Mitochondrial Cytochrome Oxidase Subunit Research Articles

Gene Sequencing Study of Mitochondrial Cytochrome Oxidase Subunit I (COI) in Aedes aegypti Mosquito

Aedes aegypti is an important vector from the public health point of view. It transmits pathogens of Dengue and various other diseases. Controlling vector populations is key to reducing disease transmission and it is possible by understanding the taxonomic characteristics of the vectors and their genetic analysis helps in the identification of the genetic variations in species of different geographical areas. The main aim of the present study is the screening of the Aedes mosquito genome of local species as genetic studies give some insights into the vaccine for the treatment of the diseases caused by mosquito species. In the present study, the adult Ae. aegypti collected from Suryapet town where Dengue fever is endemic, used for the mitochondrial Cytochrome Oxidase subunit I (COI) gene sequencing using the techniques of polymerase chain reaction, Electrophoretic separation, spectrophotometric analysis, and BLAST. The mitochondrial COI sequence of 583 bps is published in the National Centre for Biotechnology Information (NCBI) with accession number ON413769. The sequence data was matched with the database available in the NCBI to identify variation in the gene as this gene keeps changing in the organisms and may contribute to the species establishing itself in a particular region due to congenial environmental conditions. One match with the protein called haem copper oxidase subunit I. The transmission of different pathogens in different areas can be ascertained by comparing similar sequence studies documented. Genomic studies involving genetic markers are of immense help in developing the vaccine. The mitochondrial COI gene is the marker often used for evolutionary research. The single nucleotide polymorphisms (SNP) or Restricted fragment length polymorphisms (RFLPs) that occur in DNA also contribute to the vector's adaptability to the different geographical areas and also the survival benefit which are the basis for understanding the vectors thoroughly and their role in disease transmission.

Genetic diversity and species prevalence revealed by Mitochondrial COI- DNA sequence of thrips population in cotton

Aim: To elucidate the population genetic diversity and species prevalence of thrips on cotton. Methodology: Thrips species were identified utilizing both mitochondrial cytochrome oxidase subunit I (COI) sequence analysis and morphology-based keys. Results: Thrips palmi Karny, Thrips hawaiiensis (Morgan), Ayyaria chaetophora Karny, Scirtothrips dorsalis Hood and Rhipiphorothrips cruentatus Hood identified from collected samples. The distribution analysis revealed that T. palmi stands out as the most abundant and prevalent species, constituting a substantial majority (74%) and occupying a pervasive presence across most cotton-growing locations. The phylogenetic analysis, relying on COI sequences, clearly categorized these five species into four distinct major clades. T. hawaiiensis and R. cruentatus were placed within the first clade, while the second clade featured Scirtothrips dorsalis. The third clade comprised the abundant T. palmi, and the fourth clade accommodated A. chaetophora. Interpretation: The study highlighted genetic diversity of five species of thrips, accentuating the emergence of T. palmi as a dominant pest in cotton, an observation not previously reported. Key words: Cotton thrips, Genetic diversity, MtCOI, Phylogenetic analysis, Species prevalence

Morphometric trait normalization, cluster analysis and molecular phylogenetic analysis in delimiting sympatric Neolissochilus hexastichus and N. hexagonolepis in Umngi River, Meghalaya (India)

Aim: To assess the ability of an integrative approach comprising multivariate morphometric data analysis and mitochondrial cytochrome oxidase subunit I (CO I) sequence divergence analysis in delimiting sympatric Neolissochilus hexastichus (McClleland, 1839) and N. hexagonolepis (McClleland, 1839) mahseer populations in River Umngi, Meghalaya. Methodology: Authoritative descriptions of mahseers were referred during the sampling of 40 individuals from River Umngi, Meghalaya. Multivariate morphometric data were normalized with respect to standard length. The normalization procedure maintained the particular shape factor for each sample. Normalised data were subjected to hierarchical clustering on principal components Analysis (HCPC) for grouping samples according to morphometric features. Genetic affinities based on CO I sequence divergence were assessed by Bayesian Inference (BI) and Automatic Barcode Gap Discovery (ABGD) protocols. Results: The normalization protocols removed the variations in traits due to body size effects and facilitated the identification of allometric traits. HCPC analysis revealed morphometric features that had a significant impact on clustering. Nine N. hexagonolepis and 13 N. hexastichus individuals were identified by integrative morphological and molecular phylogenetic approach. The remaining 18 individuals had incongruence in their memberships in the morphometric and phylogenetic clusters. Interpretation: The integrative approach, apart from delineating N. hexagonolepis and N. hexastichus congeners, highlighted the need to include nuclear genetic markers in future studies for confirming the presence of natural hybrids in sympatric populations of Neolissochilus mahseers. Key words: Cluster, Morphometric, Neolissochilus, Normalization, Phylogenetics

Megaselia scalaris and Senotainia tricuspis Infesting Apis mellifera: Detection by Quantitative PCR, Genotyping, and Involvement in the Transmission of Microbial Pathogens.

The Megaselia scalaris and Senotainia tricuspis parasitoid flies of the honeybee Apis mellifera were found to infest apiaries of different European and Mediterranean countries but their prevalence and impact on apiary health are little known. Therefore, in this study, quantitative PCR (qPCR)-based methods were developed for their rapid detection directly in hive matrices. The newly developed qPCR assays were targeted at the mitochondrial cytochrome oxidase subunit I (COI) gene for the M. scalaris and the cytochrome B (cytB) gene for the S. tricuspis. The tests were preliminarily applied to 64 samples of adult honeybees and hive debris collected in the Abruzzo and Molise regions, Central Italy, and the Republic of Kosovo showing that both flies occur in the two countries and more frequently in Italy. The positive apiaries in Italy were re-sampled by capturing viable forager bees and isolating emerging flies to carry out the genotyping and analyses aimed at defining if these flies can transmit honeybee pathogens. Genotyping based on the COI and cytB gene sequencing for M. scalaris and S. tricuspis, respectively, identified one S. tricuspis genotype and diverse genotypes of M. scalaris highly similar to those from distant countries. Some fly isolates harbored the DNA or RNA of honeybee microbial pathogens Paenibacillus larvae, deformed wing viruses A and B (DWVA and B), black queen cell virus (BQCV), chronic paralysis virus (CBPV), and Nosema ceranae. The results indicated that these parasites should be efficiently controlled in apiaries by using rapid detection methods to facilitate the large screening studies and early detection.

Anthropogenic activities along the Lake Nyasa catchments alter the habitat and genetic diversity of a Lake Salmon, Opsaridium microlepis

The Lake Salmon, Opsaridium microlepis is an economically important fish along the Lake Nyasa. However, the species is under threat of extinction due to anthropogenic activities such as agriculture, mining, urbanization, and deforestation. Consequently, the fish species is currently regarded as a threatened species, and the International Union for Conservation of Nature (IUCN) has red-listed the species due to an apparent decline in abundance. The current study assesses the potential impact of human activities on the genetic diversity of O. microlepis using partial mitochondrial cytochrome oxidase subunit I (COI) sequences and microsatellite loci. The results indicate that genetic diversity is lower in the areas affected by human activities compared to relatively pristine areas. The results from this study may suggest that human activities taking place in the catchments are likely to contribute to the alteration of the genetic diversity of O. microlepis species. Thus, immediate measure is required to control anthropogenic activities in the areas to protect the species and other aquatic organisms from possible threats of extinction.

Relationships between crayfish population genetic diversity, species richness, and abundance within impounded and unimpounded streams in Alabama, USA.

Understanding the relationship between multi-scale processes driving community- and population-level diversity can guide conservation efforts. While the importance of population-level genetic diversity is widely recognized, it is not always assessed for conservation planning, and positive correlations with community-level diversity are sometimes assumed, such that only the latter is measured. We surveyed species richness and cumulative multispecies abundance of crayfishes in impounded and unimpounded streams in the southern Appalachian Mountains (Alabama, USA). We simultaneously assessed levels of population genetic diversity within two focal crayfishes (Faxonius validus and F. erichsonianus) using nuclear (nDNA; inter-simple sequence repeat (ISSR)) and mitochondrial DNA (mtDNA; mitochondrial DNA cytochrome oxidase subunit I (mtCOI)) markers. We then tested for species-genetic diversity correlations (SGDCs), species diversity-abundance correlations (i.e., more individuals hypothesis, MIH), and abundance-genetic diversity correlations (AGDCs) across sites. We also examined the relationship between each of the three different types of correlation (i.e., species richness, cumulative multispecies abundance, and population genetic diversity) and stream habitat characteristics and fragmentation. Surprisingly, based on F. validus mtDNA data, sites with the greatest multispecies abundance had the lowest genetic diversity, indicating a negative AGDC. However, no AGDC was evident from nDNA. There was no evidence of SGDCs for F. validus based on either of the two genetic data types. For F. erichsonianus, there was no evidence for SGDC or AGDC. When considering the community-level data only, there was no support for the MIH. Stream width was positively correlated with F. validus genetic diversity, but negatively correlated with multispecies abundance. Similarly, species richness was positively correlated with stream width in unimpounded streams but negatively correlated with width in impounded streams. These findings indicate that community-level diversity cannot be indiscriminately used as a proxy for population-level diversity without empirically testing this correlation on the focal group. As such, community- and population-level assessments for multiple crayfish species are needed to better understand drivers of diversity and eco-evolutionary processes which will aid in the conservation of this vulnerable taxonomic group.

Assessment of the Genetic Diversity of the Monogenean Gill Parasite Lamellodiscus echeneis (Monogenea) Infecting Wild and Cage-Reared Populations of Sparus aurata (Teleostei) from the Mediterranean Sea.

The diplectanid monogenean Lamellodiscus echeneis (Wagener, 1857) is a specific and common gill parasite of the gilthead seabream Sparus aurata Linnaeus, 1758, in the Mediterranean Sea. Few isolated molecular studies of this monogenean have been conducted, and its population structure and genetic diversity are poorly understood. This study represents the first analysis of the population genetics of L. echeneis, isolated from wild and cage-reared gilthead seabream from fifteen localities in both the Southern (Tunisia) and Northern (Italy and Spain) regions of the Mediterranean Sea, using nuclear ITS rDNA markers and a partial fragment of the mitochondrial gene cytochrome oxidase subunit I (COI). The phylogenetic trees based on the newly obtained dataset and the previously published sequences of L. echeneis corroborated the spread of only a single species throughout the Mediterranean Sea. The star-like haplotypes network, inferred by COI sequences, suggested a recent population expansion of L. echeneis. This is supported by the observed high haplotype diversity (Hd = 0.918) and low nucleotide diversity (Pi = 0.01595). Population structure-based AMOVA for two groups (the Adriatic Sea and the rest of the Mediterranean Sea) attributed 35.39% of the total variation to differences within populations, 16.63% to differences among populations within groups, and 47.99% to differences among groups. Fixation indices were significant, with a high FST value (0.64612), likely related to the divergence of the parasite populations from the Adriatic Sea and other Mediterranean regions. Phylogenetic analyses grouped all samples into the main clade corresponding to L. echeneis from several localities. This study provides insight into the genetic variation between L. echeneis populations, and did not show a clear genetic structure between populations of L. echeneis throughout Tunisian, Italian, and Spanish localities, which can be attributed to the considerable gene flow between the populations favoured by the potential for host dispersion within the Mediterranean Sea. Finally, haplotypes shared between wild and cage-reared hosts provided evidence for the potential for cross-infection between wild and farmed hosts in the Mediterranean Sea.

DNA barcoding and new record of Arcynopteryx dichroa (McLachlan, 1872) (Plecoptera: Perlodidae) from Serbia

Arcynopteryx dichroa (McLachlan, 1872) (Perlodidae) is recorded for the first time from southwestern Serbia. It was collected in the Djerekar Spring (the Pešter Plateau). Previously, this species was found in Timok River in the eastern part of the country, with the last known record dating back to 1995. Identification of the collected specimens was confirmed using the nucleotide sequences of the mitochondrial cytochrome oxidase subunit I (COI) gene. A diagnosis of the larval characters is provided, and species distribution is discussed.

Generalist species exhibit more genetic structure in comparison to a habitat specialist: Evidence from a phylogeographic study of two freshwater crabs (Decapoda: Potamonautidae: Potamonautes), implications for habitat conservation.

During the present study, the phylogeography of the wetland specialist, freshwater crab, Potamonautes flavusjo, was investigated and compared to that of the ubiquitous, generalist, P. sidneyi, using DNA sequence data from the mitochondrial cytochrome oxidase subunit one (COI). We inferred the evolutionary history of each species and compared their population-level genetic structure by constructing haplotype networks and using an analysis of molecular variation. Additionally, we explored the evolutionary relationship between southern African lentic and lotic freshwater crab species by examining the usefulness of carapace attributes in relation to genetic indices and a species' assumed dispersal capacity. In the lentic species, P. flavusjo, a single interconnected haplocluster characterized by shared haplotypes was observed, suggesting marked maternal dispersal, a result corroborated by the low F ST values. In contrast, for the lotic species, P. sidneyi two distinct haploclusters and marked genetic differentiation was observed indicating the absence of maternal dispersal, a result corroborated by the high F ST values. The phylogenetic relationship in P. sidneyi was further investigated using a maximum likelihood and Bayesian inference analyses with the addition of sequence data from the mitochondrial 16S rRNA locus to estimate divergence times. Three species delimitation methods (ASAP, PTP, and bGMYC) were used to explore the presence of distinct lineages in P. sidneyi. The phylogenetic results indicated that within P. sidneyi two clades were present, while divergence time estimates suggest cladogenesis during the middle to late Pleistocene. The species delimitation methods used showed moderate congruence, however all oversplit the number of putative lineages. Our results indicated that P. sidneyi is a species complex comprised of two cryptic lineages, occurring in sympatry contemporarily possibly alluding to secondary contact. Carapace height was found to have no discernible influence on the genetic indices and presumed dispersal capabilities of mainland southern African freshwater crab species. The importance of our results are discussed in terms of conservation of freshwater habitats.

Molecular genetic diversity analysis of the Nigerian laughing dove (Streptopelia senegalensis) and related species using selected mitochondrial genes

Nigerian laughing doves (Streptopelia senegalensis) are small birds with long tail and living in bushes of the Sub-Saharan regions of African continent, the middle East and Asia, especially India. They are used for food, medicinal and religious purposes in Nigeria. Despite their usage, there is a lack of information on the genetic diversity of laughing doves in Nigeria. This study investigates taxonomic order and diversity of Nigerian laughing doves based on the mitochondrial cytochrome oxidase subunit I (COI) and cytochrome B (CYTB). The results showed 20 haplotypes within the 28 Nigerian coupled with Global Streptopelia genus using concatenated sequences. The Nigerian laughing dove constitute 16 distinct haplotypes. The haplotype diversity was 0.743 ± 0.070, and nucleotide diversity 0.154 ± 0.101 within Nigerian population using COI sequences. Phylogenetic tree showed that Nigerian laughing doves were in the same monophyletic clade with other Streptopela orientalis, S. decocto and S. chinensis; and this confirmed that Nigerian laughing doves might have shared descendant. The median-joining network further grouped Nigerian laughing doves into two: the first group consisting of Nigerian populations only, while the second group are with Saudi Arabian and Djiboutian populations. Population expansion was revealed in Nigerian dove individuals. This study revealed 16 unique haplotypes among Nigerian laughing dove population using concatenated sequences. Interestingly, CYTB showed clustering in African laughing doves (For instance, Nigerian individuals shared haplotypes with Sao Tome and Principe, an island country in the Gulf of Guinea, the western equatorial coast of Central Africa). The current data is the first report on genetic diversity of Nigerian laughing dove using mitochondrial COI and CYTB genes.

Molecular identification and eco-friendly management of rice brown planthoppers in Bangladesh

Infestation by various insect pests is the main constraint for growing rice where rice brown planthopper (Nilaparvata lugens Stål) can severely damage rice plants directly through feeding. Therefore, the study aims to detect rice brown planthoppers (BPH) and provide environment-friendly management tactics to mitigate the problem which caused by brown planthoppers. The BPH samples were collected from rice fields of different locations in the Patuakhali of Bangladesh for molecular identification. A molecularly single species of rice brown planthopper, Nilaparavata lugens was identified using mitochondrial cytochrome oxidase subunit I (mtCOI) universal marker. The nucleotide sequences of collected samples were compared with other nucleotide sequences from the GenBank database of NCBI, which make single clades in the phylogenetic tree at an insignificant distance. Moreover, brown planthopper management observations were recorded in laboratory conditions after providing an artificial diet with different treatments of plant-based insecticides Neem oil (1 %, 5 %, and 10 %), Castor oil (1 %, 5 %, and 10 %) where only 20 % sucrose solution was used as negative control and Abamectin (1 %, 5 % and 10 %) were also used as a positive control for comparing the efficacy of plant-based insecticides on rice brown planthoppers. The results showed the highest mortality (100 %) of rice brown planthoppers was recorded by Abamectin 10 %, followed by Abamectin 5 %. Neem 10 % performed better than Abamectin 1 % during 1st hour. Initial after exposure of 2nd hour for Abamectin 1 % revealed greater mortality (59 %) than Neem 10 %. Neem 5 % showed less effect on mortality in brown planthopper than Neem 10 % but was higher than Neem 1 % during 6 h of observation. The Castor oil of 10 % caused higher mortality than the Castor of 5 % but not up to the marks of Abamectin and different concentrations of Neem oil. Castor oil of 1 % and control have shown no mortality of brown planthopper for 6 h of observation.

A typical coastal plain taxon in central Brazil: Relationships and description of a new species of non-annual killifish (Cyprinodontiformes; Rivulidae) from the Paraná River basin

We describe a new species of the genus Atlantirivulus from a small first-order stream on the right margin of the Paraná River basin in the Mato Grosso do Sul, Brazil. The region of occurrence of the new species is known to be a hotspot for Melanorivulus species. This area, however, is approximately 700 km away from the record of occurrence of its closest congeners, which are distributed along the Brazilian coast in different hydrographic basins. Atlantirivulus enigmaticus, new species, is distinguished from all congeners by it s unique coloration, which consists of the flank being purplish grey with light green iridescence with brownish red dots scattered along of the body, by the neuromasts of the infraorbital series being aligned around eye, and by the presence of the short ventral process of the posttemporal. The phylogenetic position of the new species within the Atlantirivulus genus was based on molecular phylogenetic analysis using sequences of the mitochondrial gene cytochrome oxidase subunit 1. The results show that the new species is closely related to Atlantirivulus luelingi from the coastal drainages of the Brazilian state Santa Catarina. The phylogenetic position of the new species among congeners endemic to coastal river basins may be related to ancestral connections between coastal drainages and the Paraná River basin. The new species constitutes the westernmost record for the genus, and the first record of Atlantirivulus in the Paraná River basin. Insufficient collection efforts in areas of potential occurrence for the group and extinction processes in the broad region between the locality of the new species and the distribution areas of the remaining species may be a possible explanation for this disjunct distribution.

Non‐destructive molecular methods to identify Monochamus alternatus (Coleoptera: Cerambycidae), a major vector of Bursaphelenchus xylophilus (Nematoda: Aphelenchoididae)

AbstractMonochamus alternatus is one of the most important borers of conifers and the main vector of the pinewood nematode, Bursaphelenchus xylophilus, the causal agent of pine wilt disease. It causes massive death of pine trees and seriously affects the health of forest ecosystems. Traditionally, adults or larvae are identified by morphological or molecular methods. However, diagnosing M. alternatus larvae at different instars collected from forests is expensive and time‐consuming. Non‐destructive molecular diagnostic protocols are, therefore, being developed to detect biological traces (i.e. exuviae, excreta) and to determine the distribution and spread of this pest. In this study, based on the alignment of mitochondrial DNA cytochrome oxidase subunit I (COI) gene sequences of M. alternatus and other insect species, we designed the primer pair of Mal‐SF/Mal‐SR and probe of Mal‐P for M. alternatus. TaqMan probe‐based qPCR was developed to identify the occurrence of M. alternatus in forests by amplifying the DNA samples obtained from its adult, larva, frass, excreta and exuviae. The amplification results were very effective. The lowest amount of M. alternatus DNA that could be detected with a Cq of 31.93 in the mixed samples was 0.64 pg, showing very high sensitivity. This assay can easily identify M. alternatus from other non‐target wood‐borer species using its frass and exuviae, providing a new diagnostic protocol for monitoring the occurrence and distribution of M. alternatus in forests.

Tiwaripotamon keeae sp. nov. (Crustacea: Decapoda: Potamidae), a new miniature species of freshwater crab from the China-Vietnam border in Napo, Guangxi

A new miniature species of freshwater crab from the genus Tiwaripotamon Bott, 1970 (Potamidae), is described from a montane karst area in southern Napo, at the China-Vietnam border. It is the twelfth known species of the genus, and is also the smallest known, with the largest adult specimen observed barely exceeding 20 mm in carapace width. Apart from the small size, features of the eyes, carapace and ambulatory legs further set Tiwaripotamon keeae sp. nov. apart from its congeners. Pairwise genetic distances of the mitochondrial cytochrome oxidase subunit I (COI) gene also support it as a distinct species.

First Report of Phytopythium litorale Causing Root Rot on Kousa Dogwood in Tennessee and the United States.

Kousa dogwood (Cornus kousa) is an economically important woody ornamental crop that exhibits creamy, white, pointed bracts in late spring, and reddish to pink drupe fruits in late summer and fall. It bears shiny dark green leaves that become reddish-purple to scarlet in the fall. In August of 2023, 3-year-old container grown C. kousa var. chinensis plants in a commercial nursery in Warren Co., Tennessee, exhibited severe yellowing, dieback and root rot symptoms (Fig. 1a and 1b). Dark brown to black lesions were observed in the root and crown region of the plants. Disease severity was 40% to 60% of root area affected, and disease incidence was approximately 40% of 1,000 plants. Surface-sterilized (10% NaOCl: 1 min) symptomatic root tissues were plated on V8-PARPH and incubated at 25°C. Sparse aerial mycelium, showing a distinct rosette or faint radiate to chrysanthemum colony pattern, was observed within four days of incubation (Fig. 2). All isolates produced ovoid or subglose, papillate, and proliferating sporangia in grass blade water cultures (Derviş et al. 2020). Sporangia measured as 19.18 to 24.80 µm X 18.08 to 22.16 µm (n = 50) with a length/width ratio of 1.06 to 1.11. Zoospores observed were between 7.07 to 9.98 µm in diameter (n = 50). Oogonia and oospores were not produced. The ribosomal internal transcribed spacer (ITS) and large subunit (LSU), as well as mitochondrial cytochrome oxidase subunit II (COX-II) genetic markers were amplified and sequenced using primer pairs ITS1/ITS4 (White et al. 1990), NL1/NL4 (Baten et al. 2014), and cox2-F/cox2-RC4 (Choi et al. 2015), respectively. The ITS, LSU, and COX-II sequences of isolates FBG6343, FBG6344 (ITS: PP458373 and PP461387; LSU: PP461390 and PP461391; COXII: PP477112 and PP477113) were 100% identical to those of MN306118, HQ643386, and MN206732, respectively. Based on the morphology (Nechwatal and Mendgen 2006) and sequence data, the isolates were identified as Phytopythium litorale (Nechw.) Abad, De Cock, Bala, Robideau, Lodhi & Lévesque. The pathogenicity test was performed on 3-year-old C. kousa var. chinensis plants grown in a 3-gal container to fulfill Koch's postulates. Kousa dogwood plants were drench inoculated (800 ml/plant) with a pathogen slurry (two plates of 7-day-old culture/liter) of isolates FBG6343 and FBG6364 (five plants per isolate). Control plants were drenched with agar slurry without the pathogen. The study was conducted in a greenhouse maintained at 21 to 23°C and 70% relative humidity with a 16-h photoperiod and irrigated twice a day for 2 min using an overhead irrigation system. Forty-five days after inoculation, plants showed dieback symptoms, and dark brown lesions developed in the roots of all inoculated plants. Isolates with morphology and sequences identical to those of FBG6343 and FBG6364 were recovered from root tissues of all inoculated plants. All control plants remained symptom-free, and P. litorale was not isolated from the root tissue. Previously, P. litorale was reported to cause disease on apple, kiwi, planatus, and rhododendron (Derviş et al. 2020; Li et al. 2021; Mert et al. 2020; Polat et al. 2023). To our knowledge, this is the first report of P. litorale causing root rot of kousa dogwood in Tennessee and the United States. Identification of this pathogen as the causal agent is crucial to developing timely management practices.

Morphology and molecular evidence reveal hidden diversity among snapping shrimp of the Alpheus obesomanus group (Decapoda: Alpheidae) with the description of a new species from Brazil

The snapping shrimp family Alpheidae Rafinesque, 1815 includes numerous species, most of which present controversial geographical distributions. The disjunct distribution of Alpheus simus Guérin-Méneville, 1856 in the western Atlantic, from Florida to the south of the Caribbean Sea and then from Rio Grande do Norte to Bahia in Brazil, suggests that Brazilian material may belong to an undescribed species. The examination of specimens previously identified as A. simus revealed the distinct morphology of material from Brazil, resulting in the description of a new species based on morphological and molecular evidence. The specimens were collected from fragments of fire coral at the Parque Municipal Marinho do Recife de Fora, Porto Seguro, Bahia, Brazil. Alpheus coralvivo sp. nov. belongs to the A. obesomanus group and is morphologically similar to A. simus (northwestern Atlantic) and A. saxidomus Holthuis, 1980 (eastern Pacific), differing mainly in the stylocerite and scaphocerite lengths and in the large gap between the cephalothorax and pleuron of the first pleonal segment. Molecular analysis using the mitochondrial cytochrome oxidase subunit I gene support the existence of the new species.

Stiphodon chlorestes, a new species of sicydiine goby (Teleostei: Gobioidei) from Taiwan and Luzon.

Stiphodon chlorestes sp. nov. is described based on seven specimens collected from Taiwan and Luzon. It is a large-sized Stiphodon species sharing the second dorsal-fin rays 9-10 and pectoral-fin rays 14-16 with similar-sized congeners. However, it differs from them by the wider interorbital width and almost complete lack of scales on the occipital region in males. In addition, the new species can be further distinguished from all congeners by seven to eight oval bands or a black longitudinal band on the lower body, black and white spots on pectoral fins, and a short red or orange line on posterior upper edge of caudal fin. Molecular analysis based on the 680-bp mitochondrial cytochrome oxidase subunit I (COI) fragments also supports it as a distinct species belonging to the "Stiphodon elegans group" and a sister group of the clade consisting of Stiphodon multisquamus and Stiphodon palawanensis.

Neurigona susanrocesae, the first Neurigona species from the Philippines

The first species of the genus Neurigona is described from the Philippines. The diagnosis, description and molecular data based on the partial DNA sequence of mitochondrial cytochrome oxidase subunit I gene were documented. The Neurigona species exhibits distinct characteristics, including a sub-apical dorsal stylus resembling an arista, a peduncle on the seventh abdominal segment, a flat dorsal postcranium, elongated legs devoid of prominent setae or bristles, and a distinctive rectangular bend in wing vein M. Based on the DNA sequence analysis, the unknown species is 90.06% similar to Cyrtona sp. and Neurigona zhejiangensis. However, 10% of its DNA sequences are significantly variable making it a totally different species. The specimen was verified by a dipterist as a species belonging to Genus Neurigona under Family Dolichopodidae. These flies are recognized for their lengthy legs and sizable yellow bodies. The species was named as N. susanrocesae as a tribute to the late Susan Roces, the “Queen of Philippine Movies”.

Morphological and molecular notes on root lesion nematode Pratylenchus coffeae and Pratylenchus neglectus (Nematoda: Pratylenchidae) from Pakistan

During a survey, a root lesion nematode Pratylenchus coffeae was recovered from the soil associated with Plantago ovata plantation in Sindh, Pakistan. The first report of P. coffeae from soil and root of Plantago ovata was examined morphologically and molecularly. The Pakistani population had few geographical variation and is characterized by body length (L = 360–450; a = 18.0–22.5, c = 7.2–16.1, c' = 1.7–3.1, V% = 72–80), a labial region with 2 annuli, a lateral field with 6 incisures, stylet 16–17 μm long with rounded knobs, spermatheca reduced or absent, post vulval uterine sac 28–35 μm long, and tail tip smooth, rounded, and containing 19–23 annuli ventrally. Morphometric and molecular features of 18 S rDNA, and mitochondrial cytochrome oxidase subunit 1 (cox1) genes are characterized. Phylogenetic analysis using cox1 of mtDNA place it close to P. coffeae, and P. speijeri with 1.00 posterior probability values. Measurements, line illustrations, and LM pictures for P. coffeae are given. Further, the detailed morphological, molecular, Discriminant analysis (DA) revealed differences between the P. coffeae and the close species morphologically and molecularly. Besides, P. neglectus verified using morphology and molecular characters. The nblast result of 18 S rDNA and cox1 of mtDNA showed 100 % similarity with the same species. Phylogenetic analysis using 18 S rDNA and cox1 placed the Pakistanis population of P. neglectus together with the other P. neglectus in a clade with 1.00 posterior probability. Population study of P. neglectus associated with grapevines (Vitis vinifera L.) revealed that this species had the most prominence value in Abbottabad of Khyber Pakhtun Khwa Province of Pakistan.

Microbiome of lovebug (Plecia longiforceps) in Seoul, South Korea.

Lovebugs appeared in large numbers across a wide area in Seoul, South Korea, in June 2023. The sudden appearance of exotic insects not only discomforts people but also fosters anxiety, as their potential for pathogen transmission would be unknown. In this study, targeted next-generation sequencing (NGS) of the 16S rRNA gene V4 region was performed using iSeq 100 to screen for bacteria in lovebugs. Forty-one lovebugs (20 females and 21 males) collected in Seoul, Korea, were identified as Plecia longiforceps based on mitochondrial cytochrome oxidase subunit 1 sequencing data using PCR. We analyzed the microbiome of the lovebugs and detected 453 species of bacteria. Among all bacteria screened based on NGS, Rickettsia was detected in all samples with an average relative abundance of 80.40%, followed by Pandoraea and Ewingella. Diversity (alpha and beta) between females and males did not differ; however, only Tumebacillus showed a higher relative abundance in females. Sequencing analysis of Rickettsia using a gltA gene-specific primer by PCR showed that it had higher sequence similarity to the Rickettsia symbiont of arthropods than to the spotted fever group rickettsiae. Eleven samples in which Pandoraea was detected by iSeq 100 were confirmed by PCR and exhibited 100% sequence identity to Pandoraea oxalativorans strain DSM 23570. Consequently, the likelihood of pathogen transmission to humans is low. The applied method may play a crucial role in swiftly identifying bacterial species in the event of future outbreaks of exotic insects that may be harmful to humans.IMPORTANCELovebugs have recently emerged in large numbers in Seoul, causing major concern regarding potential health risks. By performing the next-generation sequencing of the 16S rRNA gene V4 region, we comprehensively examined the microbiome of these insects. We identified the presence of numerous bacteria, including Rickettsia and Pandoraea. Reassuringly, subsequent tests confirmed that these detected bacteria were not pathogenic. The present study addresses health concerns related to lovebugs and shows the accuracy and efficiency of our detection technique. Such methods prove invaluable for rapidly identifying bacterial species during potential outbreaks of unfamiliar insects, thereby ensuring public safety.