Single Clade Research Articles

An Overview of Aquatica Fu et al., a Phylogeny of Aquatic Fireflies Using Mitochondrial Genomes, a Description of Two New Species, and a New Record of Aquatic Fireflies in China (Coleoptera: Lampyridae: Luciolinae).

There are now seven species in the genus Aquatica Fu & Ballantyne, with all but one known from the characteristics of males, females, and larvae. Molecular information is combined with morphological taxonomy for the delimitation of both genus and species. The monophyly of the genus Aquatica is strongly supported in six trees, and its position as sister to the genus Nipponoluciola Ballantyne Kawashima Jusoh et Suzuki is stable across maximum likelihood and Bayesian inference results. Two new species of Aquatica Fu & Ballantyne, A. qingshen sp. nov. and A. xianning sp. nov., described from the features of males, females, and larvae, are closely related within a single clade within the genus Aquatica. Females have distinctively shaped median oviduct plates. A Chinese population identified as A. lateralis was found to be morphologically similar to the Japanese population, but genetic distances suggest that it is a distinctive species. No larvae are associated with this species. Definitions of the aquatic status of Luciolinae fireflies are expanded.

Detection of foot-and-mouth disease viruses from the A/AFRICA/G-I genotype in the Sultanate of Oman

Rapid identification and characterization of circulating foot-and-mouth disease virus (FMDV) strains is crucial for effective disease control. In Oman, a few serological and molecular studies have been conducted to identify the strains of FMDV responsible for the outbreaks that have been occurring within the country. In this study, 13 oral epithelial tissue samples from cattle were collected from suspected cases of FMD in Ash Sharqiyah North, Al Batinah North, Dhofar and Ad Dhakhyilia governorates of Oman between 2018 and 2021. FMDV RNA was detected in all samples by real-time RT-PCR and viruses were isolated after one- or two-blind passages in the porcine Instituto Biologico-Rim Suino-2 cell line. Antigen capture ELISA characterized all isolates as serotype A and VP1 phylogenetic analysis placed all sequences within a single clade of the G-I genotype within the A/AFRICA topotype. These sequences shared the closest nucleotide identities to viruses circulating in Bahrain in 2021 (93.5% to 99.5%) and Kenya in 2017 (93.4% to 99.1%). To the best of our knowledge, this is the first time that A/AFRICA/G-I viruses have been detected in Oman. Together with the closely related viruses detected recently in Bahrain, these findings reinforce the importance of deploying effective quarantine control measures to minimize the risks of transboundary transmission of FMD associated with the importation of cattle from East Africa.

Molecular mechanism of antimicrobial co-resistance Colistin (mcr-1) and ESBLs genes among Escherichia coli isolates from commercial chickens in Pakistan.

Emergence of plasmid mediated colistin and extended spectrum β-lactamases (ESBL) resistant genes has been impacted the efficacy of colistin and β-lactams drugs like 3rd, 4th generation cephalosporin. Current study was aimed to investigate antimicrobial resistance genes (ARGs) among Escherichia coli isolates from meat producing commercial broilers in Pakistan. Two hundred (n=200) fecal samples were collected during January-2018 to August-2019. For isolation of E. coli, pink colonies on MacConkey agar were transferred to EMB agar. Metallic sheen color colonies were tested biochemically using API-20E kit. The molecular identification of E. coli (n=153) was targeted by amplification of uid gene through polymerase chain reaction (PCR) and different ARGs i.e. gentamicin, streptomycin, tetracycline, colistin, β-lactams drugs, quinolone and ampicillin followed by sequence analysis. Genotypically, followed by phenotypically of resistant ARGs of isolated PCR-confirmed E. coli (153) shoed resistant against gentamicin (aac(3)-IV), streptomycin (aadA1), tetracycline (tetA), colistine (mcr-1), ampicillin (bla-TEM) and bla-CTX-M were 86%, 88%, 86%, 88%, 83% & 77% respectively. 33/38 (86%) of the isolate was positive for quinolone resistance. Colistine (mcr-1), ESBLs (bla-TEM) and (bla-CTX-M) resistance genes were 88%, 83% and 77% respectively. About 33 isolated E. coli harbored the both mcr-1 and ESBLs genes. All of E. coli isolates were found sensitive to ceftriaxone (CTX-30) and imipenem (IMP-10). The Isolated E. coli showed single or multi clade decadency. The E. coli and ARGs sequences showed single or multi clade decadency. This is first comprehensive study from Pakistan that described the molecular evidences of ARGs and their co-existence in single isolates originated from commercial poultry. Commercial chicken (Broilers) can act as melting pot of antibiotic resistance genes for human being. It is alarming situation for surveillance of antibiotic resistance program because of more regulated prescription of antimicrobial agents in Pakistan.

Occurrence of Stemphylium Leaf Spot Caused by Stemphylium vesicarium in Commercial Celery Production Following an Onion Rotation

In 2022, lesions were observed on the lower leaves of ‘Fandango’ celery ( Apium graveolens L.) growing in a commercial field in Hamilton, Michigan. Brown spots were observed on the leaves’ adaxial surface and border extending to the petiole. Conidia were observed on the symptomatic tissue, and 25 fungal isolates, morphologically similar to Stemphylium vesicarium, were obtained. DNA was extracted from three isolates, and three primer sets were used to amplify and sequence the internal transcribed spacer (ITS) region and partial calmodulin ( cmdA) and glyceraldehyde-3-phosphate dehydrogenase ( gapdh) genes. The obtained sequences of the three isolates had 100% pairwise identity with S. vesicarium sequences MW798751 (ITS), MK675706 ( cmdA), and OQ925923 ( gapdh). A multilocus phylogenetic analysis (neighbor-joining tree) strongly supported clustering Michigan isolates into a single clade with S. vesicarium reference sequences. Isolates recovered from celery were pathogenic to ‘CR1’, ‘Tall Utah’, and ‘Challenger’ celery and ‘Bradley’ onion. S. vesicarium isolates isolated from symptomatic onion volunteers were pathogenic to the three celery cultivars. To our knowledge, this is the first report of S. vesicarium as a pathogen on celery causing leaf spot, and volunteer onion may be a source of Stemphylium in celery fields.

The redescription and complete mitogenomes of two Oxycarenus species (Hemiptera, Oxycarenidae) and phylogenetic implications.

In this study, the two Oxycarenidae species, O.gossypii Horváth, 1926 and Oxycarenusbicolorheraldus Distant, 1904, are redescribed, and their complete mitogenomes are sequenced and analyzed. The phylogeny of Lygaeoidea is examined using 45 complete mitogenomes of lygaeoid species and four outgroup species. The gene orientation and arrangement of the two mitogenomes are found to be consistent with typical Lygaeoidea mitochondrial features, comprising 37 genes, including 13 PCGs, 22 tRNAs, 2 rRNAs, and a control region. Nucleotide composition of the species was biased towards A and T, with the gene order identical to the putative ancestral arrangement of insects. Start codons, stop codons, RNAs, relative synonymous codon usage (RSCU), and nucleotide diversity (Pi) of Oxycarenidae exhibited characteristics similar to other families in Lygaeoidea. Bayesian-inference (BI) and maximum-likelihood (ML) methods were employed to investigate phylogenetic relationships using PCG datasets from selected species. Phylogenetic analyses reveal slightly different topologies between BI and ML methods, with variation primarily concentrated in Colobathristidae and Rhyparochromidae. Our study confirms that the two sequenced Oxycarenidae species formed a single clade, and the position of Oxycarenidae remains stable in both ML and BI phylogenetic trees. These findings expand the mitochondrial genome databases of Lygaeoidea and provide valuable insights into the phylogenetic relationships within Lygaeoidea or Pentatomomorpha.

Diversity of genes encoding immune-related GTPase B2 protein, an inherited element responsible for resistance against virulent Toxoplasma gondiistrains, among wild Mus musculus in local area of Japan.

The major genetic group of Toxoplasma gondii, known as type I, generally displays high lethality in laboratory Mus musculus (mouse) strains, with few exceptions. However, because rodents are the primary reservoir hosts for T. gondii, if this characteristic manifests in the wild, type I strains would be extinct. Therefore, we hypothesized that populations of wild rodents capable of harboring type I T. gondii asymptomatically exist globally and are not limited to a few localized areas, as previously thought. The strength of mouse resistance to T. gondii is known to depend on the affinity of the mouse-expressed immunity-related GTPases B2 (IRGB2) protein for the T. gondii-expressed rphoptry protein 5B (ROP5B) protein. Therefore, the Irgb2 gene sequences of 12 individuals mice captured at two animal farms in Gifu Prefecture, and on an island in Okinawa Prefecture, Japan were determined, and subjected to a molecular phylogenetic analysis together with those of various mouse strains worldwide. The Irgb2 gene of M. musculus individuals captured on one farm and one island showed diverse sequences. The sequences from two individual mice captured in an animal farm formed a single clade with a wild mouse derived CAST/EiJ strain, known for its exceptional resistance to type I T. gondii lethality. These results suggest that M. musuculus individuals resistant to the Type I T. gondii strain may be present in Japan, in addition to the previously known populations in South Asia, Thailand and India.

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.

Phylogenetic relationship of subterranean termite Coptotermes gestroi (Blattodea: Rhinotermitidae) inhabiting urban and natural habitats

Coptotermes termites were collected from forestry habitats at University Sains Malaysia, Penang, while urban samples were collected from residentials from Penang and Kedah, Malaysia. Mitochondrial DNA markers, Cytochrome Oxidase 1 (CO1), and 16S ribosomal RNA (16S rRNA) genes were amplified and sequenced to confirm the species of the termite samples as Coptotermes gestroi. Through Blastn, all 25 CO1 and 16S rRNA sequences, respectively from urban and natural habitats were found to be 99.54–100.00 % similar to C. gestroi reference sequences from previous studies in Peninsular Malaysia. The phylogenetic trees constructed using Neighboring-joining (NJ) and Maximum Likelihood (ML) methods resulted in CO1 sequences clustering in two clades and 16S rRNA sequences clustering in a single clade. The overall mean distance was low for the C. gestroi populations from natural habitats and urban settings (FST = 0.004). Analysis of natural habitat populations using CO1 sequences revealed two haplotypes within the population, with a haplotype diversity (Hd) of 0.045 ± 0.005, while the urban population shared a common haplotype with the natural habitat populations and there was no haplotype diversity recorded between the populations. Urban and natural habitats included only one haplotype for 16S rRNA sequences, indicating a lack of nucleotide diversity. Based on the findings, a non-significant difference between the natural habitat and urban population suggests C. gestroi inhabiting both regions likely originated from a similar source and underwent population homogeneity in different settings facilitated by anthropogenic dispersal.

Reconstructing the phylodynamic history and geographic spread of the CRF01_AE-predominant HIV-1 epidemic in the Philippines from PR/RT sequences sampled from 2008 to 2018.

The Philippines has had a rapidly growing human immunodeficiency virus (HIV) epidemic with a shift in the prevalent subtype from B to CRF01_AE. However, the phylodynamic history of CRF01_AE in the Philippines has yet to be reconstructed. We conducted a descriptive retrospective study reconstructing the history of HIV-1 CRF01_AE transmissions in the Philippines through molecular epidemiology. Partial polymerase sequences (n = 1144) collected between 2008 and 2018 from three island groups were collated from the Research Institute for Tropical Medicine drug resistance genotyping database. Estimation of the time to the most recent common ancestor (tMRCA), effective reproductive number (Re), effective viral population size (Ne), relative migration rates, and geographic spread of CRF01_AE was performed with BEAST. Re and Ne were compared between CRF01_AE and B. Most CRF01_AE sequences formed a single clade with a tMRCA of June 1996 [95 per cent highest posterior density (HPD): December 1991, October 1999]. An increasing CRF01_AE Ne was observed from the tMRCA to 2013. The CRF01_AE Re reached peaks of 2.46 [95 per cent HPD: 1.76, 3.27] in 2007 and 2.52 [95 per cent HPD: 1.83, 3.34] in 2015. A decrease of CRF01_AE Re occurred in the intervening years of 2007 to 2011, reaching as low as 1.43 [95 per cent HPD: 1.06, 1.90] in 2011, followed by a rebound. The CRF01_AE epidemic most likely started in Luzon and then spread to the other island groups of the country. Both CRF01_AE and Subtype B exhibited similar patterns of Re fluctuation over time. These results characterize the subtype-specific phylodynamic history of the largest CRF01_AE cluster in the Philippines, which contextualizes and may inform past, present, and future public health measures toward controlling the HIV epidemic in the Philippines.

The fitness landscape of a community of Darwin's finches.

Divergent natural selection should lead to adaptive radiation-that is, the rapid evolution of phenotypic and ecological diversity originating from a single clade. The drivers of adaptive radiation have often been conceptualized through the concept of "adaptive landscapes," yet formal empirical estimates of adaptive landscapes for natural adaptive radiations have proven elusive. Here, we use a 17-year dataset of Darwin's ground finches (Geospiza spp.) at an intensively studied site on Santa Cruz (Galápagos) to estimate individual apparent lifespan in relation to beak traits. We use these estimates to model a multi-species fitness landscape, which we also convert to a formal adaptive landscape. We then assess the correspondence between estimated fitness peaks and observed phenotypes for each of five phenotypic modes (G. fuliginosa, G. fortis [small and large morphotypes], G. magnirostris, and G. scandens). The fitness and adaptive landscapes show 5 and 4 peaks, respectively, and, as expected, the adaptive landscape was smoother than the fitness landscape. Each of the five phenotypic modes appeared reasonably close to the corresponding fitness peak, yet interesting deviations were also documented and examined. By estimating adaptive landscapes in an ongoing adaptive radiation, our study demonstrates their utility as a quantitative tool for exploring and predicting adaptive radiation.

Characterization of the complete chloroplast genome assembly of Amorphophallus yunnanensis Engler, Pflanzenr (Araceae) from southwestern China

Southwestern China is a biodiversity hotspot due to its diverse topography and environment. Amorphophallus yunnanensis is a species of perennial herb that is mainly distributed throughout southwestern China. The genetic diversity and divergence in this species have not been assessed largely due to a lack of genomic resources. To help with the phylogeographic study, we sequenced and assembled the first complete chloroplast genome sequence of A. yunnanensis. The length of the chloroplast genome was 164,417 bp, with an average GC content of 36% (GenBank accession no. OR400247). The genome possessed a typical quadripartite structure, and the lengths of the large single-copy (LSC), small single-copy (SSC), and two inverted repeat (IR) regions were 92,149 bp, 15,182 bp, 28,543bp, and 28,543bp, respectively. A total of 128 genes were annotated across the genome, including 82 protein-coding genes, 8 rRNAs, and 38 tRNAs. The maximum likelihood (ML) phylogeny confirmed the phylogenetic position of Amorphophallus within Araceae, with the Amorphophallus species forming a single monophyletic clade with a high bootstrap value. The ML tree also indicated that A. yunnanensis was most closely related to A. coaetaneus. This newly sequenced chloroplast genome assembly will aid in future studies of genetic diversity, historical population dynamics, and geographic differentiation patterns of A. yunnanensis.

The complete chloroplast genome assembly of Amorphophallus krausei Engler, Pflanzenr 1911 (Araceae) from southwestern China

Plants in the genus Amorphophallus, many of which possess high konjac glucomannan content, are considered important cash crops in many Asian countries. Wild relatives of cultivated Amorphophallus species are valuable resources for the genetic improvement of these crops. To aid in future genetic research of wild germplasm resources of Amorphophallus, a single individual of Amorphophallus krausei Engler, Pflanzenr 1911 was collected from southwestern China, and its chloroplast genome was sequenced using next-generation sequencing technologies. The assembled chloroplast genome was 172,418 bp in length with a GC content of 35.23% (GenBank accession no. OR416863). A typical quadripartite structure was found in the genome, which was comprised of one large single-copy (LSC), one small single-copy (SSC), and two inverted repeats (IRs), with lengths of 91,983 bp, 15,591 bp, 32,422 bp, and 32,422 bp, respectively. A total of 132 genes were annotated in the genome, including 86 protein-coding genes, 38 tRNAs, and 8 rRNAs. A maximum likelihood (ML) tree of A. krausei and 17 other species in the family Araceae suggested that all Amorphophallus species formed a single monophyletic clade. A close relationship among A. konjac, A. albus, and A. krausei was also revealed by the phylogenetic tree. The newly sequenced chloroplast genome of A. krausei will support future genetic studies, particularly the assessment of genetic diversity, resource conservation, and phylogeographic research.

Molecular characterization and phylogeny of Anaplasma marginale, A. phagocytophilum and A. bovis in livestock of Bangladesh.

The emergence of Tick-borne Anaplasma spp. poses a significant threat to humans and animals worldwide. Traditional surveys based on examining blood smears overlook the existence of emerging pathogens. This study aimed to screen Anaplasma spp. in livestock species from diverse geographies with molecular tools. We collected 276 blood samples from cattle (Bos indicus), gayals (Bos frontalis) and goats (Capra hircus) in Jhenaidah, Bogura, Sirajganj and Bandarban districts, and Naikhongchari sub-district from June 2021 to March 2022. After that, a molecular screening was conducted through polymerase chain reaction (PCR) and sequencing was done to confirm the PCR results. The PCR assays were performed based on the analyses of groEL (Anaplasma marginale) and 16S rRNA (A. phagocytophilum and A. bovis). The Anaplasma spp. detected in this study were A. marginale (10.51%), A. phagocytophilum (0.72%), and A. bovis (63.77%). However, A. platys was not detected in this study. Among the screened pathogens, the detection of A. bovis (82.86%) was significantly high in the Bandarban district, while A. marginale was found only in cattle in this location. Regarding animal species, the occurrence of A. bovis was significantly higher in cattle. Moreover, the detection rate of A. marginale was significantly higher in adult cattle (≥2 years). The phylogenetic analyses revealed that the groEL sequences of A. marginale and 16S rRNA sequences of A. bovis and A. phagocytophilum were included in a single clade in the respective phylograms, showing a single genotype of each species circulating in Bangladesh. This study reports the existence of A. phagocytophilum in Bangladesh for the first time.

Detection of a novel Babesia sp. in Amblyomma javanense, an ectoparasite of Sunda pangolins

BackgroundBabesia is a protozoal, tick-borne parasite that can cause life-threatening disease in humans, wildlife and domestic animals worldwide. However, in Southeast Asia, little is known about the prevalence and diversity of Babesia species present in wildlife and the tick vectors responsible for its transmission. Recently, a novel Babesia species was reported in confiscated Sunda pangolins (Manis javanica) in Thailand. To investigate the presence of this parasite in Singapore, we conducted a molecular survey of Babesia spp. in free-roaming Sunda pangolins and their main ectoparasite, the Amblyomma javanense tick.MethodsTicks and tissue samples were opportunistically collected from live and dead Sunda pangolins and screened using a PCR assay targeting the 18S rRNA gene of Babesia spp. DNA barcoding of the cytochrome oxidase subunit I (COI) mitochondrial gene was used to confirm the species of ticks that were Babesia positive.ResultsA total of 296 ticks and 40 tissue samples were obtained from 21 Sunda pangolins throughout the 1-year study period. Babesia DNA was detected in five A. javanense ticks (minimum infection rate = 1.7%) and in nine different pangolins (52.9%) located across the country. Phylogenetic analysis revealed that the Babesia 18S sequences obtained from these samples grouped into a single monophyletic clade together with those derived from Sunda pangolins in Thailand and that this evolutionarily distinct species is basal to the Babesia sensu stricto clade, which encompasses a range of Babesia species that infect both domestic and wildlife vertebrate hosts.ConclusionsThis is the first report documenting the detection of a Babesia species in A. javanense ticks, the main ectoparasite of Sunda pangolins. While our results showed that A. javanense can carry this novel Babesia sp., additional confirmatory studies are required to demonstrate vector competency. Further studies are also necessary to investigate the role of other transmission pathways given the low infection rate of ticks in relation to the high infection rate of Sunda pangolins. Although it appears that this novel Babesia sp. is of little to no pathogenicity to Sunda pangolins, its potential to cause disease in other animals or humans cannot be ruled out.Graphical abstract

Complete mitochondrial genome analyses confirm that bat Polychromophilus and ungulate Plasmodium constitute a distinct clade independent of other Plasmodium species

In recent phylogenetic studies, bat Polychromophilus and ungulate Plasmodium, two relatively understudied haemosporidian parasites within the Apicomplexa phylum, have often been overlooked. Instead, the focus has been primarily on haemosporidian parasites in primates, rodents, and birds. Several phylogenetic analyses of bat Polychromophilus have relied on limited datasets and short informative DNA sequences. As a result of these inherent limitations, the substantiation of their evolutionary stance has encountered a diminished degree of robust validation. This study successfully obtained complete mitochondrial genome sequences from 11 Polychromophilus parasites originating from Hipposideros gentilis and Myotis siligoensis bats for the first time. Additionally, the authors have sequenced the apicoplast caseinolytic protease C genes from Polychromophilus murinus and a potentially new Polychromophilus species. These mitochondrial genomes range in length from 5994 to 6001 bp and consist of three protein-coding genes (PCGs), seven small subunit ribosomal RNA genes (SSU rRNA), 12 large subunit ribosomal RNA genes (LSU rRNA), and seven miscellaneous RNA genes. Phylogenetic analyses using Bayesian Inference and Maximum Likelihood methods indicated robust support for the grouping of ungulate Plasmodium and bat Polychromophilus in a single clade separate from other Plasmodium spp., confirming previous reports, albeit with stronger evidence in this study. The divergence between Polychromophilus in bats and Plasmodium in ungulates occurred approximately 29.61 to 55.77 million years ago (Mya), with a node age estimated at 40.63 Mya. These findings highlight that the genus Plasmodium, which includes species found in ungulates, birds, reptiles, and other mammals, does not form a monophyletic group. By incorporating Polychromophilus in bats and Plasmodium in ungulates, this study contributes significantly to understanding the phylogenetic relationships within the Haemosporida order. It provides valuable insights into the evolutionary history and interconnections among these diverse parasites, thereby expanding knowledge in this field.

Acquired stress resilience through bacteria-to-nematode interdomain horizontal gene transfer.

Natural selection drives the acquisition of organismal resilience traits to protect against adverse environments. Horizontal gene transfer (HGT) is an important evolutionary mechanism for the acquisition of novel traits, including metazoan acquisitions in immunity, metabolic, and reproduction function via interdomain HGT (iHGT) from bacteria. Here, we report that the nematode gene rml-3 has been acquired by iHGT from bacteria and that it enables exoskeleton resilience and protection against environmental toxins in Caenorhabditis elegans. Phylogenetic analysis reveals that diverse nematode RML-3 proteins form a single monophyletic clade most similar to bacterial enzymes that biosynthesize L-rhamnose, a cell-wall polysaccharide component. C. elegans rml-3 is highly expressed during larval development and upregulated in developing seam cells upon heat stress and during the stress-resistant dauer stage. rml-3 deficiency impairs cuticle integrity, barrier functions, and nematode stress resilience, phenotypes that can be rescued by exogenous L-rhamnose. We propose that interdomain HGT of an ancient bacterial rml-3 homolog has enabled L-rhamnose biosynthesis in nematodes, facilitating cuticle integrity and organismal resilience to environmental stressors during evolution. These findings highlight a remarkable contribution of iHGT on metazoan evolution conferred by the domestication of a bacterial gene.

Mitogenome of Xya pfaendleri (Orthoptera: Caelifera): Its comparative description and phylogenetic position within Tridactylidea.

We report the comparative examination of the complete mitochondrial genome of the pygmy mole cricket Xya pfaendleri (Orthoptera: Caelifera: Tridactylidae). The mitogenome consists of 13 protein-coding regions, 22 tRNAs, two rRNAs, and one control region, following the gene order of the ancestral pancrustacean mitogenome. The length of the mitogenome in Xya pfaendleri is 15352 bp. The start and stop codons of the protein-coding genes exhibit the general pattern observed in orthopterans. The data indicate that the pattern of gene overlapping/intergenic sequences exhibits a significant phylogenetic signal. A phylogenetic tree inferred using 12 mitogenomes (seven belonging to Tridactylidea, three to Acrididea, and two to Ensifera) confirms the sister group relationship of Acrididea and Tridactylidea. The relationship among the families of Tridactylidea is Cylindrachetidae + (Ripipterygidae + Tridactylidae). The mitogenome sequences of Xya and Tridactylus constitute a single clade, sharing a last common ancestor 94 million years ago, and rendering the first genus paraphyletic. The present preliminary data suggest that we still have much to learn about the evolution and diversity of Tridactylidea.

Environmental DNA biomonitoring in biodiversity hotspots: A case study of fishes of the Okavango Delta

AbstractThe Okavango Delta is the largest freshwater wetland in southern Africa and a recognized biodiversity hotspot and UNESCO World Heritage Site. The region is extremely rich in floral and faunal diversity, including a fish fauna of ~90 species in 15 families, that also support recreational and subsistence fishing. Anthropogenic pressures and invasive species threaten the unique biodiversity and ecosystem services that the Delta provides, necessitating biomonitoring tools that can provide broad community‐level diversity insights. Here, we utilize environmental DNA metabarcoding of aquatic eDNA using the MiFish 12S rRNA primers, to investigate fish communities and also sequenced 211 mtDNA 12S barcodes for 74 species across 36 genera of fishes from the region. Metabarcoding recovered 11 of 15 families, with 40 species detected across 23 genera, representing ~50% of known diversity, with the mtDNA 12S fragment able to delineate all genera (except for the cichlid genera Serranochromis and Pharyngochromis that comprised a single clade) and most species, except for some in the Clarias, Enteromius, Labeo, Lacustricola, and Petrocephalus genera. Generally, abundant and wide‐spread taxa such as Clarias spp. and Marcusenius altisambesi, amongst others, were often detected in the surveys, with other species, including Zaireichthys kavangoensis, Schilbe intermedius, and Labeo sp. detected less frequently. Dissolved oxygen, temperature, and dissolved organic solids were positively correlated with community diversity, highlighting the influence of environmental factors in shaping fish communities in the region. Further, there was strong variability in the eDNA signal across only 1000 m, suggesting that future surveys need to consider spatio‐temporal aspects of sample collection. Our study highlights the potential of eDNA metabarcoding for surveying aquatic biodiversity in the Okavango Delta, particularly within the context of baseline biodiversity inventories, that underpin conservation and management initiatives. As such, we provide a number of recommendations that can help structure future sampling efforts in the region.

Complete chloroplast genomes of Cerastium alpinum, C. arcticum and C. nigrescens: genome structures, comparative and phylogenetic analysis

The genus Cerastium includes about 200 species that are mostly found in the temperate climates of the Northern Hemisphere. Here we report the complete chloroplast genomes of Cerastium alpinum, C. arcticum and C. nigrescens. The length of cp genomes ranged from 147,940 to 148,722 bp. Their quadripartite circular structure had the same gene organization and content, containing 79 protein-coding genes, 30 tRNA genes, and four rRNA genes. Repeat sequences varied from 16 to 23 per species, with palindromic repeats being the most frequent. The number of identified SSRs ranged from 20 to 23 per species and they were mainly composed of mononucleotide repeats containing A/T units. Based on Ka/Ks ratio values, most genes were subjected to purifying selection. The newly sequenced chloroplast genomes were characterized by a high frequency of RNA editing, including both C to U and U to C conversion. The phylogenetic relationships within the genus Cerastium and family Caryophyllaceae were reconstructed based on the sequences of 71 protein-coding genes. The topology of the phylogenetic tree was consistent with the systematic position of the studied species. All representatives of the genus Cerastium were gathered in a single clade with C. glomeratum sharing the least similarity with the others.

The Phylogenetic Relationships of Australian Species within Charopidae (Gastropoda: Punctoidea)

Charopidae is a highly speciose family of land snails mostly distributed in the main landmasses and archipelagos of the Southern Hemisphere, with a few genera in the Northern Hemisphere. DNA sequencing is making substantial contributions to understanding the phylogeography of Charopidae and the systematic relationships of its subfamilies and related families in the Punctoidea. Here, new data from 28S rRNA are combined with available GenBank sequences to investigate whether Australian Charopidae comprise a single clade, exclusive of species from other regions. Phylogenetic analyses show that this is not the case, with most species from Australia and other regions included in a bootstrap-supported clade that excludes some Australian species. Analyses of concatenated 28S rRNA data and available cytochrome c oxidase subunit I (COI) also contradicted monophyly of Australian species, as did analyses of inferred COI protein sequences. In DNA analyses, the genus Hirasea and Australian taxa with an incomplete secondary ureter were basal within Charopidae. We also examined whether known doubts about the taxonomic status of supra-generic clades in Punctoidea could be resolved by considering autapomorphic COI amino acid changes. Such variation could not generally be applied as an objective criterion for taxonomic categorization but did identify some clades for taxonomic investigation.