COI Gene Sequences Research Articles

Status of population genetic structure and distribution of Bemisia tabaci genetic groups in vegetable crops and others in India

Whitefly, Bemisia tabaci is one of the polyphagous pests infesting wide variety of crop plants and is vectors of plant viruses belonging to begomivruses whose transmission efficiencies have been reported to be varying. Therefore, we have conducted a series of extensive surveys during 2019-21 to monitor and collect B. tabaci populations across the country and studied the distribution of its genetic groups. Further, PCR amplification, purification, cycle sequencing and partial CO-I gene sequencing has been done to understand population genetic structure and the distribution of genetic groups B. tabaci in India. Data analysis revealed a total of 11 genetic groups of which Asia II-3, Asia II-6 & Asia III were new to India reported for the first time from Chhattisgarh, Andhra Pradesh and Maharashtra for the first time in India. These studies will help farmers and other stakeholders to effectively manage whitefly and whitefly borne begomoviruses for increased vegetables and in turn vegetable oil production.

Graphidessajinfoensis, a new species of longhorned beetle (Coleoptera, Cerambycidae, Lamiinae, Desmiphorini) from China.

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.

First description study on molecular characterization of Hemiclepsis khankiana (Hirudinea: Glossiphoniidae) in China

BackgroundLeeches are an integral component of aquatic biocenosis and can be found in a wide range of ecosystems such as freshwater, saltwater, flowing, and still-water ecosystems. It especially plays an important role in the freshwater benthic community and is an important part of the food web. In this study, a leech species was found in the mantle cavity of wild freshwater mussels in Zigong City, Sichuan Province, China, and its identity was determined through morphological analysis and molecular biological analysis.ResultsThe leech is Hemiclepsis khankiana, a new species of Hemiclepsis that has been discovered in Russia in recent years. Through morphological analysis, the current survey observed that the morphological characteristics of Hemiclepsis khankiana eyespots were significantly different from the first reported description. The first pair of eyespots on the leech were separated and clear, while it had been reduced to unclear shadows in the previous report. The phylogenetic tree based on the COI gene showed that the COI gene sequence obtained in this study was in the same evolutionary branch as Hemiclepsis khankiana (MN295420, MN295421). Genetically, it was most closely related to Hemiclepsis kasmiana (mean COI p-distance = 3.98%).ConclusionsThe current study reported on the new distribution range of Hemiclepsis khankiana, which was initially discovered in China. This study indicates that the distribution range of the leech species has expanded, laying a foundation for further studies in China.

First discovery of the ant genus Eburopone Borowiec, 2016 (Hymenoptera, Formicidae, Dorylinae) in the Oriental realm, with description of a new species from Vietnam.

The doryline ant genus Eburopone Borowiec, 2016 currently contains only one valid species, E.wroughtoni (Forel, 1910) from southern Africa, with a considerable number of undescribed species awaiting formal description in the Afrotropical and Malagasy regions. In the present paper, Eburoponeeasoanasp. nov. is described based on workers and dealate queens from a colony series collected in an evergreen forest on the Dak Lak Plateau of Vietnam (Ea So Nature Reserve, Dak Lak Province). The worker of the new species is morphologically clearly distinguished from E.wroughtoni by the combination of following characteristics: i) frontal line distinct, extending a little beyond mid-length of cranium; ii) anterior (frontoclypeal) margins of torulo-posttorular complex not forming conspicuous lobes protruding over anterior clypeal margin in full-face view; iii) mandibles when closed in full-face view forming only a little space between anterior clypeal margin and mandibles; iv) promesonotal suture faint and inconspicuous; v) abdominal segment III in dorsal view distinctly wider than long, with lateral margins only feebly convex. This represents the first discovery of the genus Eburopone in the Oriental realm, revealing the disjunct distribution of the genus. A partial sequence of the mitochondrial COI gene (658 bp) is provided as a DNA barcode for the new species. A worker-based key to the doryline genera of the Oriental realm is also provided.

Morphological and molecular characterisation of Longidorus patuxentensis n. sp. (Nematoda: Longidoridae) from Maryland and California, USA

Summary A new needle nematode species, Longidorus patuxentensis n. sp., was collected along the banks of the Western Branch Patuxent River from Upper Marlboro, MD, USA, and described herein. Female body length of the new species ranges from 3.8 to 5.2 mm, with a set off lip region by depression, 77-92 μm long odontostyle, 40-53 μm long odontophore, vulva located at 46.3-50.1% and tail conoid with bluntly rounded tip. The new species has four juvenile developmental stages and no males. It looks morphologically similar to L. breviannulatus, L. elongatus, L. martini, L. americanus, L. grandis, L. sabalanicus and L. sturhani by having a ventrally curved to spiral body, generally similar lip region and conoid tail with a rounded terminus, but differs from these species by the odontostyle, odontophore, total stylet length and a few other characters. Phylogenetic analysis of the D2-D3 expansion segments of 28S rRNA gene sequences placed L. patuxentensis n. sp. in a clade with L. litchi, L. fangi, L. jonesi, L. diadecturus and Longidorus sp. The D2-D3 sequence of L. patuxentensis n. sp. was identical to that of Longidorus sp.5 collected from Juglans sp. growing in Butte County, California, USA. The D2-D3 of 28S and ITS1 rRNA and COI gene sequences indicated that the Maryland and California populations belong to the same species, described herein as L. patuxentensis n. sp.

DNA barcoding, phylogeny and phylogeography of the cyst nematode species of the Schachtii group from the genus Heterodera (Tylenchida: Heteroderidae)

Summary Cyst-forming nematodes of the genus Heterodera are highly derived and economically important plant parasites. The Schachtii group of this genus is one of the largest ones with a total of 18 species parasitising dicotyledons. In this study, we provided comprehensive phylogenetic analyses of several hundred COI and ITS rRNA gene sequences of selected species from the Schachtii group, including H. betae, H. cajani, H. ciceri, H. galeopsidis, H. glycines, H. medicaginis, H. mediterranea, H. schachtii, H. sonchophila and H. trifolii, using Bayesian inference, maximum likelihood, and statistical parsimony. One hundred and twenty four new COI, 57 ITS rRNA and eight hsp90 gene sequences from 81 nematode populations collected in 19 countries were obtained in this study. Our study showed that the ITS rRNA gene has limited discrimination power compared to the COI gene. However, our analysis also revealed that partial COI gene sequences were identical for H. trifolii, H. betae and H. galeopsidis. Based on the results of phylogeographical analysis and age estimation of clades with a molecular clock approach, it was hypothesised that the majority of the Schachtii group species originated and diversified in the Mediterranean Basin biodiversity hotspot during the Pleistocene and then dispersed from this region across the world. The Sino-Japanese Floristic Region is likely one of the centres of diversification for the soybean cyst nematode, which showed distinct population structure. The possible role of hybridisation and polyploidisation in the evolution of species of the Schachtii group is discussed.

Molecular Identification of Aedes, Armigeres, and Culex Mosquitoes (Diptera: Culicidae) Using Mitochondrial Cytochrome Oxidase Subunit I Genes in Myanmar.

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. 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. 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. 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.

Establishment of Sinoxylon anale Lesne (Coleoptera, Bostrichidae) in Brazil: Identification, Host Plants, Distribution, and Damage.

Damage from Sinoxylon anale Lesne, a woodboring beetle not previously known to be established in Brazil, was observed in young jabuticaba trees (Plinia cauliflora, Myrtaceae) in a nursery in the municipality of Laranjal Paulista, state of São Paulo. We immediately advised MAPA ("Ministério da Agricultura, Pecuária e Abastecimento") and collected samples from the nursery and from different hosts in nearby areas, to identify the specimens and investigate the dynamics of the infestation in the jabuticaba trees. Sinoxylon anale was also collected in ethanol-baited and ultraviolet-light traps and in dry branches of the native species pau-jacaré (Piptadenia gonoacantha, Fabaceae) and inga (Inga vera, Fabaceae), and the exotic pau-d'água (Dracaena fragrans, Asparagaceae) in the municipality of Piracicaba, state of São Paulo. These collections established that S. anale larvae and adults develop in dead branches of four new host plants. Taxonomic studies using morphological parameters and DNA barcoding confirmed the identification of S. anale. An illustrated key to the three Sinoxylon species now recorded in Brazil is provided, and the COI gene sequences have been made available in a public database. Sinoxylon anale probably attacked the young jabuticaba trees after they were killed by larvae of long-horned beetles (Cerambycidae). So far, S. anale has been found established only in two locations in the same area of the state of São Paulo.

Clarification of the identity of deep-sea crab Tymolus uncifer (Ortmann, 1892) and validation of Tymolus melanommus (Doflein, 1904) (Decapoda: Brachyura: Cyclodorippidae).

Examination of material from Japanese waters has disclosed that two species have been confounded under Tymolus uncifer (Ortmann, 1892) (Decapoda: Brachyura: Cyclodorippidae) in previous literature. The identity of Tymolus uncifer sensu stricto is clarified in reference to the lectotype, and the second taxon is herein attributed to T. melanommus (Doflein, 1904), which was originally recognised as a forma of T. uncifer with one specimen from off Kenya and one specimen from Japan. A lectotype is herein selected for T. melanommus in the interest of nomenclatural stability. Differentiating characters between the two species are assessed in detail, and those two species are compared also with T. glaucommus (Alcock, 1894), T. similis (Grant, 1905), T. brucei Tavares, 1991, T. daviei Tavares, 1997, T. hirtipes Tan & Huang, 2000 and T. globosus Spiridonov & Trkay, 2007, all of them are considered as valid. Sexual dimorphism and ontogenetic variation are seen in the relative length and stoutness of the pereopods 2 and 3 in T. uncifer and T. melanommus, but the morphometric characters derived from the two pereopods are useful as diagnostic at comparison between same sex in adults. Comparison of partial fragments of the mitochondrial COI and 16S rRNA gene sequences strongly supports that the two taxa are distinct. Previous records of T. uncifer, including fossil records, are reviewed based on the present findings. Tymolus uncifer and T. melanommus are restricted to Japanese waters at present.

Discovery of a new Muraenichthys eel (Anguilliformes: Ophichthidae) from the Bay of Bengal, India, with its molecular characterization

Muraenichthys hibinoi sp. nov. is described herein based on two specimens collected from Shankarpur fishing harbor, West Bengal, India. The new species differs from Muraenichthys gymnopterus, Muraenichthys hattae, Muraenichthys longirostris, and Muraenichthys thompsoni by having less distance between the anus and the origin of the dorsal fin (61.8%–57.37% vs 73.8%–82% of HL). The new species has a close morphological affinity with M. gymnopterus and M. hattae with significant differences from both species. The new species differs from M. gymnopterus by having higher vertebral count, i.e., predorsal vertebrae (40– 41 vs 30), preanal vertebrae (50 vs 43), and total vertebrae (140–141 vs 130). The new species also differs from M. hattae, having fewer total vertebrae (141–142 vs 148–155) and fewer predorsal vertebrae (40–41 vs 48–53). The COI gene sequence of the new species has been generated and submitted to the NCBI database with accession number OP114397. The new species differs from all the available congeneric sequences of Myrophinae group available in NCBI or BOLD with K2P distances of 8.4% to 9.5%. Thus, both the genetic and morphometric data confirm the presence of a new species in Myrophinae from Indian waters.

Taxonomic and conservation status of Majoidea “Spider Crabs” from Indonesia: Recommendations for management

The Majoidea contains either six, seven, or eight valid families according to different sources. This has resulted in difficulties with the taxonomy and conservation of this superfamily. The Majoidea in Indonesia waters were documented in the past, however, their taxonomy and conservation status were booth poorly documented. The aims of this study were (1) to determine the taxonomic status of; and (2) to evaluate the conservation status of Indonesian majoid crabs. The IUCN red list was used to evaluate the conservation status of Majoidea identified based on DNA barcodes Mitochondrial cytochrome c oxidase I gene (COI). Considering genetic distances between specimens, genetic divergence, the phylogenetic trees and concepts of genetic species, the superfamily Majoidea is composed of multiple species or genera that belong to different subfamilies. This study differentiated all individual sequences as different species or genera except three sequences from Aceh and Solor. The results were classified into two groups based on taxonomic status. The first group comprised species, e.g. Schizophrys aspera, Tylocarcinus styx, Perinia tumida, Menaethius orientalis, and Pisa armata/Hyastenus elongatus. The second group comprise such as Majidae sp., Brachyura sp. and Decapoda sp. The Majoidea was recognized and differentiated based on data set comprising 45 haplotypes, 227 segregating sites, 0.99 haplotype diversity, and 0.17 nucleotide diversity. Minimum Spanning Tree analyses of COI gene sequences reveal 47 distinct groups of operational taxonomic units corresponding to genera or species. In 2021, all of Majoidea taxa were listed as either data deficient or not evaluated in the IUCN Red List or appendices, due to insufficient data. These results suggest that comprehensive follow-up surveys of the taxonomic and conservation status of Majoidea are still urgently required.

Phylogenetic analysis among some species of aphids (Homoptera: Aphididae) using DNA sequencing moleculartechnique

The phylogeny of aphids (Hemiptera: Aphididae) has been studied by molecular technique for the first time in Kurdistan, this study comprises relationship between six species of Aphids collected from the leaves of different plants (Chrysanthemum, Oak, Almond, Pine, Asteraceae and Herbaceous) in many localities of Erbil governorate Kurdistan region-Iraq, started in May to July 2022. For understanding about aphid evolution, phylogenetic of aphid is crucial. However, neither the phylogenetic alterations of the Aphid taxa nor their comprehensive definition have been achieved. Therefore, a unique method has been developed in this study to examine COI gene sequencing and infer the relationships between the major aphid taxa. DNA was isolated, and a band of 550 bp of mt COI gene was amplified. Then the amplicons were sequenced. The part of the COI gene of the insect samples are alignment inside of NCBI GenBank by BLAST program, were used to compare our nucleotide have sequenced with other stored species of insect sequences. This research presented that studying phylogeny of aphid species through a sequencing technique can create a phylogenetic tree for the used species with reliable results. So, our species in Kurdistan region clustered in a monophyletic clade with published from most countries with high identic value (100%). The COI sequence of aphid species were submitted to GenBank with six accessions of OP355287 - OP355292. The composition of nucleotides of the sequence of COI gene was low of G-C base pairing. We were successful in showing that the mt-COI gene can be used as a molecular marker for the identification of related species, as shown by the similarities between the phylogenetic association created by COI

The Caribbean intertidal mite Alismobates inexpectatus (Acari, Oribatida), an unexpected case of cryptic diversity?

Molecular genetic analyses of Caribbean populations of the supposedly widespread intertidal oribatid mite Alismobates inexpectatus revealed the existence of a cryptic species. The new species, Alismobates piratus sp. n., shows considerable COI and 18S rRNA gene sequence divergences and although morphometric analyses indicate considerable variation between the taxa, no distinguishing morphological feature could be detected. The extreme intertidal environment is suggested to be responsible for the observed morphological stasis of the two species and vicariance is supposed to be responsible for their speciation. Alismobates piratus sp. n. was found on Hispaniola, Guadeloupe, Barbados and Curaçao indicating a predominant distribution on the Greater and Lesser Antilles, whereas the occurrence of A. inexpectatus is primarily restricted to Central America, the northern Caribbean and the Greater Antilles. Haplotype network analyses indicate distinct geographic structuring and the absence of recent gene flow among the Antillean A. piratus sp. n. populations. Central American and Antillean populations of A. inexpectatus show similar patterns but populations from Bermuda and the Bahamas are characterized by a common origin and subsequent expansion. Genetic landscape analysis demonstrates that vast stretches of open ocean, like the Caribbean Basin and the Western Atlantic, act as rather effective barriers, whereas the continuous continental coastline of Central and North America may facilitate dispersal. Genetic data also indicates that the Gulf Stream plays an important role for the biogeography of intertidal oribatid mites as it may be responsible for the strong link between Central and North American populations as well as for the colonization of Bermuda.

Two New Species of Nipponbathynella Schminke, 1973 from South Korea, with a Brief Discussion on Korean Parabathynellid Genera (Crustacea, Bathynellacea)

Parabathynellidae is a family well known for its diversity and is endemic to the interstitial hyporheic zone of South Korea. Two new parabathynellid species, Nipponbathynella yeongchunensis sp. nov. and N. gapcheonensis sp. nov., were found through groundwater sampling on riverbanks in South Korea. Nipponbathynella yeongchunensis sp. nov. can be distinguished from its congeners by the base socket of the two dorsal setae on the antennule’s first segment. Nipponbathynella gapcheonensis sp. nov. can be distinguished by the protopod of thoracopod II–IV with hairs. We provide morphological descriptions and remarks about the two new species with molecular information based on 18S rRNA and partial COI gene sequences. In addition, phylogenetic analysis, including Nipponbathynella Schminke, 1973, which had not been previously molecularly evaluated, was inferred. The results of the 18S rRNA gene phylogeny suggest a close relationship between the genera Nipponbathynella and Arisubathynella, as well as Europe–North American genera. We have also included a brief discussion and an identification key for the five known parabathynellid genera found in South Korea.

Karyotypes and COI Gene Sequences of Chironomus annularius Sensu Lato (Diptera, Chironomidae) from Russia, Mongolia, and Armenia

The larvae of the genus Chironomus are a common object for hydrobiological studies, as well as a model object for cytogenetics. Morphologically, the species are very similar. One of these species or species complex is Chironomus “annularius”, which has a Holarctic distribution. It has chromosomal banding sequences characteristic of Nearctic and Palearctic populations. Using an integrated method that included morphology, cytogenetics, and molecular genetics, we analyzed populations from Russia, Mongolia, and Armenia. We found through cytogenetics and larval morphology that the populations have high similarity. Molecular genetic studies have shown significant differences between the populations. The genetic distances between the populations, in some cases, exceed the interspecific threshold of 3%, and are 6.5%. In the South Caucasian population (Lake Sevan), a chromosomal banding sequence, h’annD3, that was previously observed only in North America, was found for the first time. The larvae from Lake Sevan have large genetic distances from others, and are morphologically similar to the species Chironomus markosjani Shilova 1983, described from this lake without comparison with Ch. annularius nor an exact description of the karyotype. The sequences of the COI genes from Montenegro (Lake Skadar) and West Siberia (Novosibirsk) found in GenBank may belong to a new undescribed species, or a species not represented in the database. Thus, the analyzed data on Chironomus “annularius” support the presence of the complex of homosequential species under this name.

Morphological and Molecular Identification of Helopeltis Species on Cocoa from Kaliwining Experimental Station, Jember, Indonesia

Severe crop loss of cocoa in Indonesia has long been reported to be caused by Helopeltis. It has been noted that cocoa and Helopeltis have existed in Kaliwining Experimental Station of Indonesian Coffee and Cocoa Research Institute (ICCRI), Jember, Indonesia, for about 100 years, and the species have developed during that time. Our study aimed for examining the Helopeltis species that attacked cocoa in this experimental station both morphologically, morphometrically, and molecularly and to compare the species with the one attacking cocoa in Java and Nusa Tenggara, preserved at the GenBank. Helopeltis was sampled from infested cocoa pods collected representatively from cocoa at the Kaliwining ES and then reared in the Crop Protection Laboratory of ICCRI. Adults of Helopeltis laboratory-reared insects were identified, 30 females 30 males were selected. Weexamined the morphology and morphometry of the samples, and their identifica tion was based on an insect identification key. Morphometry measurement of male and female body parts includes body length, antenna segments, head, eyes, collar, wing, tibia, and femur. In addition, the molecular identification of Helopeltissp. was made through DNA extraction, amplification, and sequence analysis of DNA target regions using the COI part of mitochondrial DNA. External morphological identification, supported by morphometry of the Helopeltis specimen from Kaliwning, the white bands on all femora, body length, and the ratio of the firstantennal segment to the posterior width of the pronotum suggested that the insect belongs to Helopeltis bradyi. Similarly, molecular identification using PCR amplification through specific primers from COI gene sequences confirmed that Helopeltis specimens from Kaliwining are classified as Helopeltis bradyi.

Molecular Identification of Acrossocheilus jishouensis (Teleostei: Cyprinidae) and Its Complete Mitochondrial Genome.

Molecular identification, such as DNA barcoding, is a useful tool that is widely applied in distinguishing species. To identify the cyprinid Acrossocheilus jishouensis, which was previously known to be restricted to only its type locality, we conducted molecular identification of this species based on 23 samples in five localities. Molecular identification based on the mitochondrial COI gene sequence showed that the morphologically similar samples from the five populations were all A. jishouensis, as the mean genetic distances between populations were very small (0.1-1.6%); thus, the distribution of this species was substantially expanded. The whole mitochondrial genome of one sample was also assembled, which was 16,594bp in length and consisted of 13 protein-coding genes (PCGs), two rRNA genes, 22 tRNA genes, and one control region. All PCGs began with ATG except the COI gene, which started with GTG; seven PCGs used the complete stop codon TAA, while four terminated in T(AA) and two ended with TAG. The overall base composition reflected a higher proportion of A+T than G+C and a positive AT-skew and negative GC-skew pattern except for the opposite in ND6. Phylogenetic relationships inferred using BI and ML methods revealed that both Acrossocheilus and Onychostoma were nonmonophyletic, which indicated that the traditional diagnoses between these two genera need to be assessed further. The results of this study not only expanded the known distribution ranges of A. jishouensis, but also provided a valuable data resource for future molecular and evolutionary studies of Acrossocheilus and other cyprinids in Barbinae.

Asynchronous demographic dynamics of rodent community in low latitude Asia

Abstract There is increasing evidence that demographic history and phylogeographic consequences of past climate changes unfolded locally and varied from region to region. Despite the high rodent species diversity and endemism in low latitude Asia, how they have responded to the past climatic fluctuations remains unexplored. In the present study, we trapped 253 murine individuals and sequenced their mitochondrial COI gene sequence. A total of ten species belonging to five genera were recognized through phylogenetic analyses. The results of divergence time estimation showed that the most common ancestors for all recognized species occurred in the Pleistocene. Signals of demographic expansion were detected in six species by at least one test and the events of sharp population size increase occurred asynchronously among species. The demographic expansion during the glaciation periods was corroborated by the expanded suitable distributional areas predicted by ecological niche modelling. The diversified demographic histories of rat communities in low-latitude Asia suggested that species might have responded to past regional environmental changes in an individualistic way.

Distribution and molecular identification of ixodid ticks infesting cattle in Kilombero and Iringa Districts, Tanzania

BackgroundHard ticks infesting cattle are vectors of Tick-borne diseases that causes major public health problems and considerable socioeconomic losses to the livestock industry in tropical and subtropical countries. A repeated cross-sectional study was carried out by collecting ticks on cattle during the wet and dry seasons from January to August 2021 in order to determine hard tick prevalence, distribution, and abundance on cattle in Kilombero and Iringa Districts of Tanzania. The collected ticks were identified morphologically using published morphological keys under a stereomicroscope and confirmed by polymerase chain reaction (PCR) and sequencing.ResultsOut of 740 examined cattle, 304, (41.08%) were infested with ticks. In total, 1,780 ticks were counted on one side of the animal’s body and doubled, whereby resulting in a total of 3,560 ticks were recorded. Individual tick burden ranged from n = 2 to 82 ticks per animal, with a mean tick burden of n = 11.7 ± 0.68 per animal. A total of 1,889 ticks were collected from infected cattle including 109 additional ticks observed while collecting ticks based on the animal’s posture when restrained to the ground. Out of 1,889 ticks, nine species from three genera were identified morphologically: 1,377 fit in the genus Rhipicephalus, 459 to the genus of Amblyomma, and 53 to the genus Hyalomma. Rhipicephalus microplus was the most prevalent (n = 909, 48.1%), followed by Rhipicephalus evertsi (n = 310, 16.4%), Amblyomma lepidum (n = 310, 16.4%), Rhipicephalus appendiculatus (n = 140, 7.4%), Amblyomma gemma (n = 120, 6.4%), Hyalomma rufipes (n = 50, 2.6%), Amblyomma variegatum (n = 29, 1.5%), Rhipicephalus decoloratus (n = 18, 1.0%), while the least common was Hyalomma albiparmatum (n = 3, 0.2%). Tick diversity was higher in Iringa compared to Kilombero District. Tick prevalence was higher in wet season (n = 148, 41.11%). Ticks were widely distributed in different parts of the host body, with the highest distribution on zone 4 (n = 1,060, 56.11%), which includes groin, flank, abdomen, and around inner thigh of the hind legs and the lowest distribution on zone 2 (n = 14, 0.74%), which includes the back surface of the body. Both Amblyomma lepidum and Rhipicephalus microplus species were found in all the five body zones, and with the highest proportions recorded on zone 4: A. lepidum (n = 209, 67.42%) and R. microplus (n = 714, 78.55%). The nine tick species identified morphologically were also confirmed using molecular methods. However, during sequencing, two species (Rhipicephalus appendiculatus and R. decoloratus) had poor quality sequences and were excluded from the sequence analysis. Sequencing results indicated high nucleotide identity (96–100%) with sequences available in GenBank and Barcode of Life Database (BOLD). The phylogenetic analysis of partial mitochondrial COI and 16S rRNA gene sequences of ticks were used to confirm the morphological identification.ConclusionThe results showed a high burden of tick infestation on cattle, which could reduce animal production and potentially increase the risk of tick-borne diseases. Therefore, it is necessary to explore the epidemiological and molecular aspects of various tick species in other regions of Tanzania.

Two new species of the bathyal holothurian genus Pannychia (Elasipodida, Laetmogonidae) from Japanese waters.

Sea cucumbers are one of the most common large animals in the deep-sea benthic communities, and those of the genus Pannychia are particularly abundant in the bathyal North Pacific Ocean. In Japanese waters, three species of Pannychia have been reported, mainly from the northern and eastern parts of the country. Here, we describe two new species from the western part of Japan. These two new species were placed in Pannychia by the presence of dorsal papillae on the ventrolateral radii and the presence of large wheel ossicles with rim teeth. They are distinguished from all other nominal species of Pannychia on the basis of the morphological characteristics of their body wheel ossicles. Pannychia nagasakimaruae sp. nov. has a unique ossicle composition in its dorsal and ventral body walls: four forms of wheel ossicles. Pannychia rinkaimaruae sp. nov. differs from other congeners in its small wheel ossicles in the dorsal and ventral body walls, which have conical rim teeth. We barcoded partial COI gene sequences from holotypes and paratypes of two new Japanese species. Our molecular phylogenetic analysis showed that these two new morphologically recognized Japanese species form distinct clades separated from other Pacific Pannychia species, so that the two new Japanese species were also supported by genetic results. In addition, we describe and visualize the poorly calcified calcareous rings of the two new species using X-ray μCT system, a useful method for observing such structures, which are important structures in holothurian anatomy.