OBJECTIVE: Channidae family, are major freshwater fish species amongst the local aquatic fauna of Pakistan, while, there is limited availability of local data on their molecular identification and phylogenetic analysis.
METHODS: Channa species were collected from four different geographical sites in the tertiary of Punjab province on the Indus and Chenab rivers of Pakistan. Morphometric records and molecular techniques were used to determine the intraspecific variations among populations of Channa marulius. Mitochondrial DNA was extracted from the flesh of C. marulius, while, COI gene was used for molecular identification and variation levels were estimated by using Principal Component Analysis.
RESULTS: Data recorded on the basis of morphometric parameters clearly divided the C. marulius of different locations into two distinct categories, which accounted for a cumulative variability of 97.6%. Non-significance (P < 0.05) among the C. marulius showed that it contains a unique control haplotype localized within the sub-population. The intra-species distance ranged from 0.000 to 0.001 for four different populations, in contrast, the sequences retrieved from the NCBI database exhibited a range span of 0.000-0.003, while, sequence diversity ranged from 0.000 to 0.006 for this intra-specific comparison. The cladogram was also constructed for C. marulius of different geographical locations for observation of phylogenetic relationship. The conclusion drawn from the phylogenetic analysis of C. marulius populations used in this study, contributes significantly to the understanding of genetic variations within populations of this species. The findings provide valuable insight to devise conservation strategies in fisheries management programs in Pakistan.

The objective of the present study is to characterize the dipteran larvae species infesting the sheep being maintained at SRRC, Mannavanur, by means of COI gene based PCR. During the last week of May 2021, post mortem examination of the skull of an Avikalin male sheep (20 months old) revealed the presence of larvae in its nasal sinuses. The larvae were washed in PBS (pH 7.2) and preserved in 70% alcohol. Total genomic DNA was isolated from the larvae using an initial step of grinding with liquid Nitrogen in a sterile mortar and pestle. Using the isolated genomic DNA from the larvae as a template, Cytochrome c oxidase subunit I (COI) gene based PCR was employed using the primers designed based on the COI gene of reference isolate of Oestrus ovis available in the GenBank. Full length COI gene (1534 bp) gene of Oestrus ovis in sheep from South India was targeted in the PCR experiment. The pTZ57R/T vector was used for the cloning of the PCR amplified fragment and the confirmed recombinant plasmid was subjected to sequencing experiments. In addition to morphological examination, based on COI gene based PCR, eventual sequencing experiments and BLAST analysis, it was confirmed that the larvae in the nasal sinuses of sheep from South India were Oestrus ovis. The South Indian isolate of Oestrus ovis is sharing 100% sequence identity both at nucleotide and amino acid levels with that of O. ovis from Spain. The North Indian isolate of O. ovis (from Jammu) exhibited 92% and 99% identity at respective nucleotide and amino acid levels with South Indian isolate. With other members of the subfamily Oestrinae, the share of per cent nucleotide and amino acid identities of South Indian O. ovis ranged from 85-86% to 95-96%, respectively. O. ovis from South India was grouped with the other members of Oestrinae from different geographical areas of the globe in the analysis of phylogenetic tree based on COI amino acid sequences. Based on the research findings, it is concluded that Oestrus ovis is the dipteran species infesting the sheep at Mannavanur, Tamil Nadu, India. To our knowledge, this is the first report on full length nucleotide sequences of COI gene of O. ovis in sheep from Indian subcontinent.
UNASSIGNED: The online version contains supplementary material available at 10.1007/s12639-024-01666-2.

Based on material recently collected in northern Thailand, the present study provides an updated of the genus Baetiella, including Gratia. It comprises six species in Thailand, three of them being new species: Baetiella (Gratia) narumonae, Baetiella (Gratia) sororculaenadinae, Baetiella (Baetiella) bispinosa, Baetiella (Baetiella) baeisp. nov., Baetiella (Baetiella) lannaensissp. nov. and Baetiella (Baetiella) bibranchiasp. nov.Baetiella (Baetiella) baeisp. nov. can be distinguished from other species by the reduction of the posteromedian protuberances on abdominal tergites I-III, the asymmetrical coniform terminal segment of labial palp, the distal margin of abdominal sternites VII-X each with a row of long, spatulate setae, the dorsal margin of femur with two long, robust setae distally. Baetiella (Baetiella) lannaensissp. nov. is diagnosed by the posteromedian protuberances present on tergites I-VIII, dorsal margin of femur with a regular row of long, rounded, ciliated setae and body surface covered with numerous, dense, rounded scale-like setae. Baetiella (Baetiella) bibranchiasp. nov. can be separated from other species by coxal gills present at the base of forelegs and midlegs. The molecular study based on the mitochondrial gene COI and a larval key to species of Thai Baetiella are also provided.

Pentastiridius leporinus (Hemiptera: Cixiidae) is the main vector of an emerging and fast spreading sugar beet disease, the syndrome \'basses richesses\' (SBR), in different European countries. The disease is caused by the γ-3-proteobacterium \'Candidatus Arsenophonus phytopathogenicus\' and the phytoplasma \'Candidatus Phytoplasma solani\' which are exclusively transmitted by planthoppers and can lead to a significant loss of sugar content and yield. Monitoring of this insect vector is important for disease management. However, the morphological identification is time consuming and challenging as two additional cixiid species Reptalus quinquecostatus and Hyalesthes obsoletus with a very close morphology have been reported in sugar beet fields. Further, identification of females and nymphs of P. leporinus at species level based on taxonomic key is not possible. In this study, an isothermal nucleic acid amplification based on recombinase polymerase amplification (RPA) was developed to specifically detect P. leporinus. In addition, real-time RPA was developed to detect both adults (male and female) and nymph stages using pure or crude nucleic acid extracts. The sensitivity of the real-time RPA for detection of P. leporinus was comparable to real-time PCR, but a shorter time (< 7 min) was required. This is a first report for real-time RPA application for P. leporinus detection using crude nucleic acid templates which can be applied for fast and specific detection of this vector in the field.

In this paper, the Merodon avidus (Diptera, Syrphidae) species complex was revised, whereupon we discovered and described four new species for science: Merodon atroavidus Vujić, Radenković et Likov sp. nov., M. magnus Vujić, Kočiš Tubić et Ačanski sp. nov., M. nigroscutum Vujić, Radenković et Likov sp. nov. and M. pseudomoenium Vujić, Kočiš Tubić et Ačanski sp. nov. An integrative taxonomy approach was used to delimit species boundaries. Two molecular markers (the mitochondrial COI gene and nuclear 28S rRNA gene-newly analysed marker for the complex) and geometric morphometry of the wing shape, together with morphological data and distribution, successfully separated all species from the complex. The morphological variability of the analysed species is described and discussed and an illustrated diagnostic key for typical morpho-forms of species from the M. avidus complex is presented. A distribution map of all investigated species from the complex is provided. The level of endemicity of the M. avidus complex was discussed.

A new species of the genus Laena from Xiaolongshan in Gansu Province, China is described as Laenahuisp. nov. All Laena species known to occur in Gansu Province are reviewed, and an identification key is provided. The mitochondrial gene COI to confirm the identity of the new species, which is morphologically most similar and phylogenetically close to L.fengileana. The new species can be recognized by features of elytra and tibiae.

Graphidessajinfoensissp. nov. is described from Chongqing and Guizhou in Southwest China. The diagnostic morphological characters of the new species are described and illustrated in color plates. The distribution of all species of the genus Graphidessa Bates, 1884 is mapped and the key to all species of this genus is updated. The COI gene sequence of the new species is also provided.

Molecular examination of representatives of Balaustium from several populations in SW Poland, performed using the sequence data from the mitochondrial cytochrome c oxidase subunit I, confirmed their common specific affiliation and identity with Balaustium murorum. The potential presence of distinct species in the studied material, preliminarily inferred from the discovery of clusters as a result of Principal Component Analysis exploring the metric data sets, was rejected due to the finding of only one haplotype, at intra- and inter-population sampling. An insight into meristic traits in larvae, focused on chaetotaxy of legs, revealed wider variation than hitherto recognized for the species. The variation was higher in laboratory-reared larvae compared to field-collected ones. The overall deviations from the mean character values at intra- and interpopulation levels, higher than hitherto observed for the species, vote for the reappraisal of the criteria adopted for discrimination of members of Balaustium with the application of an integrative approach.

OBJECTIVE: Mosquitoes are important vectors that carry disease-causing agents that can affect humans and animals. DNA barcoding is a complementary identification which can be used to validate morphological characterization of mosquito species. The objectives of this study were to identify the mitochondrial sequence of the COI gene and to construct a molecular phylogeny based on the genetic divergence of the mosquito species studied.
METHODS: In this study, DNA extraction and the amplification of the mitochondrial cytochrome oxidase subunit I genes (COI) were performed on pooled mosquito samples collected in Nay Pyi Taw area, Myanmar.
RESULTS: Fragments of the COI gene showed 99-100% identity with sequences of Aedes aegypti, Armigeres subalbatus, Culex pipiens complex, and Cx. quinquefasciatus, respectively, deposited in GenBank. This study categorized two haplotypes from each Ar. subalbatus and Cx. pipiens complex COI gene sequence, as well as three haplotypes from Cx. quinquefasciatus COI gene sequences. The highest haplotype diversity and nucleotide diversity were observed in the Ar. subalbatus population (Hd = 1.0000; π = 0.0033), followed by the Cx. pipiens complex and Cx. quinquefasciatus populations.
CONCLUSIONS: This study provides useful information on the molecular identification and genetic diversity of mosquito vectors with medical and veterinary significance, which may assist in the improvement of mosquito control programs.

A new coccidian species, Isospora elliotae n. sp., from the Australian magpie Gymnorhina tibicen (Latham, 1801) in Western Australia, is described and characterized morphologically and molecularly. Microscopic analysis of a faecal sample identified subspheroidal oocysts (n = 20), 20-22 × 18-20 (20.7 × 18.7); length/width (L/W) ratio 1.05-1.14 (1.10). Wall bi-layered, 1.0-1.3 (1.2) thick, outer layer smooth, c. 2/3 of total thickness. Micropyle and oocyst residuum absent, but usually two polar granules are present. Sporocysts (n = 28) ovoidal, 12-13 × 9-11 (12.6 × 9.7); L/W ratio 1.22-1.35 (1.30). Stieda body present, flattened to half-moon-shaped, c. 0.5 deep × 2.0 wide; sub-Stieda indistinct or barely discernible, c. 1.0 deep × 2.5 wide; para-Stieda body absent; sporocyst residuum present, composed of granules dispersed among the sporozoites. Sporozoites vermiform, with anterior and posterior refractile bodies and nucleus. Segments of three gene loci (18S rRNA, 28S rRNA and COI) were sequenced and I. elliotae n. sp. exhibited 99.8% genetic similarity to Isospora sp. MAH-2013a (KF648870) followed by 99.7% genetic similarity to Isospora neochmiae (Yang, Brice & Ryan, 2016) (KT224380) at the 18S rRNA gene locus. It shared 97.0% genetic similarity with an unnamed Isospora sp. (AY283852) at the 28S rRNA gene locus and it also shared the highest genetic similarity of 99.8% with the unnamed Isospora sp. from an American crow (OL999120) at the COI gene locus. Based on morphological and molecular data, this isolate is a new species named as I. elliotae n. sp.