are a public health problem in northern and northeastern Thailand. However, the genetic epidemiology and structure of these parasites in northern Thailand are unknown. Thus, this study investigated their population genetic structure using cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1) nucleotide sequences. A study was conducted in the hill tribe regions of Chiang Mai Province, northern Thailand. Internal transcribed spacer 2 polymerase chain reaction and restriction fragment length polymorphism were used to distinguish 205 positive feces samples for Opisthorchis-like eggs. The results showed that the prevalence of O. viverrini and Haplorchis taichui was 10.5% and 38.2%, respectively, and the co-infection rate was 37.2%. To determine the genetic structure of O. viverrini and H. taichui using cox1 and nad1 genes, genetic analysis was performed using 30 randomly chosen fecal samples for Opisthorchis-like eggs. Pairwise FST analysis indicated that O. viverrini and H. taichui displayed nonsignificant genetic differentiation within Chiang Mai Province and between interpopulations from different geographic areas. Moreover, within the intrapopulation in Chiang Mai Province,cox1 presented higher gene flow than nad1 in O. viverrini, while nad1 demonstrated higher gene flow than cox1 in H. taichui. The neutrality tests based on Fu's Fs indicated population expansion and selective sweep from bottleneck or hitchhiking in O. viverrini and H. taichui populations, supported by haplotype network patterns. Phylogenetic tree analysis based on cox1 and nad1 revealed the monophyly of O. viverrini and H. taichui and genetic relationships with other isolates collected from Thailand, Lao People's Democratic Republic (PDR), and Vietnam. This study investigated the molecular discrimination and genetic structure of Opisthorchis-like eggs in northern Thailand. The genetic information derived from this study could be associated with the background, molecular epidemiology, and disease severity of these parasites.

A new species of the newt genus Hypselotriton (Amphibia, Urodela, Salamandridae) from Jiangxi Province, southeastern China.

A new newt species, Hypselotritonhuanggangensis sp. nov., is described based on nine specimens collected from Huanggangshan Mountains, Yanshan County, Jiangxi, China. Morphologically, the new species is characterized by the combination of nine external characters: (1) obvious black patches with clear boundaries on the whole body; (2) ground color of the dorsal body tan; (3) ground color of venter bright orange; (4) skin rough; (5) vertebral ridge weak; (6) fingers and toes overlapping when forelimb and hindlimb adpressed towards each other along body; (7) postocular orange spot absent; (8) small white warty glands around the eye; (9) two discontinuous longitudinal lines formed by white warty glands from neck to lateral parts of tail. Molecularly, the new species forms an independent clade with strong support in the phylogenetic trees of the genus based on the mitochondrial locus of NADH dehydrogenase subunit 2 (ND2) gene fragments. The new species distinctly differs from H.fudingensis by differences in its body measurements, vertebral ridge, dorsal black patches, and ventral black patches. Furthermore, the new species and H.fudingensis are geographically isolated by a series of high mountain ranges, including the Wuyishan and Jiufengshan Mountains. The number of Hypselotriton species is now 11.

Integrating Genetic and Cytogenetic Data: A Diversity Study of Astyanax and Psalidodon (Characidae) Species from the Paraná River Basin.

Astyanax is one of the most specious fish groups in the Neotropical region, with many cryptic species, which represents a challenge for correct identification through traditional taxonomic methods. Psalidodon is a recently resurrected genus group of species previously belonging to Astyanax, specifically those with extensive chromosomal variation of the A. scabripinnis and fasciatus complexes. In the present study, the mitochondrial genes cytochrome c oxidase subunit 1 (COI), mitochondrial ATP synthase 6 and 8 (ATPase 6/8), and NADH dehydrogenase subunit 2 (ND2) were used in conjunction with chromosomal data to characterize molecularly and cytogenetically populations of Astyanax and Psalidodon from rivers and streams of the Ivaí River Basin (Paraná Basin). The results demonstrated the effectiveness of the integrative use of molecular and cytogenetic techniques, with the confirmation of at least three species for the sampled sites: A. lacustris, P. paranae, and P. fasciatus, which showed inter- and intrapopulation karyotype variations. In addition, extensive haplotypic variation can be observed for these species within the Ivaí River Basin and throughout the Paraná River Basin. The data demonstrate a hidden diversity among the species analyzed, enrich the ichthyofaunistic knowledge of small rivers and streams, and contribute to future conservation projects in these areas.

Gene expression analyses of the ovaries in black tiger shrimp (Penaeus monodon) for reproductive trait using reference genome

The black tiger shrimp (Penaeus monodon) is widely cultured in Vietnam and worldwide. Broodstock source for post-larval production still relies on the wild shrimps due to their higher reproductive quality in comparison with the domesticated broodstock. The aims of the current study were to analyse differential gene expression in the ovaries of black tiger shrimp between the stages of reproductive maturation using the reference genome-based approaches and to compare the results generated by these approaches. cDNAs of the ovarian samples were sequenced using the Illumina platform. Genome-guided assembly (GGA) was employed to generate 71,678 contigs with an N50 length of 1,587 bp. For gene expression analysis, the newly built-transcriptome-based approach identified a total number of 288 differentially expressed genes (DEGs) when the samples between the non-mature and mature stages were compared. The gene expression analysis using the reference mapping approach resulted in only 49 DEGs. The analyses indicated that many genes involved in the reproductive maturation were up-regulated, including vitellogenin, NADH dehydrogenase subunit 2, and anti-lipopolysaccharide factor-like. Different results of gene expression from both approaches are discussed.