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 New insights into malaria vector bionomics in Lao PDR: a nationwide entomology survey

2020 ◽  
Author(s):  
Sebastien Marcombe ◽  
Santi Maithaviphet ◽  
Julie Bobichon ◽  
Nothasin Phommavan ◽  
Simone Nambanya ◽  
...  
Keyword(s):  
Molecular Identification ◽  
Sibling Species ◽  
Rural Areas ◽  
Lao Pdr ◽  
Malaria Vectors ◽  
Human Landing Catch ◽  
Morphological Identification ◽  
Anopheles Mosquitoes ◽  
Malaria Burden ◽  
Human Landing

Abstract Background . In Laos, the malaria burden remains high despite a significant reduction of cases during the last decade. In the context of the disease elimination by 2030, a nationwide entomological survey was conducted to better understand the distribution, abundance and behavior of major malaria vectors ( Anopheles spp.) in the country. Methods . Mosquito collections were implemented in ten villages from ten provinces during the rainy and dry seasons of 2014 and 2015 by using human landing catch (HLC) and cow bait collection (CBC) methods. After morphological identification in the field, molecular identification of the sibling species of Anopheles mosquitoes from the Funestus, Leucosphyrus, and Maculatus groups were determined using PCR specific alleles. A screening of Plasmodium falciparum and P. vivax infections in the vectors was carried out by quantitative PCR assays. Results. A total of 14,146 adult mosquitoes representing 25 different Anopheles species were collected and morphologically identified. Molecular identification revealed the presence of 12 sibling species within the main primary vector groups including An. maculatus s.s., An. rampae , An. sawadwongporni , An. pseudowillmori , An. dravidicus , An. minimus s.s., An. aconitus , An. pampanai , An. harrisoni , An. dirus s.s., An. baimaii , An. nemophilous . Anopheles maculatus and An. minimus were predominant during both the dry and rainy seasons, but showed highly zoophilic preferences (Zoophilic index of 98% and 95%, respectively). Overall, 22% of the total malaria vectors were collected between 10:00 PM and 5:00 AM indoors when people are sleeping. Twenty-seven percent of primary and secondary vectors were collected outdoors before 10:00 PM or after 5:00 AM, times when people are usually awake and outdoors. Only two specimens were positive for P. falciparum , one An. aconitus from Phongsaly and one An. minimus from Vientiane Province Conclusions. The results indicate that people living in rural areas in Laos are constantly exposed to malaria vectors throughout the year and specifically outdoors. The use of LLINs/IRS remains important but innovative tools and new strategies are needed to address locally, the early and outdoor malaria transmission. Lack of expertise in general entomological methods may further exacerbate the situation.

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 New insights into malaria vector bionomics in Lao PDR: a nationwide entomology survey

2020 ◽  
Author(s):  
Sebastien Marcombe ◽  
Santi Maithaviphet ◽  
Julie Bobichon ◽  
Nothasin Phommavan ◽  
Simone Nambanya ◽  
...  
Keyword(s):  
Molecular Identification ◽  
Sibling Species ◽  
Rural Areas ◽  
Lao Pdr ◽  
Malaria Vectors ◽  
Human Landing Catch ◽  
Morphological Identification ◽  
Anopheles Mosquitoes ◽  
Malaria Burden ◽  
Human Landing

Abstract Background. In Laos, the malaria burden remains high despite a significant reduction of cases during the last decade. In the context of the disease elimination by 2030, a nationwide entomological survey was conducted to better understand the distribution, abundance and behavior of major malaria vectors ( Anopheles spp.) in the country.Methods. Mosquito collections were implemented in ten villages from ten provinces during the rainy and dry seasons of 2014 and 2015 by using human landing catch (HLC) and cow bait collection (CBC) methods. After morphological identification in the field, molecular identification of the sibling species of Anopheles mosquitoes from the Funestus, Leucosphyrus, and Maculatus groups were determined using PCR specific alleles. A screening of Plasmodium falciparum and P. vivax infections in the vectors was carried out by quantitative PCR assays.Results. A total of 14,146 adult mosquitoes representing 25 different Anopheles species were collected and morphologically identified. Molecular identification revealed the presence of 12 sibling species within the main primary vector groups including An. maculatus s.s., An. rampae , An. sawadwongporni , An. pseudowillmori , An. dravidicus , An. minimus s.s., An. aconitus , An. pampanai , An. harrisoni , An. dirus s.s., An. baimaii , An. nemophilous . Anopheles maculatus and An. minimus were predominant during both the dry and rainy seasons, but showed highly zoophilic preferences (Zoophilic index of 98% and 95%, respectively). Overall, 22% of the total malaria vectors were collected between 10:00 PM and 5:00 AM indoors when people are sleeping. Twenty-seven percent of primary and secondary vectors were collected outdoors before 10:00 PM or after 5:00 AM, times when people are usually awake and outdoors. Only two specimens were positive for P. falciparum, one An. aconitus from Phongsaly and one An. minimus from Vientiane Province.Conclusions. The results indicate that people living in rural areas in Laos are constantly exposed to malaria vectors throughout the year and specifically outdoors. The use of LLINs/IRS remains important but innovative tools and new strategies are needed to address locally, the early and outdoor malaria transmission. Lack of expertise in general entomological methods may further exacerbate the situation.

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.

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 Defining species boundaries in the Merodon avidus complex (Diptera, Syrphidae) using integrative taxonomy, with the description of a new species

2016 ◽  
Author(s):  
Jelena Ačanski ◽  
Ante Vujić ◽  
Mihajla Djan ◽  
Dragana Obreht Vidaković ◽  
Gunilla Ståhls ◽  
...  
Keyword(s):  
Sibling Species ◽  
Morphological Characters ◽  
Integrative Taxonomy ◽  
Coi Gene ◽  
Species Boundaries ◽  
Geometric Morphometric ◽  
Molecular Analyses ◽  
Mtdna Coi ◽  
The Status ◽  
Coi Sequences

Several recent studies have detected and described complexes of cryptic and sibling species in the genus Merodon (Diptera, Syrphidae). One representative of these complexes is the Merodon avidus complex that contains four sibling species, which have proven difficult to distinguish using traditional morphological characters. In the present study, we use two geometric morphometric approaches, as well as molecular characters of the 5’-end of the mtDNA COI gene, to delimit sibling taxa. Analyses based on these data were used to strengthen species boundaries within the complex, and to validate the status of a previously-recognized cryptic taxon from Lesvos Island (Greece), here described as Merodon megavidus Vujić & Radenković sp. nov. Geometric morphometric results of both wing and surstylus shape confirm the present classification for three sibling species－M. avidus (Rossi, 1790), M. moenium Wiedemann in Meigen, 1822 and M. ibericus Vujić, 2015－and, importantly, clearly discriminate the newly-described taxon Merodon megavidus sp. nov. In addition to our geometric morphometric results, supporting characters were obtained from molecular analyses of mtDNA COI sequences, which clearly differentiated M. megavidus sp. nov. from the other members of the M. avidus complex. Molecular analyses revealed that the earliest divergence of M. ibericus occurred around 800 ky BP, while the most recent separation happened between M. avidus and M. moenium around 87 ky BP.

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 Amount and location of tectonic uplift in the Urmia region of northwest Iran from the Permian to the Neogene

2021 ◽  
Vol 3 (3) ◽  
Author(s):  
Akram Alizadeh
Keyword(s):  
Hypersaline Lake ◽  
Time Interval ◽  
Lake Basin ◽  
Lake Urmia ◽  
Qom Formation ◽  
Northwest Of Iran ◽  
The Subject ◽  
Orogenic Uplift ◽  
East Azerbaijan ◽  
Urmia Lake Basin

AbstractThe Urmia Lake Basin is located between the West and East Azerbaijan provinces in the northwest of Iran. Lake Urmia is the twentieth largest lake and second largest hypersaline lake in the world. Stratigraphic columns have been constructed, using published information, to compare the sedimentary units deposited from the Permian to the Neogene on the east and west sides of the lake, and to use these to quantity subsidence and uplift. East of the lake, the sedimentary section is more complete and has been the subject of detailed stratigraphic studies, including the compilation of measured sections for some units. West of the lake, the section is incomplete and less work has been done; three columns illustrate variations in the preserved stratigraphy for the time interval. In all cases, the columns are capped by the Oligocene–Miocene Qom Formation, which was deposited during a post-orogenic marine transgression and unconformably overlies units ranging from Precambrian to Cretaceous. Permian to Cretaceous stratigraphy is used to measure subsidence in the Lake Urmia basin up to the end of the Cretaceous, and then, the subsequent orogenic uplift, which was followed by further subsidence recorded by the deposition of the Qom Formation in the Oligocene–Miocene.

scholarly journals Leeches as the intermediate host for strigeid trematodes: genetic diversity and taxonomy of the genera Australapatemon Sudarikov, 1959 and Cotylurus Szidat, 1928

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Ewa Pyrka ◽  
Gerard Kanarek ◽  
Grzegorz Zaleśny ◽  
Joanna Hildebrand
Keyword(s):  
Intermediate Host ◽  
Life Cycles ◽  
Coi Gene ◽  
Small Rivers ◽  
Ecological Niches ◽  
Its2 Region ◽  
28S Rrna ◽  
Intermediate Hosts ◽  
Internal Transcribed Spacer Region ◽  
Wide Range

Abstract Background Leeches (Hirudinida) play a significant role as intermediate hosts in the circulation of trematodes in the aquatic environment. However, species richness and the molecular diversity and phylogeny of larval stages of strigeid trematodes (tetracotyle) occurring in this group of aquatic invertebrates remain poorly understood. Here, we report our use of recently obtained sequences of several molecular markers to analyse some aspects of the ecology, taxonomy and phylogeny of the genera Australapatemon and Cotylurus, which utilise leeches as intermediate hosts. Methods From April 2017 to September 2018, 153 leeches were collected from several sampling stations in small rivers with slow-flowing waters and related drainage canals located in three regions of Poland. The distinctive forms of tetracotyle metacercariae collected from leeches supplemented with adult Strigeidae specimens sampled from a wide range of water birds were analysed using the 28S rDNA partial gene, the second internal transcribed spacer region (ITS2) region and the cytochrome c oxidase (COI) fragment. Results Among investigated leeches, metacercariae of the tetracotyle type were detected in the parenchyma and musculature of 62 specimens (prevalence 40.5%) with a mean intensity reaching 19.9 individuals. The taxonomic generic affiliation of metacercariae derived from the leeches revealed the occurrence of two strigeid genera: Australapatemon Sudarikov, 1959 and Cotylurus Szidat, 1928. Phylogenetic reconstructions based on the partial 28S rRNA gene, ITS2 region and partial COI gene confirmed the separation of the Australapatemon and Cotylurus clades. Taking currently available molecular data and our results into consideration, recently sequenced tetracotyle of Australapatemon represents most probably Au. minor; however, unclear phylogenetic relationships between Au. burti and Au. minor reduce the reliability of this conclusion. On the other hand, on the basis of the obtained sequences, supplemented with previously published data, the metacercariae of Cotylurus detected in leeches were identified as two species: C. strigeoides Dubois, 1958 and C. syrius Dubois, 1934. This is the first record of C. syrius from the intermediate host. Conclusions The results of this study suggest the separation of ecological niches and life cycles between C. cornutus (Rudolphi, 1808) and C. strigeoides/C. syrius, with potential serious evolutionary consequences for a wide range of host–parasite relationships. Moreover, phylogenetic analyses corroborated the polyphyletic character of C. syrius, the unclear status of C. cornutus and the separate position of Cotylurus raabei Bezubik, 1958 within Cotylurus. The data demonstrate the inconsistent taxonomic status of the sequenced tetracotyle of Australapatemon, resulting, in our opinion, from the limited availability of fully reliable, comparative sequences of related taxa in GenBank.

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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