A recent study published in Oryx proposed that the extinct Javan tiger Panthera tigris sondaica may still survive on the Island of Java, Indonesia, based on mitochondrial DNA analysis of a single hair sample collected from a location where a tiger was reportedly encountered. However, upon reanalysing the genetic data presented in that study, we conclude that there is little support for this claim. The sequences of the putative tiger hair and Javan tiger museum specimens generated are not from tiger cytoplasmic mitochondrial DNA but more likely the nuclear pseudogene copies of mitochondrial DNA. In addition, the number of mismatches between the two Javan tiger sequences is unusually high for homologous sequences that are both from tigers, suggesting potential issues with data reliability. The paper provides insufficient details on quality control measures, making it impossible to rule out the possibility that errors were introduced during the analysis. Consequently, it is inappropriate to use the sequences presented in that study to infer the existence of the Javan tiger.

Species in the genus Engraulis show extensive intraspecific as well as interspecific morphological and genetic diversity. Since morphological differences do not necessarily correspond to genetic differences, it is necessary to clarify the relationship between morphological differences and genetic differences for a better understanding of the population structure. Fish morphology at a given standard length differs between cohorts of Japanese anchovy Engraulis japonicus during the early life stages in the Kii Channel, but it is unknown whether the differences are caused by genetic differences or not. The Kii Channel includes the boundary between the Pacific (southern side of the Kii Channel) and the Seto Inland Sea stocks (northern side), but stock separation is based primarily on demographic characteristics. In the present study, genetic analyses were conducted to examine the genetic differences among samples (month and area) based on mitochondrial DNA cytochrome b region (Cyt b), control region (CR) and microsatellite markers. AMOVA revealed that the percentage of genetic variation among samples was low at 0.11% for Cyt b, 0.30% for CR and 0.00% for microsatellite, and no significant genetic variation was observed among samples. Although two clades were identified in the unrooted neighbour-joining tree for Cyt b and CR, both Cyt b and CR sequences were similar between months and between areas. Accordingly, the morphological differences among cohorts can be attributed to phenotypic plasticity. Additionally, there were no genetic differences between samples from the southern side and the northern side of the Kii Channel, suggesting strong genetic connectivity in these areas.

Recent development in immunotherapy for cancer treatment has substantiated to be more effective than most of the other treatments. Immunity is the first line of defence of the body; nevertheless, cancerous cells can manipulate immunity compartments to play several roles in tumour progression. Tumour-associated macrophages (TAMs), one of the most dominant components in the tumour microenvironment, are recognized as anti-tumour suppressors. Unfortunately, the complete behaviour of TAMs is still unclear and understudied. TAM density is directly correlated with the progression and poor prognosis of hepatocellular carcinoma (HCC), therefore studying TAMs from different points of view passing by all the factors that may affect its existence, polarization, functions and repolarization are of great importance. Different epigenetic regulations were reported to have a direct relation with both HCC and TAMs. Here, this review discusses different epigenetic regulations that can affect TAMs in HCC whether positively or negatively.

During a 1-year study, Trichuris adults were obtained after necropsy of Arabian camels (Camelus dromedarius) from a slaughterhouse in Kuwait. Morphological and molecular identification was performed to confirm the identity of the Trichuris specimens obtained from C. dromedarius. Fifteen male Trichuris specimens were selected, and molecular identification was performed using mitochondrial cytochrome c oxidase subunit I, 12S ribosomal RNA, 16S ribosomal RNA genes and the nuclear internal transcribed spacer 2 (ITS2) region. Through phylogenetic analysis, 2 distinct groups were obtained using the mitochondrial genes, where group 1 showed a close relationship to Trichuris globulosa while group 2 showed a close relationship to Trichuris ovis, providing molecular evidence of a possible T. globulosa species complex. Additionally, the nuclear ITS2 region did not provide enough resolution to distinguish between the 2 groups of Trichuris specimens. Observation of morphological characters revealed variations in the shape of the male spicule sheath, where specimens present either a globular posteriorly truncated swelling or the absence of posteriorly truncated swelling. Moreover, the variations in male spicule sheath does not corroborate with the results of molecular data, suggesting the limited use of this character for identification of T. globulosa. In conclusion, molecular analysis suggests a possible species complex in T. globulosa, with the mitochondrial genetic markers successfully differentiating between the 2 groups. The limited use of the male spicule sheath as a diagnostic character for identification of T. globulosa is suggested.

This study aimed to investigate the structural and metabolic changes in cumulus cells of underweight women and their effects on oocyte maturation and fertilization. The cytoplasmic ultrastructure was analyzed by electron microscopy, mitochondrial membrane potential by immunofluorescence, and mitochondrial DNA copy number by relative quantitative polymerase chain reaction. The expression of various proteins including the oxidative stress-derived product 4-hydroxynonenal (4-HNE) and autophagy and apoptosis markers such as Vps34, Atg-5, Beclin 1, Lc3-I, II, Bax, and Bcl-2 was assessed and compared between groups. Oocyte maturation and fertilization rates were lower in underweight women (P < 0.05), who presented with cumulus cells showing abnormal mitochondrial morphology and increased cell autophagy. Compared with the mitochondrial DNA copies of the control group, those of the underweight group increased but not significantly. The mitochondrial membrane potential was similar between the groups (P = 0.8). Vps34, Atg-5, Lc3-II, Bax, and Bcl-2 expression and 4-HNE levels were higher in the underweight group compared with the control group (P < 0.01); however, the Bax/Bcl-2 ratio was lower in the underweight group compared with the control group (P = 0.031). Additionally, Beclin 1 protein levels were higher in the underweight group compared with the control group but without statistical significance. In conclusion, malnutrition and other conditions in underweight women may adversely affect ovulation, and the development, and fertilization of oocytes resulting from changes to the intracellular structure of cumulus cells and metabolic processes. These changes may lead to reduced fertility or unsatisfactory reproduction outcomes in women.

Squids of the family Gonatidae are key components in oceanic communities. However, issues related to correct species identification, number of species, and their genetic relatedness remain. To address these issues, sequences from three mitochondrial (cytochrome c oxidase subunit I [CO1], 16S rRNA, and 12S rRNA) and two nuclear (18S and 28S) genes were analysed in the Gonatidae. Four of the five sequences (12S rRNA, 16S rRNA, 28S, and CO1) yielded rather similar patterns of genetic relationships among the species. Molecular evidence suggested intra-familial subdivision into two major groups of species having either five or seven longitudinal rows of teeth in the radula. The former group included all species of the genus Gonatus and two sister-species of Gonatopsis s. str. suggesting that all gonatid species with five rows of radular teeth represent a single taxonomic unit of a genus or subfamily level. Species with seven rows of radular teeth formed several ‘species’ clusters. Sequence analysis also addressed species identification issues in the Gonatidae. Two genetically divergent groups were found among squid which conformed to the description of Gonatus berryi. Molecular evidence suggested sister-species relationships between ‘large’ and ‘small’ forms of Boreoteuthis borealis with size-at-maturity as the only reported difference between these two cohorts. Sequence variation was observed within Gonatus pyros. Inclusion of gonatid sequences from the GenBank into the analysis suggested probable species misidentification in several cases. Combined use of several mitochondrial and nuclear sequences served as a valuable tool for species identification and provided a solid background for unravelling molecular genetic and taxonomic relationships in the Gonatidae.

Cystic echinococcosis (CE) is considered the most severe parasitic disease that ever affected the human population in Iceland. Before the start of eradication campaign in the 1860s, Iceland was a country with very high prevalence of human CE, with approximately every fifth person infected. Eradication of CE from Iceland by 1979 was a huge success story and served as a leading example for other countries on how to combat such a severe One Health problem. However, there is no genetic information on Echinococcus parasites before eradication. Here, we reveal the genetic identity for one of the last Echinococcus isolates in Iceland, obtained from a sheep 46 years ago (1977). We sequenced a large portion of the mitochondrial genome (8141 bp) and identified the isolate as Echinococcus granulosus sensu stricto genotype G1. As G1 is known to be highly infective genotype to humans, it may partly explain why such a large proportion of human population in Iceland was infected at a time . The study demonstrates that decades-old samples hold significant potential to uncover genetic identities of parasites in the past.

Echinococcus granulosus sensu lato is a group of tapeworm species known to cause cystic echinococcosis. Within this group, the Echinococcus canadensis cluster includes genotypes G8 and G10 that have a predominantly sylvatic life cycle – transmission occurs between wild cervids and wolves. Relatively few studies have explored the genetic variation of the elusive G8 and G10, and their extent of genetic variation is yet to be investigated at the complete mitochondrial (mt) genome level. The aim was to explore the genetic variation of these 2 genotypes in Europe using complete mtDNA sequences and provide a high-quality reference dataset for future studies. Sequences of complete mt genomes were produced for 29 samples of genotype G8 and G10 from wolves, moose, reindeer and roe deer, originating from Finland, Sweden, Russia, Poland, Latvia and Estonia. Genetic variation was explored based on phylogenetic network analysis, revealing marked differences between G8 and G10 (over 400 mutations), and more detailed patterns of variability within the 2 genotypes than previously observed. Understanding the mt genetic composition of a species provides a baseline for future studies aiming to understand whether this mt distinctiveness is mirrored in the nuclear genome and whether it has any impact on any phenotypic traits or parasite transmission.

Phylogenetic analyses can be used to resolve taxonomic uncertainties and reconstruct a species’ evolutionary history. This can be combined with ecological data to predict missing life history traits which are important for creation of conservation management strategies. We investigated the evolutionary and life history of the ‘Critically Endangered’ Grenada Dove Leptotila wellsi by estimating its phylogenetic placement and using this new phylogeny to test the accuracy of phylogenetic comparative methods for estimating both documented and unknown life history traits. We extracted DNA from two Grenada Dove samples and obtained sequences from three mitochondrial markers: Cytochrome oxidase I (COI), NADH dehydrogenase 2 (ND2) and Cytochrome b (Cyt b); and one nuclear marker: β-Fibrinogen intron 7 (β-FIB). We present the first genetic data obtained for the Grenada Dove. Our data identify the Grey-Chested Dove Leptotila cassinii as the species which shares both a most recent common ancestor, with an estimated divergence of approximately 2.53 million years ago, and the smallest genetic distance (P = 0.0303) with the Grenada Dove. Life history trait values for the Grenada Dove predicted from our analyses using phylogenetic imputation are: clutch size = 2 (± 0.09) eggs; clutches per year = 1.4 (± 0.81); incubation time = 14.2 (± 0.75) days; hatching weight = 3.8 g (± 1.05) and single imputation: fledging age (genus median) = 15.5 days, longevity (genus median) = 8.6 years. This study contributes novel information regarding evolutionary history and life history characteristics to inform long-term conservation actions for a ‘Critically Endangered’ species.

DNA barcoding based on the cytochrome c oxidase subunit I (COI) gene in mitochondrial DNA has been adopted as a global biological identification system for animals due to its accuracy compared with other classical taxonomic methods. The objective of this study was to establish a reference library with generated barcodes. A total of 84 fish specimens belonging to 37 commercially important marine fish species, representing five orders, 14 families and 30 genera, were sampled along the Tunisian coast and barcoded for the first time, obtaining 637 bp sequences. The average Kimura 2-parameter (K2P) distances within species, genera and families were 0.52, 6.86 and 14.60%, respectively. The Maximum likelihood (ML) tree revealed distinct clusters in concurrence with the taxonomic status of the species. Our results confirmed the authentication of the barcode approach for the identification of the species examined and provide valuable information that would help ichthyologists to achieve better monitoring, conservation and management of fisheries in Tunisia.

We obtained new data on the complete mitochondrial DNA (mtDNA) and the ribosomal operon of the trematode Carassotrema koreanum (Digenea: Haploporata: Haploporidae), an intestinal parasite of Carassius auratus, using next-generation sequencing. The mtDNA of C. koreanum contained 13,965 bp, including 12 protein-coding genes, two ribosomal genes, 22 transport RNA (tRNA) genes and a non-coding region. The ribosomal operon of C. koreanum was 10,644 bp in length, including ETS1 (1449 bp), 18S ribosomal RNA (rRNA) gene (1988 bp), ITS1 ribosomal DNA (rDNA) (558 bp), 5.8S rRNA gene (157 bp), ITS2 rDNA (274 bp), 28S rRNA gene (4152 bp) and ETS2 (2066 bp). Phylogenetic analysis based on mtDNA protein-coding regions showed that C. koreanum was closely related to Parasaccocoelium mugili, a species from the same suborder Haploporata. Bayesian phylogenetic tree topology was the most reliable and confirmed the validity of the Haploporata. The results of sequence cluster analysis based on codon usage bias demonstrated some agreement with the results of the phylogenetic analysis. In particular, Schistosoma spp. were differentiated from the other members of Digenea and the members of Pronocephalata were localized within the same cluster. Carassotrema koreanum and P. mugili fell within different clusters. The grouping of C. koreanum and P. mugili within the same cluster was obtained on the basis of frequencies of 13 specified codons, of which three codon pairs were degenerate. A similarity was found between two haploporid species and two Dicrocoelium spp. in the presence of TTG start codon of the mitochondrial nad5 gene. Our results confirmed the taxonomical status of the Haploporata identified in the previous studies and revealed some characteristic features of the codon usage in its representatives.

Ascaridia galli (Nematoda: Ascaridiidae) is the most common intestinal roundworm of chickens and other birds with a worldwide distribution. Although A. galli has been extensively studied, knowledge of the genetic variation of this parasite in detail is still insufficient. The present study examined genetic variation in the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene among A. galli isolates (n = 26) from domestic chickens in Hunan Province, China. A portion of the cox1 (pcox1) gene was amplified by polymerase chain reaction separately from adult A. galli individuals and the amplicons were subjected to sequencing from both directions. The length of the sequences of pcox1 is 441 bp. Although the intra-specific sequence variation within A. galli is 0–7.7%, the inter-specific sequence differences among other members of the infraorder Ascaridomorpha were 11.4–18.9%. Phylogenetic analyses based on the maximum likelihood method using the sequences of pcox1 confirmed that all of the Ascaridia isolates were A. galli, and also resolved three distinct clades. Taken together, the findings suggest that A. galli may represent a complex of cryptic species. Our results provide an additional genetic marker for the management of A. galli in chickens and other birds.

Dibothriocephalus latus is the most frequent causative agent of fish-borne zoonosis (diphyllobothriosis) in Europe, where it is currently circulating mainly in the Alpine lakes region (ALR) and Russia. Three mitochondrial genes (cox1, cob and nad3) and 6 microsatellite loci were analysed to determine how is the recently detected triploidy/parthenogenesis in tapeworms from ALR displayed at the DNA level. A geographically distant population from the Krasnoyarsk Reservoir in Russia (RU-KR) was analysed as a comparative population. One or 2 alleles of each microsatellite locus was detected in plerocercoids from RU-KR, corresponding to the microsatellite pattern of a diploid organism. In contrast, 1–3 alleles were observed in tapeworms from ALR, in accordance with their triploidy. The high diversity of mitochondrial haplotypes in D. latus from RU-KR implied an original and relatively stable population, but the identical structure of mitochondrial genes of tapeworms from ALR was probably a consequence of a bottleneck typical of introduced populations. These results indicated that the diploid/sexually reproducing population from RU-KR was ancestral, located within the centre of the distribution of the species, and the triploid/parthenogenetically reproducing subalpine population was at the margin of the distribution. The current study revealed the allelic structure of the microsatellite loci in the triploid tapeworm for the first time.

We report the results of bioarchaeological, genetic, malacological, and lithic analyses of a burial located in the Calamuchita Valley in the province of Córdoba in central Argentina. We discuss these findings from an osteobiographical perspective that considers the physical body from three spheres of analysis: biological, social, and political. The skeletal remains correspond to an adult male who died 3770 ± 90 14C years BP (4404–3850 cal BP). His mitochondrial lineage A2+16192-16248 could have originated in central Argentina; this lineage subsequently became extinct in modern populations. In association with the human bones, there were two lithic tools, ornamental shells, and a stone pendant arranged as a necklace. This type of context is infrequently found in the region and the period studied. Thus, this burial represents a rare and relevant record for regional and South American archaeology, providing the oldest direct evidence of a burial with grave goods in hunter-gatherer societies of the province of Córdoba, central region of Argentina.

Dietary l-carnitine (LC) is a nutritional factor that reduces liver lipid content. However, whether dietary LC can improve lipid metabolism via simultaneous activation of mitochondrial fatty acid (FA) β-oxidation and suppression of endoplasmic reticulum (ER) stress is still unknown. Large yellow croaker were fed with a high-fat diet (HFD) supplemented with dietary LC at 0, 1·2 or 2·4 ‰ for 10 weeks. The results indicated that a HFD supplemented with LC reduced the liver total lipid and TAG content and improved serum lipid profiles. LC supplementation administered to this fish increased the liver antioxidant capacity by decreasing serum and liver malondialdehyde levels and enhancing the liver antioxidant capacity, which then relieved the liver damage. Dietary LC increased the ATP dynamic process and mitochondrial number, decreased mitochondrial DNA damage and enhanced the protein expression of mitochondrial β-oxidation, biogenesis and mitophagy. Furthermore, dietary LC supplementation increased the expression of genes and proteins related to peroxisomal β-oxidation and biogenesis. Interestingly, feeding fish with LC-enriched diets decreased the protein levels indicative of ER stress, such as glucose-regulated protein 78, p-eukaryotic translational initiation factor 2a and activating transcription factor 6. Dietary LC supplementation downregulated mRNA expression relative to FA synthesis, reduced liver lipid and relieved liver damage through regulating β-oxidation and biogenesis of mitochondria and peroxisomes, as well as the ER stress pathway in fish fed with HFD. The present study provides the first evidence that dietary LC can improve lipid metabolism via simultaneously promoting FA β-oxidation capability and suppressing the ER stress pathway in fish.