scholarly journals Molecular and morphological characterization of three new species of avian Onchocercidae (Nematoda) with emphasis on circulating microfilariae

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Rasa Binkienė ◽  
Carolina Romeiro Fernandes Chagas ◽  
Rasa Bernotienė ◽  
Gediminas Valkiūnas
Keyword(s):  
Molecular Characterization ◽  
Peripheral Blood ◽  
Phylogenetic Analyses ◽  
Dna Barcode ◽  
Morphological Characters ◽  
Buffy Coat ◽  
Blood Parasites ◽  
Species Identity ◽  
Blood Samples ◽  
Characterization Methods

Abstract Background Blood parasites have been the subject of much research, with numerous reports of the presence of microfilariae in the peripheral blood (circulating microfilariae) of birds belonging to many orders. Current limitations in molecular characterization methods and species identification using morphological characters of circulating microfilariae are major obstacles to improving our understanding the biology of Filarioidea species, particularly in wildlife. The aim of this study was to partially fill these gaps, with particular emphasis on morphological features of microfilariae, which are the most readily accessible stages of these pathogens. Methods Peripheral blood samples of 206 birds belonging to genera Acrocephalus (five species) and Sylvia (five species) were examined using the buffy coat method to process the blood samples for the presence of microfilariae. Positive birds were dissected to collect adult nematodes. Microfilariae and adult nematodes were described, and sequences of their mitochondrial cytochrome c oxidase subunit I and nuclear 28S rDNA gene fragments were obtained and used for molecular characterization and Bayesian phylogenetic inferences. Results Overall prevalence of microfilariae was 2.9%. Microfilariae were found in the blood samples from six birds (2 Acrocephalus scirpaceus and 1 each of A. arundinaceus, Sylvia atricapilla, S. borin and S. curruca), which were dissected. All parasite species observed were new. Eufilaria acrocephalusi sp. n. and Eufilaria sylviae sp. n. were present in subcutaneous, peritracheal and periesophageal connective tissues in A. scirpaceus and S. borin, respectively. Splendidofilaria bartletti sp. n. was found in finger joins of S. atricapilla. Illustrations of microfilariae and adult nematodes are shown, and morphological and phylogenetic analyses identified the DNA barcode haplotypes that are associated with these species. Phylogenetic analysis places the parasites of different genera in different closely related clades. Conclusions Adult nematode morphological characters, which have been traditionally used in the taxonomy of Filarioidea species, have a phylogenetic value. Importantly, in our study parasites of different genera were readily distinguishable based on the morphology of their microfilariae. The link between molecular and morphology data requires more study in Filarioidea species research, particularly because this approach provides new knowledge on species identity using only readily accessible blood stages (microfilariae), thereby avoiding host dissection and thus minimizing harm to wildlife during research.

scholarly journals Morphometry and Phylogeny of the Different Populations of Selaginella tamariscina (P. Beauv.) Spring and S. pulvinata (Hook. & Grev.) Maxim. in Northern Thailand

2021 ◽  
Vol 20 (4) ◽  
Author(s):  
Udon Pongkawong ◽  
◽  
Jatupol Kampuansai ◽  
Rossarin Pollawatn ◽  
Arunothai Jampeetong ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Southern China ◽  
Phylogenetic Analyses ◽  
Dna Barcode ◽  
Morphological Characteristics ◽  
Taxonomic Status ◽  
Morphological Characters ◽  
The Philippines ◽  
Northern Thailand ◽  
Selaginella Tamariscina

Abstract “Dok Hin” is the Thai local name for Selaginella species that form rosettes. They commonly distributes in Siberia, Manchuria, southern China, Japan, the Philippines and Thailand. Morphology of Dok Hin is very resemble leading to misidentification. So, exactly number of species of Dok Hin in Thailand and their differences in morphological characteristics is not well understood. Thus, revision of morphological characters and phylogenetic confirmation of the taxonomic identification are needed. This study aims to examine morphological charateristics and phylogenetic patterns in eight populations of the Dok Hin in Northern Thailand. Morphology of Dok Hin from each populations was quantitatively examined using 15 vegetative and 6 reproductive characters meanwhile phylogenetic analyses was explored by DNA barcode ITS2. The results of the phylogenetic analysis revealed the existence of two species of Dok Hin, S. tamariscina and S. pulvinata. Selaginella tamariscina can be distinguished from S. pulvinata by its presence of a pseudotrunk above ground and ridges of dorsal leaves. On the other hand, the results of phylogenetic analysis indicated the differences among populations of S. pulvinata as well. Chiang Mai populations of S. pulvinata was characterized by peculiar set of characters long leaves and leaf apices look like caudate, while the rest of their populations have shorter leaves and leaf apices look like aristate. It indicates that S. pulvinata has genetic and phenotypic divergence among populations. However, additional studies of Dok Hin populations in other parts of Thailand and studies on different genetic markers are necessary to confirm the taxonomic status of S. pulvinata. Keywords: Dok Hin, Morphometric, Phylogeny, Pseudotrunk, Resurrection plant

scholarly journals QBC® for the diagnosis of human and canine american visceral leishmaniasis: preliminary data

2001 ◽  
Vol 34 (6) ◽  
pp. 577-581 ◽  
Author(s):  
Daniel B. Liarte ◽  
Ivete L. Mendonça ◽  
Francisco C.O. Luz ◽  
Elza A.S. de Abreu ◽  
Gustavo W.S. Mello ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Visceral Leishmaniasis ◽  
Preliminary Data ◽  
Peripheral Blood ◽  
Buffy Coat ◽  
Promising Method ◽  
Blood Parasites ◽  
Leishmania Chagasi ◽  
Asymptomatic Carriers ◽  
Fluorescent Method

"Quantitative Buffy Coat" (QBC®) is a direct and fast fluorescent method used for the identification of blood parasites. Since Leishmania chagasi circulates in blood, we decided to test it in American visceral leishmaniasis (AVL). Bone marrow (BM) and peripheral blood (PB) of 49 persons and PB of 31 dogs were analyzed. QBC® was positive in BM of 11/11 patients with AVL and in 1/6 patients with other diseases. Amastigotes were identified in PB of 18/22 patients with AVL and in none without AVL. The test was positive in 30 out of the 31 seropositive dogs and in 28/28 dogs with Leishmania identified in other tissues. QBC® is a promising method for diagnosis of human AVL, and possibly for the exam of PB of patients with AVL/AIDS, for the control of the cure and for the identification of asymptomatic carriers. Because it is fast and easy to collect and execute, QBC® should be evaluated for programs of reservoir control.

Preparation of the Buffy Coat From Small-Volume Blood Samples For Ultrastructural Study

1985 ◽  
Vol 43 ◽  
pp. 712-713
Author(s):  
M. J. Mitchell ◽  
E. J. Mirro ◽  
H. E. Black ◽  
E. Schwartz
Keyword(s):  
Peripheral Blood ◽  
Small Volume ◽  
Electron Microscopic Examination ◽  
Buffy Coat ◽  
Electron Microscopic ◽  
Blood Samples ◽  
Toxicological Studies ◽  
Volume Blood ◽  
Toxicological Research ◽  
Early Indication

Electron microscopic examination of peripheral blood is a valuable technique in toxicological research. Blood can be collected for examination several times during a toxicological study. Compound-induced cellular changes detected in peripheral blood cells prior to the terminal necropsy may be an early indication of potential compound-related toxicity.The routine use of small rodents in toxicological studies presents a problem; the amount of blood available at interim collections is minimal. The following technique has been refined for the purpose of examining leukocytes in small blood samples (1 cc or less). Leukocytes are concentrated so that a maximal number per sample can be examined.

scholarly journals DNA barcode of the Lanzones scale insect, Unaspis mabilis Lit & Barbecho (Hemiptera: Diaspididae)

2020 ◽  
Vol 14 (3) ◽  
Keyword(s):  
Cytochrome C ◽  
Cytochrome C Oxidase ◽  
Phylogenetic Analyses ◽  
Dna Barcode ◽  
Scale Insect ◽  
Coi Gene ◽  
Species Identity ◽  
Management Options ◽  
Nucleotide Sequence Alignment ◽  
Coi Sequences

The mitochondrial cytochrome c oxidase subunit 1 (COI) nucleotide sequences of Unaspis mabilis Lit & Barbecho (Hemiptera: Diaspididae), are provided for the first time. The total genomic DNA of each scale insect was extracted from individuals infesting lanzones leaves from three selected sites in Los Baños, Laguna. A partial COI gene amplicon with approximately 750 bp was obtained using the primer pair PcoF1 and LepR1. Nucleotide sequence alignment showed no variation among the COI sequences from all the samples. BLASTn search yielded no significant match with any of the available sequences for Unaspis species. The closest hit was Aulacaspis tubercularis Newstead (GenBank Accession No. HM474091) with 87.4% nucleotide similarity. Nonetheless, phylogenetic analyses revealed that generated COI sequences from U. mabilis form a monophyletic clade with U. yanonensis and U. euonymi, with closer proximity to the former. These findings also strengthen the species status of U. mabilis under the genus Unaspis. The DNA barcodes generated from this study (GenBank Acc. Nos. MN114099, MN14101, and MN114102), could, therefore, be used to verify the species identity of other lanzones scale accessions, as well as monitor the distribution and spread of U. mabilis, which would greatly influence possible pest management options. KEYWORDS: cytochrome C oxidase I, COI, Lansium domesticum Correa, lanzones

scholarly journals Resolution of the Portunus gladiator species complex: taxonomic status and identity of Monomia gladiator (Fabricius, 1798) and Monomia haanii (Stimpson, 1858) (Brachyura, Decapoda, Portunidae)

ZooKeys ◽  
2019 ◽  
Vol 858 ◽  
pp. 11-43 ◽  
Author(s):  
Amanda M. Windsor ◽  
Jose Christopher E. Mendoza ◽  
Jonathan R. Deeds
Keyword(s):  
Phylogenetic Analyses ◽  
Dna Barcode ◽  
Taxonomic Status ◽  
The United States ◽  
Zoological Nomenclature ◽  
Swimming Crab ◽  
Species Identity ◽  
Commercial Products ◽  
Seafood Industry ◽  
Seafood Products

The United States Food and Drug Administration (FDA) has recently adopted DNA barcoding for the purpose of determining the species identity of commercial seafood products. This effort has revealed instances of incongruence between current scientifically accepted taxon names and those utilized by the seafood industry in product labelling. One such case is that of “Portunushaanii”, a name utilized by the seafood industry to label commercial products under the market name “red swimming crab.” However, carcinologists currently regardP.haaniias synonym ofPortunusgladiatorFabricius, 1798, which itself is the subject of debate over whether it is a secondary homonym ofCancer gladiatorFabricius, 1793. Further complicating matters, DNA barcode sequences from commercial products match GenBank sequences identified asPortunuspseudoargentatusStephenson, 1961. Here the complicated taxonomic history of thePortunusgladiatorcomplex is reviewed and a resolution proposed based on combined morphological descriptions and molecular phylogenetic analyses. It is demonstrated that, given the provisions of the International Code of Zoological Nomenclature and the current elevation ofMonomiaGistel, 1848, to full genus rank, its type species,PortunusgladiatorFabricius, 1798, should be treated as a valid and available taxon name. It is also shown, upon examination and comparison of types and topotypic material thatMonomiahaanii(Stimpson, 1858) is a distinct taxon fromM.gladiator, andPortunuspseudoargentatusStephenson, 1961, is a junior subjective synonym ofM.haanii(Stimpson, 1858). Furthermore, it is shown that crab meat sold in the US currently labeled as “Portunushaanii” and/or “red swimming crab” is in factM.haaniiusing comparative analysis of DNA barcode sequences between museum-vouchered reference specimens, whole crabs provided directly by a seafood importer, and processed commercial products purchased at retail.

Diversity of Karyolysus and Schellackia from the Iberian lizard Lacerta schreiberi with sequence data from engorged ticks

Parasitology ◽  
2019 ◽  
Vol 146 (13) ◽  
pp. 1690-1698
Author(s):  
Kristína Zechmeisterová ◽  
Joëlle Goüy de Bellocq ◽  
Pavel Široký
Keyword(s):  
Sequence Data ◽  
Phylogenetic Analyses ◽  
Molecular Genetic ◽  
Blood Parasites ◽  
Mixed Infections ◽  
Blood Samples ◽  
Non Invasive ◽  
Blood Smears ◽  
The World ◽  
Life Cycle Patterns

AbstractApicomplexan haemoparasites of the genera Schellackia Reichenow, 1919, and Karyolysus Labbé, 1894, seem to be common in lizards and widespread across the world. For decades, their identification has been based on morphological descriptions and life cycle patterns, with molecular characterizations, applied only recently. We used molecular characterization to confirm the identification of haemoparasites detected by microscopy in blood smears of Lacerta schreiberi Bedriaga, 1878, a lizard of the Iberian Peninsula. Since blood samples other than blood smears were not available from the studied lizards, 264 engorged ticks Ixodes ricinus (Linneaus, 1758) collected from them were used as an alternative non-invasive source of haemoparasite DNA for molecular genetic analyses. Of the 48 blood smears microscopically examined, 31 were positive for blood parasites (64.6% prevalence). We identified trophozoites and gamonts similar to Karyolysus lacazei (Labbé, 1894) (24/48; 50%) and Schellackia-like sporozoites (20/48; 41.7%). Mixed infections with both species occurred in 13 blood smears (27.1%). Sequence data were obtained for both parasites from engorged ticks. Phylogenetic analyses placed our unique haemogregarine sequence within the Karyolysus clade, nevertheless, within substantial polytomy. Thus, according to its morphology and effect on the host cell, we refer to this haemogregarine as Karyolysus cf. lacazei. Besides the Schellackia sequences being identical to a previously identified haplotype, we also obtained sequences of three new closely related haplotypes.

scholarly journals Functional and Molecular Characterization of the Halomicrobium sp. IBSBa Inulosucrase

2021 ◽  
Vol 9 (4) ◽  
pp. 749
Author(s):  
Gülbahar Abaramak ◽  
Jaime Ricardo Porras-Domínguez ◽  
Henry Christopher Janse van Rensburg ◽  
Eveline Lescrinier ◽  
Ebru Toksoy Öner ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Phylogenetic Analyses ◽  
Halophilic Archaea ◽  
Maximum Activity ◽  
Physiological Relevance ◽  
Health Related ◽  
Specific Alternative ◽  
Binding Groove ◽  
Native Host

Fructans are fructose-based (poly)saccharides with inulin and levan being the best-known ones. Thanks to their health-related benefits, inulin-type fructans have been under the focus of scientific and industrial communities, though mostly represented by plant-based inulins, and rarely by microbial ones. Recently, it was discovered that some extremely halophilic Archaea are also able to synthesize fructans. Here, we describe the first in-depth functional and molecular characterization of an Archaeal inulosucrase from Halomicrobium sp. IBSBa (HmcIsc). The HmcIsc enzyme was recombinantly expressed and purified in Escherichia coli and shown to synthesize inulin as proven by nuclear magnetic resonance (NMR) analysis. In accordance with the halophilic lifestyle of its native host, the enzyme showed maximum activity at very high NaCl concentrations (3.5 M), with specific adaptations for that purpose. Phylogenetic analyses suggested that Archaeal inulosucrases have been acquired from halophilic bacilli through horizontal gene transfer, with a HX(H/F)T motif evolving further into a HXHT motif, together with a unique D residue creating the onset of a specific alternative acceptor binding groove. This work uncovers a novel area in fructan research, highlighting unexplored aspects of life in hypersaline habitats, and raising questions about the general physiological relevance of inulosucrases and their products in nature.

scholarly journals Comparison of capillary, venous and buffy coat blood samples in detecting Plasmodium species among malaria suspected patients attending at Hamusite health center. A cross-sectional study

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Getu Abeje ◽  
Woyneshet Gelaye ◽  
Getaneh Alemu
Keyword(s):  
Health Center ◽  
Detection Rate ◽  
Venous Blood ◽  
Cross Sectional Study ◽  
Buffy Coat ◽  
Blood Film ◽  
Capillary Blood ◽  
Sectional Study ◽  
Cross Sectional ◽  
Blood Samples

Abstract Background Both capillary and venous blood samples have been interchangeably used for the diagnosis of malaria in Ethiopia. However, Plasmodium parasites are thought to be more concentrated in capillary than in venous blood. Hence, selecting a sample source where parasites are more concentrated is indispensable approach in order to maximize the accuracy of blood film microscopy. Therefore, the present study aimed to compare the detection rate and the parasitemia level of Plasmodium species from conventional capillary and venous blood films, and buffy coat preparations. Methods A facility based cross-sectional study was conducted from Feburary to March 2020 among 210 febrile patients attending Hamusite health center, northwest Ethiopia. Capillary and venous blood samples were collected and buffy coat was prepared from each sample. Thin and thick blood films were prepared, stained, and examined microscopically following standard protocol. Data were analysed using Statistical Package for Social Sciences Software version 20 and Med-Calc software version 19.3. Results Capillary blood buffy coat (61/210, 29.0%) had significantly higher detection rate as compared to capillary (48/210, 22.9%) and venous (42/210, 20.0%) blood films (p < 0.001). However, no significant difference was observed between capillary and venous blood films (p = 0.070) in detecting Plasmodium species. The highest and the lowest mean asexual stage parasite counts were found in capillary blood buffy coat (4692.88) and venous blood (631.43) films, respectively showing significant variations (p < 0.001). Mean gametocyte count was also highest in capillary blood buffy coat (3958.44). As compared to capillary blood buffy coat, the sensitivity of venous blood buffy coat, capillary blood film and venous blood film were 73.8, 78.7, 68.9%, respectively. Conclusion Capillary blood buffy coat samples showed the highest sensitivity in detecting and quantitating malaria parasites that its use should be promoted in clinical settings. However, conventional capillary and venous blood films could be used interchangeably.

scholarly journals Evaluation of qPCR on blood and skin microbiopsies, peripheral blood buffy coat smear, and urine antigen ELISA for diagnosis and test of cure for visceral leishmaniasis in HIV-coinfected patients in India: a prospective cohort study

BMJ Open ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. e042519
Author(s):  
Sophie I Owen ◽  
Sakib Burza ◽  
Shiril Kumar ◽  
Neena Verma ◽  
Raman Mahajan ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Visceral Leishmaniasis ◽  
Hiv Infection ◽  
Peripheral Blood ◽  
Medical Science ◽  
Bone Marrow Aspiration ◽  
Buffy Coat ◽  
Tropical Medicine ◽  
Urine Antigen ◽  
Antigen Elisa

IntroductionHIV coinfection presents a challenge for diagnosis of visceral leishmaniasis (VL). Invasive splenic or bone marrow aspiration with microscopic visualisation of Leishmania parasites remains the gold standard for diagnosis of VL in HIV-coinfected patients. Furthermore, a test of cure by splenic or bone marrow aspiration is required as patients with VL-HIV infection are at a high risk of treatment failure. However, there remain financial, implementation and safety costs to these invasive techniques which severely limit their use under field conditions.Methods and analysisWe aim to evaluate blood and skin qPCR, peripheral blood buffy coat smear microscopy and urine antigen ELISA as non-invasive or minimally invasive alternatives for diagnosis and post-treatment test of cure for VL in HIV-coinfected patients in India, using a sample of 91 patients with parasitologically confirmed symptomatic VL-HIV infection.Ethics and disseminationEthical approval for this study has been granted by The Liverpool School of Tropical Medicine, The Institute of Tropical Medicine in Antwerp, the University of Antwerp and the Rajendra Memorial Research Institute of Medical Science in Patna. Any future publications will be published in open access journals.Trial registration numberCTRI/2019/03/017908.

scholarly journals Disentangling relationships among eastern Mediterranean Cymbalaria including description of a novel species from the southern Peloponnese (Greece)

2021 ◽  
Vol 307 (2) ◽  
Author(s):  
Pau Carnicero ◽  
Núria Garcia-Jacas ◽  
Llorenç Sáez ◽  
Theophanis Constantinidis ◽  
Mercè Galbany-Casals
Keyword(s):  
Eastern Mediterranean ◽  
Phylogenetic Analyses ◽  
Taxonomic Revision ◽  
Distribution Patterns ◽  
Morphological Characters ◽  
Molecular Data ◽  
Morphological Data ◽  
Long Distance ◽  
Aegean Islands ◽  
Molecular Phylogenetic

AbstractThe eastern Mediterranean basin hosts a remarkably high plant diversity. Historical connections between currently isolated areas across the Aegean region and long-distance dispersal events have been invoked to explain current distribution patterns of species. According to most recent treatments, at least two Cymbalaria species occur in this area, Cymbalaria microcalyx and C. longipes. The former comprises several intraspecific taxa, treated at different ranks by different authors based on morphological data, evidencing the need of a taxonomic revision. Additionally, some populations of C. microcalyx show exclusive morphological characters that do not match any described taxon. Here, we aim to shed light on the systematics of eastern Mediterranean Cymbalaria and to propose a classification informed by various sources of evidence. We performed molecular phylogenetic analyses using ITS, 3’ETS, ndhF and rpl32-trnL sequences and estimated the ploidy level of some taxa performing relative genome size measures. Molecular data combined with morphology support the division of traditionally delimited C. microcalyx into C. acutiloba, C. microcalyx and C. minor, corresponding to well-delimited nrDNA lineages. Furthermore, we propose to combine C. microcalyx subsp. paradoxa at the species level. A group of specimens previously thought to belong to Cymbalaria microcalyx constitute a well-defined phylogenetic and morphological entity and are described here as a new species, Cymbalaria spetae. Cymbalaria longipes is non-monophyletic, but characterized by being glabrous and diploid, unlike other eastern species. The nrDNA data suggest at least two dispersals from the mainland to the Aegean Islands, potentially facilitated by marine regressions.

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