Search

Fish substitution in fish products is an important issue in fish markets, as it is a widespread practice. An authentication protocol for Rohu, Thaila and Tilapia was developed by multiplex PCR. Three species-specific and one degenerate common forward primer were designed using the Cytb gene of the mitochondrial genome. These primers for Labeo rohita, Labeo catla and Oreochromis niloticus showed the fragment size of 235 bp, 186 bp and 506 bp on the agarose gel, respectively. The primers for L. rohita and L. catla were sensitive to 0.1 ng of DNA template, while for O. niloticus this value was 1 ng of DNA template. A total of 230 commercial samples (160 fried and 70 processed fish products) were screened, where 60% mislabeling in fried and 30% mislabeling in processed fish were found. This multiplex PCR protocol could give useful insights for food inspection and enforcement of regulatory food control. [ABSTRACT FROM AUTHOR]

Background: Ariidae species play a significant role as fishing resources in the Amazon region. However, the family's systematic classification is notably challenging, particularly regarding species delimitation within certain genera. This difficulty arises from pronounced morphological similarities among species, posing obstacles to accurate species recognition. Methods: Following morphological identification, mitochondrial markers (COI and Cytb) were employed to assess the diversity of Ariidae species in the Amazon. Results: Our sampling efforts yielded 12 species, representing 92% of the coastal Amazon region's diversity. Morphological identification findings were largely corroborated by molecular data, particularly for species within the Sciades and Bagre genera. Nonetheless, despite morphological support, Cathorops agassizii and Cathorops spixii displayed minimal genetic divergence (0.010). Similarly, Notarius quadriscutis and Notarius phrygiatus formed a single clade with no genetic divergence, indicating mitochondrial introgression. For the majority of taxa examined, both COI and Cytb demonstrated efficacy as DNA barcodes, with Cytb exhibiting greater polymorphism and resolution. Consequently, the molecular tools utilized proved highly effective for species discrimination and identification. [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]

Abstract 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 ( 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.

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]

The monogeneric family Calomyscidae includes the brush-tailed mice, genus Calomyscus , which have a Palearctic distribution ranging from the Hindu Kush Mountains in western Pakistan to the eastern Mediterranean region. Zagros Mountains—stretching from northwest to south of Iran—was assumed for long as the range of a single species, Calomyscus bailwardi ; however, recent studies revealed considerable heterogeneities among the geographical populations presumably referring to the persistence of undescribed diversity. One such group from the western Zagros Mountains has recently been recognized as C. behzadi Akbarirad, Dezhman, Aliabadian, Siahsarvie, Shafaeipour, and Mirshamsi, 2021. Using molecular, morphometric, geometric morphometric, and karyotypic data, we examine the divergence of 2 additional groups from western Iran and name these as new species. The first of these species is distributed in the northern Zagros and western Alborz Mountains in northwestern Iran was recovered as the sister species to C. urartensis from the South Caucuses. The second species from a single locality in the central Zagros Mountains was clustered closely with C. grandis from the western Alborz Mountains. Morphologic analyses, karyological features, and genetic distances disentangled these 2 new species from their sister species. [ABSTRACT FROM AUTHOR]

[Objective] Sichuan province has abundant rivers and rich resources of wild Macrobrachium nipponense. Understanding the genetic diversity of M. nipponense in different river basins within Sichuan province is significant. [Method] In the present study, the genetic diversity of wild M. nipponense populations in the Bahe river (BH), Fujiang river (FJ), Jialingjiang river (JLJ), Minjiang river (MJ) and Simenghe river (SMH) in Sichuan province was analyzed based on the mitochondrial Cytb sequence. [Result] The study detected 36 haplotypes among 201 M. nipponense individuals from five populations. The average haplotype diversity (Hd) for all populations ranged from 0. 345 to 0.959, with an average Hd value of 0.802. The JLJ population had the highest Hd value (0.959), while the MJ, SMH and BH populations had relatively low Hd value, all of which were less than 0. 5. The nucleotide diversity (Pi) values of all populations ranged from 0.002 25 to 0.015 47, and the mean Pi value was 0.015 83. The Pi value of JLJ population was the highest (0.01547), and the Pi value of BH population was the lowest (0.002 25). The genetic variation of M. nipponense was mainly due to differences among populations (68.20%), indicating significant genetic differentiation. The phylogenetic tree analysis revealed three distinct branches among all populations. The MJ network map of haplotypes revealed that all haplotypes could be grouped into three distinct lineages. Lineage A contained the dominant haplotype Hap 14, lineage B contained the dominant haplotype Hap 16, and lineage C contained the dominant haplotype Hap 1. [Conclusion] The genetic diversity of M. nipponense populations is generally high, but varies greatly among different basins. The populations in MJ, BH and SMH have relatively low genetic diversity. Additional efforts are needed to enhance the conservation of the genetic resources of M. nipponense in MJ, BH and SMH. The study provides a reference basis for the conservation and sustainable development of wild M. nipponense, and is conducive to the sustainable development of aquaculture in Sichuan province. [ABSTRACT FROM AUTHOR]

Background: We aimed to determine the common Echinococcus granulosus genotypes in Ağrı, Türkiye and to obtain information on the transmission of this parasite. Methods: Cystic echinococcosis samples from 100 slaughtered cattle and 100 slaughtered sheep and faecal samples from 200 stray dogs were included in 2021. Collected cyst fluid samples and faces were examined microscopically. DNA was isolated from the germinal membrane of the cysts and from the parasite eggs in the stool samples. The mitochondrial cytb gene region of the parasite was amplified by PCR. Genotypes were determined using the Basic Local Alignment Search Tool (BLAST) after sequence analysis of PCR amplicons. Results: The highest percentage of cysts was found in the lungs of sheep and the liver of cattle. In addition, 75% of sheep cysts and 25.6% of cattle cysts were fertile. Taenia spp./Echinococcus spp. eggs were found in 6% of the faeces of 200 dogs examined microscopically. E. granulosus eggs were detected in 4 out of 50 stool samples analysed by PCR. All samples analysed by sequence analysis were identified as E. granulosus s.s. G1 genotype. Sequence comparison revealed revealed one or more-point mutations in different regions of the five samples. Conclusion: E. granulosus s.s. G1 genotype, known as sheep strain, is common in the Ağrı, Türkiye. The controlled slaughter of livestock, especially sheep, and the avoidance of feeding hydatid cyst organs to dogs, together with public education, were necessary to prevent the spread of the disease. [ABSTRACT FROM AUTHOR]

Background Ariidae species play a significant role as fishing resources in the Amazon region. However, the family’s systematic classification is notably challenging, particularly regarding species delimitation within certain genera. This difficulty arises from pronounced morphological similarities among species, posing obstacles to accurate species recognition. Methods Following morphological identification, mitochondrial markers (COI and Cytb) were employed to assess the diversity of Ariidae species in the Amazon. Results Our sampling efforts yielded 12 species, representing 92% of the coastal Amazon region’s diversity. Morphological identification findings were largely corroborated by molecular data, particularly for species within the Sciades and Bagre genera. Nonetheless, despite morphological support, Cathorops agassizii and Cathorops spixii displayed minimal genetic divergence (0.010). Similarly, Notarius quadriscutis and Notarius phrygiatus formed a single clade with no genetic divergence, indicating mitochondrial introgression. For the majority of taxa examined, both COI and Cytb demonstrated efficacy as DNA barcodes, with Cytb exhibiting greater polymorphism and resolution. Consequently, the molecular tools utilized proved highly effective for species discrimination and identification.

Kawakawa (Euthynnus affinis) is a widely distributed epipelagic tuna, considered among the most economically significant species in the subtropical and tropical waters of the Indo-Pacific region. However, the lack of comprehensive information on the stock structure, management, and conservation of E. affinis populations in Malaysia raises concerns of potential overfishing and depletion of this marine fish. To ensure effective and successful species management, it is imperative to conduct a molecular- based assessment of the stock structure. In the present study, the genetic diversity and population structure of E. affinis in Malaysia and its surrounding waters were investigated using mitochondrial DNA cytochrome oxidase I (COI) and cytochrome b (Cyt b) genes. A total of 390 (COI) and 414 (Cyt b) individuals of E. affinis were sampled from 20 distinct locations situated within five geographic regions: the Straits of Malacca, South China Sea, Sulu Sea, Celebes Sea, and the Arabian Sea. The results revealed that the genetic divergence among 20 populations of E. affinis varied from low to high. The average haplotype diversity (Hd) and nucleotide diversity (π) for COI were calculated to be 0.5131 and 0.0043, respectively. On the other hand, the average haplotype diversity (Hd) and nucleotide diversity (π) for Cyt b were found to be 0.7450 and 0.0162, respectively. The analysis of haplotype distribution revealed that a total of 20 distinct haplotypes were identified for both the COI and Cyt b genes across 20 populations of E. affinis. The analysis of the Neighbor-Joining (NJ) tree and the Minimum Spanning Network (MSN) revealed the formation of three distinct clades among the samples. AMOVA analysis showed a significant genetic structure among the 20 populations of E. affinis, as indicated by the COI: FST = 0.5364 (P

Pakistan has natural freshwater resources acting as a hotspot for diverse fish fauna. However, this aquatic fauna is declining at an alarming rate due to over-exploitation, habitat degradation, water pollution, climate change, and certain anthropogenic activities. The freshwater shark, Wallago attu, is a popular edible catfish inhabiting these freshwater ecosystems. Habitat degradation, overfishing, and human activities are heavily impacting the natural population of this species. So, sound knowledge about its population structure is necessary for its proper management in natural waters. The current study involves utilizing two mtDNA markers (COI, Cytb) to assess the genetic structure and differentiation among W. attu populations of Pakistani Rivers. Genetic variability analysis indicated a high haplotype (0.343 ± 0.046–0.870 ± 0.023) and low nucleotide diversity (0.0024 ± 0.012–0.0038 ± 0.018) among single and combined gene sequences, respectively. Overall, River Indus was populated with more diverse fauna of Wallago attu as compared to River Chenab and River Ravi. Population pairwise, Fst values (0.40–0.61) were found to be significantly different (p < 0.01) among three Riverine populations based upon combined gene sequences. The gene flow for the combined gene (COI + Cytb) dataset among three populations was less than 1.0. The transition/transversion bias value R (0.58) was calculated for testing of neutral evolution, and it declared low genetic polymorphism among natural riverine populations of Wallago attu. The current study's findings would be meaningful in planning the management and conservation of this economically important catfish in future. [ABSTRACT FROM AUTHOR]

We reassessed the taxonomic status of lizards of the Iphisa elegans complex, a monotypic genus of Gymnophthalmidae, based on the congruence between mitochondrial DNA (Cytb) and nuclear DNA (c-mos and prlr) and characters of internal and external morphology, both quantitative and qualitative (meristic, morphometric and hemipenial), throughout its wide Amazonian distribution. The congruence of three species delimitation analyses (mPTP, bGMYC and ASAP), nuclear DNA topology and morphological evidence resulted in nine operational taxonomic units in the genus Iphisa. Besides being recovered as monophyletic groups in mitochondrial and nuclear phylogenetic trees, these nine operational taxonomic units can be distinguished by external and hemipenial morphology. Finally, we propose a new taxonomic rearrangement of the genus, whereby we: (i) restrict I. elegans to north-east Amazonia in the Guiana Shield and southwards, west from the Purus–Madeira interfluvium, on the right bank of the Amazon River; (ii) elevate Iphisa elegans soinii Dixon, 1974 to the specific level Iphisa soinii Dixon, 1974 comb. nov.; and (iii) name and describe five new species. [ABSTRACT FROM AUTHOR]

Copyright of Firat Universitesi Saglik Bilimleri Veteriner Dergisi is the property of Firat Universitesiu, Saglik Bilimleri Enstitusu and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Simple Summary: The tundra vole's (Alexandromys oeconomus) distribution in Lithuania has been documented for 70 years, yet its genetic diversity remains unexplored. We analyzed vole samples from northern and western sites in Lithuania, using mtDNA sequence analysis. Despite landscape barriers, our phylogenetic analyses placed Lithuanian voles in the Central European phylogroup, suggesting an origin from northeastern Poland. Genetic diversity in Lithuanian A. oeconomus at the mtDNA loci was low compared to other European samples, revealing distinctions from Poland and Northern Europe. Genetic divergence among western and northern Lithuanian samples, coupled with low variability, provides novel insights into species phylogeography and the influence of barriers on colonization dynamics. The distribution and spread of the tundra vole (Alexandromys oeconomus) in Lithuania have been documented over the last 70 years, but the genetic diversity of the species has not been studied. In this study, we examined A. oeconomus trapped in three sites in northern and western Lithuania using mtDNA sequence analysis of the cytb and control region. The western and northern sites are separated by anthropogenic landscape barriers. The western site is subject to regular spring flooding. Phylogenetic analyses of the studied individuals placed them in the Central European phylogroup, suggesting that Lithuanian A. oeconomus originated from northeastern Poland. In Lithuania, the genetic diversity of A. oeconomus at both mtDNA loci was relatively low (Hd < 0.6, π < 0.002) compared to that found in other European samples (Hd = 0.833–0.958; π = 0.00402–0.01552). Individuals analyzed in Lithuania were genetically different from samples collected in Poland and Northern Europe (ΦST > 0.15, p < 0.05). The genetic divergence between the western and northern samples of A. oeconomus in Lithuania, together with the low genetic variability among the voles studied, provides new insights into the phylogeography of the species and the influence of barriers on the colonization of the country. [ABSTRACT FROM AUTHOR]

A study of taxonomic differentiation and transition bias of the cytb gene, focusing on the early stages of speciation, was carried out in 15 subfamilies/families of five orders of Palearctic mammals. It was confirmed that the genetic differentiation among taxa of small and large mammals is shifted by the same taxonomic level, in which connection the period of cryptic speciation of Micromammalia (semi- and allospecific levels) corresponds to the divergence of type species in their genus within Macromammalia. In all cases, both transition bias and its evolutionary compensation took place. The novelty of the study also lies in the proof that the alignment of the transitions and transversion rates in evolutionary lineages has the pattern of a phase transition, and the frequency of transversions in short-cycle species of the orders Insectivora and Rodentia is higher than in long-cycle orders Artiodactyla, Carnivora and Chiroptera while with relative equality of transitions. The latter circumstance might be associated with the traits of metabolism and determines the characteristics of the molecular evolution of mammalian groups with short and long life cycles. [ABSTRACT FROM AUTHOR]

One specimen of frog was collected from Mohan Town, Mengla County, Xishuangbanna Prefecture, Yunnan Province, China, which is the type locality of Rana bannanica Rao & Yang, 1997, currently considered as a junior synonym of Hylarana milleti (Smith, 1921). This specimen well agrees with the original description of R. bannanica. In regard to morphology, R. bannanica and H. milleti are, indeed, very similar, but can still be distinguished from each other. Phylogenetic analysis, based on mitochondrial gene sequences, showed that this specimen and H. milleti are different species. Hence, we remove R. bannanica from the synonymy of H. milleti. [ABSTRACT FROM AUTHOR]

Background: We aimed to determine the common Echinococcus granulosus genotypes in Ağrı, Türkiye and to obtain information on the transmission of this parasite. Methods: Cystic echinococcosis samples from 100 slaughtered cattle and 100 slaughtered sheep and faecal samples from 200 stray dogs were included in 2021. Collected cyst fluid samples and faces were examined microscopically. DNA was isolated from the germinal membrane of the cysts and from the parasite eggs in the stool samples. The mitochondrial cytb gene region of the parasite was amplified by PCR. Genotypes were determined using the Basic Local Alignment Search Tool (BLAST) after sequence analysis of PCR amplicons. Results: The highest percentage of cysts was found in the lungs of sheep and the liver of cattle. In addition, 75% of sheep cysts and 25.6% of cattle cysts were fertile. Taenia spp./Echinococcus spp. eggs were found in 6% of the faeces of 200 dogs examined microscopically. E. granulosus eggs were detected in 4 out of 50 stool samples analysed by PCR. All samples analysed by sequence analysis were identified as E. granulosus s.s. G1 genotype. Sequence comparison revealed revealed one or more-point mutations in different regions of the five samples. Conclusion: E. granulosus s.s. G1 genotype, known as sheep strain, is common in the Ağrı, Türkiye. The controlled slaughter of livestock, especially sheep, and the avoidance of feeding hydatid cyst organs to dogs, together with public education, were necessary to prevent the spread of the disease.

Haemosporidian parasites that infect birds (Apicomplexa: Haemosporida) are blood parasites that require an invertebrate host (vector) and a vertebrate host for their lifecycle and cause malaria-like diseases. This group of parasites has provided valuable insights into host specificity, virulence, and parasite dispersal. Additionally, they have played a significant role in reshaping our understanding of the evolutionary history of apicomplexans. In order to accurately identify species and to address phylogenetic questions such as the timing of the haemosporidian radiation, the use of a sufficiently large genetic data set is crucial. However, acquiring this genetic data poses significant challenges. In this research, a sensitive nested PCR assay was developed. This assay allows for the easy amplification of complete mitochondrial genomes of haemosporidian parasites in birds, even during the chronic stage of infection. The effectiveness of this new nested PCR assay was evaluated using blood and tissue samples of birds with verified single parasite infections from previous studies. The approach involves amplifying four overlapping fragments of the mitochondrial genome and requires DNA extracts from single-infected samples. This method successfully amplified the complete mitochondrial genomes of 24 distinct haemosporidian parasite lineages found in various bird species. This data is invaluable for conducting phylogenetic analyses and accurately defining species. Furthermore, this study proposes the existence of at least 15 new haemosporidian parasite species based on the genetic information obtained. Data regarding pGRW04, previously categorized as Plasmodium relictum like pSGS1 and pGRW11, indicates that the pGRW04 lineage is actually a separate, hidden Plasmodium species. [ABSTRACT FROM AUTHOR]

Background: The time required for PCR detection of DNA in human blood meals in vector mosquitoes may vary, depending on the molecular markers used, based on the size and copy number of the amplicons. Detailed knowledge of the blood-feeding behavior of mosquito populations in nature is an essential component for evaluating their vectorial capacity and for assessing the roles of individual vertebrates as potential hosts involved in the transmission of vector-borne diseases. Methods: Laboratory experiments were conducted to compare the time course of PCR detection of DNA in human blood meals from individual blood-fed Anopheles stephensi mosquitoes, using loci with different characteristics, including two mitochondrial DNA (mtDNA) genes, cytB (228 bp) and 16S ribosomal RNA (rRNA) (157 bp) and nuclear Alu-repeat elements (226 bp) at different time points after the blood meal. Results: Human DNA was detectable up to 84–120 h post-blood-feeding, depending on the length and copy number of the loci. Our results suggest that 16S rRNA and Alu-repeat markers can be successfully recovered from human DNA up to 5 days post-blood-meal. The 16S rDNA and Alu-repeat loci have a significantly (P = 0.008) slower decline rate than the cytB locus. Median detection periods (T50) for the amplicons were 117, 113 and 86.4 h for Alu-repeat, 16S rDNA and cytB, respectively, suggesting an inverse linear relationship between amplicon size/copy number and digestion time. Conclusion: This comparative study shows that the Alu-repeat locus is the most efficient marker for time-course identification of human DNA from blood meals in female mosquitoes. It is also a promising tool for determining the anthropophilic index (AI) or human blood index (HBI), i.e. the proportion of blood meals from humans, which is often reported as a relative measure of anthropophagy of different mosquito vectors, and hence a measure of the vector competence of mosquito species collected in the field. [ABSTRACT FROM AUTHOR]

Cobia (Rachycentron canadum) is a coastal pelagic migratory fish species of tropical and subtropical waters, where it is an important game fish and it has been commercially expanded in offshore aquaculture systems. Understanding population connectivity is of utmost importance to the sustainable use and conservation of aquatic resources, and information on genetic diversity and structure is key element in unraveling differentiation when no clear physical barriers exist. In the present study, cobia genetic diversity and structure were depicted using mitochondrial DNA cytochrome b sequencing and microsatellite genotyping in samples from the Southwestern Atlantic and showed that a major single population inhabits the southern hemisphere. Cytochrome b sequencing also suggested that the Indian Ocean is the center of origin for this species' diversification. A hierarchical analysis of AMOVA compared sampling locations from the Northwestern Atlantic (from a previous study) with the Southwestern ones using nine shared microsatellite markers. Differentiation among groups (FCT = 0.41), Bayesian clustering analysis, and complementary ordination analyses (by discriminant analysis of principal components [DAPC] and factorial correspondence analysis [3D‐FCA]) presented a clear separation between the two hemispheres, supported by a Lagrangian model that explained the ocean dynamics over larval retention on the Western Atlantic. Another genetic subgroup intermingled with the main Southwestern group may also exist further south, probably associated with the Vitória‐Trindade Ridge and the local current systems. The distribution of this species in metapopulations is of extreme relevance for fisheries and fish hatcheries management in the Atlantic Ocean. [ABSTRACT FROM AUTHOR]

Acclimatized fungal diseases in wheat crops pose a severe threat to world wheat production. Zymoseptoria tritici, which causes Septoria tritici blotch (STB), is known as a devastating latent necrotrophic fungal pathogen that causes a significant yield loss, especially in Europe and in other wheat-growing countries. This fungal pathogen developed rapid resistance against new and old fungicides and has become a major threat to future wheat crops. Strobilurines and Azoles are the main group of contact and systemic fungicides along with succinate dehydrogenase inhibitors with reliable results to control the Z. tritici all over the world. However, many reports from Europe described the presence of resistance in Z. tritici against these fungicides. In the present study, a total of 102 isolates of Z. tritici were collected from five regions (Central Anatolia, Southeastern Anatolia, Marmara, Aegean, and the Mediterranean) of 19 provinces of Turkey to check the presence of target-site mutation at a molecular level to these two main fungicides. Azole target-site mutation in the CYP51 gene was checked by sanger sequencing while the strobilurin resistance mutation in the Cytb gene was confirmed by using PCR–RFLP using the FnuHI restriction enzyme. The obtained results from the two genes revealed some target-site mutations, however no known amino acid changes that cause resistance against these two main group fungicides in Turkish isolates have been detected. We concluded that the severe infection of STB pathogen in Turkish wheat growing may be still manageable with these fungicides although the resistance tests must be conducted for certainity. [ABSTRACT FROM AUTHOR]

Current genetic patterns of marine organisms are the result of various historical and environmental factors working together on different spatial scales. As a unique economic fish species in the Northwestern Pacific Ocean, Psenopsis anomala is ideal for studying the molecular phylogenetic geography of the Northwestern Pacific Ocean. To understand the genetic background of P. anomala, we analyzed a total of 98 samples from the East and South China Seas and found a total of 18 haplotypes. The overall haplotype diversity (0.3991 ± 0.0641) and nucleotide diversity (0.0005 ± 0.0005) were low. An analysis of molecular variance, haplotype network analysis, interpopulation Fst values, and maximum likelihood phylogenetic trees indicated that there was no significant difference between P. anomala populations, which could be due to frequent gene exchange (spawning migration and ocean circulation) and recent population expansion events. Overall Tajima’s D and Fu’s Fs values of P. anomala were significantly negative. The mismatch analysis diagram showed an obvious single peak, and the haplotype network diagram had a star-shaped structure, indicating that the P. anomala population has experienced population expansion. It is estimated that the expansion occurred approximately 17,377–13,901 years ago and might have been caused by the alternating rise and fall of the sea level during the Pleistocene. In addition, the haplotype diversity (0.1000) and nucleotide diversity (0.0001) of the Zhoushan population were significantly lower than those of the other geographic populations, and thus, this population urgently needs to be protected. [ABSTRACT FROM AUTHOR]

Abstract Background The time required for PCR detection of DNA in human blood meals in vector mosquitoes may vary, depending on the molecular markers used, based on the size and copy number of the amplicons. Detailed knowledge of the blood-feeding behavior of mosquito populations in nature is an essential component for evaluating their vectorial capacity and for assessing the roles of individual vertebrates as potential hosts involved in the transmission of vector-borne diseases. Methods Laboratory experiments were conducted to compare the time course of PCR detection of DNA in human blood meals from individual blood-fed Anopheles stephensi mosquitoes, using loci with different characteristics, including two mitochondrial DNA (mtDNA) genes, cytB (228 bp) and 16S ribosomal RNA (rRNA) (157 bp) and nuclear Alu-repeat elements (226 bp) at different time points after the blood meal. Results Human DNA was detectable up to 84–120 h post-blood-feeding, depending on the length and copy number of the loci. Our results suggest that 16S rRNA and Alu-repeat markers can be successfully recovered from human DNA up to 5 days post-blood-meal. The 16S rDNA and Alu-repeat loci have a significantly (P = 0.008) slower decline rate than the cytB locus. Median detection periods (T50) for the amplicons were 117, 113 and 86.4 h for Alu-repeat, 16S rDNA and cytB, respectively, suggesting an inverse linear relationship between amplicon size/copy number and digestion time. Conclusion This comparative study shows that the Alu-repeat locus is the most efficient marker for time-course identification of human DNA from blood meals in female mosquitoes. It is also a promising tool for determining the anthropophilic index (AI) or human blood index (HBI), i.e. the proportion of blood meals from humans, which is often reported as a relative measure of anthropophagy of different mosquito vectors, and hence a measure of the vector competence of mosquito species collected in the field. Graphical Abstract

Abstract A new species of the genus Amolops, Amolops ailao sp. nov., is described from central Yunnan, China. The new species belongs to the A. mantzorum species group. Phylogenetic analyses based on the combination of mitochondrial 16S rRNA, COI, and cytb genes revealed that the new species is the sister taxon to Amolops ottorum with strong support. Genetically, the new species differs from A. ottorum by 5.0% in cytb sequences. Morphologically, the new species can be distinguished from known congeners by the combination of the following characters: true dorsolateral folds absent, but dorsolateral folds formed by series of glands present; circummarginal groove on tip of first finger absent; body size small (males SVL 33.0–35.1 mm and female SVL 41.3 mm); HW/SVL 0.32‒0.35; UEW/SVL 0.08‒0.10; THL/SVL 0.52‒0.56; vomerine teeth absent; interorbital distance narrower than internarial distance; tympanum distinct, less than half eye diameter; supratympanic fold present, indistinct; a pair of large tubercles on sides of cloaca; tibiotarsal articulation reaching beyond anterior corner of eye; and vocal sac absent. The cladogenesis events within the A. mantzorum group rapidly occurred from Pliocene 4.23 Mya to Pleistocene 1.2 Mya, coinciding with the recent intensive uplift of the Qinghai-Tibetan Plateau since the Pliocene. Combining findings in this study with the most recent taxonomic progress, we consider that there are 20 known Amolops species in Yunnan, China, accounting for the highest proportion of amphibian diversity of Yunnan, and five of them belong to the A. mantzorum group. Among different subfauna and water systems in Yunnan, the species diversity of Amolops in northwestern Yunnan and Nu River Basin is highest.

Geothelphusa dehaani, a freshwater crab species endemic to Japan, has the largest distribution range amongst the 19 known species in the country. Due to its low dispersal capability and restricted habitat to freshwater, it serves as an excellent model for understanding gene flow between geographically isolated populations. In this study, we analysed the genetic relationships of 26 G. dehaani populations collected from different locations in the Japanese archipelago using two mitochondrial DNA regions - cytochrome oxidase subunit I (COI) and cytochrome b (cytB). Our results from the analysis of molecular variance (AMOVA) revealed high genetic variation amongst populations and the phylogenetic analysis identified four geographical groups: Clade I - Honshu and Shikoku, Clade II - north-eastern Kyushu, Clade III - southern Kyushu and Clade IV - north-western Kyushu. Notably, Clade IV exhibited the highest genetic distance amongst the observed groupings. These findings highlight the need for further examination of G. dehaani in Kyushu, including morphological and behavioural traits, to better understand the observed diversity within the species in the region.

A study of taxonomic differentiation and transition bias of the cytb gene, focusing on the early stages of speciation, was carried out in 15 subfamilies/families of five orders of Palearctic mammals. It was confirmed that the genetic differentiation among taxa of small and large mammals is shifted by the same taxonomic level, in which connection the period of cryptic speciation of Micromammalia (semi- and allospecific levels) corresponds to the divergence of type species in their genus within Macromammalia. In all cases, both transition bias and its evolutionary compensation took place. The novelty of the study also lies in the proof that the alignment of the transitions and transversion rates in evolutionary lineages has the pattern of a phase transition, and the frequency of transversions in short-cycle species of the orders Insectivora and Rodentia is higher than in long-cycle orders Artiodactyla, Carnivora and Chiroptera while with relative equality of transitions. The latter circumstance might be associated with the traits of metabolism and determines the characteristics of the molecular evolution of mammalian groups with short and long life cycles.

Abstract Sapajus libidinosus members of the Pedra Furada group, living in the Serra da Capivara National Park, use stone tools in a wider variety of behaviors than any other living animal, except humans. To rescue the evolutionary history of the Caatinga S. libidinosus and identify factors that may have contributed to the emergence and maintenance of their tool-use culture, we conducted fieldwork seasons to obtain biological samples of these capuchin monkeys. UsingCYTBsequences, we show a discrete but constant population growth from the beginning of the Holocene to the present, overlapping the emergence of the Caatinga biome. Our habitat suitability reconstruction reports the presence of plants whose hard fruits, seeds, or roots are processed by capuchins using tools. TheS. libidinosusindividuals in the Caatingawere capable of dynamically developing and maintaining their autochthonous culture thanks to: a) cognitive capacity to generate and execute innovation under selective pressure; b) tolerance favoring learning and cultural inheritance; c) an unknown genetic repertoire that underpins the adaptive traits; d) a high degree of terrestriality; e) presence and abundance of natural resources, which makes some places “hot spots” for innovation, and cultural diversification within a relatively short time.

In a recent survey of populations of the Korean torrent catfish Liobagrus, a distinctive species was discovered from the Geum River and its tributaries flowing into the western coast of Korea, and here described as a new species, L. geumgangensis sp. nov. It is distinguishable from other congeners by a combination of the following characters: I, 8 pectoral fin-rays; 52-56 caudal-fin rays; a relatively short occiput to dorsal-fin origin distance (6.9-9.8% SL); a short pelvic-fin insertion to anal-fin origin distance (11.9-17.3% SL); a long dorsal-fin base (10.6-13.5% SL); 8-9 gill rakers; 5-8 serrations on the pectoral fin; the body and fins are dark yellow, the margins of the dorsal, anal, and caudal fins are dark brown, but the outermost rim is faintly yellow. Analysis of the cytb gene also confirmed that L. geumgangensis is a monophyletic lineage distinct from other congeners. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the association between polymorphisms of ND1 and CYTB genes and in vitro early embryo development of Sanjabi sheep. Blood and ovarian samples were collected from a local slaughterhouse. The cumulus-oocyte complexes with a diameter greater than 3 mm were aspirated from follicles, and in vitro maturation (IVM) and in vitro culture (IVC) rates of them were recorded. A respective 1200 bp and 980 bp fragments of ND1 and CYTB genes were genotyped using a modified single strand conformation polymorphism (SSCP) method. The results of this study revealed that four different patterns, named as A, B, C, and D were observed for both ND1 and CYTB genes. The ND1 gene polymorphisms had significant effects on the IVM and IVC rate (p < 0.05). The pattern C of the ND1 gene significantly increased the IVM rate compared to the patterns A, B and D. For the IVC, the highest and lowest means were related to the C and B patterns, respectively. The CYTB gene polymorphisms also had significant effects on IVC (p < 0.01), but the IVM did not affected (p = 0.07). Here, the pattern D had the highest and the pattern C had the lowest means for both IVM and IVC rates. [ABSTRACT FROM AUTHOR]

Summary: In 2022, during potato cyst nematode surveys in several departments of Bolivia, the potato cyst nematode Globodera ellingtonae Handoo et al. , 2012 was detected for the first time in Bolivia from potato fields located in the Department of Tarija. Cysts and second-stage juveniles of the Bolivian populations were morphologically and morphometrically similar to those of the USA (Oregon and Idaho) and Argentina. Nine new ITS rRNA, 15 COI , 13 cytb gene sequences of G. ellingtonae from ten Bolivian populations were obtained in this study. The phylogenetic analyses of the ITS rRNA and cytb genes showed that all Bolivian sequences clustered together and formed a major clade with other G. ellingtonae sequences from Argentina, Chile and the USA. Molecular results confirmed the hypothesis that the mountain region in southern Bolivia, northwest Argentina and northern Chile could be considered as an ancient centre of origin of G. ellingtonae. The present molecular results suggested that G. ellingtonae was likely introduced into the USA from Chile, rather than from Bolivia and Argentina. [ABSTRACT FROM AUTHOR]

A new species of the genus Amolops, Amolops ailao sp. nov., is described from central Yunnan, China. The new species belongs to the A. mantzorum species group. Phylogenetic analyses based on the combination of mitochondrial 16S rRNA, COI, and cytb genes revealed that the new species is the sister taxon to Amolops ottorum with strong support. Genetically, the new species differs from A. ottorum by 5.0% in cytb sequences. Morphologically, the new species can be distinguished from known congeners by the combination of the following characters: true dorsolateral folds absent, but dorsolateral folds formed by series of glands present; circummarginal groove on tip of first finger absent; body size small (males SVL 33.0–35.1 mm and female SVL 41.3 mm); HW/SVL 0.32‒0.35; UEW/SVL 0.08‒0.10; THL/SVL 0.52‒0.56; vomerine teeth absent; interorbital distance narrower than internarial distance; tympanum distinct, less than half eye diameter; supratympanic fold present, indistinct; a pair of large tubercles on sides of cloaca; tibiotarsal articulation reaching beyond anterior corner of eye; and vocal sac absent. The cladogenesis events within the A. mantzorum group rapidly occurred from Pliocene 4.23 Mya to Pleistocene 1.2 Mya, coinciding with the recent intensive uplift of the Qinghai-Tibetan Plateau since the Pliocene. Combining findings in this study with the most recent taxonomic progress, we consider that there are 20 known Amolops species in Yunnan, China, accounting for the highest proportion of amphibian diversity of Yunnan, and five of them belong to the A. mantzorum group. Among different subfauna and water systems in Yunnan, the species diversity of Amolops in northwestern Yunnan and Nu River Basin is highest. [ABSTRACT FROM AUTHOR]

The distribution and spread of the tundra vole (Alexandromys oeconomus) in Lithuania have been documented over the last 70 years, but the genetic diversity of the species has not been studied. In this study, we examined A. oeconomus trapped in three sites in northern and western Lithuania using mtDNA sequence analysis of the cytb and control region. The western and northern sites are separated by anthropogenic landscape barriers. The western site is subject to regular spring flooding. Phylogenetic analyses of the studied individuals placed them in the Central European phylogroup, suggesting that Lithuanian A. oeconomus originated from northeastern Poland. In Lithuania, the genetic diversity of A. oeconomus at both mtDNA loci was relatively low (Hd < 0.6, π < 0.002) compared to that found in other European samples (Hd = 0.833–0.958; π = 0.00402–0.01552). Individuals analyzed in Lithuania were genetically different from samples collected in Poland and Northern Europe (ΦST > 0.15, p < 0.05). The genetic divergence between the western and northern samples of A. oeconomus in Lithuania, together with the low genetic variability among the voles studied, provides new insights into the phylogeography of the species and the influence of barriers on the colonization of the country.

The red porgy (Pagrus pagrus) and the common dentex (Dentex dentex) are Sparidae species of high commercial value, traded in the Greek market. In some cases, fish species identification from Greek fisheries is difficult for the consumer due to the strong morphological similarities with their imported counterparts or closely related species such as Pagrus major, Pagrus caeroleustictus, Dentex gibbosus and Pagellus erythrinus, especially when specimens are frozen, filleted or cooked. Techniques based on DNA sequencing, such as COI barcoding, accurately identify species substitution incidents; however, they are time consuming and expensive. In this study, regions of mtDNA were analyzed with RFLPs, multiplex PCR and HRM in order to develop a rapid method for species identification within the Sparidae family. HRM analysis of a 113 bp region of cytb and/or a 156 bp region of 16s could discriminate raw or cooked samples of P. pagrus and D. dentex from the aforementioned closely related species and P. pagrus specimens sampled in the Mediterranean Sea when compared to those fished in the eastern Atlantic. HRM analysis exhibited high accuracy and repeatability, revealing incidents of mislabeling. Multiple samples can be analyzed within three hours, rendering this method a useful tool in fish fraud monitoring. [ABSTRACT FROM AUTHOR]

The Chinese banyan, Ficus microcarpa is an ornamental fig tree, widely planted throughout Southern Greece. Native to Asian - Australasian regions, it can be found in plazas, parks, roadsides as well as private gardens. Along with its host, various insect species have managed to follow and establish themselves outside of their native range. In Rhodes, the mass defoliation of an emblematic fig tree, led to the subsequent finding of Josephiella microcarpae infestation. Repetitive samplings in the following years confirmed the establishment of the species on the island, which constitutes the first record of this species in Greece. Collected specimens from Rhodes and Cyprus were sequenced for the COI DNA barcoding region. Further investigation of fig trees throughout various locations in Southern Greece, proved fruitless. Based on our findings, the distribution of J. microcarpae in Greece seems restricted in the city of Rhodes. [ABSTRACT FROM AUTHOR]

Plasmodium spp. and some other blood parasites belonging to the order Haemosporida are the focus of many epidemiological studies worldwide. However, haemosporidian parasites from wild animals are largely neglected in scientific research. For example, Polychromophilus parasites, which are exclusive to bats, are described in Europe, Asia, Africa, and Oceania, but little is known about their presence and genetic diversity in the New World. In this study, 224 samples of bats from remaining fragments of the Atlantic Forest and Pantanal biomes, as well as urbanized areas in southern and southeastern Brazil, were analyzed for the presence of haemosporidian parasites by PCR of the mitochondrial gene that encodes cytochrome b (cytb). The PCR fragments of the positive samples were sequenced and analyzed by the Bayesian inference method to reconstruct the phylogenetic relationships between Polychromophilus parasites from bats in Brazil and other countries. Sequences from Brazilian lineages of Polychromophilus were recovered in a clade with sequences from Polychromophilus murinus and close to the one Polychromophilus sequence obtained in Panama, the only available sequence for the American continent. This clade was restricted to bats of the family Vespertilionidae and distinct from Polychromophilus melanipherus, a parasite species mainly found in bats of the family Miniopteridae. The detection of Polychromophilus and the genetic proximity to P. murinus were further confirmed with the amplification of two other genes (clpc and asl). We also found a Haemosporida parasite sequence in a sample of Noctilio albiventris collected in the Pantanal biome, which presents phylogenetic proximity with avian Haemoproteus sequences. Morphological and molecular studies are still needed to conclude and describe the Polychromophilus species in Brazilian Myotis bats in more detail and to confirm Haemoproteus parasites in bats. Nevertheless, these molecular results in Brazilian bats confirm the importance of studying these neglected genera. [ABSTRACT FROM AUTHOR]

Geothelphusa dehaani, a freshwater crab species endemic to Japan, has the largest distribution range amongst the 19 known species in the country. Due to its low dispersal capability and restricted habitat to freshwater, it serves as an excellent model for understanding gene flow between geographically isolated populations. In this study, we analysed the genetic relationships of 26 G. dehaani populations collected from different locations in the Japanese archipelago using two mitochondrial DNA regions - cytochrome oxidase subunit I (COI) and cytochrome b (cytB). Our results from the analysis of molecular variance (AMOVA) revealed high genetic variation amongst populations and the phylogenetic analysis identified four geographical groups: Clade I - Honshu and Shikoku, Clade II - north-eastern Kyushu, Clade III - southern Kyushu and Clade IV - north-western Kyushu. Notably, Clade IV exhibited the highest genetic distance amongst the observed groupings. These findings highlight the need for further examination of G. dehaani in Kyushu, including morphological and behavioural traits, to better understand the observed diversity within the species in the region. [ABSTRACT FROM AUTHOR]

Nearctic migratory songbirds have demonstrated low levels of genetic differentiation and weak phylogeographical structure in mitochondrial DNA lineages compared with resident species. The common grackle, Quiscalus quiscula , is a widespread, partially migratory, North American icterid composed of three currently recognized subspecies. In this study, mensural characters (external and skeletal measurements) and the complete mitochondrial genome together with two mitochondrial genes, Cytb and ND2 , were used to investigate subspecific differentiation and demographic history of the common grackle. The results showed substantial variation in body size among subspecies, mostly distributed between the 'Florida grackle', Quiscalus quiscula quiscula , and the two other subspecies. Analysis of mitochondrial DNA indicated low levels of genetic variation, but we found distinct haplotypes in Florida that form a clade in the phylogenetic tree. This suggests that the nominate subspecies in Florida is a distinct evolutionary unit. The sharing of haplotypes among the other subspecies (Quiscalus quiscula versicolor and Quiscalus quiscula stonei) in the north suggests high levels of gene flow, making the status of these two subspecies equivocal. Gene flow between nominate Q. q. quiscula , Q. q. versicolor and putative Q. q. stonei is probably attributable to historical changes in distribution and abundance following climate change events. We therefore recognize only two subspecies in the common grackle complex. [ABSTRACT FROM AUTHOR]

Zacco tiaoxiensis, a new species of cyprinid fish from Tiaoxi river system in Zhejiang Province, southern China, is described. With morphological comparison, the new species is similar to Z. acanthogenys, which is widely distributed in Zhejiang Province. However, the new species could be distinguished from Z. acanthogenys by the following combination of characters: lateral- line scales comprising 41-44 (vs. 44-47), scales above lateral-line 8 (vs. 8-10), scales below lateral-line 3-4 (vs. 4-5), scales surrounding caudal peduncle 12-15 (vs. 14-17), for mature male individuals, the ventral fin reaching (vs. exceeding) the anus, etc. The morphological differences among the new species and the allopatric congener species of Z. sinensis and Z. platypus are mainly in the number of scales under the lateral line, comparable characters such as the ratio of head length to body length, and the color of the upper edge of the eyes when living. Moreover, the genetic analysis based on mitochondrial cytochrome b gene also supports that Zacco tiaoxiensis sp. nov. is a valid species. [ABSTRACT FROM AUTHOR]

The authentication of food products and the verification of their identity are of major importance for consumers. Food fraud through mislabeling is an illegal practice consisting of the substitution of an expensive food product by a relatively cheaper one, misleading false labelling of their origin and adulteration in processed or frozen products. This issue is particularly of high importance concerning fish and seafood, which are easily adulterated primarily due to difficult morphological identification. Fish species of the Mullidae family are considered among the most high-valued seafood products traded in Greece and Eastern Mediterranean in general, in terms of the price and demand. Specifically, the red mullet (Mullus barbatus) and the striped red mullet (Mullus surmuletus) are both indigenous in the Aegean (FAO Division 37.3.1) and the Ionian (FAO Division 37.2.2) Seas, with high levels of consumers' preferences. However, they could be easily adulterated or misidentified by the invasive Aegean Sea Lessepsian migrator goldband goatfish (Upeneus moluccensis) as well as by the imported West African goatfish (Pseudupeneus prayensis). Keeping this in mind, we designed two novel, time-saving and easy-to-apply multiplex PCR assays and one multiple Melt–Curve analysis real-time PCR for the identification of these four species. These methodologies are based on species-specific primers targeting single nucleotide polymorphisms (SNPs) detected via sequencing analysis of the mitochondrial cytochrome C oxidase subunit I (CO1) and of the cytochrome b (CYTB) genes in newly collected individuals, with additional comparison with congeneric and conspecific haplotypes obtained from the GenBank database. Both methodologies, targeting CO1 or CYTB, utilize one common and four diagnostic primers, producing amplicons of different length that are easily and reliably separated on agarose gel electrophoresis, yielding a single clear band of diagnostic size for each species or a certain Melt–Curve profile. The applicability of this cost-effective and fast methodology was tested in 328 collected specimens, including 10 cooked samples obtained from restaurants. In the vast majority (327 out of the 328) of the specimens tested, one single band was produced, in agreement with the expected products with a single exception a M. barbatus sample that was identified as M. surmuletus, the identity of which was confirmed using sequencing, indicating erroneous morphological identification. The developed methodologies are expected to contribute to the detection of commercial fraud in fish authentication. [ABSTRACT FROM AUTHOR]

Trinomys , one of the most species-rich spiny rat genera in Brazil, is widely distributed in Caatinga, Cerrado and Atlantic Forest biomes, and currently includes ten recognized species, three of which are polytypic. Although some studies employing molecular data have been conducted to better characterize phylogenetic relationships among species, 19 nominal taxa have been suggested, implying considerable incongruence regarding species boundaries. We addressed this incongruence by intensively sampling all species across the geographic distribution of the genus. In addition to publicly available data, we generated 182 mt-Cytb gene sequences, and employed phylogenetic and computational species delimitation methods to obtain a clearer picture of the genus diversity. Moreover, we evaluated populational diversity within each accepted species, considering their geographical distribution and a timescale for the evolution of the genus. Beyond confirming the general patterns described for the evolution of the group, this new analysis suggests that Trinomys is comprised of at least 16 evolutionary lineages, 13 of them recognized as species or subspecies, and three never before characterized. This study highlights the importance of increased sample sizes and computational species delimitation methods in uncovering hidden diversity in Trinomys. [ABSTRACT FROM AUTHOR]

We describe a population of pipistrelle-like bats from Príncipe Island (Gulf of Guinea, Western Central Africa) as a new species based on the molecular and morphological characteristics of six specimens collected more than 30 years ago. The description of this new species was not possible until the traditionally entangled systematics of the whole pipistrelle group was clarified in recent years with the inclusion of molecular techniques and adequate species sampling. In this new taxonomic framework, the new species was clearly included within the dark-winged group of the recently described genus Pseudoromicia. The pipistrelles from Príncipe Island present a moderately inflated skull in lateral view with inner upper incisors that are moderately bicuspids and a baculum distinctly long with expanded tips. Besides these morphological characters, the new bat species is distinguished by its dwarfism, being the smallest species recognized within the genus. The ecology and conservation status of this endemic island species are unknown and field studies are urgently needed to evaluate the situation and conservation threats to this new species in its natural habitat. [ABSTRACT FROM AUTHOR]

Taxonomic decisions made by palaeontologists are often based on a few morphological features preserved in the fossil material. This practice may sometimes lead to the description of new species based on single specimens, which are, in fact, extreme or aberrant morphological variants of known taxa. Ancient DNA (aDNA) analysis of the Late Pleistocene specimens from the archaeological site Petits Guinards (Creuzier-la-Vieux, Allier, France), described as a new vole Microtus (Sumeriomys) bifrons, did not confirm the species distinctness of the studied population. The genetically examined specimens belonged to Stenocranius anglicus and/or Microtus arvalis, the dominant species at the site. Our findings show that it is risky to describe new fossil taxa on the basis of phenotypic outliers or morphologically aberrant, rare specimens that do not fall within the previously known population variability. We also highlight the importance of ancient DNA in resolving taxonomic and nomenclature problems and classifying fossil mammals of the Late Pleistocene age. [ABSTRACT FROM AUTHOR]

The author examined the occurrence pattern of mitochondrial DNA (mtDNA) haplotypes of house mouse (Mus musculus) subspecies musculus and castaneus using 61 individuals caught mainly in the northern area of Tochigi Prefecture, and previously unclarified genealogical discordance on the Pacific Ocean side of the Tohoku region (eastern Tohoku) was characterized. The musculus and castaneus mtDNA haplotypes were found to be widespread, respectively, in the southern and northern areas of the study area, and their geographic ranges are separated by a narrow boundary in Nasushiobara. On the northern side of the boundary, all 27 individuals carrying the castaneus mtDNA haplotypes proved to be homozygous for musculus-type alleles of nuclear genes Tcf25 and Dbndd1, certainly indicating 'mito-nuclear discordance' in eastern Tohoku. All 34 individuals on the boundary's southern side concordantly carry the musculus mtDNA haplotype and are homozygous for the same nuclear musculus-type alleles as is the population on the northern side of the boundary. The occurrence pattern of the mtDNA haplotypes differed from that seen in a hybrid zone (their mixture, i.e., around the Yangtze River in South China), suggesting that extensive introgression across the boundary is unlikely. As an alternative hypothesis, it is explained that the mito-nuclear discordance has arisen before the migration to eastern Tohoku. One individual carries a domesticus-type allele of the nuclear genes, suggesting that the mouse has been affected by introgression of the domesticus genome. These findings should help to clarify some issues in the colonization history of M. musculus on the Japanese islands. [ABSTRACT FROM AUTHOR]

Seven different methods of DNA extraction were performed and compared in order to have the better DNA recovery in terms of quality and quantity. A PCR assay was realised based on the Cytb mitochondrial gene to identify 53 commercial food products of wide range diversity (Scombridae Thunnus species) commercialised in the Tunisian market, and we checked whether these products were correctly labelled under European and national legislations. Phylogenetic analyses on Cytb mitochondrial gene were used to study the relationships among the considered species. Tuna species was easily differentiated and confirmed by direct sequencing. The results showed a high deficiency in labelling: 96% of canned tuna did not specify the species, although the name of the species had to be mentioned. Partial least squares regression (PLS) showed relatively high values (45%) in all analysed data, suggesting substantial difference between the extraction procedures and matrices with SDS/PCI method being the most convenient.

Abstract Understanding environmental DNA (eDNA) dynamics in the major developmental stages of fish species is fundamental for developing eDNA methods to aid conservation and management of fish species and/or populations. In this study, we used the model fish species, the rare minnow (Gobiocypris rarus), to assess the temporal changes of its mitochondrial and nuclear eDNA concentrations during its embryonic development in a culture dish experiment. Mitochondrial cytochrome b (Cytb) and nuclear internal transcribed spacer‐1 (ITS1), considered as representative DNA fragments, were amplified by quantitative PCR (qPCR). In this embryo culture trial, spikes in mitochondrial (mt‐) and nuclear (nu‐) eDNA concentrations were found just after egg fertilization and after embryo hatching, and a modest but positive relationship was found between the eDNA concentration and the number of hatched individuals, illustrating eDNA may be used to monitor the spawning and hatching events of externally fertilizing fishes. These findings provided basic information on eDNA dynamics during fish embryonic development and a new insight to design eDNA methods to monitor early developmental stages of fish species.

Abstract Background Avian haemosporidia infect both domestic and wild birds, causing anemia, acute tissue degeneration, and depopulation in wild birds. Poultry and wild birds have been reported as common reservoirs of haemosporidia, but limited information is available for red junglefowl (Gallus gallus) in China. The present study investigated the prevalence and molecular characterization of haemosporidia in red junglefowl. Methods Blood samples were collected from 234 red junglefowl from Jinghong City of Yunnan Province, and genomic DNA was extracted from these samples. The prevalence of haemosporidia was determined by nested PCR targeting the mitochondrial cytochrome b (cytb) gene. Molecular characterization was investigated based on phylogenetic analysis of cytb sequences, and associated risk factors were analyzed using the Chi-square (χ 2) test. Results The overall prevalence of haemosporidia was 74.8% (175/234), and three species were identified, namely Haemoproteus enucleator, Leucocytozoon californicus, and Plasmodium juxtanucleare. The prevalence of haemosporidia in adult fowl (81.1%, 107/132) was significantly higher (χ 2 = 6.32, df = 1, P = 0.012) than that in juveniles (66.7%, 68/102). Three novel haemosporidian lineages were revealed. Conclusions This study examined the prevalence and identified species of avian haemosporidians in red junglefowl, providing new information on the molecular epidemiology and geographical distribution of haemosporidian parasites. Our results indicated high prevalence and diverse species distribution of these haemosporidians in red junglefowl. To the best of our knowledge, this is the first record of haemosporidian infection in red junglefowl in China. Graphical Abstract

Recent studies have revealed that the Kerivoula depressa complex should be divided into two species, K. depressa distributed mainly in Myanmar, Vietnam, Laos and Cambodia, and K. dongduongana found only in the Annamite Mountains of Vietnam, Laos and Cambodia. In November 2018 and April 2019, 24 woolly bats were collected by two-band harp traps in Xishuangbanna, Yunnan, China. Based on morphological, morphometric, and phylogenetic (COI, Cytb, and RAG2 gene sequences) analyses, these bats were identified as K. depressa and K. dongduongana, representing two new species records for the country. Including the new records, six Kerivoula species have been recorded in China, namely K. depressa, K. dongduongana, K. furva, K. kachinensis, K. picta and K. titania. To facilitate their identification and biological research in the future, we have provided an up-to-date key to all Kerivoula species occurring in China. [ABSTRACT FROM AUTHOR]

The European eel (Anguilla anguilla) has declined by over 90% since the early 1980s and has been listed as critically endangered. Yet, despite strict export bans from the European Union, the European eel is still sold illegally in many countries. Efforts to monitor the trade of European eels have been primarily concentrated in Asian markets where concerningly high rates of European eel have been reported. Comparably fewer studies have assessed the identities of eel samples from the United States (US), despite the obvious implications for eel conservation. To address this knowledge gap, we purchased 137 eel products (134 freshwater eels and three saltwater eels) from grocers, sushi bars, and restaurants in nine states across the US from 2019 to 2021. Seven samples (5.2%) labeled as freshwater eels (or "unagi") were identified as European eels using a combination of mitochondrial (cytochrome b) and nuclear (18S rRNA) restriction digestion assays, a fast and inexpensive molecular tool for seafood identification that can identify hybrids between European eels (A. anguilla) and American eels (A. rostrata). No hybrids between European and American eels were found and all seven samples identified with restriction digestion as European eels were confirmed by sequencing of cytochrome b and 18S rRNA. Frequency of European eels in US markets did not significantly correlate with state or retail type. Although illegal eel exports are likely reaching US consumers, the frequency of European eel samples in this study of the US market is much lower than found in other non-European countries. [ABSTRACT FROM AUTHOR]

The bearded fireworm Hermodice carunculata is a conspicuous omnivorous benthic invertebrate distributed in the wider Atlantic Ocean, Mediterranean Sea and the Red Sea. Questions were raised in the past regarding the taxonomic status of amphiatlantic fireworm specimens but subsequent analysis of morphological data with new molecular data reconfirmed the presence of only one species, H. carunculata with amphiatlantic distribution. Hermodice carunculata has been the subject of several taxonomic and genetic studies but there are still questions regarding the genetic diversity and population structure of the species throughout its range. We contributed to the genetic studies of H. carunculata with ~200 new samples from over 20 locations in the Caribbean, Eastern Atlantic Ocean and the Mediterranean Sea. We sequenced the mitochondrial markers Cytochrome c oxidase Subunit I (COI) and Cytochrome b (Cytb) from each polychaete to examine the patterns of genetic diversity of H. carunculata. Our data revealed a significant population structure between the Caribbean H. carunculata including those from Brazil compared with those from the eastern Atlantic (Tenerife, Canary Islands) and eastern Mediterranean (Greece and Malta). We inferred rates of gene flow between eastern and western Atlantic populations by estimating the bidirectional effective migration rate (Nem). We corroborate previous studies indicating the existence of one species with genetically distinct populations. However, the biological significance of the observed population divergence should be evaluated with cross-breeding experiments. [ABSTRACT FROM AUTHOR]

Cystic echinococcosis (CE) is a neglected zoonotic disease caused by Echinococcus granulosus (sensu stricto). The parasite affects a wide range of livestock and wild animals. In this study, the population diversity of the Echinococcus species was investigated based on mitochondrial cytochrome b (cytb) and NADH dehydrogenase subunit 5 (nad5) genes. In addition to this, β-tubulin gene isoforms of Echinococcus granulosus were amplified to determine the resistance against benzimidazoles. For this purpose, 40 cyst samples from cattle (n = 20) and buffaloes (n = 20) were collected from the main abattoir of Sialkot. DNA extraction was performed using Qiagen Blood and Tissue Kits. Amplification was performed through PCR. Each amplicon was confirmed by GelRed™ stained agarose gel (2%). Samples were sequenced in a DNA analyzer and viewed for any misread nucleotide by using MEGA (v.11). Corrections in nucleotide sequence and multiple sequence alignment were made through the same software. NCBI-BLAST was used for sample specific sequences to identify them as belonging to a particular species. Diversity indices were estimated using DnaSP (v.6) while phylogenetic analysis was inferred using the Bayesian method using MrBayes (v.1.1). β-tubulin gene isoforms sequence analysis was performed to find out the candidate gene causing benzimidazole resistance. All 40 isolates were found positive for E. granulosus. BLAST-based searches of sequences of each isolate for each gene (nad5 and cytb) confirmed their maximum similarity with the G1 genotype. Overall, high haplotype diversity (Hd nad5 = 1.00; Hd cytb = 0.833) and low nucleotide diversity (π nad5 = 0.00560; π = cytb = 0.00763) was identified based on diversity indices. For both the genes, non-significant values of Tajima’s D (nad5 = −0.81734; cytb = −0.80861) and Fu’s Fs (nad5 = −1.012; cytb = 0.731) indicate recent population expansion. Bayesian phylogeny-based results of nad5 and cytb sequences confirmed their genotypic status as distinct from other Echinococcus species. This study shed light on the status of benzimidazole resistance in Echinococcus granulosus for the very first time from Pakistan. The findings of this study will significantly add in the information available on genetic diversity of Echinoccous granulosus based on cytb and nad5 genes sequences.