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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]

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]

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.

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]

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

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.

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: 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]

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]

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 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]

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]

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]

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]

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

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]

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]

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.

The Glossogobius species play an essential role in food supply and are distributed widely from the marine to freshwater, especially in the Mekong Delta, Vietnam (VMD). Some of their morphometrics and meristics are observed to vary with species and sampling sites. Therefore, the present study aims to verify if their mitochondrial Cytochrome b (Cytb) gene, one of the popular gene sequences used in fish phylogenic variation assessment, varies with species and sampling sites in the VMD. The Cytb gene size was 1300 bp for GcytbH/GcytbL primer pair and 1045 bp for GluMuq1-F/Mixcyto937-2R. The genetic distances within and among these three fish species groups were 0–11%. The Cytb gene sequences’ similarity between this study and the NCBI database was 85.84–100%. The Glossogobius specimens were observed to disperse in small branches of the phylogenetic tree with a low K2P value, suggesting that the Cytb genetic diversity may be low among species.

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 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.

Old world monkeys comprise 28 Langur species which belong to subfamily Colobinae under family Cercopithecidae. Nilgiri Langurs (Trachypithecus johnii) are endemic to the rain forests of the Western Ghats. The current study is an attempt at comparative phylogeny based on mitochondrial CYTB (mtCYTB) gene of Nilgiri Langur with other langurs. Faecal sample was collected from Nilgiri langur and the genomic DNA was isolated. The 1140 bp mitochondrial CYTB was amplified and sequenced using Sanger’s di-deoxy method. The amplified sequence along with the 27 sequences of Trachypithecus and Semnopithecus that were retrieved from the GenBank database were used for analysis. The phylogenetic tree was constructed by the maximum likelihood method in MEGA X. The analysis showed the clustering of Nilgiri langur with other langurs of the Semnopithecus sp. as a single clade.

Bombus terrestris dalmatinus is naturally common in many countries, including Turkey, and is also used commercially for the pollination of greenhouse plants. Intensive commercial production and international trade in many countries are considered as reasons for the disappearance of some natural populations. Hybridization of native bumble bees with those produced commercially, but having escaped from greenhouses and colonization of these commercial bees in natural habitats are cause for concern. In order to assess this concern, B. t. dalmatinus workers were collected from twelve different populations: five commercial producers, three surrounding greenhouse centers, three natural areas at least 30 km away from greenhouses, and one more recent greenhouse zone in Antalya, Turkey. The genetic variations and relationships among the twelve populations were estimated using SNP haplotypes determined in mitochondrial COI and CytB. Twenty and sixteen haplotypes were obtained for COI and CytB, respectively. A single haplotype, H1, was widespread with a high frequency in all individuals for both genes. Individuals collected from around greenhouse centers and commercial companies had more common haplotypes. The genetic variations of intra-populations were higher than the interpopulations in both COI (65.41%>34.59%) and CytB (72.47%>27.53%). The natural and commercial populations were genetically more distant from each other considering Fst values. However, samples from near the greenhouses had a higher similarity with the commercially produced samples, while the natural populations far away from greenhouses still retained their genetic distinctiveness.

Abstract Background African trypanosomiasis, caused by protozoa of the genus Trypanosoma and transmitted by the tsetse fly, is a serious parasitic disease of humans and animals. Reliable data on the vector distribution, feeding preference and the trypanosome species they carry is pertinent to planning sustainable control strategies. Methodology We deployed 109 biconical traps in 10 villages in two districts of northwestern Uganda to obtain information on the apparent density, trypanosome infection status and blood meal sources of tsetse flies. A subset (272) of the collected samples was analyzed for detection of trypanosomes species and sub-species using a nested PCR protocol based on primers amplifying the Internal Transcribed Spacer (ITS) region of ribosomal DNA. 34 blood-engorged adult tsetse midguts were analyzed for blood meal sources by sequencing of the mitochondrial cytochrome c oxidase 1 (COI) and cytochrome b (cytb) genes. Results We captured a total of 622 Glossina fuscipes fuscipes tsetse flies (269 males and 353 females) in the two districts with apparent density (AD) ranging from 0.6 to 3.7 flies/trap/day (FTD). 10.7% (29/272) of the flies were infected with one or more trypanosome species. Infection rate was not significantly associated with district of origin (Generalized linear model (GLM), χ2 = 0.018, P = 0.895, df = 1, n = 272) and sex of the fly (χ2 = 1.723, P = 0.189, df = 1, n = 272). However, trypanosome infection was highly significantly associated with the fly’s age based on wing fray category (χ2 = 22.374, P

The phylogeographic analysis of Stylodipus telum was performed based on cytochrome b mitochondrial gene. Two westernmost subspecies, S.t. falzfeini and S.t. turovi, both formed independent monophyletic clades, but the genetic distance between them clearly indicates that neither should be treated as a separate species. Individuals from remote populations of S.t. falzfeini mix together and do not have a clear phylogeographic structure, which could indicate a recent panmixia within the subspecies. Populations in Kherson Region are under threat due to the ongoing Russian invasion.

Due to environmental pollution and overfishing, the germplasm resources of wild loach (Misgurnus anguillicaudatus) have been degraded. To explore the genetic diversity of different geographical populations of loach in the Pearl River Basin, a total of 184 M. anguillicaudatus individuals from six dojo loach natural populations sampled from Yingde (Beijiang), Qingyuan (Beijiang), Liannan (Beijiang), Nanxiong (Dongjiang), Meizhou (Dongjiang), and Liuzhou (Liujiang) were investigated based on microsatellite DNA markers and mitochondrial DNA markers (cytb and D-loop). The results showed that a total of 71 alleles (Number of alleles, Na) were amplified in the six M. anguillicaudatus populations by 10 pairs of microsatellite primers. The average observed heterozygosity (Ho), expected heterozygosity (He), and polymorphic information content (PIC) among were 0.3892, 0.5101, and 0.4781, respectively. The average genetic differentiation coefficient (Fst) and number of migrants per generation (Nm) of the six M. anguillicaudatus populations were 0.3395 and 0.4863, respectively, indicating a low level of genetic exchange among the six populations and a high degree of genetic differentiation. Moreover, based on cytb, the number of haplotypes (h), the haplotype diversity (Hd), and nucleotide diversity (π) of the six M. anguillicaudatus populations were 34, 0.925, and 0.0539, respectively, and were 36, 0.879, and 0.0160 based on D-loop sequences. The Fst values among the six populations were 0.4963 (cytb) and 0.4833 (D-loop), and they all showed a high degree of genetic differentiation. The above results indicate that there is abundant genetic diversity of M. anguillicaudatus in the Pearl River Basin in China, and the fish could be used as excellent materials for breeding. The study provides a theoretical basis for the protection and utilization of M. anguillicaudatus natural resources in the Pearl River Basin of China.

The phylogeography of the filed vole (Microtus agrestis) in Eurasia was thoroughly examined using mitochondrial DNA (mtDNA) of the cytochrome b (cytb) gene. However, the former conclusions about genetic variability and the contact zone of eastern and western genetic lineages in Lithuania were based on the analysis of a very limited number of individuals. In the present study, we examined 74 M. agrestis individuals trapped in four sites in the eastern, northern, and western parts of the country using sequence analysis of cytb and D-loop. Totals of 25 new cytb haplotypes and 19 new D-loop haplotypes were identified for this species. Higher nucleotide diversity was observed for D-loop (π = 0.01147 ± 0.00070) as compared to cytb (π = 0.00694 ± 0.00039). The phylogenetic analysis based on both loci revealed the presence of two genetic lineages, i.e., the eastern and western ones, which were mixed in Lithuanian samples, with the exception of the Rusnė site in the west of the country. Only the western lineage was observed in this island population of M. agrestis; the sample differed in low genetic variability and genetic differentiation from other investigated samples. We found D-loop to be an appropriate locus for the evaluation of the genetic variability of M. agrestis. [ABSTRACT FROM AUTHOR]

An integrative analysis of an isolated population of Harting's vole (Microtus hartingi) from the Eastern Rhodope Mountains (Bulgaria) was carried out by morphological and morphometric methods, computed tomography, Cytb variation data, and experimental hybridization. Substantial changes in the development of the skull and teeth were found. Nevertheless, those voles can live to the senex stage. A phylogenetic reconstruction based on Cytb sequences showed that M. hartingi from the Eastern Rhodopes forms a separate clade, which is a sister clade to the voles from Northeastern Greece (also from the foothills of the Rhodopes). M. hartingi from the Rhodopes is mostly isolated reproductively from M. h. ankaraensis from Anatolia (Turkey), as evidenced by a decrease in the proportion of pairs that started breeding, relatively high mortality of pups, and increased sterility of hybrid males. Possible time of isolation of the Rhodopean population and consequences of the "bottleneck" effect on its current state are discussed. At the same time, we believe that, at present, it is impossible to determine its taxonomic status. It is necessary to recognize M. hartingi from the Bulgarian Rhodopes as an endangered population and to include it in regional Red Lists of Bulgaria. [ABSTRACT FROM AUTHOR]

Widespread surveys of avian haemosporidia (Plasmodium, Haemoproteus, and Leucocytozoon) in wild birds have substantially advanced information on the haemosporidian fauna of Japan. However, many areas and bird species remain insufficiently investigated. Bird carcasses collected for personal specimen collection seldom reach academic audience particularly in the veterinary field. The presence of avian haemosporidia was investigated in these personally collected bird carcasses, in order to better understand the avian haemosporidian fauna in Japan. Bird carcasses were donated through personal contact upon approval of the study. Tissue samples were collected from the birds and examined for haemosporidian parasites using nested-PCR targeting the cytochrome b gene. One hundred and forty-three birds of 85 species were donated, including 34 species and two subspecies that were molecularly or collectively investigated for the first time in Japan. Avian haemosporidian DNA was detected from 37 of the 134 tested birds (27.61%). In 8 bird species, avian haemosporidia was detected for the first time. Twenty-nine lineages were detected, including 8 novel and 9 known lineages detected in Japan for the first time. Furthermore, 16 lineages were detected from novel host species. While information that could be drawn was limited and risk management of zoonotic diseases needs re-consideration, these findings expanded information on the host range and distribution of several lineages. Collectively, this method of investigation using personally collected bird samples can provide important additions to more fully understand the avian haemosporidian fauna of Japan, as well as other areas with limited investigations. [ABSTRACT FROM AUTHOR]

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Flies (Diptera) play a significant role in the ecosystem as pollinators and decomposers, and they are also important vermin and disease vectors. Studies on the dipteran species are still lacking in Malaysia; therefore, the dipteran species' biology, morphology, distribution, and abundance are necessary. The objectives of this study were to identify dipteran species using a molecular approach, determine flies' Bovidae hosts, and investigate the diversity of the fly's species at three different cattle farms purposively selected in Selangor, Malaysia. The fly species were identified using cytochrome oxidase subunit I (COI) (Haematopota javana, Tabanus rubidus, Tabanus fontinalis, Iranihindia martellata, Musca domestica, and Chrysomya megacephala), while another six species only up to genus level (Haematopota sp. 1, Musca sp. 1, Asilus sp., Metopia sp., Anasillomos sp., and Ommatius sp.). In addition, two species of hosts: Bos indicus and Bos taurus, were proven to be the associated host species for the dipteran species based on molecular data of cytochrome b (cytb). However, there were no significant differences between farms in species diversity and richness (F = 1.262, df = 2, p = 0.2459 >0.05). Interestingly, the two most dominant dipteran genera collected from the cattle farms were Musca and Chrysomya. At the same time, its abundance may have been influenced by the structure of the cattle cage flooring, which serves as a breeding site and food source. These findings contribute to fundamental epidemiological data in developing control strategies for dipteran species and are of great economic and health importance to livestock production in Malaysia. [ABSTRACT FROM AUTHOR]

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 (χ2) 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 (χ2 = 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. [ABSTRACT FROM AUTHOR]

Ants of the «Formica picea—F. candida» complex are widespread across Eurasia. However, it is still a matter of debate if it constitutes one or two species. In this study, we collected a sample of specimens from different parts of Eurasia, sequenced the mitochondrial cox1 and cytb genes, as well as three nuclear loci—wg, Top1, and ITS2—and analyzed the available published data. We found this complex to contain a new, yet undescribed, taxon that has a large distribution in Siberia and East Asia. Thus, the «Formica picea—F. candida» complex consists of at least three taxa with distinct distributions. [ABSTRACT FROM AUTHOR]

HCO: untrained group receiving high-dose cocoa at 24.6 mg/kg b.w.; CONTEx: control exercise-trained group; COEx: exercise-trained group supplemented with cocoa at 8.2 mg/kg; HCOEx: exercise-trained group supplemented with high-dose cocoa at 24.6 mg/kg b.w. The performance of an exercise bout led to an elevation in oxidative stress, as supported by the TBARS and SOD data, and this was attenuated after 10 weeks in the both CT and CO groups by the practice of physical exercise and the increased physical condition, as expected [[47]], but this was more marked in athletes supplemented with cocoa, which may diminish the adaptations to exercise [[48]]. The period in which the supplementation occurred, during which time great adaptations to exercise were taking place, the framework of the athletes' diets, which were rich in fruits and vegetables, and the type of cocoa used in the study, could be determining factors in the observed effects of cocoa on athletes. (CO), (3) untrained group receiving high-dose cocoa at 24.6 mg/kg b.w (HCO), (4) control exercise-trained group (CONTEx), (5) exercise-trained group supplemented with cocoa at 8.2 mg/kg b.w. [Extracted from the article]

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.

We demonstrated the genetic variation and population structure of a direct developing marine gastropod, Littorina horikawai which is endemic species distributed in the western coast of Kyushu, Japan and is found mainly on the adjacent islands. We conducted the field survey and sampling covering most of the distribution area of L. horikawai. In total, we collected 173 individuals from 13 localities (between Tsushima Island and Yakushima Island) and determined the sequences of mitochondrial DNA, the cytochrome b (CYTB) gene. We detected a significant genetic population differentiation in L. horikawai despite the limited distribution area of the western coast of Kyushu. Also, its haplotype distribution is mainly consistent with the geographical structure: the northern part of Kyushu, the central islands, and the southern islands. However, some haplotypes obtained from the Goto Islands which is located in the border between the northern part of Kyushu and the central islands do not correspond to the above geographic distribution. The high level of genetic differentiation of L. horikawai compared to other low dispersal Littorina species despite its narrow distribution range may be due to its predominantly fragmented island habitats. Although it is necessary for understanding the detailed evolutionary history of L. horikawai to perform the high-resolution genetic analysis, our present findings indicated that a high level of genetic differentiation of L. horikawai occurred even in a narrow distribution range because of not only the ecological traits linking to dispersal ability but also its discontinuous pattern such as the island inhabitation. [ABSTRACT FROM AUTHOR]

In this study, the phylogeographical pattern of the Amur minnow (Rhynchocypris lagowskii) widely distributed in the cold freshwaters of the Qinling Mountains was examined. A total of 464 specimens from 48 localities were sequenced at a 540‐bp region of the mitochondrial cytochrome b (Cytb) gene, and 69 haplotypes were obtained. The mean ratio of the number of synonymous and nonsynonymous substitutions per site (dN/dS) was 0.028 and indicated purifying selection. Haplotype diversity (h) and nucleotide diversity (π) of natural populations of R. lagowskii varied widely between distinct localities. Phylogenetic trees based on Bayesian inference (BI), maximum likelihood (ML), and maximum parsimony (MP) methods, and network analysis showed five well‐differentiated lineages, but these did not completely correspond to localities and geographic distribution. Meanwhile, analysis of molecular variances (AMOVA) indicated the highest proportion of genetic variation was attributed to the differentiation between populations rather than by our defined lineages. In addition, there was no significant correlation between the pairwise Fst values and geographic distance (p >.05). Based on the molecular clock calibration, the time to the most recent common ancestor (TMRCA) was estimated to have emerged from the Late Miocene to the Early Pleistocene. Finally, the results of demographic history based on the neutrality test, mismatch distribution, and Bayesian skyline plot (BSP) analyses showed that collectively, the populations were stable during the Pleistocene while one lineage (lineage E) probably underwent a slight contraction during the Middle Pleistocene and a rapid expansion from the Middle to the Late Pleistocene. Therefore, the study suggests the current phylogeographical pattern of R. lagowskii was likely shaped by geological events that led to vicariance followed by dispersal and secondary contact, river capture, and climatic oscillation during the Late Miocene to the Early Pleistocene in the Qinling Mountains. [ABSTRACT FROM AUTHOR]