Phylogeography and population structure of Lagocephalus spadiceus (Richardson, 1845) (Tetraodontiformes, Tetraodontidae) in the South China Sea.

The climate fluctuations during the Late Pleistocene significantly influenced the phylogeographic structure and historical dynamics of marine fishes in the marginal seas of the western Pacific Ocean. The puffer fish, Lagocephalus spadiceus, holds substantial nutritional and economic value in the South China Sea. To investigate the demographic history and population structure of the L. spadiceus, the mitochondrial DNA COI and Cyt b gene datasets from 300 individuals across eight populations in the South China Sea were sequenced. Our findings revealed high haplotype diversity (0.874 ± 0.013) and low nucleotide diversity (0.00075 ± 0.00058). The phylogenetic tree and haplotype networks revealed no significant genetic differentiation along the northern coast of South China Sea. Neutrality tests, mismatch distribution analyses, and Bayesian skyline plots suggested that L. spadiceus underwent population expansion during the Late Pleistocene. Both ocean currents and climate change significantly influenced the geographical distribution and genetic population structure of L. spadiceus.

Population genetic characterization of Theileria annulata based on the cytochrome b gene, with genetic insights into buparvaquone susceptibility in Haryana (India).

The present investigation was aimed at population genetic characterization of Theileria annulata on the basis of the cytochrome b (cyt b) gene along with the evaluation of status of buparvaquone resistance in Haryana (India). The sequences originating from China, Egypt, India, Iran, Iraq, Tunisia, Turkey and Sudan were included in the analysis. The maximum likelihood tree based on the Tamura-Nei (TN93+G) model placed all the sequences of T. annulata into a single clade. The median-joining haplotype network exemplified geographical clustering between T. annulata haplotypes originating from each country. Only five haplotypes (7.81 %) were shared between any two countries, while the remaining 59 haplotypes (92.19 %) were singleton and unique to one country. The values of pairwise genetic distance (FST) between all the populations indicated huge genetic differentiation (> 0.25) between different T. annulata populations, barring the FST value between Iraq and Turkey (0.14454) which suggested a moderate differentiation. Contrary to the FST index, the values of gene flow (Nm) between T. annulata populations were very low. The neutrality indices and mismatch distributions indicated a population expansion in the Indian T. annulata population. Furthermore, the secondary structure and homology modeling of the partial cyt b protein is also reported. The molecular analysis of newly generated sequences for buparvaquone resistance revealed that all the isolates were susceptible to buparvaquone treatment. However, two novel mutations at positions V203I and V219I in between the Q01 and Q02 drug-binding regions of the cyt b gene were observed for the first time.

Confirmation of the existence of Himalayan long-eared bats, Plecotushomochrous (Chiroptera, Vespertilionidae), in China.

The existence of Himalayan long-eared bats, Plecotushomochrous (Chiroptera, Vespertilionidae), in China has not been previously confirmed. In this study, four bats captured with harp traps from two sites in the Maoershan National Nature Reserve in Guangxi, China were investigated. These bats have long, wide auricles, each with a prominent tragus. The length of each auricle is about the same as that of a forearm. Hairs on the ventral fur have a dark base with mixed grey and yellowish tips; those on the dorsal fur also have a dark base and are bicolored with brown tips. The thumbs are very short. A concavity is present in the front of the dorsal side of the cranium. Based on morphological characteristics and phylogeny using Cyt b gene sequences, these bats were identified as P.homochrous, thus confirming the existence of Himalayan long-eared bats in China.

Genetic diversity and relationship among Indonesian local sheep breeds on Java Island based on mitochondrial cytochrome b gene sequences.

BACKGROUND: The cytochrome b (Cyt b) gene is one of the most studied mitochondrial DNA (mtDNA) genes to determine sheep's genetic profile. This study aimed to determine the genetic diversity and relationships of several Indonesian local sheep populations on Java Island, Indonesia, based on the mtDNA Cyt b gene sequences. Blood samples were collected from forty-one individual sheep in seven populations of Indonesia local sheep breeds on Java Island (Priangan = 6, Garut = 6, Batur = 7, Wonosobo = 5, Javanese Thin-Tailed/JTT = 7, Javanese Fat-Tailed/JFT = 5, and Sapudi = 5). DNA extraction was performed on blood samples, and the mtDNA Cyt b gene was amplified using specific primers (Alek-CBF: 5'-CAACCCCACCACTTACAA-3' and Alek-CBR: 5'-CCTTGAGTCTTAGGGAGGTT-3'). The polymerase chain reaction (PCR) products were then sequenced, and data were analyzed using the MEGA version 7.0, DNA SP version 6.0, and NTSYS-pc version 2.11 software. RESULTS: A total of 1140 bp complete mtDNA Cyt b gene sequences in this study obtained 1134 monomorphic sites (I), six polymorphic sites (V), one segregation site (S), and five parsimony informative sites (P) with a nucleotide diversity (Pi), the average number of nucleotide differences (K), and sequence conservation (SC) were 0.00119, 1.35610, and 0.9947, respectively. There were six haplotypes consisting of two unique haplotypes and four shared haplotypes with a haplotype diversity (Hd) of 0.5415. The genetic distance within and between populations ranged from 0.0000 to 0.0016 and 0.0000 to 0.0020, respectively. Wonosobo, JFT, and Sapudi sheep have the closest relationship, and then these three breeds were close to JTT sheep, followed by Batur, Priangan, and Garut sheep. Two haplogroups have been found based on the Ovine haplogroup clustering. All Wonosobo, JTT, JFT, Sapudi sheep, and most Batur sheep were clustered into haplogroup B. In contrast, Garut sheep were mostly clustered into haplogroup A, while Priangan sheep were clustered into both haplogroups with the same percentage. CONCLUSION: Seven Indonesian local sheep breeds on Java Island have a close relationship clustered into two haplogroups, namely haplogroups A and B. Most Indonesian local sheep breeds on Java Island in this study were clustered into haplogroup B, except for Garut and Priangan sheep.

Temperature-Specific and Sex-Specific Fitness Effects of Sympatric Mitochondrial and Mito-Nuclear Variation in Drosophila obscura.

The adaptive significance of sympatric mitochondrial (mtDNA) variation and the role of selective mechanisms that maintain it are debated to this day. Isofemale lines of Drosophila obscura collected from four populations were backcrossed within populations to construct experimental lines, with all combinations of mtDNA Cyt b haplotypes and nuclear genetic backgrounds (nuDNA). Individuals of both sexes from these lines were then subjected to four fitness assays (desiccation resistance, developmental time, egg-to-adult viability and sex ratio) on two experimental temperatures to examine the role of temperature fluctuations and sex-specific selection, as well as the part that interactions between the two genomes play in shaping mtDNA variation. The results varied across populations and fitness components. In the majority of comparisons, they show that sympatric mitochondrial variants affect fitness. However, their effect should be examined in light of interactions with nuDNA, as mito-nuclear genotype was even more influential on fitness across all components. We found both sex-specific and temperature-specific differences in mitochondrial and mito-nuclear genotype ranks in all fitness components. The effect of temperature-specific selection was found to be more prominent, especially in desiccation resistance. From the results of different components tested, we can also infer that temperature-specific mito-nuclear interactions rather than sex-specific selection on mito-nuclear genotypes have a more substantial role in preserving mtDNA variation in this model species.

Development of loop-mediated isothermal method and comparison with conventional PCR assay for rapid on spot identification of tissue of cattle origin.

Loop-mediated isothermal amplification (LAMP) is a diagnostic method for meat speciation with rapid and minimal equipment requirements. In this study, we developed cattle-specific tube-based LAMP assays targeting mitochondrial Cyt b gene sequence, compared with conventional PCR assay for specificity, sensitivity, and validation of the assay was made. The LAMP reaction was carried at 64 °C for 45 min, and results were confirmed by SYBR Green I dye and agarose gel-electrophoresis. The specificity of the assays was cross-tested with DNA of buffalo, goat, sheep, and pork. The amplification was observed with samples from cattle only without cross-reactivity with other meat species. The analytical sensitivity of LAMP and PCR method for cattle DNA detection was 0.0001 ng and 1 ng, respectively. Repeatability of the assay was achieved on samples from known/blind and admixture meat with other than cattle at the relative percentage of 20%, 10%, 5%, and 1%. The study concluded that the developed assay can be easily employed for the rapid identification of tissue of cattle origin in meat and meat products in low resource areas.

Vanmaneniamarmorata, a new species of loach (Teleostei: Gastromyzontidae) from the middle Chang-Jiang Basin in Guizhou Province, south China.

BACKGROUND: The gastromyzontid genus Vanmanenia was established by Hora in 1932, based on the type species Vanmaneniastenosoma. The genus is a loach group adapted to running waters of streams from southern China, northern Vietnam and Laos. Currently, 19 valid species of the genus have been recognised. The northernmost distribution of the genus is the Yangtze River (= Chang-Jiang in Chinese) Basin and five species (V.maculata, V.intermedia, V.stenosoma, V.pseudostriata and V.gymnetrus) have been reported from the Basin. NEW INFORMATION: Vanmaneniamarmorata, a new hillstream species of loach, is here described from the middle Chang-Jiang Basin in Guizhou Province, south China. It is distinguished from its congeners by having a combination of the following characters: three triangular-shaped rostral lobules; postdorsal saddles wider than interspaces; a more backwards-placed anus (the vent to anal distance 30.5-36.9% of the pelvic to anal distance); a larger gill opening with its upper extremity reaching the level of the middle of the orbit; anal-fin base length 5.6-6.4% of SL; caudal-peduncle length 11.6-12.9% of SL; prepelvic length 51.1-53.4% of SL. Its validity is also affirmed by its distinct cyt b gene sequence divergence with all sampled congeners and its monophyly recovered in a cyt b gene-based phylogenetic analysis.

CRISPR-Cas12-Based Rapid Authentication of Halal Food.

The halal food market is globally growing along with the increased risk of adulteration. We proposed an amplification-free and mix-to-read CRISPR-Cas12-based nucleic acid analytical strategy allowing rapid identification and analysis of pork components, thus enriching the toolbox for ensuring halal food authenticity. We designed and optimized guide RNA (gRNA) targeting the pork cytochrome b (Cyt b) gene. gRNA allowed specific identification of the target Cyt b gene from pork components followed by activation of Cas12 protein to abundantly cleave single-stranded DNA probes with terminally labeled fluorophore and quencher groups, thus turning on fluorescence. The presence of the pork Cyt b gene thus can be mix-and-read- and only-one-step-detected, which may indicate the risk of halal food adulteration. The method allowed specific discrimination of pork meat from beef, mutton, and chicken and yielded a detection limit of 2.7 ng/µL of total DNA from pork meat. The reliability of the method was tested using the following processed meat products: halal foods beef luncheon meat and spiced beef and non-halal foods sausage and dried pork slices. The CRISPR-Cas12-based nucleic acid test strategy is promising for rapid food authentication.

Genetic diversity of wild wintering red-crowned crane (Grus japonensis) by microsatellite markers and mitochondrial Cyt B gene sequence in the Yancheng reserve.

The red-crowned crane (Grus japonensis) is one of the most endangered cranes in the world, and its wild population is still declining. To characterize the genetic resources of East Asian migratory populations, we studied the genetic variation in wild red-crowned cranes at the Yancheng reserve. Based on a partial Cyt b gene sequence, 32 wild red-crowned crane samples were screened from 100 feathers with unknown sample information. Twelve haplotypes were detected using 32 wild red-crowned crane samples. Six pairs of published microsatellite primers were selected for genotyping. A total of 47 alleles were obtained, with an average of 7.8 alleles per locus. All microsatellite loci were highly polymorphic; the average polymorphic information content and expected heterozygosity were 0.721 ± 0.080 and 0.768 ± 0.071, respectively. These results show that the East Asian migratory population of wild red-crowned cranes exhibits high polymorphism. These data are useful for informing reintroduction efforts. The study results provide a basis for understanding the population genetic properties of an endangered crane.

Vanmanenia intermedia Fang, 1935, a valid hill-stream species of loach (Teleostei: Gastromyzontidae) from the middle Yangtze River basin, Southwest China.

Vanmanenia intermedia, which has long been synonymized with V. pingchowensis since 1980, is here revalidated using morphological characters and molecular evidences, and redescribed on the basis of topotypic specimens. This species is distinguished from all other congeneric species by the following combination of characters: triangular-shaped rostral lobules without secondary rostral barbels; distal edge of dorsal-fin slightly concave; no longitudinal black stripe on the flank; a longer snout (length 52.6-64.5 % of HL); a more backwards-placed anus (the vent to anal distance 34.7-46.1% of the pelvic to anal distance); a slender caudal peduncle (depth 9.1-11.3 % of SL); a longer anal-fin base (length 7.5-9.5 % of SL); and a smaller gill opening with its upper extremity closer to the level of the lower margin of the orbit. The validity of V. intermedia is also confirmed by its significant cyt b gene sequence divergence with all sampled congeneric species and its monophyly recovered in the cyt b gene-based phylogenetic analysis.

Study on Identification Methods of Bullwhip by PCR Technology Based on Cyt b Gene / 中国药房

OBJECTIVE： To establish the method for PCR identification of bullwhip， and to identify the authenticity of bullwhip at the molecular level. METHODS： DNA samples of bullwhip and its counterfeits （donkey whip， pig whip， sheep whip） were extracted and their integrity， purity and concentration were detected. Using GenBank related information， using mitochondrial cytochrome b （Cyt b） gene of bullwhip as target gene， Primer-BLAST online software was used to design specific primer. PCR amplification was performed for whips of different species， and electrophoretic analysis was conducted for the product. PCR products of bullwhip samples were cloned and confirmed by DNA sequencing. The specificity and repeatability of the established PCR method were verified. RESULTS： DNA purity of the bullwhip and its counterfeits was high， and there was no protein or RNA pollution. 1.5％ agarose gel electrophoresis showed that there were obvious target gene bands of bullwhip samples at 200-300 bp， while no corresponding bands appeared in other counterfeit products. The results of DNA sequencing showed that the nucleotide sequence of the gene fragment of bullwhip was 100％ similar to that of the bullwhip in GeneBank. Results of methodological validation showed that established method was specific and reproducible. CONCLUSIONS： The established PCR identification method based on Cyt b gene in the study is simple， rapid， accurate， specific and reproducible， and can meet the requirements of analysis and identification of bullwhip and its counterfeits.

Garra dengba, a new species of cyprinid fish (Pisces: Teleostei) from eastern Tibet, China.

Garra dengba is here described from the Chayu-Qu, a tributary flowing into the Brahmaputra River, in Chayu County, eastern Tibet, China. It shares the presence of an incipient proboscis on the snout with G. arupi, G. elongata, G. gravelyi, G. kalpangi, and G. rotundinasus, but is distinguished from these five species in having, among other features, fewer branched dorsal- and anal-fin rays and more perforated lateral-line scales. Its validity was also confirmed by a molecular phylogenetic analysis based on the cytochrome b gene.

Species adulteration in raw milk samples using polymerase chain reaction-restriction fragment length polymorphism.

BACKGROUND AND AIM: Milk adulteration is pivotal because it leads to worse effects in public health as human adverse reactions with clinical signs ranged from gastrointestinal signs to anaphylactic shock. This study was carried out to estimate the prevalence of adulteration in buffalo's milk sold in Assiut City, Egypt. MATERIALS AND METHODS: A total of 50 raw buffalo's milk samples were collected and examined for adulteration by addition of cow's milk. The examination carried out by applying polymerase chain reaction-restriction fragment length polymorphism technique using cytochrome b (cyt b) gene primers and Hinf I enzymes. The size of target gene was 360 bp in both animal species and amplicon can be digested using Hinf I enzyme, this restriction enzyme divided the essential band to clear three bands at 360, 210, and 150 bp in cows' milk, while, the enzyme could not be cleaved the amplicon in buffalo's samples. RESULTS: The obtained results cleared that the incidence of adulteration of buffalo's milk very high percentage reaches 90%. CONCLUSION: It could be concluded that the raw buffalo's milk sold in Assiut City subject to fraudulent practice and thus can lead to public health hazards.

Two new species of Garra (Cypriniformes: Cyprinidae) from the lower Yarlung Tsangpo River drainage in southeastern Tibet, China.

Two new species of the fish genus Garra (Cypriniformes: Cyprinidae) are described from the lower Yarlung Tsangpo River drainage in southeastern Tibet, China. Garra motuoensis, a member of the proboscis species group, is primarily distinguished from the members of this group by having a prominent, quadrate, and slightly bilobed proboscis; 6-20 small to middle-sized unicuspid tubercles on the anterior region of the proboscis, some middle-sized unicuspid tubercles on the transverse lobe of the snout, and several small unicuspid tubercles on the lateral surface of snout; the anus situated slightly closer to the anal-fin origin than to the pelvic-fin origin (distance from anus to anal fin 36-46% of pelvic-anal distance); 36-37 lateral-line scales; and 12 circumpeduncular scales. Garra yajiangensis, a member of the proboscis species group, is primarily distinguished from the members of this group by having a prominent, quadrate, and slightly bilobed proboscis, covered with 2-7 middle-sized unicuspid tubercles, including two large-sized tubercles on anterior margin of each lobe; some small unicuspid tubercles on the transverse lobe and the lateral surface of snout; the anus situated substantially closer to the anal-fin origin than to the pelvic-fin origin (distance from anus to anal fin 19-24% of pelvic-anal distance); 34-36 lateral-line scales; and 12 circumpeduncular scales. The validities of these two new species were also corroborated by molecular phylogenetic analysis based on Cyt b gene sequences.

Exostoma tibetana, a new glyptosternine catfish from the lower Yarlung Tsangpo River drainage in southeastern Tibet, China (Siluriformes: Sisoridae).

Exostoma tibetana, new species, is described from the lower Yarlung Tsangpo River drainage in southeastern Tibet, China. It is distinguished from congeners by having an adipose fin separate from the upper procurrent caudal-fin rays and incised at the posterior extremity of its base; the tip of the pelvic fin not reaching to the anus when adpressed; 10 branched pectoral-fin rays; and morphometric features associated with the body depth, adipose-fin base length, caudal-peduncle depth, snout length, and eye diameter. The validity of this new species was also supported by molecular phylogenetic analysis based on Cyt b gene sequences.

Genetic characterization of wild swamp deer populations: ex situ conservation and forensics implications.

Swamp deer (Rucervus duvaucelii) is an endemic, Scheduled I species under the Wildlife (Protection) Act 1972, India. According to variations in antler size, it has been classified into three subspecies, namely Western (R. duvaucelii duvaucelii), Central (R. duvaucelii branderi), and Eastern (R. duvaucelii ranjitsinhii). For planning effective ex situ and in situ conservation of a wide-ranging species in different bioclimatic regions and in wildlife forensic, the use of genetic characterization in defining morpho/ecotypes has been suggested because of the geographic clines and reproductive isolation. In spite of these morphotypes, very little is known about the genetic characteristics of the three subspecies, hence no strict subspecies-based breeding plan for retaining the evolutionary characteristics in captive populations for subsequent re-introduction is available except for a few studies. We describe the genetic characteristics of these three subspecies using cytochrome b of the mtDNA genome (400 bp). The DNA sequence data indicated 11 variable sites within the three subspecies. Two paraphyletic clades, namely the Central India and Western-Eastern populations were found, whereas the Western and Eastern populations are monophyletic with a bootstrap value of 69% within the clade. We suggest the need of sorting these three subspecies using different molecular mtDNA markers in zoos for captive breeding purposes so as to retain the genetic diversity of the separate geographic clines and to use a subspecies-specific fixed-state nucleotide to assess the extent of poaching to avoid any population demography stochastically in India.

Genetic characteristics of Spermophilus in Shaanxi Province, China / 中国人兽共患病学报

We studied the genetic characteristics of Spermophilus in Shaanxi Province,China.The COI,Cyt-b gene were sequenced and the results were compared with those of dauricus from Inner Mongolia Keyouzhong Banner and Zhengxiangbai Banner,and S.alaschanicus from Haiyuan County of Ningxia.And genetic distance was analyzed and Neighbor-Joining tree was built.Results showed that the genetic distance of COI gene sequences between Spermophilus from Dingbian County in Shaanxi and S.alaschanicus in Ningxia was ≤0.5％,and the genetic distance was ranged from 7.9％ to 9.3％ with Citellus dauricus from Inner Mongolia.The genetic distance of Cyt-b gene between Spermophilus from Dingbian County in Shaanxi and S.alaschanicus in Ningxia was ≤2.2％,and ranged from 8.9％ to 11.2％ with Citellus dauricus from Inner Mongolia.The Neighbor-Joining tree of COI,Cyt-b gene showed two major clusters.One of them were clustered by Spermophilus from Dingbian County in Shaanxi and S.alaschanicus in Ningxia,and another one was Citellus dauricus from Inner Mongolia.The Neighbor-Joining tree of COI gene showed that all samples from Shaanxi Province clustered in a group.In conclusion,the Spermophilus in Shaanxi Province were S.alaschanicus.

A comparative molecular survey of malaria prevalence among Eastern chimpanzee populations in Issa Valley (Tanzania) and Kalinzu (Uganda).

BACKGROUND: Habitat types can affect vector and pathogen distribution and transmission dynamics. The prevalence and genetic diversity of Plasmodium spp. in two eastern chimpanzee populations-Kalinzu Forest Reserve, Uganda and Issa Valley, Tanzania-inhabiting different habitat types was investigated. As a follow up study the effect of host sex and age on infections patterns in Kalinzu Forest Reserve chimpanzees was determined. METHODS: Molecular methods were employed to detect Plasmodium DNA from faecal samples collected from savanna-woodland (Issa Valley) and forest (Kalinzu Forest Reserve) chimpanzee populations. RESULTS: Based on a Cytochrome-b PCR assay, 32 out of 160 Kalinzu chimpanzee faecal samples were positive for Plasmodium DNA, whilst no positive sample was detected in 171 Issa Valley chimpanzee faecal samples. Sequence analysis revealed that previously known Laverania species (Plasmodium reichenowi, Plasmodium billbrayi and Plasmodium billcollinsi) are circulating in the Kalinzu chimpanzees. A significantly higher proportion of young individuals were tested positive for infections, and switching of Plasmodium spp. was reported in one individual. Amongst the positive individuals sampled more than once, the success of amplification of Plasmodium DNA from faeces varied over sampling time. CONCLUSION: The study showed marked differences in the prevalence of malaria parasites among free ranging chimpanzee populations living in different habitats. In addition, a clear pattern of Plasmodium infections with respect to host age was found. The results presented in this study contribute to understanding the ecological aspects underlying the malaria infections in the wild. Nevertheless, integrative long-term studies on vector abundance, Plasmodium diversity during different seasons between sites would provide more insight on the occurrence, distribution and ecology of these pathogens.

Genetic dynamics in the sand fly (Diptera: Psychodidae) nuclear and mitochondrial genotypes: evidence for vector adaptation at the border of Iran with Iraq.

BACKGROUND: Our investigation uses nucleotide variations of the genera Phlebotomus and Sergentomyia using the EF-1α and Cyt b genotype regions to describe the sand fly fauna and genetic aspects collected at war-torn sites of the Khuzestan boundary between Iraq and Iran. METHODS: All sand fly species were characterized using molecular genetics. The field work was conducted in six districts including 24 locations in remote areas for three years at the peak of sand fly activity during cutaneous leishmaniasis (CL) transmission seasons. The distribution of CL vectors was determined based on the climatic regionalization using the kriging method in ArcGIS model. DNA of sand fly pools were screened via polymerase chain reaction (PCR) using neutrality (Tajima's D) and neutral allele frequency (Fu's F s) tests to measure the effect of randomly evolving DNA sequence on the genetic diversity of sand fly populations in response to habitat fragmentation and landscape modification. RESULTS: Among the 1213 specimens, ten species were identified based on morphology. The non-native species Phlebotomus sergenti was unequivocally found for the first time in the studied regions. Nucleotide substitutions of sand fly sequences varied most in the most disrupted districts (Dashte-Azadegan and Abadan; disparity index test: P < 0.05). The haplotypes of Cyt b from the subgenus Sergentomyia and P. papatasi revealed more heterogeneity (Tajima's D > +2) than P. alexandri (D > +1), which suggests widespread heteroplasmic mitochondrial DNA mutations in the same mtDNA gene among different sand fly species. Subgenus  Sintonius exhibited greater fitness (D = 0) and (neutrality test; P > 0.05) no evidence of selection. The sequence of the nuclear gene EF-1α indicated similar nucleotide differences, as observed for the Cyt b gene, in all sand fly species, but lower levels of polymorphisms (D > +1) were observed compared with the mitochondrial Cyt b gene (D > +2) in the subgenus  Sergentomyia. CONCLUSION: Our findings describe random nucleotide diversity in the Phlebotomus and Sergentomyia population gene pools due to recent anthropogenic influence. A phylogenetic analysis showed that the closely related species are positioned in monophyletic clades, except for the subgenus Sergentomyia and P. sergenti, and highlights the importance of haplotype variations for the development of adaptability.

Mitochondrial phylogeny of the Eurasian/African reed warbler complex (Acrocephalus, Aves). Disagreement between morphological and molecular evidence and cryptic divergence: A case for resurrecting Calamoherpe ambigua Brehm 1857.

A tree based on the mitochondrial cyt b gene for 278 samples from throughout the range of the Eurasian Reed Warbler Acrocephalus scirpaceus - African Reed Warbler A. baeticatus complex shows well-supported geographically structured divergence for eight distinct lineages. The phylogenetic structuring together with the clarification of priority, provided by sequence data from seven type specimens, suggests that both taxonomy and distribution boundaries are in need of revision. The Iberian and Moroccan populations form a well-supported clade, and we propose that these are treated as taxonomically distinct, under the name ambiguus (Brehm, 1857). We propose that the names scirpaceus, fuscus, avicenniae, ambiguus, minor, cinnamomeus, hallae and baeticatus are used for the well supported clades in the complex, which we recommend to treat as one polytypic species, A. scirpaceus, pending studies of gene flow and assortative mating in the contact zones.