scholarly journals Molecular evidence confirms occurrence of Rhipicephalus (Boophilus) microplus in Kenya and suggests that an undifferentiated genotype is prevalent in the African continent

2020 ◽  
Author(s):  
ESTHER GATHONI KANDUMA ◽  
David Emery ◽  
Naftaly W. Githaka ◽  
Edward K. Nguu ◽  
Richard P. Bishop ◽  
...  
Keyword(s):  
Vector Competence ◽  
Genetic Relationships ◽  
Boophilus Microplus ◽  
Babesia Bovis ◽  
Taqman Probe ◽  
Coi Gene ◽  
Molecular Evidence ◽  
Morphological Identification ◽  
Genome Sequences ◽  
Coi Sequences

Abstract Background The tick vector Rhipicephalus (Boophilus) microplus which transmits Babesia and rickettsial pathogens has not been reported in Kenya since 1998. More recently, the pathogenic Babesia bovis has been detected in cattle blood DNA. The status of R. microplus in Kenya remains unknown. This study employed morphological and molecular tools to characterize R. microplus originating from Kenya and assess the genetic relationships between Kenyan and other African R. microplus genotypes. Methods Morphological identification and differentiation of 35 Kenyan and 11 reference tick specimens was implemented by following standard tick reference keys and identification guides. Genetic and phylogenetic relationships between the Kenyan and other annotated R. microplus reference sequences was investigated by analysis of the cytochrome c oxidase subunit I (COI) gene. The complete mitochondrial (mt) genome of four tick specimens which included two R. microplus was assembled from whole genome data to further characterize the Kenyan ticks. A B. bovis specific Taqman probe qPCR assay was used to detect B. bovis in gDNA from R. microplus ticks. Results Of the 35 Kwale tick specimens analysed, 23 were confirmed to be R. microplus by both morphology and molecular characterizations. The Kenyan R. microplus COI sequences showed very high pairwise identities (>99%) and clustered very closely with reference African sequences. We also found a low differentiation and lack of geographical sub-structuring among the African COI sequences. The mitochondrial genome sequences of the two Kenyan R. microplus ticks also clustered closely with reference genome sequences from Brazil, USA, Cambodia and India. No B. bovis was detected in the Kwale R. microplus DNA. Conclusions These findings confirm the presence of R. microplus in Kenya and suggest that a common undifferentiated genotype is prevalent in cattle in Africa. These and other recent findings of widespread occurrence of R. microplus in Africa provide a strong justification for urgent surveillance to determine and monitor the spread of R. microplus and vector competence of Rhipicephalid ticks for B. bovis in Africa, with the ultimate goal of strategic control.

scholarly journals Molecular Characterization of Anopheles sacharovi Based on Sequences of ITS2-rDNA Region and COI Gene in North of Iran

2019 ◽  
Author(s):  
Sahereh Gholami ◽  
Hasan Bakhshi ◽  
Seyyed Hassan Moosa-Kazemi ◽  
Alireza Zahraei-Ramazani ◽  
Alireza Chavshin ◽  
...  
Keyword(s):  
Sibling Species ◽  
Coi Gene ◽  
Its2 Region ◽  
Morphological Identification ◽  
Anopheles Mosquitoes ◽  
North Of Iran ◽  
Northwest Of Iran ◽  
Coi Sequences ◽  
West Azerbaijan ◽  
East Azerbaijan

Background: Malaria is an important mosquito-borne disease considered as one of the public health concerns across many countries. Anopheles mosquitoes are the main vectors of Plasmodium parasites, which cause malaria. Some of these vectors such as Anopheles maculipennis s.l. and Anopheles sacharovi are considered as complex of sibling species distributed in north of Iran. Methods: This study was conducted in north and northwest of Iran including East Azerbaijan, West Azerbaijan, Ar­dabil, Golestan and North Khorasan provinces with emphasis on the northern borders of the country during 2015–2016. Adult specimens were collected and subjected to morphological identification as well as molecular analysis. Results: Overall, 10405 mosquitoes were collected comprising 21 species. Culex pipiens and Cx. theileri were found as the most frequent species in whole study area. Morphological identification showed that out of 1455 female Anoph­eles specimens, 77% belonged to An. maculipennis Group. Out of the identified species, ITS2 region and COI gene sequences of 8 An. maculipennis s.s. and 31 An. sacharovi representing all provinces were obtained and submitted to GenBank. The COI sequences for An. sacharovi revealed the presence of 9 haplotypes with similarity of 98.17–100%. Conclusion: Some investigations have reported An. martinius as a member of sibling species of An. sacharovi among Iranian Anopheles genus; while based on our study, there was no evidence of the presence of this species in north and northwest of Iran.  

scholarly journals Combining old and new evidence to increase the known biodiversity value of the Sahamalaza Peninsula, Northwest Madagascar

2017 ◽  
Vol 86 (4) ◽  
pp. 273-295 ◽  
Author(s):  
Samuel G. Penny ◽  
Angelica Crottini ◽  
Franco Andreone ◽  
Adriana Bellati ◽  
Lovasoa M.S. Rakotozafy ◽  
...  
Keyword(s):  
16S Rrna ◽  
Molecular Taxonomy ◽  
Coi Gene ◽  
Morphological Identification ◽  
Rrna Gene ◽  
Taxonomic Identification ◽  
Pitfall Trapping ◽  
Wet Season ◽  
Mitochondrial Coi ◽  
Coi Sequences

Prior herpetological surveys in 1996 and 2000 identified 14 species of amphibians and 32 species of reptiles from the Sahamalaza Peninsula. This work increases the total number of amphibian and reptile species known from this area to 20 and 43 respectively. To maximise our chances of species detection, survey effort covered the entire wet season and part of the dry season, and utilised a combination of opportunistic searching, transect searching, pitfall trapping, and acoustic recording. We identified species through an integrative taxonomic approach, combining morphological, bioacoustic and molecular taxonomy. Together, this enabled the detection of cryptic and seasonally inactive species that were missed in the shorter prior surveys that relied on morphological identification alone. The taxonomic identification of amphibians utilised a fragment of the mitochondrial 16S rRNA gene; taxonomic identification of reptiles utilised a fragment of the mitochondrial COI gene, and when necessary, also mitochondrial fragments of the 16S rRNA ND1, ND2, ND4 genes. All sequences were deposited in Genbank and COI sequences were also deposited in the BOLD database to foster taxonomic identification of malagasy reptiles. We report two new taxa: a species of Boophis, since described as B. ankarafensis, and a candidate new species of microhylid (genus: Stumpffia). We document range expansions of Boophis tsilomaro, Cophyla berara, Blaesodactylus ambonihazo beyond their type localities. Along with significant range expansions across a range of taxa, including Blommersia sp. Ca05, Boophys brachychir, Brookesia minima, Ebenavia inunguis, Geckolepis humbloti, Madascincus stumpffi, Pelomedus subrufa and Phelsuma kochi. Forest in the peninsula is under extreme pressure from human exploitation. Unless unsustainable agricultural and pastoral practices encroaching on these habitats halt immediately, both forest and the species that occur there, several of which appear to be local endemics, may be irreversibly lost.

scholarly journals Using high-resolution melting to identify Calliphoridae (blowflies) species from Brazil

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e9680
Author(s):  
Pablo Viana Oliveira ◽  
Francine Alves Nogueira de Almeida ◽  
Magda Delorence Lugon ◽  
Karolinni Bianchi Britto ◽  
Janyra Oliveira-Costa ◽  
...  
Keyword(s):  
High Resolution ◽  
Dna Sequencing ◽  
High Resolution Melting ◽  
Coi Gene ◽  
Morphological Identification ◽  
Mitochondrial Coi ◽  
New Methods ◽  
Degraded Samples ◽  
Sequencing Technique ◽  
Coi Sequences

Forensic entomology is the study of insects and other arthropods used in the solution of crimes. Most of entomological evidences strongly depend on accurate species identification. Therefore, new methods are being developed due to difficulties in morphological identification, including molecular methods such as High-Resolution Melting. In this study, we reported a new HRM primer set to identify forensically important Calliphoridae (blowflies) from Brazil. For such purpose, Calliphoridae species of forensic importance in Brazil were listed and confirmed by specialists. Mitochondrial COI sequences of those species were downloaded from databases and aligned, and polymorphic variations were selected for distinction between species. Based on it, HRM primers were designed. Forty-three fly samples representing six species were tested in the HRM assay. All samples had the COI gene sequenced to validate the result. Identifying and differentiating the six species proposed using a combination of two amplicons was possible. The protocol was effective even for old insect specimens, collected and preserved dried for more than ten years, unlike the DNA sequencing technique that failed for those samples. The HRM technique proved to be an alternative tool to DNA sequencing, with advantage of amplifying degraded samples and being fast and cheaper than the sequencing technique.

DNA barcoding and taxonomic validation of Caranx spp. larvae

2021 ◽  
pp. 1-9
Author(s):  
Claudia A. Silva-Segundo ◽  
René Funes-Rodríguez ◽  
Jaime Gómez-Gutiérrez ◽  
Griselda Gallegos-Simental ◽  
Sergio Hernández-Trujillo ◽  
...  
Keyword(s):  
Dna Barcoding ◽  
Genetic Similarity ◽  
Fish Larvae ◽  
Coi Gene ◽  
Molecular Evidence ◽  
Nominal Species ◽  
Central Pacific ◽  
Diagnostic Characteristics ◽  
Coi Sequences ◽  
First Time

Abstract Of the five nominal species in the genus Caranx Lacepède 1801 distributed throughout the Eastern Central Pacific, Caranx caballus and Caranx sexfasciatus are the only two that have formal fish larval descriptions based on diagnostic characteristics (morphology, meristics and pigmentation). In this study, the diagnostic characteristics of three Caranx species larvae were validated using DNA barcoding analysis cytochrome c oxidase subunit I (COI; 651 bp). For the first time, the morphological taxonomic assignation of C. caballus fish larvae was confirmed using COI gene partial sequences of adults, with a genetic similarity between 99.8–100%. However, molecular evidence demonstrated that fish larvae previously described as C. sexfasciatus had high genetic similarity (99.7–100%) and low genetic distance (<1%) to Caranx caninus adults. An undescribed larval morphotype collected in the present study genetically matched (100%) with COI sequences of C. sexfasciatus adults. The diagnostic characteristics of this new morphotype were a lack of pigmentation in the supraoccipital crest, over the gut, and at the terminal region of the gut. The combination of diagnostic characteristics and DNA barcoding evidence allowed the discrimination and validation of C. caballus, C. caninus and C. sexfasciatus larvae. The diagnostic characteristics and COI sequences of Caranx lugubris and Caranx melampygus larvae, which are also distributed in the Eastern Central Pacific, remain to be investigated.

scholarly journals Systematic revision of the limestone karst-restricted land snail genus Aenigmatoconcha (Eupulmonata: Helicarionidae), with description of a new species

2021 ◽  
Vol 767 ◽  
pp. 55-82
Author(s):  
Arthit Pholyotha ◽  
Chirasak Sutcharit ◽  
Piyoros Tongkerd ◽  
Somsak Panha
Keyword(s):  
New Species ◽  
Phylogenetic Analyses ◽  
Comparative Morphology ◽  
Land Snail ◽  
Land Snails ◽  
Coi Gene ◽  
Molecular Evidence ◽  
C Subunit ◽  
Coi Sequences ◽  
A New Species

Thai limestone karsts are known to contain a rich biodiversity of animals, especially terrestrial snails, but still require further intensive exploration to evaluate their biodiversity. To date, only a few studies on the limestone karst-inhabiting land snail genera have been published. The present work focuses on the species diversity and phylogenetic relationships of the limestone karst-restricted land snail genus Aenigmatoconcha from Thailand, based on comparative morphology and molecular evidence. The results yielded three known species (A. clivicola Tumpeesuwan & Tumpeesuwan, 2017, A. sumonthai Tumpeesuwan & Tumpeesuwan, 2018, and A. mitis (Pfeiffer, 1863) comb. nov.), plus a new species (A. eunetis Pholyotha & Panha sp. nov). The phylogenetic analyses of partial fragments of the mitochondrial cytochrome oxidase c subunit I (COI) gene confirmed the monophyly of all recognized species and congruence with the traditional morphology-based species designations. Average uncorrected p-distances of COI sequences between species were 9.7–12.0% and within species were 0.2–4.2%. This study also provides the re-description of penial sculpture, penial sheath, flagellum, penial caecum, and mantle lobe morphology that were neglected from the type species description. The present discovery of a new species increases the known diversity of Thai land snails and will support the conservation planning to protect karst biodiversity.

scholarly journals Molecular analysis of the mitochondrial markers COI, 12S rDNA and 16S rDNA for six species of Iranian scorpions

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Parisa Soltan-Alinejad ◽  
Javad Rafinejad ◽  
Farrokh Dabiri ◽  
Piero Onorati ◽  
Olle Terenius ◽  
...  
Keyword(s):  
16S Rdna ◽  
Phylogenetic Analyses ◽  
Coi Gene ◽  
Diagnostic Tools ◽  
Morphological Identification ◽  
Species Determination ◽  
Important Species ◽  
Mitochondrial Markers ◽  
12S Rdna ◽  
Rdna Sequences

Abstract Objectives Annually, 1.2 million humans are stung by scorpions and severely affected by their venom. Some of the scorpion species of medical importance have a similar morphology to species with low toxicity. To establish diagnostic tools for surveying scorpions, the current study was conducted to generate three mitochondrial markers, Cytochrome Oxidase I (COI gene), 12S rDNA and 16S rDNA for six species of medically important Iranian scorpions: Androctonus crassicauda, Hottentotta saulcyi, Mesobuthus caucasicus, M. eupeus, Odontobuthus doriae, and Scorpio maurus. Results Phylogenetic analyses of the obtained sequences corroborated the morphological identification. For the first time, 12S rDNA sequences are reported from Androctonus crassicauda, Hottentotta saulcyi, Mesobuthus caucasicus and M. eupeus and also the 16S rDNA sequence from Hottentotta saulcyi. We conclude that the mitochondrial markers are useful for species determination among these medically important species of scorpions.

scholarly journals Draft Genome Sequences of Four Clinical Legionella pneumophila Isolates from Ontario, Canada

2018 ◽  
Vol 6 (15) ◽  
pp. e00295-18
Author(s):  
Alexander Fortuna ◽  
Ricardo Ramnarine ◽  
Aimin Li ◽  
Nahuel Fittipaldi ◽  
Christine Frantz ◽  
...  
Keyword(s):  
Legionella Pneumophila ◽  
Genetic Relationships ◽  
Draft Genome ◽  
Genome Sequences ◽  
Content Type ◽  
Outbreak Investigations ◽  
Typing Methods

ABSTRACT Legionella pneumophila outbreak investigations require the development of reliable typing methods to better understand the genetic relationships of the isolates involved. Here, we report the draft genome sequences of four clinical Legionella pneumophila isolates obtained between 2000 and 2012 in Ontario, Canada.

DNA barcodes provide evidence for the independent origin of the cultivated silkworms Samia ricini and Samia cynthia

2021 ◽  
pp. 1-8
Author(s):  
J.-C. Huang ◽  
X.-Y. Li ◽  
Y.-P. Li ◽  
R.-S. Zhang ◽  
D.-B. Chen ◽  
...  
Keyword(s):  
Genetic Distance ◽  
Phylogenetic Relationship ◽  
Dna Barcode ◽  
Distinct Species ◽  
Coi Gene ◽  
Molecular Evidence ◽  
Dna Barcodes ◽  
Close Relationship ◽  
Molecular Phylogenetic ◽  
Phylogenetic Framework

Samia ricini (Wm. Jones) and Samia cynthia (Drury) (Lepidoptera: Saturniidae) have been used as traditional sources of food as well as silk-producing insects. However, the phylogenetic relationship between the two silkworms remains to be addressed. In this study, the mitochondrial cytochrome c oxidase subunit 1 (COI) gene sequences corresponding to DNA barcodes from 13 Samia species were analysed, and a DNA barcode-based phylogenetic framework for these Samia species was provided. Phylogenetic analysis showed that multiple individuals of a species could be clustered together. Our analysis revealed a close relationship among Samia yayukae Paukstadt, Peigler and Paukstadt, Samia abrerai Naumann and Peigler, Samia kohlli Naumann and Peigler, Samia naessigi Naumann and Peigler, Samia naumanni Paukstadt, Peigler and Paukstadt, and Samia kalimantanensis Paukstadt and Paukstadt. The mixed clustering relationship and low Kimura-2-parameter (K2P) genetic distance (0.006) between individuals of S. ricini and Samia canningi (Hutton) indicated that the cultivated silkworm S. ricini was derived from the non-cultivated silkworm S. canningi. The remote phylogenetic relationship and high K2P genetic distance (0.039) indicated that S. ricini and S. cynthia are distinct species, thus providing solid molecular evidence that they had entirely independent origins. The relationships between S. kalimantanensis and S. naumanni and between S. cynthia and Samia wangi Naumann and Peigler, as well as the potential cryptic species within S. abrerai were also discussed. This is the first study to assess the DNA barcodes of the genus Samia, which supplements the knowledge of species identification and provides the first molecular phylogenetic framework for Samia species.

scholarly journals A new miniature species of Priocharax (Characiformes: Characidae) from the upper rio Ipixuna, Purus drainage, Brazil

2021 ◽  
Vol 19 (2) ◽  
Author(s):  
George Mendes Taliaferro Mattox ◽  
Camila da Silva de Souza ◽  
Mônica Toledo-Piza ◽  
Claudio Oliveira
Keyword(s):  
New Species ◽  
Lower Number ◽  
Valid Species ◽  
Molecular Evidence ◽  
Evidence Based ◽  
Fin Rays ◽  
Coi Sequences ◽  
Amazonas State ◽  
A New Species ◽  
Pelvic Fin

Abstract A new species of miniature fish of the characid genus Priocharax is described from a small lake near the rio Ipixuna, rio Purus drainage, Amazonas State, Brazil. It is distinguished from all congeners except P. pygmaeus by the lower number of teeth on the maxilla and dentary. It differs from P. pygmaeus by the presence of two postcleithra and 22–27 branched anal-fin rays (vs absence and 19–22). The new species is further distinguished from other species of Priocharax by a combination of characters involving the number of pelvic-fin rays and branched anal-fin rays, the number of postcleithra, the shape of postcleithrum 3, and the absence of the claustrum. Molecular evidence based on COI sequences of all valid species of Priocharax also corroborates the validity of this new species.

scholarly journals Molecular Characterisation using 16S rRNA and COI Gene Sequences in Hard Ticks of Gwalior, India

2020 ◽  
Vol 5 (3) ◽  
pp. 173-179
Author(s):  
Pooja Ghosh ◽  
Sachin Tikar ◽  
Mahendra K. Gupta ◽  
D Sukumaran
Keyword(s):  
16S Rrna ◽  
Phylogenetic Trees ◽  
Tick Infestation ◽  
Rhipicephalus Sanguineus ◽  
Total Sample ◽  
Domestic Animals ◽  
Coi Gene ◽  
Morphological Identification ◽  
Surveillance Studies ◽  
Global Threat

Tick infestation in humans and animals represents a global threat for different tick-borne diseases. In the present study, the ticks from the Gwalior region of India have been mapped to create a database of tick diversity. We explored 773 ticks collected from domestic animals and vegetation in Gwalior. Animals were screened visually, and ticks were collected manually, whereas the flag-drag method was used to collect ticks from the vegetation. The 16S rRNA and cytochrome oxidase I (COI) genes of tick samples were amplified and purified for sequencing and respective phylogenetic trees were constructed. The ticks were morphologically identified using taxonomical keys, revealing the presence of five genera in the region: Hyalomma, Haemaphysalis, Rhipicephalus, Boophilus, and Nosomma. Hyalomma spp. (Hy. annatolicum and Hy. marginatum) were the most abundant accounting for 69.598% of the total sample, followed by Rhipicephalus sanguineus (17.335%), Rhipicephalus microplus (7.115%), Haemaphysalis sp. (5.692%), and Nosomma monstrotum (0.258%). The tick sequences were submitted to the GenBank database. Phylogenetic analysis confirmed the morphological identification at the species level. The combination of molecular and morphological analyses of the ticks supported the result obtained with each method, thus providing more reliable estimates for continued surveillance studies.

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