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ABSTRACT Objective Methods Results Conclusion The aim of this study is to explore the mechanism of benzylurea in the inflammatory injury of human periodontal ligament fibroblasts (hPDLFs).An inflammation model of hPDLFs was established using LPS. Nuclear transport of nuclear transcription factor‐κB (NF‐κB), secretion of cytokines, and the morphology and distribution of F‐actin were determined. Mitochondrial function was assessed by measuring mitochondrial membrane potential (MMP), mitochondrial permeability transition pore (mPTP), and reactive oxygen species (ROS) levels. The expression of mitochondrial carrier homolog 2 (MTCH2) and Cytochrome b5 type B (CYB5B) was detected.Benzylurea alleviated the effects of lipopolysaccharide (LPS) on the proliferation and apoptosis of hPDLFs. It reduced the release of inflammatory cytokines and inhibited NF‐κB nuclear translocation. Benzylurea improved mitochondrial function by regulating MMP and preventing excessive mPTP opening. Furthermore, LPS elevated the expression of MTCH2 and reduced the expression of CYB5B in hPDLFs. However, these effects can be inhibited by benzylurea. The altered expression of MTCH2 directly affected CYB5B expression, the release of inflammatory cytokines, and the activation of nuclear translocation of NF‐κB.CYB5B may act as an effector of MTCH2, with benzylurea enhancing mitochondrial function and protecting hPDLFs from LPS‐induced injury through MTCH2. [ABSTRACT FROM AUTHOR]

The Silver-backed Chevrotain (Tragulus versicolor), a mammal endemic to the unique coastal arid ecosystem in south-central Vietnam, is an enigmatic mouse deer with little information available about its ecology and evolutionary history. The recent rediscovery of the species provides a rare opportunity to clarify its taxonomy and phylogenetic relationship within the genus Tragulus. In this study, we sequenced the complete mitochondrial cytochrome b gene for samples of T. versicolor and T. kanchil from Vietnam. Our molecular results show that the species forms a distinct evolutionary lineage sister to all other species in the genus, confirming the earlier hypothesis based on morphological evidence. However, further research is needed to assess the genetic diversity of the species and design appropriate conservation measures. As the species has not been secured over much of its range, it is recommended that the population in Nui Chua National Park, the only one covered by a protected area, be prioritized for conservation actions. Conserving the species in this unique and imperiled ecosystem will also benefits a suite of locally endemic and threatened species of fauna and flora. [ABSTRACT FROM AUTHOR]

Accurate information on species range contraction is the cornerstone of effective biodiversity conservation. The eastern indigo snake (Drymarchon couperi) is an apex predator in Florida and, similar to many species native to Florida, is threatened by widespread habitat destruction. Environmental deoxyribonucleic acid (eDNA) monitoring of this elusive snake would provide a non‐invasive approach to improve our knowledge of the species' range and distribution. We designed and tested an eDNA assay that can detect the presence of D. couperi from soil samples from their natural scrub habitat in Florida. We validated our assay in silico, in vitro, and in situ. Furthermore, we investigated the influence of temperature and humidity on the degradation rate of eDNA over time. We successfully amplified the cytochrome b gene for D. couperi at concentrations as low as 3 × 10−3 ng/μL and successfully detected the presence of D. couperi in 2 of 30 in situ field soil samples. The degradation experiment resulted in detectable DNA for 10 days. Interestingly, temperature and humidity had no effect on the degradation rate of eDNA in our experimental conditions. This study provides support for soil eDNA applications to detect the presence of a federally threatened species in their natural environment bolstering our ability to monitor the conservation and management of imperiled species. Environmental DNA provides an additional conservation tool to quickly and effectively monitor species range shifts driven by multiple anthropogenic stressors to promote the persistence of imperiled species. [ABSTRACT FROM AUTHOR]

Thomasomys is the most diverse genus of the Sigmodontinae subfamily, comprising at least 51 species. Despite recent systematic advances, the monophyly of this genus, the proposed species groups and their content, and the species limits are not yet firmly established. Using a well‐sampled mitochondrial dataset, we aim to test the monophyly of the genus, infer phylogenetic relationships among species and species groups and test the limits between valid species and candidate species. For this, we used a large matrix of 272 partial non‐redundant sequences of the cytochrome b mitochondrial gene from 40 valid and 20 candidate species. Using probabilistic approaches on this dataset, we inferred Thomasomys phylogenetic relationships and explored species boundaries using four unilocus species delimitation methods (ABGD, ASAP, bPTP and GMYC). Thomasomys sensu stricto was recovered monophyletic and well‐supported, excluding a Peruvian lineage misidentified as part of the genus. Analyses consistently recovered 10 well‐supported major clades and several paraphyletic or polyphyletic species. Delimitation methods and genetic divergences estimated that Thomasomys comprises between 81 and 93 putative species, thus potentially doubling the species diversity of Thomasomys. Such a result clearly calls for an urgent taxonomic revision of this genus and the use of further molecular loci within an integrative taxonomic approach to describe its diversity and understand its evolutionary history. [ABSTRACT FROM AUTHOR]

Background: Sand flies, belonging to the Psychodidae family, represent small, hairy insects that serve as significant vectors in various important medical and veterinary diseases. Despite being recognized by the World Health Organization as an endemic area for leishmaniasis, Southeast Asia lacks comprehensive information on the species composition and biology of sand flies. To address this, the current study aimed to survey sand fly biodiversity. Methods: Sand flies from six provinces in Southern Vietnam were collected using CDC light traps. Sand flies were subsequently identified morphologically and confirmed molecularly using mitochondrial cytochrome oxidase c subunit I (COI) and cytochrome b (cytb) sequences. BLASTN searches were conducted, and the species identity of sand flies was further confirmed through a Barcode of Life Database (BOLD) search utilizing COI sequences. Subsequently, nucleotide sequences were subjected to a panel of analyses including intraspecific variation, phylogenetic relationships and haplotype network. The average densities of collected sand flies (sand flies/trap/night) and species richness were also recorded. Results: A total of 753 sand flies were collected. After excluding damaged specimens, six sand fly species, namely Phlebotomus stantoni, Sergentomyia khawi, Se. silvatica, Se. barraudi, Se. bailyi and Grassomyia indica, were identified. All conspecific sand fly sequences, including Ph. stantoni, Se. barraudi, Gr. indica, Se. bailyi, Se. khawi and Se. silvatica, clustered with their reference sequences, corroborating the results of morphology-based identification, BLASTN analysis and BOLD search. For intraspecific variation of sand flies obtained from the current study, COI diversity indices were consistently higher than those of cytb. Conclusions: This study provides the first updates on morphological and molecular characterization of sand flies in Southern Vietnam. This acquired knowledge on sand fly species composition is essential for controlling sand fly-borne diseases in this potentially endemic region. [ABSTRACT FROM AUTHOR]

The authentication of milk and dairy products has great significance for food fraud. The present investigation entailed the development of a novel method that amalgamates the double-tube approach with multiplex real-time polymerase chain reaction (PCR) technology, incorporating TaqMan probes, to facilitate the high-throughput screening and detection of animal-derived constituents within milk and dairy products. Eight dairy-derived animal-specific primers and probes were designed for the mitochondrial cytochrome b (Cytb) gene of eight dairy products, including cow, buffalo, yak, goat, sheep, horse, donkey, and camel. Through the developed double-tube detection assays, the above eight targets could be simultaneously identified with a detection limit of 0.00128–0.0064 ng/μL. The multiplex qPCR assay was effectively validated using simulated adulterated samples with different mixing ratios and demonstrated a detection limit of 0.1%. Upon analysis of 54 commercially available dairy products, a mislabeling rate of 33% was revealed. This method affords an efficacious means of detecting dairy product ingredients, thereby offering robust technical backing for market oversight and regulatory enforcement of milk and dairy products. [ABSTRACT FROM AUTHOR]

The distribution of species and the way that lineages are structured are the result of intrinsic historical processes of the species and their relationships with landscape features. Paleosuchus palpebrosus is one of the smallest crocodilians in the world and has a wide geographic distribution in South America, occurring in different habitats. Here, we analyzed mitochondrial cytochrome b sequences of 227 individuals, investigated how elements of landscape heterogeneity impact genetic differentiation, and evaluated the evolutionary and phylogeographic history of the species. Thus, we hypothesized that landscape structures, such as aridity and slope, should act as resistance surfaces to population connectivity, while rivers could mediate the dispersion of the species as a conductor of the gene flow. The analyses recovered three main lineages of P. palpebrosus. While most of the observed genetic variation was explained by geographic resistance distance, river connectivity had a smaller contribution for the observed variation. We also found a recent history with limited genetic divergence throughout the wide distribution of the species. Our findings highlight the main drivers for the evolutionary history of the species and how landscape features can shape the diversification, especially if we consider rivers as a facilitator of gene flow. [ABSTRACT FROM AUTHOR]

Clarifying the genetic structure and population history of a species can reveal the impacts of historical climate and geological changes, providing critical insights for developing effective conservation strategies for ecologically significant fish. The Markakol grayling (Thymallus brevicephalus), an endangered species found in the Altai‐Sayan Mountain region of Central Asia, serves as an ideal model for studying these factors. In this study, populations of a grayling (Thymallus) species discovered in the upper Irtysh River headwaters in Xinjiang, China, were analyzed to assess genetic diversity and population structure. Mitochondrial DNA sequences (cytochrome b and control region), along with 10 microsatellite markers, were used to examine genetic variation. Phylogenetic and genetic distance analyses confirmed the species, long misidentified as Arctic grayling (T. arcticus), as T. brevicephalus. This species can be divided into two distinct geographic groups: eastern and western, with the Crane River acting as the boundary. The divergence between these groups likely corresponds to refugia formed during the Pleistocene glaciation of the Altai Mountains, approximately 0.48 MA (million years ago) (range: 0.30 to 0.71 Ma). High haplotype diversity (Hd > 0.5) and low nucleotide diversity (π < 0.005) suggest that, despite the species' genetic richness, T. brevicephalus remains vulnerable to genetic drift, which could threaten its long‐term survival. This vulnerability may stem from inbreeding within small refugial populations during the glacial period, followed by gradual population expansion. Our study offers novel insights into grayling populations, with results that have direct implications for management by serving as a tool for the identification of conservation units. This study reveals the impact of past climate events on a fish species (Thymallus brevicephalus) in China. Genetic analysis suggests the species split into eastern and western populations during a past glaciation event. While the fish show genetic diversity, specific management plans are needed to conserve both eastern and western populations due to their unique evolutionary histories. [ABSTRACT FROM AUTHOR]

Diabetes mellitus is a metabolic disease characterized by hyperglycemia. Type 2 diabetes mellitus patients are known to have mutations in their mitochondrial DNA (mtDNA). mtDNA easily undergoes mutations because it does not have histone proteins for protection and lacks a proofreading mechanism during its replication. The mtDNA mutations T15663C found in type 2 diabetes mellitus patients sufferers with a cataract disease. This mutation was found to occur in the cytochrome b gene, one of the gene in the complex III respiratory chain. In this study we investigate the effect of this mutation on the structure of cytochrome b using molecular dynamics simulations using PDB ID: 5XTE as a template and carrying out simulations for 250 ns. The models of wild type and mutant protein structure were constructed using CHARMM-Gui so that, the protein was looked like in the mitochondrial membrane. The results of the molecular dynamics simulation show that the potential energy of the wild type model is lower than the mutant model, the RMSD value of the wild type model is more stable at 114 - 180 ns, and the RMSF value in the wild type model does not fluctuate as much as the mutant model. In addition, the hydrogen bond (H bond) analysis of the mutant model has a higher H bond number than the wild type model, which means that the mutant model is more rigid. The simulation results were visualized using VMD, and new hydrogen bonds were found in the mutant model. [ABSTRACT FROM AUTHOR]

Simple Summary: The taxonomy of the sinica group of macaques has been unresolved due to inconsistencies between their physical traits and genetic relationships. To clarify this taxonomic issue, we analyzed DNA from previously unsampled populations of the macaques in the Himalayas. Our results revealed that the sinica group consists of seven distinct species, rather than the previously considered six species as the two subspecies of Assamese macaques (Macaca assamensis assamensis and M. a. pelops) are separate candidate species with strong genetic variations. The study also identified Arunachal macaques (M. munzala) in the Shannan area of Xizang Zizhiqu in China. Multiple analyses revealed complex historical patterns of species dispersal and separation, mostly linked to climatic changes during the Quaternary period. We propose a new hypothesis about the sinica group's evolutionary history. Owing to the taxonomic incongruence between the morphological features and genetic relationships of the sinica group of macaques (genus Macaca), the taxonomy of this macaque group has remained inconclusive. We aimed to resolve the taxonomic quandary and improve our understanding of the historical biogeography of the group by including macaque DNA samples from previously unsampled areas in the Himalayas. We sequenced and analyzed three mitochondrial DNA loci [cytochrome b (CYTB), cytochrome oxidase subunit 1 (COI) and D-loop; 2898 bp] for sequence polymorphism, phylogenetics, species delimitation, and ancestral area reconstruction. We confirmed the occurrence of Arunachal macaque (Macaca munzala) on the southern slopes of the Eastern Himalayas in the Xizang Zizhiqu (Tibet Autonomous Region) of China. The results revealed that the sinica group of macaques is a parapatric species group composed of seven distinct species. Phylogenetic and species delimitation analyses revealed that the two previously considered subspecies of Assamese macaques (the eastern subspecies M. assamensis assamensis and the western subspecies M. a. pelops) are two distinct species. The eastern Assamese macaque is a sister species to the Tibetan macaque, whereas the western Assamese macaque and Arunachal macaque are the closest genetic sister species. The sinica group of macaques underwent five vicariance and seven dispersal radiations in the past, which mainly coincided with the Quaternary climatic oscillations between the late Pliocene and the late Pleistocene. By integrating our phylogenetic and ancestral area reconstruction results with findings from previous paleontological and molecular studies, we propose a robust hypothesis about the phylogeography of the sinica group of macaques. [ABSTRACT FROM AUTHOR]

Simple Summary: Among the pests infesting the honeybee Apis mellifera, the parasitoid flies Megaselia scalaris and Senotainia tricuspis are not yet well characterized for the occurrence and harmfulness to bee colonies. These flies can attack the forager bee and lay eggs or larvae on its body, which will develop by feeding on the host tissues until it is killed. Their prevalence in different countries is still not well defined, so, in this study, rapid molecular methods were developed to facilitate their detection. The new methods were applied in a preliminary analysis of adult bees and hive debris from parts of Central Italy and the Republic of Kosovo. Both flies were detected in the two countries and more frequently in Italy. The M. scalaris flies isolated from Italian apiaries were genotypically related to biotypes from distant countries. Single flies were shown to possibly transmit honeybee microbial pathogens, so their presence should be efficiently contrasted in apiaries. The Megaselia scalaris and Senotainia tricuspis parasitoid flies of the honeybee Apis mellifera were found to infest apiaries of different European and Mediterranean countries but their prevalence and impact on apiary health are little known. Therefore, in this study, quantitative PCR (qPCR)-based methods were developed for their rapid detection directly in hive matrices. The newly developed qPCR assays were targeted at the mitochondrial cytochrome oxidase subunit I (COI) gene for the M. scalaris and the cytochrome B (cytB) gene for the S. tricuspis. The tests were preliminarily applied to 64 samples of adult honeybees and hive debris collected in the Abruzzo and Molise regions, Central Italy, and the Republic of Kosovo showing that both flies occur in the two countries and more frequently in Italy. The positive apiaries in Italy were re-sampled by capturing viable forager bees and isolating emerging flies to carry out the genotyping and analyses aimed at defining if these flies can transmit honeybee pathogens. Genotyping based on the COI and cytB gene sequencing for M. scalaris and S. tricuspis, respectively, identified one S. tricuspis genotype and diverse genotypes of M. scalaris highly similar to those from distant countries. Some fly isolates harbored the DNA or RNA of honeybee microbial pathogens Paenibacillus larvae, deformed wing viruses A and B (DWVA and B), black queen cell virus (BQCV), chronic paralysis virus (CBPV), and Nosema ceranae. The results indicated that these parasites should be efficiently controlled in apiaries by using rapid detection methods to facilitate the large screening studies and early detection. [ABSTRACT FROM AUTHOR]

Here we present, through field collections conducted via monitoring with Sherman traps and phylogenetic analyses of the mitochondrial cytochrome b gene obtained through Sanger sequencing, records worth recognizing of rodent species from Las Joyas Scientific Station (LJSS) within the Sierra de Manantlán Biosphere Reserve. We report for the first time the presence of Nyctomys sumichrasti colimensis in this locality. Additionally, we present new records of Reithrodontomys wagneri and specimens belonging to Reithrodontomys sumichrasti clade IV, a clade recognized as an undescribed species within the R. sumichrasti species complex; the latter expands the geographic distribution of this new species. Finally, we include records of Microtus specimens forming a sister clade with M. mexicanus / M. mogollonensis, albeit with genetic distance values exceeding 9%. Our findings underscore the evolutionary significance of the apparent isolation of Sierra de Manantlán, Jalisco, particularly in influencing the speciation process of various rodent species. These findings underscore the importance of continuing biological inventories in less explored areas of Mexico. [ABSTRACT FROM AUTHOR]

Because of their cryptic morphology, pocket gophers of the genus Geomys are difficult organisms for examining species boundaries and taxonomic classification. However, this difficulty allows us to examine mechanisms of speciation more thoroughly. The cryptic morphology and complicated taxonomic history of pocket gophers also make identifying species distributions difficult. The use of molecular techniques has allowed for cryptic species to be recognized, including the recently proposed species G. jugossicularis (formally included in G. lutescens and G. bursarius). Previous work identifying the proposed distribution of G. jugossicularis was based on low sample numbers and limited geographical representation, and the data showed evidence of hybridization, further complicating relationships within the Geomys bursarius-lutescens-jugossicularis species complex. Samples suggested to represent G. jugossicularis have been identified in southwestern Nebraska, southeastern Colorado, southwestern Kansas, and, most recently, the Texas panhandle. Its distribution has been proposed to include the Oklahoma panhandle; however, gophers in this region currently are recognized as G. bursarius major. Samples were collected throughout western Oklahoma and the Oklahoma and Texas panhandles to determine identification and evaluate boundaries and distribution of gopher species in this region, as well as to compare our findings with previous work. Using the mitochondrial cytochrome b gene and nuclear microsatellite markers, we showed some support for the recognition of the proposed species G. jugossicularis and determined it occurs in the Oklahoma panhandle. However, our findings indicate the need for additional research to resolve species delimitation within the Geomys bursarius-lutescens-jugossicularis species complex. [ABSTRACT FROM AUTHOR]

Combating wildlife crimes in South Africa requires accurate identification of traded species and their products. Diagnostic morphological characteristics needed to identify species are often lost when specimens are processed and customs officials lack the expertise to identify species. As a potential solution, DNA barcoding can be used to identify morphologically indistinguishable specimens in forensic cases. However, barcoding is hindered by the reliance on comprehensive, validated DNA barcode reference databases, which are currently limited. To overcome this limitation, we constructed a barcode library of cytochrome c oxidase subunit 1 and cytochrome b sequences for threatened and protected mammals exploited in southern Africa. Additionally, we included closely related or morphologically similar species and assessed the database's ability to identify species accurately. Published southern African sequences were incorporated to estimate intraspecific and interspecific variation. Neighbor-joining trees successfully discriminated 94%–95% of the taxa. However, some widespread species exhibited high intraspecific distances (>2%), suggesting geographic sub-structuring or cryptic speciation. Lack of reliable published data prevented the unambiguous discrimination of certain species. This study highlights the efficacy of DNA barcoding in species identification, particularly for forensic applications. It also highlights the need for a taxonomic re-evaluation of certain widespread species and challenging genera. [ABSTRACT FROM AUTHOR]

Simple Summary: African tropical regions have a remarkably high bird diversity, yet few studies have tried to unravel the presence of blood parasites in birds found in conservation areas. Knowing which blood parasites are present will help us to prepare for potential disease outbreaks. We test the hypothesis that conservation regions have a high diversity of parasites. Molecular methods were used to screen 1035 blood samples from 55 bird species for blood infections on sites inside and adjacent to the Kruger National Park in South Africa. Overall, 28.41% of birds were found infected with at least one type of blood parasites. Bird malaria of the type Haemoproteus and Plasmodium was found in 17.39% and 4.64% of the birds respectively. Leucocytozoon blood parasite was found in 9.24% of birds. One hundred distinct blood parasite types were detected, of which 56 were new types. Similar bird malaria (Haemoproteus and Plasmodium) infections were found in closely related birds, while Leucocytozoon was found in almost every bird type. Sites with a high bird diversity also had a high parasite diversity. These findings provide insight of how birds can gradually survive their blood parasite infections. Afrotropical regions have high bird diversity, yet few studies have attempted to unravel the prevalence of avian haemosporidia in conservation areas. The diversity and host specificity of parasites in biodiversity hotspots is crucial to understanding parasite distribution and potential disease emergence. We test the hypothesis that biodiverse regions are associated with highly diverse parasites. By targeting the cytochrome b (Cytb) gene, we molecularly screened 1035 blood samples from 55 bird species for avian haemosporidia infections to determine its prevalence and diversity on sites inside and adjacent to the Kruger National Park. Overall infection prevalence was 28.41%. Haemoproteus, Leucocytozoon, and Plasmodium presented prevalences of 17.39%, 9.24%, and 4.64%, respectively. One hundred distinct parasite lineages were detected, of which 56 were new lineages. Haemoproteus also presented the highest diversity compared to Leucocytozoon and Plasmodium with varying levels of specificity. Haemoproteus lineages were found to be specialists while Plasmodium and Leucocytozoon lineages were generalists. We also found a positive relationship between avian host diversity and parasite diversity, supporting an amplification effect. These findings provide insight data for host–parasite and co-evolutionary relationship models. [ABSTRACT FROM AUTHOR]

Simple Summary: The present study aimed to assemble and annotate the mitochondrial genome of Keenocardium buelowi and conducted an in-depth analysis to investigate the molecular features of this mitochondrial genome. Our findings established phylogenetic relationships within the Cardiidae family. Additionally, evidence of selection pressure on the cytochrome b gene was detected, suggesting its important role in the evolution of K. buelowi. The mitochondrial genome provides valuable data for phylogenetic analysis and evolutionary research. In this study, we sequenced, assembled, and annotated the mitochondrial genome of Keenocardium buelowi using the Illumina platform. The genome spanned 16,967 bp and included 13 protein-coding genes (PCGs), two ribosomal RNAs, and 22 transfer RNAs. All PCGs utilized standard ATN start codons and TAN stop codons. The phylogenetic tree based on maximum likelihood and Bayesian inference analyses revealed Clinocardiinae as the sister group to Trachycardiinae, with the estimated divergence time being 44.5 million years ago (MYA) between K. buelowi and Vasticardium flavum. Notably, the cytochrome b gene (cob) exhibited a positive selection signal. Our findings provide valuable insights into the evolutionary history and molecular phylogeny of K. buelowi. [ABSTRACT FROM AUTHOR]

The common opossum (Didelphis marsupialis) is a marsupial widely distributed in the Neotropics, where it lives in urbanized environments. The apparent scarcity and fragmentation of available habitat, and high rates of vehicle collisions, may represent barriers to dispersal. To assess the functional connectivity of this species and its potential use as a model in urban ecology, we evaluated the structure and genetic diversity of opossums in the metropolitan area of the Aburrá Valley (AMVA), Colombia, using sequences of the mitochondrial Cytochrome B (Cytb) gene and nine autosomal microsatellite loci. Although Cytb presented low levels of variation, microsatellite markers revealed high genetic diversity (He = 0.852 and Ho = 0.698). The geographic distribution of mitochondrial lineages and a spatial principal component analysis (sPCA) of microsatellites showed the absence of genetic structure and effective barriers to dispersal of opossums in the study area. This scenario suggests that hostile landscapes such as those in urban environments may be suitable enough to allow the effective connectivity of some species such as the common opossum and others with high dispersal ability in urbanized areas. Nonetheless, it is necessary to evaluate the impact of releases of rescued fauna made by the environmental authority on the population structure of urban opossums. [ABSTRACT FROM AUTHOR]

Populations of plants and animals at the edge of a species' range may exhibit lower genetic diversity due to a combination of historical (e.g., postglacial dispersal) and contemporary (e.g., low effective population size, strong genetic drift or bottlenecks) processes. In this study, we analysed the genetic structure of populations of the European pond turtle (Emys orbicularis) in the northern part of the Pannonian Basin (Slovakia, Hungary, Austria), which represents the edge of the species' range. Using mitochondrial DNA (mtDNA) and microsatellite markers, we aimed to assign individuals and populations into phylogeographic lineages and to determine genetic variation and population structure. Our study revealed that most individuals from natural populations belong to mtDNA lineage II (widely distributed haplotype IIa, and likely endemic haplotypes IIl and IIo), which is typical for the whole Pannonian region. However, non-native haplotypes were detected in one Slovak population (haplotype Ib) and in the Austrian Donau Auen National Park (haplotype IVa). Microsatellite markers revealed high variability comparable to that in other parts of the range, suggesting that marginal populations may not necessarily have reduced genetic diversity. A genetically mixed population was found in the Donau Auen National Park, consisting of turtles belonging to mtDNA lineage II (taxonomically assessed as subspecies E. orbicularis orbicularis) and lineage IV (taxonomically assessed as E. orbicularis hellenica), which is native to the circum-Adriatic region. The admixture suggests intense hybridization and introgression between the two lineages (subspecies). [ABSTRACT FROM AUTHOR]

Barramundi, or Asian seabass Lates calcarifer, is a commercially important food fish with a wide geographic distribution in the Indo-West Pacific region. Despite its economic significance, particularly in Taiwan, information on the sources of fertilized eggs, fry and stock management practices remains largely unavailable. To ensure sustainable exploitation and effective selective breeding programs, assessing the genetic diversity and population genetic structure of barramundi is crucial. This study investigated the genetic diversity and population genetic structure of three cultured populations (Chiayi A, Chiayi B, and Tainan) and one wild-caught population in Taiwan using mitochondrial DNA cytochrome b and eight microsatellite loci as genetic markers. The analysis of haplotype network revealed that all populations shared an ancestral haplotype, yet each population had its own unique haplotype. Notably, the wild and cultured populations showed the largest genetic distance. However, overall, the genetic distances among all populations were small and similar. This finding suggests potential gene exchange, possibly through broodstock exchange practices between farms. The valuable genetic information obtained from this study holds the potential to improve aquaculture programs through targeted breeding initiatives and inform the development of sustainable management policies for barramundi. [ABSTRACT FROM AUTHOR]

Genetic connectivity and historical population expansion are characteristics that have largely been studied and observed in several members of the Lutjanidae family throughout the Western South Atlantic, particularly in Brazilian waters. In this study, the population genetic structure of Lutjanus jocu was investigated using four genomic regions: two from mitochondrial DNA (Control Region and Cytochrome B) and two nuclear genes (Delta 6 desaturase—intron 8 and Ribosomal Protein S7—intron 1), in 144 specimens distributed in six locations on the Atlantic coast of Brazil. The results revealed high levels of genetic diversity for both mtDNA and nuDNA markers, and intense gene flow, indicating, consistent with other studies, that this species is represented by a single genetic stock along the Brazilian coast. This is likely due to the high dispersal capacity of L. jocu larvae, driven by the main equatorial currents, as well as biological and ecological factors, contributing to a genetically homogeneous population. Historical demographic analysis and neutrality tests suggest that the species experienced population expansion, possibly during the Pleistocene. Information regarding the genetic structure of populations is extremely important, especially for species of high economic and fishing importance, and can be very useful in guiding stock management and management measures. [ABSTRACT FROM AUTHOR]

BACKGROUND: Wheat crown rot (WCR), primarily caused by Fusarium pseudograminearum has become more and more prevalent in winter wheat areas in China. However, limited fungicides have been registered for the control of WCR in China so far. Pyraclostrobin is a representative quinone outside inhibitor (QoI) with excellent activity against Fusarium spp. There is currently limited research on the resistance risk and resistance mechanism of F. pseudograminearum to pyraclostrobin. RESULTS: Here, we determined the activity of pyraclostrobin against F. pseudograminearum. The EC50 values ranged from 0.022 to 0.172 μg mL−1 with an average EC50 value of 0.071 ± 0.030 μg mL−1. Four highly pyraclostrobin‐resistant mutants were obtained from two sensitive strains by ultraviolet (UV) mutagenesis in the laboratory. The mutants showed decreased mycelial growth rate and virulence as compared with the corresponding wild‐type strains, indicating that pyraclostrobin resistance suffered a fitness penalty in F. pseudograminearum. It was found that the high resistance of four mutants was caused by the G143S mutation in Cytb. Molecular docking analysis also further confirms that the G143S mutation in Cytb decreased the binding affinity between pyraclostrobin and Cytb. CONCLUSION: The resistance risk of F. pseudograminearum to pyraclostrobin could be low to medium. Although a mutation at the G143S position of Cytb could potentially occur, this mutation decreases the fitness of the mutant, which may reduce its survival in the environment. Therefore, the negative consequences of a possible mutation are lower. This makes pyraclostrobin a good candidate for controlling crown rot in wheat. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Background: Southeast Asia is well known as a hotspot of biodiversity. However, very little is known about cave-dwelling hematophagous insects that are medically important. Taxonomic knowledge and ecology of phlebotomine sand flies are very poorly studied in Laos, as well as in other countries in the region. Herein, we report species diversity data and some notes on the ecology of the detected species from these karstic limestone areas of Laos. Methods: Phlebotomine sand flies were collected using Centers for Disease Control and Prevention (CDC) light traps from limestone cave locations in three districts of Vientiane Province, Laos. Both morphological and molecular techniques were used for sand fly identification. Species diversity and abundance were analyzed according to sites, locations, collection seasons, and trapping positions. Results: A total of 6564 sand flies, of which 5038 were females and 1526 were males, were morphologically identified into 20 species belonging to five genera (Chinius, Idiophlebotomus, Phlebotomus, Sergentomyia, and Grassomyia). The most abundant species were Chinius eunicegalatiae, Phlebotomus stantoni, Sergentomyia hivernus, Se. siamensis, and Idiophlebotomus longiforceps. Cytochrome b analysis results supported the morphological identification and revealed that Se. siamensis was separated from other members of the Se. barraudi group. Two new species, Se. dvoraki n. sp. and Se. marolii n. sp., were described. Sand fly density was generally high except in a cave in Vangvieng, with species richness ranging from 14 to 18 across different caves. Outside caves had higher species richness (R = 20) and diversity (H = 2.50) than cave entrances (R = 18, H = 2.41) and interiors (R = 16, H = 2.13). Seasonal variations showed high sand fly density in Feung and Hinheup during both dry and rainy seasons, while Vangvieng had a notable decrease in density during the dry season (D = 6.29). Conclusions: This study revealed that the diversity of phlebotomine sand fly fauna in Laos, particularly in karstic limestone areas, is greater than previously known. However, the taxonomic status of many species in Laos, as well as Southeast Asia, still needs more in-depth study using both morphological characters and molecular methods. Many species could be found from inside, at the entrance, and outside of caves, indicating a wide range of host-seeking behavior or possible natural breeding in the karstic cave areas. [ABSTRACT FROM AUTHOR]

In this work, we describe a new species of the genus Oreonectes, Oreonectes yuedongensis sp. nov., collected from the Lianhua Mountains in eastern Guangdong, China. Phylogenetic trees constructed based on the mitochondrial cytochrome b (Cyt b) gene showed that this new species represents an independent evolutionary lineage, with uncorrected genetic distances (Kimura 2‐parameter model) from congeners ranging from 5.1% to 8.3%. In addition, nuclear DNA analysis indicated O. yuedongensis as an independent lineage separate from its closely related species. Morphologically, the new species can be distinguished from other six species in the genus Oreonectes by a combination of serial characters. The description of this new species suggests that it is necessary to reassess the biodiversity of Oreonectes platycephalus as a complex, especially in the middle reaches of the Pearl River near the border between Guangdong and Guangxi. Morphological and genetic evidence supports O. yenlingi as a synonym of O. platycephalus. [ABSTRACT FROM AUTHOR]

Tenualosa ilisha is an anadromous fish from the Clupidae family that is distributed in several Asian countries. The relationship between T. ilisha from Indonesian waters and other waters in Asia is unknown, therefore this study was conducted to provide an overview of the relationship between T. ilisha from Indonesia and other waters in Asia based on the Cytochrome b gene (Cyt b) of mitochondrial DNA (mtDNA). The base sequence of T. ilisha from Labuhanbatu with a length of 568-572 base pairs (bp) was analyzed, then compared with Genbank data. All sequences compared showed that the composition of A+T (52.5%) was greater than G+C (47.5%). Genetic distance was calculated using Kimura two parameter (K2P); the lowest genetic distance of 0.000 was obtained between T. ilisha Labuhanbatu and Iraq with accession numbers LC619671.1; LC619673.1; LC619674.1. Meanwhile, the highest genetic distance is known to be 0.021 between T. ilisha Labuhanbatu and Bangladesh with the accession numbers MN748964.1, and MN748966.1. Neighbor-joining and maximum likelihood with 1000x repetition were the phylogenetic methods used to analyze the evolutionary relationships of T. ilisha. In general, both methods showed the same result that T. ilisha from Indonesia and other Asian waters are the same population. [ABSTRACT FROM AUTHOR]

Neotoma magister (Allegheny Woodrat) has a conservation status of vulnerable or at even greater risk across 13 eastern US states. A lack of recent museum vouchers at the southern limit of its distribution, where its sister species Neotoma floridana (Eastern Woodrat) occurs, creates uncertainty as to the current range of the Allegheny Woodrat in Alabama. We opportunistically collected woodrat scat samples for species identification using mitochondrial cytochrome b comparisons that provided genetic evidence of Allegheny Woodrats in Alabama north of the Tennessee River, consistent with its recognized distribution. These new samples of Allegheny Woodrats are most closely related to Allegheny Woodrats from Pennsylvania, Virginia, and West Virginia, and with greater intraspecific cyt b variation (5%) than previously reported. Comparisons between our woodrat samples from Alabama (2 north and 2 south of the Tennessee River), and published Allegheny and Eastern Woodrat sequences revealed a clear distinction between species even at small geographic distances, with mean interspecific divergence values of over 8%. We add to the known genetic variation of Allegheny Woodrats, and further characterize their genetic differentiation from Eastern Woodrats. Additional data are needed to delimit the distributional boundaries of the 2 species in Alabama and beyond. [ABSTRACT FROM AUTHOR]

Significance: Hyperspectral imaging sensors have rapidly advanced, aiding in tumor diagnostics for in vivo brain tumors. Linescan cameras effectively distinguish between pathological and healthy tissue, whereas snapshot cameras offer a potential alternative to reduce acquisition time. Aim: Our research compares linescan and snapshot hyperspectral cameras for in vivo brain tissues and chromophore identification. Approach: We compared a linescan pushbroom camera and a snapshot camera using images from 10 patients with various pathologies. Objective comparisons were made using unnormalized and normalized data for healthy and pathological tissues. We utilized the interquartile range (IQR) for the spectral angle mapping (SAM), the goodness-of-fit coefficient (GFC), and the root mean square error (RMSE) within the 659.95 to 951.42 nm range. In addition, we assessed the ability of both cameras to capture tissue chromophores by analyzing absorbance from reflectance information. Results: The SAM metric indicates reduced dispersion and high similarity between cameras for pathological samples, with a 9.68% IQR for normalized data compared with 2.38% for unnormalized data. This pattern is consistent across GFC and RMSE metrics, regardless of tissue type. Moreover, both cameras could identify absorption peaks of certain chromophores. For instance, using the absorbance measurements of the linescan camera, we obtained SAM values below 0.235 for four peaks, regardless of the tissue and type of data under inspection. These peaks are one for cytochrome b in its oxidized form at λ = 422 nm, two for HbO2 at λ = 542 nm and λ = 576 nm, and one for water at λ = 976 nm. Conclusion: The spectral signatures of the cameras show more similarity with unnormalized data, likely due to snapshot sensor noise, resulting in noisier signatures post-normalization. Comparisons in this study suggest that snapshot cameras might be viable alternatives to linescan cameras for real-time brain tissue identification. [ABSTRACT FROM AUTHOR]

Background: Porcine cysticercosis, caused by Cysticercus cellulosae a larval stage of adult Taenia solium, is a zoonotic parasitic disease where pigs harbour intermediate stage and human being acts as a definite host. The people of north eastern region of India are mostly non-vegetarian and consumption of pork is very much preferred by the people of this region. Hence, it is essential to detect C. cellulosae infections in pork. But traditional method of detection of C. cellulosae by post mortem examination of pork has disadvantages like need of expert and may be over lookedin case of light infections. Molecular diagnosis have been reported to be highly specific and sensitive for its diagnosis. Keeping in view of the above, the present study on molecular detection of larval cestode C. cellulosae of pigs was undertaken. A study on phylogenetic relation of C. cellulosae of pigs or human Taenae solium of this region was done to know its relation with other parts of the world, as not yet done so far. Methods: A total of 654 pig carcasses in 17 market places of three prime districts of state Arunachal Pradesh, India were examined to detect Cysticercus cellulosae of pigs. The cysticerci samples were obtained manually from the infected muscles and organs of the infected pigs that were preserved in phosphate buffer saline until DNA extraction. Stool samples of human patients who attended out-patient department (OPD) of Community Health Centers, Nursing homes and District etc. of study area of Aunachal Pradesh, India were collected randomly and examined by salt flotation technique for the presence of T. solium eggs. The segments of tapeworm voided by patientswere then identified for species identification and T. solium segments were collected in normal saline solutions (NSS) after clearing the debris and faecal materials. Genomic DNA extraction from 3-4 numbers of cysticerci and T. solium segments collected from affected human being were extractedusing a spin column kit (D Neasy tissue kit: QUIGEN). The technique polymerase chain reaction (PCR) with published primers were used for molecular detection of C. cellulosae and to get molecular (PCR) products of T. solium for further study. The mitochondrial gene cytochrome b oxidase subunit was amplified by PCR. The PCR products were purified, sequenced and phylogenetic tree was constructed using the neighbor-joining method. Result: The present study recorded a PCR amplification of cytochrome b oxidase genes with a definite product size of 1068 bp from DNA extracted from C. cellulosae and T. solium. The product size obtained from C. cellulosae will be helpful for meat inspection by molecular detection of C. cellulosae infections in pork. The present finding signifies that the same genomic isolate of both the larval cestode and adult parasite of T. solium is prevailing in the study areas. The neighbors-joining phylogenic tree shows close similarity of the present isolates with that prevailing in other South East Asian countries and thus it can be assumed from the present finding that the same genotypic isolate of T. solium parasite is prevalent in the whole of South East Asian region. [ABSTRACT FROM AUTHOR]

The variability of the cytochrome b gene in 11 individuals of Korean field mouse from the Ussuri Nature Reserve located on the southern spurs of the Sikhote-Alin in the Przewalski Mountains has been analyzed. In the population, a high level of genetic diversity was identified owing to the discovery of individuals with haplotypes of three phylogenetic lineages, i.e., Amur, Korea, and Manchuria. The majority of individuals (72.73%) in the population had Amur haplotypes. A significantly lower frequency of occurrence of individuals with Korea haplotypes in the Ussuri Nature Reserve (9.09%) compared to the previously analyzed population of the Khasansky district (38.46%) located in the very south of the Primorsky krai was noted. In the Ussuri Nature Reserve, for the first time in the Russian Far East, two individuals with Manchuria haplotypes were discovered. Previously, only one finding of an individual with a similar haplotype was noted in the literature in the Heilongjiang province of Northeast China. It was suggested that individuals with haplotypes of the Korea phylogroup penetrate into the south of Sikhote-Alin from South Korea and with haplotypes of the Manchuria phylogroup from Northeast China. [ABSTRACT FROM AUTHOR]

The Caucasian pine vole Microtus daghestanicus was characterized by high karyotypic and molecular genetic variability. However, in the eastern part of the Greater Caucasus, the populations of this species were poorly studied. To reveal the genetic characteristics of these populations and their phylogenetic relationships with other conspecific populations, a sample of Caucasian pine voles from the south of the Chechen Republic (Daikhokh Mount) was analyzed. Immunocytochemical analysis of the synaptonemal complexes at meiotic prophase I showed the belonging of the studied population to the most widespread 54-chromosomal form of M. daghestanicus. Analysis of the complete sequence of the mitochondrial cytochrome b gene showed that on the phylogenetic tree, specimens from the Chechen Republic and the previously studied Caucasian pine vole from North Georgia grouped into one compact cluster. It indicates that they belong to a distinct genetic form. Within the studied sample from the Chechen Republic, variation of the BRCA1 and XIST nuclear genes was revealed that probably reflects complex history of this population associated with alternating events of its isolation from neighboring populations and restoration of contacts between them. [ABSTRACT FROM AUTHOR]

Alternaria brown spot (ABS), caused by Alternaria alternata, is one of the main citrus diseases that causes heavy production losses and reductions in fruit quality worldwide. The application of chemical fungicides has a key role in the management of ABS. In this study, 48 isolates of A. alternata collected from citrus orchards since 2014 were tested in vitro for their sensitivity to pyraclostrobin and fludioxonil, the latter being temporarily registered in Italy since 2020. Pyraclostrobin sensitivity was determined using spore germination and mycelial growth assays. The effective concentration inhibiting 50% of fungal growth (EC50) was determined for each isolate. The sensitivity assays showed that the majority of A. alternata isolates tested were sensitive to pyraclostrobin. EC50 values of fludioxonil in a mycelial growth assay indicated that 100% of isolates were sensitive to this fungicide. The analysis of the cytochrome b gene showed that none of the 40 isolates with a different sensitivity profile had the G143A mutation, and the subgroup of 8 isolates analyzed by real-time PCR did not carry the G137R and F129L mutations. A subset of four more sensitive and two reduced-sensitive isolates was chosen to assess sensitivity on detached citrus leaves treated with pyraclostrobin at the maximum recommended label rate. Disease incidence and symptom severity were significantly reduced, with a small reduction reported in leaves inoculated with the reduced-sensitive isolates. Furthermore, there was no correlation between sensitivity and fitness parameters evaluated in vitro (mycelium growth and sporulation rate). These findings help the development of monitoring resistance programs and, consequently, set up effective anti-resistance strategies for managing ABS on citrus orchards. [ABSTRACT FROM AUTHOR]

The house mouse is the best-studied mammal species after humans, yet our understanding of its evolutionary history remains incomplete. Here, we focused on the colonisation of Europe by two subspecies and formation of a hybrid zone between them. We carried out a large-scale study of ~7000 mice sampled across an area embracing an ~900 km long portion of the zone, supplemented with cytochrome b and D-loop sequencing of ~1200 individuals collected worldwide. We demonstrate that the course of the mtDNA contact front is intricate and highly discordant with the consensus front for autosomal and X-linked markers, suggesting that local snapshots of mtDNA introgression may be misleading when treated in isolation. While multiple unrelated M. m. domesticus haplotypes occur in the zone area, the diversity of M. m. musculus haplotypes is limited. Moreover, we uncovered a vast region of domesticus introgression into musculus territory in northern areas and showed this introgression is unlikely to be driven by positive selection. We consider two previously published hypotheses explaining this phenomenon and put forth an alternative scenario assuming associations between mice and humans in northern Europe during the Nordic Bronze Age and subsequent zone movement associated with the expansion of Balto-Slavic peoples. [ABSTRACT FROM AUTHOR]

Simple Summary: Correct taxonomic identification is essential for conducting successful biological research, especially with regard to economically important insects, such as jumping plant lice or psyllids. In the present study, we identify and diagnose the morphologically characterised aphalarid species from Bulgaria using two molecular markers, cytochrome c oxidase I and cytochrome b. A total of 80 sequences of 25 Aphalaridae species were obtained and included in the BOLD and GenBank databases. This should enable even non-experts to identify these species quickly and accurately. The results of the current study show that two barcode genes are sufficient to distinguish most aphalarid species. Psyllids (Hemiptera: Psylloidea) are plant sap-sucking insects whose identification is often difficult for non-experts. Despite the rapid development of DNA barcoding techniques and their widespread use, only a limited number of sequences of psyllids are available in the public databases, and those that are available are often misidentified. Here, we provide 80 sequences of two mitochondrial genes, cytochrome c oxidase I (COI) and cytochrome b (Cytb), for 25 species of Aphalaridae, mainly from Bulgaria. The DNA barcodes for 15 of these species are published for the first time. In cases where standard primers failed to amplify the target gene fragment, we designed new primers that can be used in future studies. The distance-based thresholds for the analysed species were between 0.0015 and 0.3415 for COI and 0.0771 and 0.4721 for Cytb, indicating that the Cytb gene has a higher interspecific divergence, compared to COI, and therefore allows for more accurate species identification. The species delimitation based on DNA barcodes is largely consistent with the differences resulting from morphological and host plant data, demonstrating that the use of DNA barcodes is suitable for successful identification of most aphalarid species studied. The phylogenetic reconstruction based on maximum likelihood and Bayesian inference analyses, while showing similar results at high taxonomic levels to previously published phylogenies, provides additional information on the placement of aphalarids at the species level. The following five species represent new records for Bulgaria: Agonoscena targionii, Aphalara affinis, Colposcenia aliena, Co. bidentata, and Craspedolepta malachitica. Craspedolepta conspersa is reported for the first time from the Czech Republic, while Agonoscena cisti is reported for the first time from Albania. [ABSTRACT FROM AUTHOR]

The European eel (Anguilla anguilla (L.)) is a unique catadromous euryhaline fish and the only anguillid eel species classified as 'critically endangered'. The Baltic Lakeland area contains naturally recruited and introduced eels. There is great uncertainty regarding a baseline for the location and number of naturally dispersed eels in the region. Little is known about the genetic structure of the European eel populations in Baltic countries. The estimation of population genetic structure is important for the efficient management of naturally recruited and introduced eels. Two mitochondrial regions were used to investigate the genetic structure within and between eel samples from 11 waterbodies. In this study, new, unique, and widely distributed haplotypes were revealed. The studied eel population in the Baltic Lakeland shows high genetic diversity, which is possibly a result of intensive restocking programs. Sequences characterized for Anguilla rostrata were revealed in both mitochondrial regions. Understanding the genetic structure of eel populations worldwide is crucial for conservation efforts. Eel restocking in waterbodies where natural migration is restricted contributes to diversity loss for the world gene pool of eels. [ABSTRACT FROM AUTHOR]

The hypervariable region (HVR) in the control region of the mitochondrial DNA has frequently been used for population genetics and phylogeographic studies because of its highly variable nature. Although the HVR is beneficial for evaluating recent evolutionary history, including population demography, recent studies have implied the incidence of homoplasy in this region. To assess the accuracy of relying solely on the HVR for population genetics studies, molecular evolutionary analysis of the HVR, NADH-dehydrogenase subunit 2 (ND2), and cytochrome b genes were performed using 120 individuals of marbled flounder Pseudopleuronectes yokohamae. The HVR exhibited the highest genetic variability among the three regions, with sites showing high site-specific substitution rates. Considering the reticulate haplotype network structure and evolutionary linkages between regions, homoplastic mutations were indicated in the HVR in addition to ND2, underestimating genetic diversity. We found that homoplasy was less likely to affect coalescent-based demographic inferences in the population; however, there is still a potential risk of misinterpretation of population demography when solely using the HVR owing to its hypervariable nature. Collectively, we suggest analyzing other regions in addition to the HVR in fish population genetic research to improve accuracy and eliminate biases caused by homoplasy. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The taxonomy of the freshwater featherback fish genus Chitala (Osteoglossomorpha: Notopteridae) remains unsettled because delimiting and naming Chitala species based solely on morphology led to different hypotheses. In this study, we aim to determine the number and identity of Chitala species present in Peninsular Malaysia by employing an integrative taxonomic approach that combines morphology and two mitochondrial markers: the cytochrome c oxidase subunit I (COI) and cytochrome b genes. A total of 60 specimens of Chitala were collected throughout Peninsular Malaysia. Initially, all specimens were sorted into three distinct categories based on their body marking patterns. A molecular phylogenetic analysis revealed that each of these groups was monophyletic, and an automated species partition method further recognized them as distinct species. These results support the presence of three Chitala species in Peninsular Malaysia, even though a multivariate morphometric analysis failed to fully differentiate each of these three species. Comparison with original species descriptions and genetic data from an additional 70 Chitala specimens from neighbouring regions allowed us to name these species as Chitala lopis, Chitala borneensis, and Chitala ornata. We also showed the presence of two undescribed species in Indonesia. [ABSTRACT FROM AUTHOR]

The current study aimed to detect the genetic variability of mitochondrial DNA (mtDNA) cytochrome B (CYTB) gene in 80 native Egyptian rabbits (NER) belonging to three populations in three different agriculture regions (Delta, Middle, and Upper Egypt) and also investigate the origin of these breeds by phylogenetic relationship analysis. A total of 62 haplotypes were recorded among the three NER populations. The native Upper Egypt rabbits (NUER) expressed the highest number of haplotypes, mutations, polymorphic sites, and haplotype diversity (23, 55, 51, and 1.00, respectively). While it gave the lowest values of nucleotide diversity (0.0262) and Tajima’s D (− 0.01435), the percentage of mutual haplotypes was 4.8 % between the Middle and Delta Egypt populations. The phylogenetic analysis showed that there were more separated haplotypes of NUER compared to other populations. The same finding was also observed when supported by 31 sequences of different rabbit breeds retrieved from the GeneBank database. A mutual haplotype was observed between native Delta Egyptian rabbits (NDER) and New Zealand (NWZ) rabbits. The results of this study shed light on the importance of indigenous breeds in rural areas through mtDNA, which contributes to finding sustainable strategies to conserve and improve genetic resources in Egyptian rural areas. [ABSTRACT FROM AUTHOR]

The evolutionary dynamics of the ecoregions of southern South America and the species that inhabit them have been poorly studied, and few biogeographic hypotheses have been proposed and tested. Quaternary climatic oscillations are among the most important processes that have led to the current distribution of genetic variation in different regions of the world. In this work, we studied the evolutionary history and distribution of the Córdoba vesper mouse (Calomys venustus), a characteristic rodent of the region of which little is known about its natural history. Since the population dynamics of this species are influenced by climatic factors, this rodent is a suitable model to study the effects of Quaternary climatic oscillations in central Argentina. The mitochondrial cytochrome b gene was sequenced to analyze the phylogeography of C. venustus, and ecological niche modeling tools were used to map its potential distributions. The results of these approaches were combined to provide additional spatially explicit information about this species' past. Our results suggest that the Espinal was the area of origin of this species, which expanded demographically and spatially during the last glacial period. A close relationship was found between the Espinal and the Mountain Chaco. These results are consistent with previous studies and emphasize the role of the Espinal in the biogeographic history of southern South America as an area of origin of several species. [ABSTRACT FROM AUTHOR]

The Eichwald Toad, Bufo eichwaldi is one of the endemic amphibians distributed throughout Hyrcanian forests, marshes, wetlands, and streams in the north of Iran and the south of Azerbaijan. Populations of B. eichwaldi have previously been studied based on a few molecular and morphological data. Here, we undertake taxonomic and phylogenetic analyses based on morphological and morphometric features for 56 specimens of B. eichwaldi from Iran, and mitochondrial cyt b marker for 20 specimens from the whole species range in the west and east Caspian Sea lowland. Molecular analyses based on 794 bp of cyt b gene demonstrated two clusters: the east Caspian Sea lowland (A1) and the west Caspian Sea lowland (A2) clusters. Univariate analyses indicated six out of 25 morphometric variables in male B. eichwaldi specimens from the east (A1) are significantly different from male specimens of B. eichwaldi from the west Caspian Sea (A2; P≤0.05). Females have been also significantly different between two clusters in one character (distance between lids; P≤0.05). Sexual size dimorphism was also significantly different within and between populations in some variables. However, the PCA analyses show no differentiation among B. eichwaldi populations in the east and west Caspian Sea lowland. Based on p-distance parameter, the east and west groups showed a low level of genetic differentiation (0.2%). [ABSTRACT FROM AUTHOR]

The clinical implications of Theileria sp. Yokoyama, a recently identified Theileria species in cattle, remain uncertain. The objective of the present study was to evaluate the anemia status in cattle infected with Theileria sp. Yokoyama. Blood samples were collected from 206 cattle across seven Veterinary Ranges in Sri Lanka and analyzed for red blood cell (RBC) indices, including hemoglobin concentration, hematocrit, and RBC counts. Additionally, DNA was extracted from the samples and screened with a newly developed Theileria sp. Yokoyama-specific PCR assay targeting the cytochrome b gene. The PCR results revealed that 60 (29.1%) of the surveyed cattle tested positive for Theileria sp. Yokoyama, with 47 (78.3%) of them being co-infected with other hemopathogen species. Our findings revealed that the cattle breeds, management systems, and tick infestations are potential risk factors for the Theileria sp. Yokoyama infection. Next, we evaluated the anemia status among the surveyed cattle based on the RBC indices. We found that all non-infected cattle were non-anemic. By contrast, anemia was observed in 15 Theileria sp. Yokoyama-infected cattle, including 3 singly infected (anemia rate 3/13, 23.1%) and 12 co-infected cattle (12/47, 25.5%). Our findings suggest that Theileria sp. Yokoyama causes anemia in infected cattle. [ABSTRACT FROM AUTHOR]

The Chimanimani Mountains of Mozambique and Zimbabwe harbour diverse and unique flora and fauna. Because of these unique floral characteristics, this region has received considerable attention by botanists. In contrast, the vertebrates occurring here have received little attention. The aim of this paper was to synthesise data collected on multiple recent surveys into the first annotated checklist of the mammals of the Mozambican side of the Chimanimani Mountains. We identified medium-sized and large mammals by exterior appearance, mostly as captured on camera traps. We combined morphological and molecular methods to identify small mammals, and we report on echolocation calls of some of the poorly known bat species. In total, we recorded 69 species, including 23 species of bats (Chiroptera), 15 species of rodents (Rodentia), 11 species of carnivores (Carnivora), nine species of ungulates (Cetartiodactyla), and the rest comprising Primates, Eulipotyphla, Lagomorpha, Proboscidea, and Pholidota. Of these, five species are listed as threatened, demonstrating the importance of the Chimanimani Mountains for mammalian biodiversity conservation in South Eastern Africa.

In addition to the issue of pork contamination, processed meats frequently contain traces of rat meat. Therefore, detection and quantification of the pork and rat DNA in cases of meat and processed meat adulteration are necessary. In the current study, two gene targets of the cytochrome b for pigs and the Mt‐atp6 of Rattus norvegicus for rats were used in the absolute multiplex quantitative real‐time PCR (m‐qPCR). The sample DNA was amplified with a standard as positive control in the various concentration of 1000 pg, 100 pg, 10 pg, 0.1 pg, 0.01 pg, and 0.001 pg. There were 25 processed meat samples and 5 fresh meat samples identified in this study. Among the total of 30 samples assessed, 6 samples were successfully detected and quantified their pork and rat DNA contamination. One sample was contaminated with pork DNA with a concentration of 2.451×10‐4 pg (“Meatball 3). Five samples were contaminated with rat DNA with a concentration of 3.603×10‐11 pg (“Sempol 3”), 2.196×10‐10pg (“Meatball 6”), 4.908×10‐11 pg (“Siomay 3”), 1.489×10‐10 pg (“Grinding 2”), and 3.564×10‐10 pg (“Grinding 4”). In this study, we have discovered that the contamination of pork and rat were detected in the samples. It suggested that this method is applicable for detecting the contaminant in processed meat samples

The current study aimed to detect the genetic variability of mitochondrial DNA (mtDNA) cytochrome B (CYTB) gene in 80 native Egyptian rabbits (NER) belonging to three populations in three different agriculture regions (Delta, Middle, and Upper Egypt) and also investigate the origin of these breeds by phylogenetic relationship analysis. A total of 62 haplotypes were recorded among the three NER populations. The native Upper Egypt rabbits (NUER) expressed the highest number of haplotypes, mutations, polymorphic sites, and haplotype diversity (23, 55, 51, and 1.00, respectively). While it gave the lowest values of nucleotide diversity (0.0262) and Tajima's D (−0.01435), the percentage of mutual haplotypes was 4.8 % between the Middle and Delta Egypt populations. The phylogenetic analysis showed that there were more separated haplotypes of NUER compared to other populations. The same finding was also observed when supported by 31 sequences of different rabbit breeds retrieved from the GeneBank database. A mutual haplotype was observed between native Delta Egyptian rabbits (NDER) and New Zealand (NWZ) rabbits. The results of this study shed light on the importance of indigenous breeds in rural areas through mtDNA, which contributes to finding sustainable strategies to conserve and improve genetic resources in Egyptian rural areas.

Investigations of host feeding behaviour in haematophagous insects are critical to assess transmission routes of vector‐borne diseases. Understanding if a certain species has ornithophilic or mammalophilic feeding behaviour can facilitate future studies focused on pathogens transmission to and from certain host species. Culicoides Latreille (Diptera: Ceratopogonidae) are vectors of several pathogens, which include arboviruses, bacteria and parasites to a considerable diversity of vertebrate hosts. However, most of the studies focused on feeding habits target Culicoides species that could transmit the Bluetongue virus, consequently with a mammalophilic feeding behaviour, leaving aside the Culicoides species that are involved in the transmission of vector‐borne parasites to birds, such as Haemoproteus Kruse (Haemosporida: Haemoproteidae). This study aimed to investigate the source of blood meals of wild‐caught Culicoides using molecular‐based methods and to correlate our findings with the reports of Haemoproteus parasites in Culicoides species. Engorged Culicoides females were collected using ultraviolet (UV)‐light traps at seven different localities in Lithuania in 2021–2023. Biting midges were dissected, and the abdomens of engorged females were used for molecular investigation of the blood meal source. A polymerase chain reaction (PCR) protocol that amplifies a fragment of the Cytochrome B gene of vertebrates was used. Obtained sequences were compared to available information in GenBank database to confirm the source of the blood meal. In total, 258 engorged Culicoides females, representing nine different species, were analysed. The source of blood meal was identified in 29.1% of them with most of the insects having fed on birds (74.7%) and the remaining on mammals (25.3%). Culicoides segnis Campbell, Pelham‐Clinton was the only species to feed exclusively on birds; Culicoides from the Obsoletus group, C. pallidicornis Kieffer and C. punctatus Latreille were found to feed exclusively on mammals; C. festivipennis Kieffer, C. kibunensis Tokunaga and C. pictipennis Staeger had an opportunistic feeding behaviour, with the first two preferably feeding on birds. Due to their feeding behaviour and the presence of Haemoproteus parasites reported in the literature, C. festivipennis, C. kibunensis, C. pictipennis, and C. segnis play an important role in the transmission of those avian vector‐borne parasite in the wild. These Culicoides species were already confirmed as being able to support the development of several Haemoproteus species and lineages. Future studies focused on understanding the epidemiology of avian pathogens transmitted by Culicoides should target these species. [ABSTRACT FROM AUTHOR]

Research voyages onboard RV ‘Investigator’ in 2021 (IN2021_V08) and 2022 (IN2022_V04) sampled benthic communities of seamounts off Christmas Island and Cocos (Keeling) Islands, also known as Indian Ocean Australian Territories (IOT). Over 150 specimens of the family Serpulidae collected during the voyages were deposited in the Australian Museum. The animals belonged to the filogranin genera Apomatus, Bathyvermilia, Bathyvermiloides gen. nov., Filogranula, and Protis, as well as to serpulin genera Hyalopomatus, Placostegus and Vitreotubus. In total, 12 species were identified, including three named, but very poorly known serpulids Bathyvermilia challengeri, Filogranula stellata, and Vitreotubus digeromimoi. Nine species Apomatus nishii n. sp., Bathyvermilia rolandobastidai n. sp., Bathyvermiloides juliebrockae n. sp., Protis melmackenzieae n. sp., Protis perneti n. sp., Hyalopomatus nogueirai n. sp., Hyalopomatus rossanae n. sp., Hyalopomatus suelindsayae n. sp., and Placostegus leslieharrisae n. sp. were described. All species descriptions are accompanied by DNA sequence data and their phylogenetic positions in the family Serpulidae have been assessed.

The Javan tiger Panthera tigris sondaica and the Bali tiger P. tigris balica were categorized as Extinct on the IUCN Red List in both 2003 and 2008, leaving only the Sumatran subspecies P. tigris sumatrae extant in Indonesia. There have, however, been occasional, more recent reports of the Javan tiger but without conclusive evidence. Here, a potential observation in 2019 of a Javan tiger in a community plantation near the village of Cipendeuy in the forest of South Sukabumi, West Java, and a single hair found on a fence nearby, are assessed. The cytochrome b mitochondrial DNA (mtDNA) gene sequence of the putative Javan tiger hair were compared with that of a Javan tiger specimen in Museum Zoologicum Bogoriense, collected in 1930; hair samples of several tiger subspecies and the Javan leopard Panthera pardus melas were used as controls. The results showed that the genetic distances (d) of the putative Javan tiger hair with the Sumatran, Bengal P. tigris tigris and Amur P. tigris altaica tigers and the Javan leopard are 0.074 ± SE 0.009, 0.071 ± SE 0.009, 0.072 ± SE 0.009 and 0.088 ± SE 0.010, respectively, whereas the genetic distance of the putative Javan tiger hair with the Javan tiger museum specimen is 0.040 ± SE 0.006. In addition, phylogenetic trees showed that the putative Javan tiger hair sample belongs to the same group as the museum specimen of the Javan tiger, but is differentiated from other tiger subspecies and the Javan leopard. Whether the Javan tiger still occurs in the wild needs to be confirmed with further genetic and field studies.

Nile tilapia Oreochromis niloticus is one of the most important fishery resources and a valuable fish species for aquaculture programmes. It is found in almost all waters and is widely dispersed. Several natural populations of this species have been impacted by genetic pollution despite their significant economic relevance. Understanding population structure is a crucial first step in protecting this species in its native habitats as well as in choosing which wild stocks to use in hatchery initiatives. To demonstrate the genetic-population structure of this species, genetic differences among three geographically secluded populations of O. niloticus were investigated utilizing mitochondrial DNA cytochrome b gene sequences. The results were used to estimate the levels of genetic variability within and among the populations. The 56 cyt b (821 bp) sequences analysis revealed 21 haplotypes, with a nucleotide diversity of 0.0510 and a haplotype diversity of 0.881. In each of the populations, seven (7) singleton variable sites and 19 informative-parsimony sites, genetic diversity could be identified and few population haplotypes were found, indicating a minor genetic distinction between them. For the purpose of conservation and/or Nile tilapia breeding programmes, this information would assist in choosing the fish populations that maintain greater genetic variation in O. niloticus.

Background: Amblyomma spp. ticks, known for their long mouthparts, bright ornate appearance and aggressive hunting behaviour, are vectors of a number of important pathogens. In southern Africa, 17 Amblyomma spp. are currently documented. Of these species, Amblyomma hebraeum and Amblyomma variegatum have been well studied due to their wide geographical range and their status as competent vectors of pathogens that are of veterinary and medical importance. Studies on other Amblyomma spp. in southern Africa have been neglected, fostering ongoing debates on the validity of certain species such as Amblyomma pomposum. This study investigated the inter- and intra-species variation of Amblyomma ticks collected in southern Africa, focusing on resolving the dispute about A. pomposum and A. variegatum being distinct species. Methods: Four Amblyomma tick species were collected from Angola, Mozambique, South Africa, Zambia and Zimbabwe, and were identified morphologically as Amblyomma eburneum (208), A. hebraeum (4758), A. pomposum (191) and A. variegatum (2577) using identification keys. Gene amplification targeting the 12S and 16S rRNA, cytochrome oxidase I, cytochrome B and internal transcribed spacer-2 genes was conducted for 204 ticks, for which varying success was achieved during amplification for each of the markers. Maximum likelihood analyses were performed in IQ-TREE. Results: The phylogenetic topologies and ABGD analyses of each individual gene clustered A. pomposum within the A. variegatum clade, while clearly separating A. eburneum and A. hebraeum from all other species. None of the genetic markers indicated intraspecific structuring on the basis of geographical origin, despite great distances between sampling sites. Conclusion: Our study concludes that there is insufficient molecular evidence to differentiate A. pomposum and A. variegatum from each other. We highlight the need for whole mitochondrial genome sequencing of these two species to resolve the ongoing controversies. Furthermore, we propose mating and hybrid viability studies between the two species to confirm their reproductive isolation. [ABSTRACT FROM AUTHOR]

Introduction: Leucocytozoon is an intracellular blood parasite that affects various bird species globally and is transmitted by blackfly vectors. This parasite is responsible for leucocytozoonosis, a disease that results in significant economic losses due to reduced meat and egg production. There is limited knowledge about the epidemiological pattern of leucocytozoonosis and its causative species in Egypt, particularly in pigeons. Methods: The current study involved the collection of 203 blood samples from domestic pigeons from various household breeders and local markets across Qena Province, Upper Egypt. Samples were initially examined for potential Leucocytozoon infection using blood smears, followed by an evaluation of associated risk factors. Molecular identification of the parasite in selected samples (n = 11), which had initially tested positive via blood smears, was further refined through nested PCR and sequence analysis of the mitochondrial cytochrome b gene to ascertain the Leucocytozoon species present. Additionally, histopathological examination of the liver, spleen, and pancreas was conducted on animals that tested positive by blood smears. Results: Interestingly, 26 out of 203 samples (12.08%) had confirmed Leucocytozoon infections based on microscopic analysis. Additionally, all 11 samples that initially tested positive via blood smears were confirmed positive through nested PCR analysis, and their sequencing revealed the presence of Leucocytozoon sabrazesi, marking the first report of this parasite in Egypt. The study into potential risk factors unveiled the prevalence of Leucocytozoon spp. seems host gender-dependent, with males exhibiting a significantly higher infection rate (33.33%). Additionally, adult birds demonstrated a significantly higher infection prevalence than squabs, suggesting an age-dependent trend in prevalence. Seasonality played a significant role, with the highest occurrence observed during summer (37.25%). Histopathological examination revealed the presence of numerous megaloschizonts accompanied by lymphocytic infiltration and multiple focal areas of ischemic necrosis. Conclusion: To our knowledge, this is the first study to shed light on the epidemiological characteristics and molecular characterization of leucocytozoonosis in pigeons in Egypt. Further research endeavors are warranted to curb the resurgence of Leucocytozoon parasites in other avian species across Egypt, thereby refining the epidemiological understanding of the disease for more effective control and prevention measures. [ABSTRACT FROM AUTHOR]