Abstract:
:Standardization of the methods for extraction of DNA from sand flies is essential for obtaining high efficiency during subsequent molecular analyses, such as the new sequencing methods. Information obtained using these methods may contribute substantially to taxonomic, evolutionary, and eco-epidemiological studies. The aim of the present study was to standardize and compare two methods for the extraction of genomic DNA from sand flies for obtaining DNA in sufficient quantities for next-generation sequencing. Sand flies were collected from the municipalities of Campo Grande, Camapuã, Corumbá and Miranda, state of Mato Grosso do Sul, Brazil. Three protocols using a silica column-based commercial kit (ReliaPrep™ Blood gDNA Miniprep System kit, Promega®), and three protocols based on the classical phenol-chloroform extraction method (Uliana et al., 1991), were compared with respect to the yield and quality of the extracted DNA. DNA was quantified using a Qubit 2.0 fluorometer. The presence of sand fly DNA was confirmed by PCR amplification of the IVS6 region (constitutive gene), followed by electrophoresis on a 1.5% agarose gel. A total of 144 male specimens were analyzed, 72 per method. Significant differences were observed between the two methods tested. Protocols 2 and 3 of phenol-chloroform extraction presented significantly better performance than all commercial kit extraction protocols tested. For phenol-chloroform extraction, protocol 3 presented significantly better performance than protocols 1 and 2. The IVS6 region was detected in 70 of 72 (97.22%) samples extracted with phenol, including all samples for protocols 2 and 3. This is the first study on the standardization of methods for the extraction of DNA from sand flies for application to next-generation sequencing, which is a promising tool for entomological and molecular studies of sand flies.
journal_name
Exp Parasitoljournal_title
Experimental parasitologyauthors
Casaril AE,de Oliveira LP,Alonso DP,de Oliveira EF,Gomes Barrios SP,de Oliveira Moura Infran J,Fernandes WS,Oshiro ET,Ferreira AMT,Ribolla PEM,de Oliveira AGdoi
10.1016/j.exppara.2017.04.010subject
Has Abstractpub_date
2017-06-01 00:00:00pages
66-72eissn
0014-4894issn
1090-2449pii
S0014-4894(16)30323-Xjournal_volume
177pub_type
杂志文章abstract::The ring stages of Plasmodium falciparum within red blood cells cultured with complete medium stop growing when transferred to a basic medium containing RPMI plus fatty acid-free bovine serum albumin and dialyzable factors from human serum. Growth and multiplication can be partially restored by the addition of lipopro...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/0014-4894(83)90008-5
更新日期:1983-02-01 00:00:00
abstract::Complex life cycle parasites can manipulate the behavior of intermediate hosts in order to reduce their fitness and increase the chance of completing life cycle. In order to understand the effects of the trematode parasites of the genus Clinostomum on host fish Loricariichthys platymetopon, a filmed experiment was car...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/j.exppara.2020.107916
更新日期:2020-09-01 00:00:00
abstract::A series of novel hybrid 4-aminoquinoline 1,3,5-triazine derivatives was synthesized in a five-steps reaction and evaluated for their in vitro antimalarial activity against chloroquine-sensitive (3D7) and chloroquine-resistant (RKL-2) strains of Plasmodium falciparum. Entire synthetic derivatives showed higher antimal...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/j.exppara.2015.06.016
更新日期:2015-10-01 00:00:00
abstract::The tick Rhipicephalus microplus is an ectoparasite harmful to livestock, a vector of disease agents that affects meat and milk production. However, resistance to acaricides reflects the need for alternative tick control methods, among which vaccines have gained increasing relevance. In this scenario, monoclonal antib...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/j.exppara.2012.02.006
更新日期:2012-04-01 00:00:00
abstract::The study of the encystation process of Entamoeba histolytica has been hampered by the lack of experimental means of inducing mature cysts in vitro. Previously we have found that cytoplasmic vesicles similar to the encystation vesicles of Entamoeba invadens are present in E. histolytica trophozoites only in amebas rec...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/j.exppara.2011.05.001
更新日期:2011-09-01 00:00:00
abstract::It is generally accepted that a majority of individuals infected by Entamoeba histolytica do not develop symptomatic disease. However, the parasite and the host factors contributing to the development of the disease, remain undetermined. It is also unclear why certain individuals develop extra-intestinal amebiasis wit...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/j.exppara.2005.02.015
更新日期:2005-07-01 00:00:00
abstract::Complement activation by protoscoleces of Echinococcus granulosus was studied by analyzing the damage to their tegumental membrane produced by incubation in both normal and hydatid human sera. The state of the apical tegumental membrane was evaluated by measuring the electric potential difference with microelectrodes....
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/0014-4894(92)90211-r
更新日期:1992-11-01 00:00:00
abstract::The biochemical properties of the enzyme responsible for nematode "activated L-serine sulphydrase" activity (L-cysteine + R-SH----cysteine thioether + H2S) have been investigated using primarily the gastro-intestinal nematodes Nippostrongylus brasiliensis and Haemonchus contortus. The activated L-serine sulphydrase en...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/0014-4894(92)90048-f
更新日期:1992-03-01 00:00:00
abstract::Calfluxin (CaFl), one of the gonadotropic hormones of Lymnaea stagnalis, stimulates the influx of Ca2+ into the mitochondria of the cells of the albumen gland, one of the accessory sex organs of the snail. This effect is suppressed in glands of noninfected snails by an agent (schistosomin) present in the hemolymph of ...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/0014-4894(88)90110-5
更新日期:1988-02-01 00:00:00
abstract::Human cysticercosis caused by Taenia crassiceps is unusual; however, it is an useful experimental model for cysticercosis studies. Benzimidazole derivatives are important antihelminthic drugs widely used against helminths. A novel compound 6-chloro-5-(1-naphthyloxy) -2-(trifluoromethyl)-1H-benzimidazole (RCB20) is a b...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/j.exppara.2016.11.002
更新日期:2017-01-01 00:00:00
abstract::The codon for serine residue 108 of the Plasmodium falciparum dihydrofolate reductase gene was replaced with those for the other 19 amino acids. Except for the Lys108 mutant, which was not expressed, all other substitutions yielded DHFR mutants which were expressed in Escherichia coli as inactive inclusion bodies. Nin...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1006/expr.1997.4221
更新日期:1997-11-01 00:00:00
abstract::The tegumental alterations in adult Fasciola hepatica induced by the experimental fasciolide OZ78 were investigated utilizing scanning electron microscopy (SEM). Twelve weeks post-infection with F. hepatica, rats were treated with a single 100mg/kg oral dose of OZ78 and flukes were recovered from the bile ducts after ...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/j.exppara.2007.12.015
更新日期:2008-05-01 00:00:00
abstract::Shigellosis is a serious public health problem in Korea, because large outbreaks of Shigella sonnei infections were recorded in many parts of the country during the period 1998-2000. However, the epidemiological features of shigellosis are not well known. In this study, we devised conditions suitable for the growth an...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/j.exppara.2006.08.002
更新日期:2007-02-01 00:00:00
abstract::This study examines the effects of ricinoleic acid esters from Ricinus communis castor oil on the vitellogenesis of Rhipicephalus sanguineus ticks attached to hosts that were fed with commercial rabbit food containing these esters. The oocytes of ticks from the treatment group (TG) showed cytoplasmic changes that inhi...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/j.exppara.2010.10.006
更新日期:2011-02-01 00:00:00
abstract::Giardia duodenalis is one of the most important intestinal parasites globally, especially in children, and in Cuba is the leading cause of chronic paediatric diarrhoea in this population. G. duodenalis is composed of eight genetic groups (or assemblages), two of which (A and B) are apparently zoonotic, occurring in bo...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/j.exppara.2019.107814
更新日期:2020-02-01 00:00:00
abstract::Shiraz district in south of Iran is a classical focus of Cutaneous leishmaniasis (CL) and previous research has consistently documented the etiologic agent to be Leishmania tropica and Leishmania major in urban and rural areas, respectively. However, none of the Phlebotomus sergenti, a known vector for L. tropica, of ...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/j.exppara.2010.05.020
更新日期:2010-12-01 00:00:00
abstract::Extracting genomic DNA of pathogenic agents from formalin-fixed specimens is inherently difficult. Storage of samples in formalin results in nucleic acid cross-linking and DNA fragmentation. In this study, DNA was extracted from 45 Giardia-positive stool samples stored in formalin and subjected to PCR amplification ta...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/j.exppara.2019.01.010
更新日期:2019-03-01 00:00:00
abstract::Three studies were conducted to examine the function of ecdysteroids in the development of parasitic nematodes. Ecdysone and 20-hydroxyecdysone were extracted, separated chromatographically, and measured in the reproductive tracts of adult female Ascaris suum. Perienteric fluid and the body wall did not contain measur...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/0014-4894(85)90024-4
更新日期:1985-10-01 00:00:00
abstract::Cryptosporidiosis, caused by the protozoan parasite Cryptosporidium, represents the major public health concern of water utilities in developed nations due to its small size, resistance to disinfection and ability to be shed in large numbers in faeces. In Australia, recreational access is not allowed on direct supply ...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/j.exppara.2012.04.015
更新日期:2012-08-01 00:00:00
abstract::Trichomonas vaginalis is a human urogenital pathogen that causes trichomoniasis, the most common nonviral, parasitic sexually transmitted infection in the world. At present, little is known regarding the degree of strain variability of T. vaginalis. A classification method for T. vaginalis strains would be a useful to...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/j.exppara.2015.10.011
更新日期:2015-12-01 00:00:00
abstract::A mode of action of the inducible treatment with trypsin for the development of Mesocestoides lineatus tetrathyridium to adult was analyzed by administering various agents effective on Ca2+-dependent metabolic pathways in the cells: protein kinase C activators such as a synthetic diacylglycerol, 1-oleoyl-2-acetylglyce...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/0014-4894(86)90036-6
更新日期:1986-12-01 00:00:00
abstract::This study investigated the genetic characteristics of Toxoplasma gondii samples collected from 62 patients with toxoplasmosis in Sao Paulo State, Brazil. DNA samples were isolated from blood, cerebrospinal fluid and amniotic fluids of 25 patients with cerebral toxoplasmosis and AIDS, two patients with acute toxoplasm...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/j.exppara.2011.06.002
更新日期:2011-10-01 00:00:00
abstract::The protozoan Trypanosoma evansi is described as presenting high morphological and genetic similarities among the isolates despite its biological heterogeneity and wide geographical distribution. PCR amplification of the internal transcribed spacers of the ribosomal gene in combination with the coding region of the 5....
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/j.exppara.2007.10.003
更新日期:2008-03-01 00:00:00
abstract::An active Trypanosoma cruzi transmission cycle maintained by wild rodents in the Andean valleys of Cochabamba Bolivia is described. Wild and domestic Triatoma infestans with 60% infection with T. cruzi were found and was evidenced in 47.5% (rodents) and 26.7% (marsupial) by parasitological and/or serologycal methods. ...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/j.exppara.2006.04.010
更新日期:2006-12-01 00:00:00
abstract::We studied the effects of tick saliva on cell migration, cell signaling, phagocytosis, and gene expression in the murine macrophage cell line, IC-21. Saliva increased both basal- and platelet-derived growth factor (PDGF)-stimulated migration in IC-21 cells. However, saliva did not affect PDGF-stimulated extracellular ...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/j.exppara.2010.11.012
更新日期:2011-03-01 00:00:00
abstract::Purine uptake has been studied in many protozoan parasites in the last few years, and several of the purine transporters have been cloned. In contrast, very little is known about the salvage of preformed pyrimidines by protozoa, and no pyrimidine transporters have been cloned, yet chemotherapy based on pyrimidine nucl...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/j.exppara.2006.02.018
更新日期:2006-10-01 00:00:00
abstract::Giardia lamblia, an early diverging eukaryote that infects several species including humans and a major agent of water-borne diarrhea throughout the world, can be infected with a double-stranded RNA virus, giardiavirus (GLV). A chimeric GLV cDNA and green fluorescent protein (GFP) according to the cis-acting signals o...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/j.exppara.2004.12.006
更新日期:2005-03-01 00:00:00
abstract::Crohn's disease (CD) is considered to be caused by a disorder of the immune system and helminth infections may interact with development of the disease. We induced colitis in mice by trinitrobenzenesulfonic acid (TNBS) and observed the effects of intraperitoneally injected eggs of Schistosoma japonicum on the course o...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/j.exppara.2007.02.009
更新日期:2007-08-01 00:00:00
abstract::Schistosomiasis is a chronic disease caused by trematode flatworms of the genus Schistosoma; it accounts for more than 280,000 deaths annually. In this work we investigated the effect of the alkaloidic extract obtained by acid-base extraction of the dried fruits of Solanum lycocarpum on schistosomiasis. We used this e...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1016/j.exppara.2012.12.012
更新日期:2013-04-01 00:00:00
abstract::A murine model system was developed to study the induction and mechanism of protective immunity to L3 of Strongyloides stercoralis. L3 were implanted in BALB/cByJ mice in diffusion chambers constructed with 0.1- or 2.0-microns-pore-size membranes. Parasites survived equally well regardless or membrane type for 7 days,...
journal_title:Experimental parasitology
pub_type: 杂志文章
doi:10.1006/expr.1995.1036
更新日期:1995-03-01 00:00:00