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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]

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]

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]

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]

Background: Rift Valley fever virus (RVFV) is a zoonotic mosquito-borne virus with serious implications for livestock health, human health, and the economy in Africa, and is suspected to be endemic in north-eastern KwaZulu-Natal (KZN), South Africa. The vectors of RVFV in this area are poorly known, although several species, such as Aedes (Neomelaniconion) mcintoshi, Aedes (Neomelaniconion) circumluteolus, Aedes (Aedimorphus) durbanensis, and Culex (Lasioconops) poicilipes may be involved. The aim of the study was to determine the vertebrate blood meal sources of potential RVFV mosquito vectors in north-eastern KZN and to characterize the host-biting network. Methods: Blood-fed mosquitoes were collected monthly from November 2019 to February 2023 using a backpack aspirator, CO2-baited Centers for Disease Control and Prevention (CDC) miniature light traps and tent traps, in the vicinity of water bodies and livestock farming households. The mosquitoes were morphologically identified. DNA was extracted from individual mosquitoes and used as templates to amplify the vertebrate cytochrome c oxidase I (COI) and cytochrome b (cytb) genes using conventional polymerase chain reaction (PCR). Amplicons were sequenced and queried in GenBank and the Barcode of Life Data systems to identify the vertebrate blood meal sources and confirm mosquito identifications. All mosquitoes were screened for RVFV using real time reverse transcription (RT)-PCR. Results: We identified the mammalian (88.8%) and avian (11.3%) blood meal sources from 409 blood-fed mosquitoes. Aedes circumluteolus (n = 128) made up the largest proportion of collected mosquitoes. Cattle (n = 195) and nyala (n = 61) were the most frequent domestic and wild hosts, respectively. Bipartite network analysis showed that the rural network consisted of more host-biting interactions than the reserve network. All mosquitoes tested negative for RVFV. Conclusions: Several mosquito species, including Ae. circumluteolus, and vertebrate host species, including cattle and nyala, could play a central role in RVFV transmission. Future research in this region should focus on these species to better understand RVFV amplification. [ABSTRACT FROM AUTHOR]

Aim: The central and western Amazonia underwent several landscape changes during the Quaternary. Whereas the Riverine Barrier Hypothesis is traditionally used to explain the influence of rivers on speciation, processes such as river rearrangements have been overlooked to explain the geographic distribution and evolutionary history of Amazonia biota. Here, we tested how river rearrangements influenced the evolutionary history of uakari monkeys, genus Cacajao, a primate genus primarily associated with seasonally flooded forests in central and western Amazonia. Location: Central and Western Amazonia. Taxon: The genus Cacajao, including the black uakaris (C. melanocephalus, C. ayresi, C. hosomi); and the bald‐headed uakaris (C. calvus, C. amuna, C. rubicundus, C. ucayalii, C. novaesi). Methods: We performed a continuous phylogeographic analysis using 77 cytochrome b sequences to identify the origin and dispersal of Cacajao lineages. We used genome‐wide SNP variation (ddRADseq) to investigate population structure, gene flow and demographic history in Cacajao populations and used digital elevation models to identify landscape and riverscape characteristics that may have influenced the geographic distribution of Cacajao. Results: Our continuous phylogeographic reconstruction pointed out that the ancestral Cacajao lineage occupied the flooded forests of the Solimões River, in central Amazonia, at ~1.7 Mya and descendant lineages dispersed throughout central and western Amazonia more recently. We identified gene flow in both black and bald‐headed uakari populations, even across rivers considered barriers (e.g. the Negro River). Landscape analysis showed that river rearrangements influenced the geographic distribution and population structure in Cacajao. Historical demographic analyses suggest varied scenarios of population size changes among Cacajao monkeys consistent with periods of intense dynamism in flooded habitats and the formation of non‐flooded upland forests. Main Conclusion: Our results support that the river rearrangements have shaped the geographic distribution and divergence of recently diverged Cacajao lineages. Landscape and riverscape changes, along with retractions of the flooded forests, isolated some Cacajao populations in floodplain areas. Our study also suggests that these events led to the recent changes in demographic histories in species with a restricted geographic distribution. [ABSTRACT FROM AUTHOR]

The complex III of the mitochondrial respiration chain is an effective fungicidal target in plant pathogenic fungi. Especially the QoIs (FRAC Group 11 and 11A) fungicides cover a broad pathogen spectrum, that includes ascomycetes, basidiomycetes and oomycetes, by inhibiting the complex III at the outer binding site. Many of the target pathogens have developed resistance mediated by the target site mutation G143A. All fungicides that belong to Group 11 are affected by this mutation. Metyltetraprole has been recently developed, and it also binds at the Qo-site but is not affected by the G143A mutation and has therefore been classified differently (FRAC Group 11A). In experiments on solid medium with sequential exposure to complex III inhibitors, laboratory mutants of Zymoseptoria tritici with a resistance to metyltetraprole could be selected. Resistance to metyltetraprole was mediated by the target site mutation in the cytochrome B gene leading to the aa change Y132C. Metyltetraprole resistant isolates could only be generated from wildtype parents, but not from G143A parents. Further experiments used serial batch cultivations in liquid media with increasing fungicide concentrations. These were aimed to select double mutants that harboured both G143A and Y132C. Several different parental strains were treated with increasing concentrations of metyltetraprole, but no Y132C mutants were found when G143A parents were used. When Y132C mutants were used as the initial inoculum, and these were exposed to increasing concentrations of pyraclostrobin, the parental mutation was replaced by G143A within a few cycles; this could suggest that both mutations are incompatible. [ABSTRACT FROM AUTHOR]

Chemical control of grape black rot, caused by Phyllosticta ampelicida, relies mainly on the use of demethylation inhibitors (DMIs) and quinone outside inhibitors (QoIs). The effectiveness of QoI fungicides is influenced by alternative respiration activity, and the exon/intron structure and point mutations in the target protein's gene, the cytochrome b (cytb) gene. Our study aims to investigate the QoI fungicide sensitivity of 48 P. ampelicida isolates in vitro by measuring EC50 and the molecular characteristics of the cytb gene and its mRNA in P. ampelicida and other Phyllosticta species. Mycelial growth tests revealed that the P. ampelicida isolates were sensitive to both azoxystrobin and trifloxystrobin; baseline EC50 values were 0.029 and 0.022 μg/mL, respectively. Addition of salicylhydroxamic acid (SHAM) resulted in lower EC50 values (0.024 and 0.017 μg/mL, respectively). None of the typical point mutations conferring resistance to QoIs in some fungi were detected. A group I intron was present right after the 143rd codon in the cytb gene in four of the six Phyllosticta species examined. The sequence and exon/intron structure of the cytb gene of P. ampelicida isolated from Vitis vinifera is studied in detail and published here. Our results indicate a low risk of QoI resistance development via the G143A mutation in P. ampelicida. [ABSTRACT FROM AUTHOR]

Species of the Rhodnius genus have a complex taxonomy because the events of phenotypic plasticity and cryptic speciation make it difficult to correctly classify these vectors. During the taxonomic history of the genus, five synonymization events occurred. Additionally, some authors suggest that R. milesi possibly represent only phenotypic polymorphisms of R. neglectus. Thus, we analyzed the specific status of R. milesi in relation to R. neglectus using phylogenetic studies with the mitochondrial gene cytochrome B and the study of reproductive barriers. The phylogenetic reconstruction grouped R. milesi together with R. neglectus from different localities, demonstrating that these taxa represent the same species based on the phylogenetic species concept. Experimental crosses demonstrate the absence of pre- and postzygotic barriers under laboratory conditions. Additionally, when the hatch rates of crosses are compared to intraspecific crosses, it can be noted that they are high and very similar. Finally, the mortality rate of the hybrids does not indicate hybrid inviability, the absence of chromosome pairing errors does not indicate hybrid sterility, and the proportion between male and female hybrids demonstrates that Haldane's rule was not acting. Therefore, we perform the formal synonymization of R. milesi with R. neglectus. [ABSTRACT FROM AUTHOR]

Small and isolated populations are at risk of local extinction, either due to the reduction of genetic diversity or due to stochastic events. We assessed genetic diversity in populations of the red-spotted (Luscinia svecica svecica) and white-spotted bluethroat (L. s. cyanecula) subspecies from six European sites, focusing on a peripheral and declining, red-spotted population from Central Europe (Krkonoše Mountains, Czech Republic). Analysis of population structure using mitochondrial sequences (cytochrome b) and ddRAD genomic data revealed that the two subspecies form clearly defined clusters, but traces of possible admixture were found in three populations of the white-spotted bluethroat. Demographic history reconstruction indicates past population range expansion in both subspecies, while the occurrence of short (0.4 Mbp) runs of homozygosity (ROH) segments suggests possible inbreeding 50 to 200 generations ago. Interestingly, although established by a small number of individuals, the Krkonoše population show reduced genetic diversity in only one measure of summary statistics, possibly due to sustained gene flow. Correspondingly, nine highly homozygous genes were recovered within a 2.6-Mbp long ROH region on the Chr 5. Thus, we found only a small reduction of genetic diversity in a population facing extinction, contrary to the expectations for a geographically distant and small population. [ABSTRACT FROM AUTHOR]

Characterizing genetically distinct populations of primates is important for protecting biodiversity and effectively allocating conservation resources. Skywalker gibbons (Hoolock tianxing) were first described in 2017, with the only confirmed population consisting of 150 individuals in Mt. Gaoligong, Yunnan Province, China. Based on river geography, the distribution of the skywalker gibbon has been hypothesized to extend into Myanmar between the N'Mai Kha and Ayeyarwaddy Rivers to the west, and the Salween River (named the Thanlwin River in Myanmar and Nujiang River in China) to the east. We conducted acoustic point-count sampling surveys, collected noninvasive samples for molecular mitochondrial cytochrome b gene identification, and took photographs for morphological identification at six sites in Kachin State and three sites in Shan State to determine the presence of skywalker gibbons in predicted suitable forest areas in Myanmar. We also conducted 50 semistructured interviews with members of communities surrounding gibbon range forests to understand potential threats. In Kachin State, we audio-recorded 23 gibbon groups with group densities ranging between 0.57 and 3.6 group/km2. In Shan State, we audio-recorded 21 gibbon groups with group densities ranging between 0.134 and 1.0 group/km2. Based on genetic data obtained from skin and saliva samples, the gibbons were identified as skywalker gibbons (99.54–100% identity). Although these findings increase the species' known population size and confirmed distribution, skywalker gibbons in Myanmar are threatened by local habitat loss, degradation, and fragmentation. Most of the skywalker gibbon population in Myanmar exists outside protected areas. Therefore, the IUCN Red List status of the skywalker gibbon should remain as Endangered. [ABSTRACT FROM AUTHOR]

Background: West Nile virus (WNV), Everglades virus (EVEV), and five species of Orthobunyavirus were isolated from mosquitoes collected in the Everglades in 2016–2017. Prior studies of blood meals of mosquitoes in southern Florida have related findings to acquisition and transmission of EVEV, St. Louis encephalitis virus, and WNV, but not the Orthobunyavirus viruses associated with the subgenus Melanoconion of the genus Culex. Materials and Methods: In the present study, blood-fed mosquitoes were collected in the Everglades in 2016, 2017, 2021, and 2022, and from an industrial site in Naples, FL in 2017. Blood meals were identified to host species by PCR assays using mitochondrial cytochrome b gene. Results: Blood meals were identified from Anopheles crucians complex and 11 mosquito species captured in the Florida Everglades and from 3 species collected from an industrial site. The largest numbers of blood-fed specimens were from Culex nigripalpus, Culex erraticus, Culex cedecei, and Aedes taeniorhynchus. Cx. erraticus fed on mammals, birds, and reptiles, particularly American alligator. This mosquito species could transmit WNV to American alligator in the wild. Cx. nigripalpus acquired blood meals primarily from birds and mammals and frequently fed on medium-sized mammals and white-tailed deer. Water and wading birds were the primary avian hosts for Cx. nigripalpus and Cx. erraticus in the Everglades. Wading birds are susceptible to WNV and could serve as reservoir hosts. Cx. cedecei fed on five species of rodents, particularly black and hispid cotton rats. EVEV and three different species of Orthobunyavirus have been isolated from the hispid cotton rat and Cx. cedecei in the Everglades. Cx. cedecei is likely acquiring and transmitting these viruses among hispid cotton rats and other rodents. The marsh rabbit was a frequent host for An. crucians complex. An. crucians complex, and other species could acquire Tensaw virus from rabbits. Conclusions: Our study contributes to a better understanding of the host and viral associations of mosquito species in southwestern Florida. [ABSTRACT FROM AUTHOR]

Most Pamphagidae species are at risk of being endangered due to their flightlessness, low mobility, and small habitat range. For the same reason, Haplotropis brunneriana (Orthoptera: Pamphagidae) has been designated an endangered species in South Korea. Endangered species with small populations are difficult to observe and investigate because any damage or disturbance to them are prohibited. To address these problems, we first performed non-invasive genomic DNA (gDNA) extraction using exuviae to identify H. brunneriana. Second, we tested the limit of detection of gDNA in the exuviae and how long it could persist when exposed to artificial environmental conditions. Using exuviae, we identified cytochrome b as a species-specific marker of H. brunneriana that could distinguish them from other grasshoppers with sufficient specificity. In the limit of detection test, gDNA could persist and be amplified from exuviae for up to 40 days. Our results demonstrated that fewer than 10 days is recommended for amplifying gDNA from exuviae to obtain reproducible results. In conclusion, this non-invasive identification method using exuviae can be used as an alternative to species identification when it is difficult to find H. brunneriana in the habitat and has the advantage of allowing genetic research to be conducted without harming the endangered species. Implications for insect conservation: Our study used a non-invasive genomic DNA extraction method to conserve endangered grasshoppers. Using exuviae, endangered grasshopper species could be identified, thus confirming their presence or absence in the habitat. This identification method can be an alternative method that can be used when conducting genetic analyses accompanied by physical harm is difficult due to small populations, such as endangered and rare species. [ABSTRACT FROM AUTHOR]

Beluga sturgeon (Huso huso Linnaeus, 1758, acipenseridae) and Adriatic sturgeon (Acipenser naccarii, Bonaparte, 1836, acipenseridae) within the Po River basin have been recently assessed for the IUCN Red List of Threatened Species and were found to be Extinct in the Wild and Critically Endangered, respectively. Significant declines in both species' abundance have spurred major research efforts and management actions in recent decades. Recently, specific actions have been conducted to recover habitat connectivity through projects of river defragmentation and reintroduction plans have been implemented for both sturgeon species. To manage effective conservation efforts, knowledge of a species' distribution and abundance is critical, especially for adult sturgeon that are able to move hundreds of kilometers away from release sites. Here, two new quantitative PCR (qPCR) assays to detect beluga sturgeon and Adriatic sturgeon environmental DNA (eDNA) in water samples have been developed with the goal of providing an alternative method to monitor their presence. Two Taqman-based assays targeting the mitochondrial cytochrome b region were developed and showed no amplification of other related and co-occurring fishes. A mesocosm within the Ticino Park on the Ticino River (a main tributary of the Po River), where both species are bred, was used to develop and validate the assays. The LOQ for H. huso assay corresponded to Ct = 41 (7.33 × 107 DNA counts/µL of reaction) and for A. naccarrii it was Ct = 37 (2.23 × 1016 DNA counts/µL of reaction). Additionally, water samples were taken from the discard drainage, which flows directly into the Ticino River, testing positive detection of eDNA within a distance of up to 2 km. Overall, the results suggested that the two assays developed in this study could represent a promising new tool for monitoring both beluga and Adriatic sturgeon. [ABSTRACT FROM AUTHOR]

Aim: Hyperglycemia damages mitochondria and downregulates transcription of mtDNA-encoded genes and the long noncoding RNA LncCytB, causing mitochondrial genomic instability. The genes encoded by mtDNA are transcribed as large polycistronic transcripts, and the 5′ ends of precursor tRNAs are processed by mitochondrial-targeted ribonuclease P (MRPPs). Our aim was to investigate the role of MRPP1 in the downregulation of LncCytB in diabetic retinopathy. Methods: Using human retinal endothelial cells incubated in 20 mM D-glucose for 96 h, the gene expression and mitochondrial localization (immunofluorescence) of MRPP1 and the interaction between MRPP1 and LncCytB (determined by RNA-FISH and RNA immunoprecipitation) were quantified. The results were confirmed in retinal microvessels from streptozotocin-induced diabetic mice and from human donors with documented diabetic retinopathy. Results: Compared to normal glucose, high glucose decreased mRNA and mitochondrial localization of MRPP1 and its interaction with LncCytB. While MRPP1 overexpression prevented glucose-induced decrease in MRPP1–LncCytB interaction, LncCytB expression and mitochondrial damage (reduction in protective nucleoids in mtDNA), MRPP1-siRNA further worsened them. Similar results were obtained from retinal microvessels from diabetic mice and from human donors with diabetic retinopathy. Conclusions: Downregulation of MRPP1 in diabetes suppresses LncCytB transcription, resulting in mitochondrial functional and genomic instability, ultimately leading to the development of diabetic retinopathy. Thus, preventing MRPP1 downregulation has the potential to inhibit retinopathy and prevent the fear of vision loss in diabetic patients. [ABSTRACT FROM AUTHOR]

We report Bungarus suzhenae Chen, Shi, Vogel, Ding & Shi, 2021 and Ovophis zayuensis (Jiang, 1977) for the first time from India. Specimens of B. suzhenae and O. zayuensis were collected during our field surveys in north (Arunachal Pradesh) and south (Nagaland-Manipur border) of the river Brahmaputra. Species identity was supported by partial cytochrome b (cyt b), and 16s mitochondrial gene. We provide a detailed morphological description and a key to the two genera of this region. This report extends the westernmost distribution of B. suzhenae by ca. 300 km from Myanmar, and the southernmost range of O. zayuensis by 170 km from Tibet. Until now eight species of Bungarus and only one Ovophis species have been reported from India. Ovophis species are recently reported to be medically important venomous snakes whose venom properties have not been investigated in depth. [ABSTRACT FROM AUTHOR]

Background: Babesia spp. are protozoan parasites that infect the red blood cells of domesticated animals, wildlife and humans. A few cases of giant pandas (a flagship species in terms of wildlife conservation) infected with a putative novel Babesia sp. have been reported. However, comprehensive research on the morphological and molecular taxonomic classification of this novel Babesia sp. is still lacking. This study was designed to close this gap and formally describe this new Babesia sp. infecting giant pandas. Methods: Detailed morphological, molecular and phylogenetic analyses were conducted to characterise this Babesia sp. and to assess its systematic relationships with other Babesia spp. Blood samples from giant pandas infected with Babesia were subjected to microscopic examination. The 18S ribosomal RNA (18S rRNA), cytochrome b (cytb) and mitochondrial genome (mitogenome) of the new Babesia sp. were amplified, sequenced and assembled using DNA purified from blood samples taken from infected giant pandas. Based on the newly generated 18S rRNA, cytb and mitogenome sequences, phylogenetic trees were constructed. Results: Morphologically, the Babesia sp. from giant pandas exhibited various forms, including round to oval ring-shaped morphologies, resembling those found in other small canine Babesia spp. and displaying typical tetrads. Phylogenetic analyses with the 18S rRNA, cytb and mitogenome sequences revealed that the new Babesia sp. forms a monophyletic group, with a close phylogenetic relationship with the Babesia spp. that infect bears (Ursidae), raccoons (Procyonidae) and canids (Canidae). Notably, the mitogenome structure consisted of six ribosomal large subunit-coding genes (LSU1-6) and three protein-coding genes (cytb, cox3 and cox1) arranged linearly. Conclusions: Based on coupled morphological and genetic analyses, we describe a novel species of the genus Babesia, namely, Babesia ailuropodae n. sp., which infects giant pandas. [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]

Introduction: Globally, rodents and shrew populations constitute crucial elements of diverse environments and animal communities. It is imperative to study their population dynamics to mitigate any potential negative impact on humans, as they can be involved in the transmission of critical zoonotic agents, such as Blastocystis. Therefore, this study aimed to identify the prevalence and genetic composition of Blastocystis in wild rodents and shrews residing in the Zhejiang provinces of China. Methods: A total of 652 wild rodents and and shrews were captured from three different regions in Zhejiang Province from April 1st to October 31, 2023. The DNA was isolated by collecting fresh feces from the intestines of each rodent or and shrew. Rodent and shrew species were examined by vertebrate cytochrome b (cytb) analysis and PCR amplification. Blastocystis was also found in all fecal samples using PCR analysis and sequencing of the partial small subunit of ribosomal RNA (SSU rRNA) gene. Results: Among all the samples, 6.6% (43/652) showed a positive result for Blastocystis. In the results, 6 species of rodent and shrew were identified with Blastocystis, including Apodemusagrarius (n = 36) (2.8%), Niviventer confucianus (n = 75) (17.3%), Rattus losea (n = 18) (5.6%), R. norvegicus (n = 155) (2.6%), R. tanezumi (n = 86) (3.5%), and Suncus murinus (n = 282) (7.4%). The existence of 6 Blastocystis subtypes, ST4 (n = 33), ST1 (4), ST7 (n = 3), ST2 (n = 1), ST3 (n = 1), and ST5 (n = 1), were confirmed by sequence analysis. Discussion: Based on the molecular data obtained, the wild rodents and shrews under investigation were found to be concurrently infected with zoonotic subtypes of Blastocystis, including ST1 to ST5 and ST7. This suggests that these animals could potentially pose a zoonotic threat to humans and other animals susceptible to Blastocystis infection. [ABSTRACT FROM AUTHOR]

This study aimed to determine the coexistence and food sources of adult mosquitoes (Diptera: Culicidae) in a rural health center in Piura, Peru by using a descriptive cross-sectional design. Entomological techniques were used to capture and identify mosquitoes, and molecular biotechnology techniques were used to identify food sources. A total of 793 specimens of the Culex and Aedes genera were found coexisting, 789 (99.5%) were Culex quinquefasciatus, 607 (76.9%) were males and 182 (23.1%) were females. Likewise, 4 (100%) corresponded to Aedes aegypti females. The food sources of Aedes aegypti were Homo sapiens sapiens, and Homo sapiens sapiens and Canis familiaris were the food sources of Culex quinquefasciatus. This study provides evidence that rural health centers could be acting as foci of arbovirosis, with the risk that people who come for different ailments could contract diseases transmitted by C. quinquefasciatus and A. aegypti. [ABSTRACT FROM AUTHOR]

Three nominal species of the killifish genus Aplocheilus are reported from the lowlands of Sri Lanka. Two of these, Aplocheilus dayi and Aplocheilus werneri, are considered endemic to the island, whereas Aplocheilus parvus is reported from both Sri Lanka and Peninsular India. Here, based on a collection from 28 locations in Sri Lanka, also including a dataset of Asian Aplocheilus downloaded from GenBank, we present a phylogeny constructed from the mitochondrial cytochrome b (cytb), mitochondrial cytochrome c oxidase subunit 1 (cox1), and nuclear recombination activating protein 1 (rag1), and investigate the interrelationships of the species of Aplocheilus in Sri Lanka. The endemic Sri Lankan aplocheilid clade comprising A. dayi and A. werneri is recovered as the sister group to the clade comprising A. parvus from Sri Lanka and Aplocheilus blockii from Peninsular India. The reciprocal monophyly of A. dayi and A. werneri is not supported in our molecular phylogeny. A. dayi and A. werneri display strong sexual dimorphism, but species‐level differences are subtle, explained mostly by pigmentation patterns. Their phenotypes exhibit a parapatric distribution and may represent locally adapted forms of a single species. Alternatively, the present study does not rule out the possibility that A. dayi and A. werneri may represent an incipient species pair or that they have undergone introgression or hybridization in their contact zones. We provide evidence that the Nilwala‐Gin region of southwestern Sri Lanka may have acted as a drought refugium for these fishes. [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]

Human‐facilitated introductions of nonnative populations can lead to secondary contact between allopatric lineages, resulting in lineage homogenisation or the formation of stable hybrid zones maintained by reproductive barriers. We investigated patterns of gene flow between the native Sacramento Valley red fox (Vulpes vulpes patwin) and introduced conspecifics of captive‐bred origin in California's Central Valley. Considering their recent divergence (20–70 kya), we hypothesised that any observed barriers to gene flow were primarily driven by pre‐zygotic (e.g. behavioural differences) rather than post‐zygotic (e.g. reduced hybrid fitness) barriers. We also explored whether nonnative genes could confer higher fitness in the human‐dominated landscape resulting in selective introgression into the native population. Genetic analysis of red foxes (n = 682) at both mitochondrial (cytochrome b + D‐loop) and nuclear (19,051 SNPs) loci revealed narrower cline widths than expected under a simulated model of unrestricted gene flow, consistent with the existence of reproductive barriers. We identified several loci with reduced introgression that were previously linked to behavioural divergence in captive‐bred and domestic canids, supporting pre‐zygotic, yet possibly hereditary, barriers as a mechanism driving the narrowness and stability of the hybrid zone. Several loci with elevated gene flow from the nonnative into the native population were linked to genes associated with domestication and adaptation to human‐dominated landscapes. This study contributes to our understanding of hybridisation dynamics in vertebrates, particularly in the context of species introductions and landscape changes, underscoring the importance of considering how multiple mechanisms may be maintaining lineages at the species and subspecies level. [ABSTRACT FROM AUTHOR]

Genetic adaptation is the change of a population toward a phenotype that best fits the present ecological conditions of the environment it inhabits. As environmental conditions change, allele frequencies shift, resulting in different populations of the same species possessing genetic variation and divergent phenotypes. Cooperatively breeding common mole-rats (Cryptomys hottentotus hottentotus) inhabit environments along an aridity gradient in South Africa, which provides an opportunity for local genetic adaptations to occur. Using one mitochondrial gene (cytochrome b) and 3,540 SNP loci across the whole genome, we determined the phylogenetic relationship, population structure and genetic diversity of five populations of C. h. hottentotus located along an aridity gradient. Mitochondrial data identified population-specific clades that were less distinct in the two mesic populations, potentially indicating historical or recent gene flow, or the retention of ancestral haplotypes. Arid and semi-arid populations formed a distinct cluster from the non-arid populations. Genetic diversity and gene flow were higher in arid-dwelling individuals, suggesting greater connectivity and interactions between colonies in arid regions in comparison to mesic ones. Using an Aridity Index, we determined that isolation by environment, rather than isolation by geographical distance, best explains the genetic distance between the populations. Further analyses using target loci may determine if there are differing underlying genetic adaptations among populations of C. h. hottentotus. These analyses could help unravel population differences in response to environmental factors within a subspecies of bathyergid mole-rat and determine the adaptive capacity of this small nonmigratory subterranean rodent species in response to aridification in the face of climate change. [ABSTRACT FROM AUTHOR]

In Brazil, the Whitespotted Eagle Ray Aetobatus narinari is considered data-deficient due to the scarcity of basic information on its biology, ecology, and vulnerability to capture. Despite this, the species is caught by artisanal fishing along most of the coast, especially in the northeast of Brazil. This study analyzed mitochondrial DNA data in specimens of A. narinari caught by artisanal fishing in the northeast coast of Brazil to understand their population structure. For this, 42 individuals were sequenced at three mitochondrial genes: cytochrome oxidase 1 (COI), cytochrome b (Cytb), and NADH dehydrogenase subunit 4 (ND4). Concatenated COI-Cytb-ND4 sequences yielded 14 haplotypes, with moderate haplotype diversity (h = 0.646), low nucleotide diversity (π = 0.00087), and low fixation index ΦST values, indicating no population structure. Our results suggest that there is only one population of A. narinari in the study area. Genetic studies can contribute to improving management plans in these areas, avoiding the overexploitation of this and other species. [ABSTRACT FROM AUTHOR]

Aim To explore the mechanism of oxidized lipoprotein(a) (oxLp(a)) inducing pyroptosis of vascular endothelial cells. Methods After incubating human umbilical vein endothelial cells (HUVEC) with 100 mg / L oxLp( a) for 24 hours, Western blot and RT-qPCR was used to detect pyroptosis related proteins, pro-inflammatory cytokines, mitochondrial related proteins NRF1, NRF2, PGC-1α and mitochondrial gene cytochrome b (CYTB), ELISA was used to detect the levels of inflammatory factors, scanning electron microscopy was used to detect cell membrane rupture, transmission electron microscopy was used to detect mitochondrial morphology, Hoechst33342 / PI staining was used to detect cell apoptosis, MitoSOX probe was used to detect mitochondrial reactive oxygen species (mtROS), Flu-4AM probe was used to detect calcium ions, JC-1 probe was used to detect mitochondrial membrane potential (MMP), and Calcein AM staining was used to detect mitochondrial permeability transition pore ( mPTP). Transfecting HUVEC with CYTB overexpressing lentivirus and analyzing its effects on oxLp(a) induced pyroptosis and mitochondrial function. Results After treatment with oxLp(a), the expression of NLRP3, pro-Caspase-1, Caspase-1, GSDMD and GSDMD-N proteins related to pyroptosis were significantly increased (P<0. 05); the protein and mRNA levels of CYTB and pro-inflammatory cytokine IL-1β, IL-18 were significantly increased (P<0. 05). Small pores appeared on the cell membrane, the percentage of PI stained positive cells significantly increased (P<0. 05). OxLp( a) significantly inhibited the expression of mitochondrial related proteins NRF1, NRF2 and PGC-1α, and the expression of mitochondrial gene CYTB, promoted an increase in mtROS generation, Ca2+ overload, a decrease in ATP levels, a decrease in MMP, an increase in mPTP values, and abnormal mitochondrial morphology. After transfection with pHelper 2. 0 lentivirus vector overexpressing CYTB, it was found that oxLp(a) induced HUVEC pyroptosis and mitochondrial morphological and functional abnormalities were partially reversed by overexpression of CYTB. Conclusion oxLp ( a ) promotes mitochondrial morphological and functional abnormalities and induces HUVEC pyroptosis by downregulating CYTB. [ABSTRACT FROM AUTHOR]

Yak (Bos grunniens) is a unique livestock animal originating from the Qinghai-Tibet Plateau in China. In the current study, we investigated the maternal genetic diversity, differentiation and phylogeny of wild yak population and four domestic yak breeds (Qinghai-Gaoyuan, Huanhu, Xueduo, and Yushu) in Qinghai, China by analyzing 166 mitochondrial cytochrome b (Cytb) gene sequence variations. The results showed that the five yak breeds/populations had high genetic diversity (Hd = 0.501 ± 0.088–0.770 ± 0.053) and each yak breed/population owned unique haplotypes. Estimates of FST values showed a moderate genetic differentiation between wild yak and Huanhu yak (FST = 0.0577) as well as that between Huanhu yak and Yushu yak breeds (FST = 0.0520), but a weak genetic differentiation was observed between the other yak breeds/populations (–0.0209 < FST < 0.0372). Additionally, the clustering analysis based on RST values showed that Xueduo yak and Huanhu yak were clustered into one group, and each of the other three yak breeds/populations was separated into one group, respectively. Overall, the clustering relationship between wild yak and Yushu yak was closer. Maternal phylogenetic analysis showed that wild yak and four domestic yak breeds/populations in Qinghai represented in three maternal lineages (Mt-I, Mt-II, and Mt-III), indicating three maternal origins in yak. Our study would provide valuable information for the conservation and utilization of wild yak and Qinghai domestic yak breeds. [ABSTRACT FROM AUTHOR]

Based on the analysis of a fragment of the cytochrome b (cytb) gene of mitochondrial DNA (mtDNA), the genetic diversity of the little ground squirrel Spermophilus pygmaeus Pallas, 1779 of the Central and Eastern Caucasus was studied. Phylogenetic analysis revealed the existence of two clusters A and B within the western clade of S. pygmaeus 2. Cluster A is formed by haplotypes of ground squirrels from the Eastern Caucasus and the right bank of the Volga River, while B, only haplotypes of Central Caucasian animals. The distance between clusters A and B reaches 1.3%. The relatively isolated position on the phylogenetic tree of the ground squirrel population of the Central Caucasus, the absence of identical haplotypes in Central and Eastern Caucasian animals, and the distances obtained indicate genetic heterogeneity of the ground squirrel in the North Caucasus. The results of molecular dating showed that the evolutionary age of S. pygmaeus mtDNA haplotypes from the studied areas of the Eastern Caucasus was approximately 260 000 years, and that of the Central Caucasus, 163 000 years. A decrease in haplotypic and nucleotide variability was noted in the Central Caucasian populations of the little squirrel as compared to those from the Eastern Caucasus, which in general indicates the low viability of S. pygmaeus inhabiting the mountains of the Central Caucasus. [ABSTRACT FROM AUTHOR]

Members of the genus Lanius, known as shrikes, are economically important song birds distributed widely in several Asian countries. DNA sequencing, based on a partial segment of the cytochrome oxidase b (Cyt b) gene, was used to determine genetic variations and population genetic structure in four populations of shrike samples from north, east, and west Pakistan. We observed nucleotide diversities in L. isabellinus (= 0.02), L. schach (= 0.005), L. vittatus (= 0.004) and L. lahtora (= 0.008). FST values were significant (P < 0.05) in all species populations apart from L. isabellinus, which were non-significant. The negative values of Fu's Fs and Tajima's D showed significant population expansion in three species, L. schach, L. vittatus and L. Lahtora, whereas they were insignificant in populations of L. isabellinus. Fu's Fs and Tajima's D values explain that a strong genetic structure is present among members of L. isabellinus. On this basis, it is inferred that strong genetic structuring in L. isabellinus populations within this region was largely shaped by a common origin, with secondary influences from geographical factors and isolation. [ABSTRACT FROM AUTHOR]

Recent genetic studies have revealed that the Japanese dojo loach populations include a cryptic biological species (Misgurnus sp. Type I sensu Okada et al. 2017, hereafter called Type I loach), which is distributed in discontinuous restricted areas in certain parts of northeastern Japan, including the Hokkaido Island. The current exceptional distribution of the Type I loach beyond the major faunal boundary line between the Honshu and Hokkaido Islands (Blakiston's Line) may provide novel and vital perspectives on the formation of the Japanese archipelago and distribution patterns of primary freshwater fishes. In this study, we reconstructed the native distribution range of the Type I loach based on the nucleotide sequence of the cytochrome oxidase b (cytb) gene in mitochondrial DNA (mtDNA) from samples covering its entire current distribution range. Three major clades consisting of eight subclades were identified among the Type I loach haplotypes. The Hokkaido population comprised haplotypes belonging to only two subclades and had some common haplotypes with the Japanese Sea side of Honshu Island. Considerably low genetic divergence and unclear geographic genetic structures in Hokkaido haplotypes indicate their origin of artificial introduction. This speculation was further supported by the cultural background and recorded history, accompanied by human migration from the Honshu to Hokkaido Islands. Our reconstruction of the native distribution of the Type I loach by removing the non-indigenous populations revealed geographically cohesive haplotype distribution patterns. It provided some divergent scenarios for each clade and subclade. [ABSTRACT FROM AUTHOR]

BACKGROUND: The two‐spotted spider mite Tetranychus urticae causes significant damage to ornamental, cotton, sugarcane and horticultural crops in Australia. It has a long history of developing resistance to many acaricides including bifenazate. A mutation in the conserved cd1‐ and ef‐helices of the Qo pocket of cytochrome b is recognized as the primary mechanism of bifenazate resistance. To investigate the resistance mechanisms against bifenazate in Australian two‐spotted spider mite, we sequenced the complete mitochondrion genome of five mite strains including a susceptible and bifenazate‐resistant strain. RESULTS: We identified a novel mutation D252N in the G126S background at cytochrome b being the cause of bifenazate resistance in a bifenazate‐resistant strain, Bram. We validated the role of this mutation combination by reciprocal crosses between a bifenazate resistant and susceptible strain. By doing these crosses we confirmed the pattern of inheritance was maternal. Additionally, mitochondrial heteroplasmy was not observed by single mite genotyping of the mutations in cytb in a known bifenazate‐resistant strain Bram. The phylogenetic analysis with the complete mitochondrion genome sequences revealed that Australian two‐spotted spider mite strains are closely related to the green form of T. urticae found in China. CONCLUSIONS: The novel mutation D252N found in the cytochrome b in the G126S background was revealed to be the main cause of bifenazate resistance in the Australian T. urticae strain Bram. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Seafood is a prime target for fraudulent activities due to the complexity of its supply chain, high demand, and difficult discrimination among species once morphological characteristics are removed. Instances of seafood fraud are expected to increase due to growing demand. This manuscript reviews the application of DNA‐based methods for commercial fish authentication and identification from 2000 to 2023. It explores (1) the most common types of commercial fish used in assay development, (2) the type of method used, (3) the gene region most often targeted, (4) provides a case study of currently published assays or primer‐probe pairs used for DNA amplification, for specificity, and (5) makes recommendations for ensuring standardized assay‐based reporting for future studies. A total of 313 original assays for the detection and authentication of commercial fish species from 191 primary articles published over the last 23 years were examined. The most explored DNA‐based method was real‐time polymerase chain reaction (qPCR), followed by DNA sequencing. The most targeted gene regions were cytb (cytochrome b) and COI (cytochrome c oxidase 1). Tuna was the most targeted commercial fish species. A case study of published tuna assays (n = 19) targeting the cytb region found that most assays were not species‐specific through in silico testing. This was conducted by examining the primer mismatch for each assay using multiple sequence alignment. Therefore, there is need for more standardized DNA‐based assay reporting in the literature to ensure specificity, reproducibility, and reliability of results. Factors, such as cost, sensitivity, quality of the DNA, and species, should be considered when designing assays. [ABSTRACT FROM AUTHOR]

Introduction: Wild rodents can serve as reservoirs or carriers of E. bieneusi, thereby enabling parasite transmission to domestic animals and humans. This study aimed to investigate the prevalence of E. bieneusi in wild rodents from the Inner Mongolian Autonomous Region and Liaoning Province of China. Moreover, to evaluate the potential for zoonotic transmission at the genotype level, a genetic analysis of the isolates was performed. Methods: A total of 486 wild rodents were captured from two provinces in China. Polymerase chain reaction (PCR) was performed to amplify the vertebrate cytochrome b (cytb) gene in the fecal DNA of the rodents to detect their species. The genotype of E. bieneusi was determined via PCR amplification of the internal transcribed spacer (ITS) region of rDNA. The examination of genetic characteristics and zoonotic potential requires the application of similarity and phylogenetic analysis. Results: The infection rates of E. bieneusi in the four identified rodent species were 5.2% for Apodemus agrarius (n = 89), 4.5% for Cricetulus barabensis (n = 96), 11.3% for Mus musculus (n = 106), and 38.5% for Rattus norvegicus (n = 195). Infection was detected at an average rate of 17.4% among 486 rodents. Of the 11 identified genotypes, nine were known: SHR1 (detected in 32 samples), D (30 samples), EbpA (9 samples), PigEbITS7 (8 samples), HNR-IV (6 samples), Type IV (5 samples), HNR-VII (2 samples), HNH7 (1 sample), and HNPL-V (1 sample). Two novel genotypes were also discovered, NMR-I and NMR-II, each comprising one sample. The genotypes were classified into group 1 and group 13 via phylogenetic analysis. Discussion: Based on the initial report, E. bieneusi is highly prevalent and genetically diverse in wild rodents residing in the respective province and region. This indicates that these animals are crucial for the dissemination of E. bieneusi. Zoonotic E. bieneusi-carrying animals present a significant hazard to local inhabitants. Therefore, it is necessary to increase awareness regarding the dangers presented by these rodents and reduce their population to prevent environmental contamination. [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.

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.

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.

Deltacoronavirus, widely distributed among pigs and wild birds, pose a significant risk of cross-species transmission, including potential human epidemics. Metagenomic analysis of bird samples from Qinghai Lake, China in 2021 reported the presence of Deltacoronavirus. A specific gene fragment of Deltacoronavirus was detected in fecal samples from wild birds at a positive rate of 5.94% (6/101). Next-generation sequencing (NGS) identified a novel Deltacoronavirus strain, which was closely related to isolates from the United Arab Emirates (2018), China (2022), and Poland (2023). Subsequently the strain was named A/black-headed gull/Qinghai/2021(BHG-QH-2021) upon confirmation of the Cytochrome b gene of black-headed gull in the sample. All available genome sequences of avian Deltacoronavirus, including the newly identified BHG-QH2021 and 5 representative strains of porcine Deltacoronavirus (PDCoV), were classified according to ICTV criteria. In contrast to Coronavirus HKU15, which infects both mammals and birds and shows the possibility of cross-species transmission from bird to mammal host, our analysis revealed that BHG-QH2021 is classified as Putative species 4. Putative species 4 has been reported to infect 5 species of birds but not mammals, suggesting that cross-species transmission of Putative species 4 is more prevalent among birds. Recombination analysis traced BHG-QH-2021 origin to dut148cor1 and MW01_1o strains, with MW01_1o contributing the S gene. Surprisingly, SwissModle prediction showed that the optimal template for receptor-binding domain (RBD) of BHG-QH-2021 is derived from the human coronavirus 229E, a member of the Alphacoronavirus, rather than the anticipated RBD structure of PDCoV of Deltacoronavirus. Further molecular docking analysis revealed that substituting the loop 1–2 segments of HCoV-229E significantly enhanced the binding capability of BHG-QH-2021 with human Aminopeptidase N (hAPN), surpassing its native receptor-binding domain (RBD). Most importantly, this finding was further confirmed by co-immunoprecipitation experiment that loop 1–2 segments of HCoV-229E enable BHG-QH-2021 RBD binding to hAPN, indicating that the loop 1–2 segment of the RBD in Putative species 4 is a probable key determinant for the virus ability to spill over into humans. Our results summarize the phylogenetic relationships among known Deltacoronavirus, reveal an independent putative avian Deltacoronavirus species with inter-continental and inter-species transmission potential, and underscore the importance of continuous surveillance of wildlife Deltacoronavirus. [ABSTRACT FROM AUTHOR]

Introduction: Wild rodents are key hosts for Cryptosporidium transmission, yet there is a dearth of information regarding their infection status in the Inner Mongolian Autonomous Region and Liaoning Province of China. Therefore, the present study was conducted to determine the prevalence and genetic characteristics of Cryptosporidium among wild rodents residing in these two provinces. Methods: A total of 486 rodents were captured, and fresh feces were collected from each rodent's intestine for DNA extraction. Species identification of rodents was performed through PCR amplification of the vertebrate cytochrome b (cytb) gene. To detect the presence of Cryptosporidium in all fecal samples, PCR analysis and sequencing of the partial small subunit of the ribosomal RNA (rRNA) gene were performed. Results: Four species of rodents were identified: Rattus norvegicus, Mus musculus, Apodemus agrarius, and Cricetulus barabensis. Positive results for Cryptosporidium were obtained for 9.2% (18/195), 6.6% (7/106), 5.6% (5/89), and 6.3% (6/96) of these rodents, respectively, with an average infection rate of 7.4% (36/486). The identification revealed the presence of five Cryptosporidium species, C. ubiquitum (n = 8), C. occultus (n = 5), C. muris (n = 2), C. viatorum (n = 1), and C. ratti (n = 1), along with two Cryptosporidium genotypes: Rat genotype III (n = 10) and Rat genotype IV (n = 9). Discussion: Based on the molecular evidence presented, the wild rodents investigated were concurrently infected with zoonotic (C. muris, C. occultus, C. ubiquitum and C. viatorum) as well as rodent-adapted (C. ratti and Rat genotype III and IV) species/genotypes, actively participating in the transmission of cryptosporidiosis. [ABSTRACT FROM AUTHOR]

Background: Trichuris spp. (whipworms) are soil-transmitted helminths distributed worldwide, parasitizing several mammalian hosts such as ruminants, primates, and rodents. Trichuris spp. is one of the most common intestinal parasites affecting both humans and animals, and it can spread directly through the fecal-oral route, resulting in severe illness and financial loss. So, this work aims to detect the frequency of Trichuris spp. in camels in Beheira Governorate, Egypt, and to identify Trichuris spp. through morphometrical studies, molecular analysis, and phylogenetic analysis. Results: A total of 35 dromedaries out of 127 investigated had Trichuris spp. infection, meaning that the overall prevalence was 27.56%. The age of the camel affected the infection rate, older animals (> 5 years) having a higher prevalence of infection (24%) than animals of ages (< 3 years) (20%) than animals of ages (3–5 years) (19.14%). According to season: Trichuris spp. showed a unique pattern in camels in different seasons: summer (31.25%) > autumn (28.13%) > spring (25.8%) > winter (25%) indicating year-round infection. T. globulosa was identified morphometrically from camels in Beheira Governorate, Egypt. The BLAST analysis revealed the presence of T. globulosa isolate from camels using the Genbank database depending on nuclear small subunit ribosomal RNA (18s) and cytochrome b (Cytb) genes. Conclusion: A high prevalence of T. globulosa was found in camels in Beheira Governorate, Egypt. This is the first report to confirm the identification of T. globulosa from camel based on morphometrical studies and molecular and phylogenetic analysis in Egypt. More thorough studies on the incidence, molecular, and genetic analysis of Trichuris spp. in Egypt are required in addition to camel control programs. [ABSTRACT FROM AUTHOR]

Bats from the genus Vampyressa Thomas, 1900 are known from a relatively small number of individuals in Mexico. Through recent collecting efforts in southwestern Mexico, we detected several previously unknown populations of Vampyressa , which appeared to represent a lineage independent from but closely related to V. thyone. Here, we describe this lineage as a new species of the genus Vampyressa from the Mexican states of Guerrero and Oaxaca. The new species is currently known from at least 8 collected specimens and 3 released individuals from 8 localities, and can be differentiated from V. pusilla and V. thyone based on pelage and skull characters, size, mitochondrial Cytochrome b, and nuclear Recombination-activating gene 2 sequence data. The new species has been captured mainly in the lowlands but occurs along a wide altitudinal range from 150 to 2,200 m above sea level. Most of the records of the new species are from west of the Isthmus of Tehuantepec, which evidences a new case of a phylogenetic break at this biogeographic barrier. The genus Vampyressa now includes 6 species, and Mexico expands its position as the country with the largest number of endemic bats in the Neotropics, totaling 20 endemics. [ABSTRACT FROM AUTHOR]

Membrane proteins carrying redox cofactors are key subunits of respiratory chain complexes, yet the exact path of their folding and maturation remains poorly understood. Here, using cryo‐EM and structure prediction via Alphafold2, we generated models of early assembly intermediates of cytochrome b (Cytb), a central subunit of complex III. The predicted structure of the first assembly intermediate suggests how the binding of Cytb to the assembly factor Cbp3‐Cbp6 imposes an open configuration to facilitate the acquisition of its heme cofactors. Moreover, structure predictions of the second intermediate indicate how hemes get stabilized by binding of the assembly factor Cbp4, with a concomitant weakening of the contact between Cbp3‐Cbp6 and Cytb, preparing for the release of the fully hemylated protein from the assembly factors. [ABSTRACT FROM AUTHOR]

Crithidia mellificae and Lotmaria passim are trypanosomatids that infect honeybees, and many studies suggest that both parasites contribute to the decline in the honeybee population. The Japanese honeybee (Apis cerana japonica) is a native honeybee that inhabits various areas of Japan and is one of the most important pollinators. Both parasites have been well studied in the western honeybee (Apis mellifera) but have been poorly investigated in A. c. japonica. The present study investigated the presence of C. mellificae and L. passim in one feral and six managed A. c. japonica colonies at three different institutes. Five out of seven colonies were polymerase chain reaction‐positive for L. passim (71.4%); however, C. mellificae was not detected. Four of the five colonies were positive in both the midgut and hindgut, but one was positive only in the hindgut. A homology search and phylogenetic tree of the glycosomal glyceraldehyde‐3‐phosphate dehydrogenase (gGAPDH) and cytochrome b (Cytb) genes obtained in this study indicated that the sequences belonged to L. passim. The results of this study emphasize concerns regarding the health of this ecologically important pollinator and the importance of further investigation of the prevalence of L. passim in A. c. japonica in a wider area of Japan. [ABSTRACT FROM AUTHOR]

Large-bodied mole rats (Spalax) are a speciosus genus among obligate subterranean rodents, with seven currently recognized species, ranging from the Carpathians to the North Caucasus and further into the Caspian basin. Several conflicting hypotheses were proposed to explain the phylogenetic relationship among these taxa, mostly based on the subjective interpretation of the importance of certain morphologic characters in species delineation. We sequenced one mitochondrial (cytb) and one nuclear (IRBP) gene in six Spalax species, representing the most complete molecular dataset up to date. Both resulting phylogenies placed S. graecus, S. antiquus and S. giganteus at the base of the tree, while S. microphtalmus, S. zemni and S. arenarius appeared to have differentiated later in the evolutionary history of the genus. Cytb phylogeny supports monophyletic positions of all currently recognized species. According to the nuclear IRBP gene S. zemni and S. arenarius share similar haplotypes, which may represent either hybridization or recent separation from a common gene pool. The westernmost species S. antiquus and S. graecus represent the earliest split within the genus Spalax, indicating the possible origin of large-bodied blind mole rats from the South-West Europe. S. giganteus may represent the eastern relic of the ancient Spalax population. The central part of the genus distribution is inhabited by the most derived species: S. zemni + S. arenarius + S. microphthalmus. Large rivers of the Eastern Europe might have played a limited role in the distribution and speciation of mole rats and were crossed regularly by various genotypes. [ABSTRACT FROM AUTHOR]