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1.
Exp Parasitol ; 206: 107771, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31585116

RESUMO

A PCR targeting mitochondrial cytochrome oxidase subunit III (cox3) for molecular detection of Babesia gibsoni infection in dogs has been developed in this study. Fifty blood samples from suspected clinical cases from dogs, brought to the veterinary college clinics, were examined for presence of B. gibsoni using conventional diagnosis by microscopic examination of Giemsa stained thin blood smears. In addition, species specific PCRs targeting ITS-1 region (BgITS-1 PCR) and nested PCR targeting 18S ribosomal RNA gene (Bg18SnPCR) were carried out. A 634 bp PCR fragment of B. gibsoni cox3 gene was amplified in positive samples from three geographical locations of Satara, Wai and Pune in Maharashtra state of India. From analysis of the sequence of the B. gibsoni cox3 gene, we found that the Indian isolate had 96-98% similarity to the isolate from Japan and China. Post sequencing, de-novo diagnostic primer pair for species specific amplification of 164 bp fragment of B. gibsonicox3 was designed and the PCR was standardized. The diagnostic results of de-novo Bgcox3 PCR were compared with BgITS-1 PCR and Bg18S nPCR. Thin blood smears detected 22% (11/50) samples positive for small form of Babesia species. The BgITS-1 PCR detected 25% samples (15/50) as positive and Bg18S nPCR detected 80% (40/50) B. gibsoni positive samples. The de-novo Bgcox3 PCR detected 66% (33/50) samples positive for B. gibsoni (at 95% CI). The analytical sensitivity of cox3 PCR was evaluated as 0.000003% parasitaemia or 09 parasites in 100  µl of blood. The de-novo diagnostic cox3 PCR did not cross react with control positive DNA from other haemoprotozoa and rickettsia like B. vogeli, Hepatozoon canis, Trypanosoma evansi, Ehrlichia canis and Anaplasma platys. Statistically, cox3 PCR had better diagnostic efficiency than ITS-1 PCR in terms of sensitivity (p = 0.0006). No statistically significant difference between results of cox3 PCR and 18S nPCR was observed (p = 0.1760). Kappa values estimated for each test pair showed fair to moderate agreement between the observations. Specificity of Bgcox3 PCR was 100% when compared with microscopy or BgITS-1 PCR. Sensitivity of Bgcox3 PCR was 100% when compared with that of Bg18S nPCR.


Assuntos
Babesia/isolamento & purificação , Babesiose/diagnóstico , Doenças do Cão/diagnóstico , Complexo IV da Cadeia de Transporte de Elétrons/genética , Mitocôndrias/enzimologia , Animais , Babesia/classificação , Babesia/genética , Babesiose/parasitologia , Sequência de Bases , Reações Cruzadas , DNA Espaçador Ribossômico/química , Doenças do Cão/parasitologia , Cães , Eritrócitos/parasitologia , Funções Verossimilhança , Filogenia , Reação em Cadeia da Polimerase/veterinária , Valor Preditivo dos Testes , RNA Ribossômico 18S/análise , Sensibilidade e Especificidade , Alinhamento de Sequência/veterinária
2.
Rev Inst Med Trop Sao Paulo ; 61: e45, 2019 Sep 12.
Artigo em Inglês | MEDLINE | ID: mdl-31531623

RESUMO

Myiasis caused by Dermatobia hominis , the human botfly, is frequent in the Americas, however, scarce morphological and molecular information exist regarding this dipteran. We describe three cases in urban areas of Mexico were D. hominis is not endemic. Morphological and genetic identification were performed using the cytochrome oxidase I as a molecular marker. The mitochondrial cytochrome oxidase I gene is useful for inferring the genetic divergence of D. hominis .


Assuntos
Dípteros/enzimologia , Dípteros/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Miíase/parasitologia , Adulto , Animais , Feminino , Humanos , Masculino , México , Pessoa de Meia-Idade , Miíase/diagnóstico , Filogenia , População Urbana
3.
Phytopathology ; 109(11): 1900-1907, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31369362

RESUMO

Floricolous downy mildews (Peronospora, oomycetes) are a small, monophyletic group of mostly inconspicuous plant pathogens that induce symptoms exclusively on flowers. Characterization of this group of pathogens, and information about their biology, is particularly sparse. The recurrent presence of a disease causing flower malformation which, in turn, leads to high production losses of the medicinal herb Matricaria chamomilla in Serbia has enabled continuous experiments focusing on the pathogen and its biology. Peronospora radii was identified as the causal agent of the disease, and morphologically and molecularly characterized. Diseased chamomile flowers showed severe malformations of the disc and ray florets, including phyllody and secondary inflorescence formation, followed by the onset of downy mildew. Phylogeny, based on internal transcribed spacer and cox2, indicates clustering of the Serbian P. radii with other P. radii from chamomile although, in cox2 analyses, they formed a separate subcluster. Evidence pointing to systemic infection was provided through histological and molecular analyses, with related experiments validating the impact of soilborne and blossom infections. This study provides new findings in the biology of P. radii on chamomile, thus enabling the reconstruction of this floricolous Peronospora species' life cycle.


Assuntos
Camomila , Peronospora , Camomila/microbiologia , DNA Espaçador Ribossômico/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Genes Fúngicos/genética , Peronospora/classificação , Peronospora/genética , Peronospora/fisiologia , Filogenia , Doenças das Plantas/microbiologia
4.
Onderstepoort J Vet Res ; 86(1): e1-e11, 2019 Jun 18.
Artigo em Inglês | MEDLINE | ID: mdl-31291732

RESUMO

Fasciola spp. are the causative agents of fascioliasis in humans and livestock. Before the development of control and management measures, the geographical distribution of the species and patterns of infection must be considered. Because of difficulties in the phenotypic differentiation and morphometric classification of Fasciola spp., DNA molecular markers have become more useful for fluke differentiation and description of phylogenetic patterns. This study aimed to differentiate and describe the phylogenetic background of Fasciola spp. isolated from cattle slaughtered at three abattoirs in the Mpumalanga and KwaZulu-Natal provinces of South Africa. The cytochrome c oxidase I (COI) - FHCO1 (forward: 5'-TTGGTTTTTTGGGCATCCT-3') and FHCO1 (reverse: 5' -AGGCCACCACCAAATAAAAGA3') - marker was sequenced from 55 Fasciola flukes that were collected from abattoirs in catchment areas of the KwaZulu-Natal and Mpumalanga provinces. Fasciola hepatica was demonstrated to have 100% prevalence in KwaZulu-Natal and Mpumalanga (highveld), respectively, and 76% prevalence in the lowveld (Belfast area) of Mpumalanga. Two animals from the Belfast metapopulation were co-infected with both Fasciola gigantica and F. hepatica. DNA sequence analysis of all the isolates demonstrated a sequence conservation of 0.472, nucleotide diversity of 0.082 and Tajima's D of -1.100; however, it was not statistically significant (p > 0.05). Twenty-two haplotypes were identified, with 18 novel haplotypes being unique to the isolates from South Africa. Within the study samples, 12 haplotypes were isolated to a few individuals, with a haplotype diversity of 0.8957 indicating high genetic diversity. Principal coordinate analysis supported the clustering and distribution of the haplotypes, with 11.38% of the variation being attributed to coordinate 2 and 55.52% to coordinate 1. The distribution of Fasciola spp. has been demonstrated to be related to the distribution of the freshwater intermediate host snails, Lymnaea spp., as well as the relative altitude of the localities in South Africa. Information provided by this study serves as preliminary evidence for further studies on the mapping of the distribution of F. gigantica and F. hepatica in South Africa, which is key in designing control programmes for fascioliasis in humans and livestock.


Assuntos
Doenças dos Bovinos/epidemiologia , Complexo IV da Cadeia de Transporte de Elétrons/genética , Fasciola/isolamento & purificação , Fasciolíase/veterinária , Microbiologia de Alimentos , Matadouros , Animais , Bovinos , Doenças dos Bovinos/parasitologia , DNA de Helmintos/análise , Fasciola/classificação , Fasciola/genética , Fasciolíase/epidemiologia , Marcadores Genéticos , Filogenia , África do Sul/epidemiologia
5.
Rev Bras Parasitol Vet ; 28(2): 258-265, 2019 Jun 27.
Artigo em Inglês | MEDLINE | ID: mdl-31271641

RESUMO

Cysticercus ovis or sheep measles is the larval stage of Taenia ovis, which is the intestinal tapeworm of dogs. It is found in the cardiac and skeletal muscles of sheep and can be the cause of partial or total condemnation of carcasses at abattoirs. The aim of the current work was to determine the prevalence of C. ovis among sheep in Upper Egypt and to present the molecular and phylogenetic analysis of this using the amplified Mitochondrial Cytochrome Oxidase subunit 1 (MT-CO1) gene. A total of 1885 sheep slaughtered at local abattoirs of 4 different governorates of Upper Egypt (Asuit, Sohag, Qena and Aswan) were carefully examined for C. ovis. The overall prevalence of infection was 2.02%. The highest rate of infection was observed in adult animals over 4 years of age (44.73%). There was no significant effect of animal sex on infection rates. The phylogenic analysis of C. ovis Egyptian isolates showed very close similarity to the New Zealand isolate (AB731675). This is the first report showing the genetic analysis of C. ovis in Egypt, which provides a very powerful tool for taxonomy and definitive diagnosis of C. ovis, which could be helpful for preventive and control programs.


Assuntos
Cisticercose/veterinária , Cysticercus/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Doenças dos Ovinos/parasitologia , Ovinos/parasitologia , Matadouros , Animais , Cisticercose/epidemiologia , Cysticercus/isolamento & purificação , Egito/epidemiologia , Perfilação da Expressão Gênica , Filogenia , Prevalência , Fatores de Risco , Doenças dos Ovinos/epidemiologia
6.
BMC Evol Biol ; 19(1): 118, 2019 06 11.
Artigo em Inglês | MEDLINE | ID: mdl-31185884

RESUMO

BACKGROUND: There is a biogeographic break located at 30°S in the southeast Pacific, in a coastal area of strong environmental discontinuities. Several marine benthic taxa with restricted dispersal have a coincident phylogeographic break at 30°S, indicating that genetic structure is moulded by life history traits that limit gene flow and thereby promote divergence and speciation. In order to evaluate intraspecific divergence at this biogeographic break, we investigated the genetic and morphological variation of the directly developing beach isopod Excirolana hirsuticauda along 1900 km of the southeast Pacific coast, across 30°S. RESULTS: The COI sequences and microsatellite data both identified a strong discontinuity between populations of E. hirsuticauda to the north and south of 30°S, and a second weaker phylogeographic break at approximately 35°S. The three genetic groups were evidenced by different past demographic and genetic diversity signatures, and were also clearly distinguished with microsatellite data clustering. The COI sequences established that the genetic divergence of E. hirsuticauda at 30°S started earlier than divergence at 35°. Additionally, the three groups have different past demographic signatures, with probable demographic expansion occurring earlier in the southern group (south of 35°S), associated with Pleistocene interglacial periods. Interestingly, body length, multivariate morphometric analyses, and the morphology of a fertilization-related morphological character in males, the appendix masculina, reinforced the three genetic groups detected with genetic data. CONCLUSIONS: The degree of divergence of COI sequences, microsatellite data, and morphology was concordant and showed two geographic areas in which divergence was promoted at differing historical periods. Variation in the appendix masculina of males has probably promoted reproductive isolation. This variation together with gene flow restrictions promoted by life history traits, small body size, oceanographic discontinuities and sandy-beach habitat continuity, likely influenced species divergence at 30°S in the southeast Pacific coast. The degree of genetic and morphological differentiation of populations to the north and south of 30°S suggests that E. hirsuticauda harbours intraspecific divergence consistent with reproductive isolation and an advanced stage of speciation. The speciation process within E. hirsuticauda has been shaped by both restrictions to gene flow and a prezygotic reproductive barrier.


Assuntos
Isópodes/anatomia & histologia , Isópodes/genética , Filogeografia , Análise de Variância , Animais , Teorema de Bayes , Tamanho Corporal , DNA Mitocondrial/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Fluxo Gênico , Loci Gênicos , Variação Genética , Haplótipos/genética , Isópodes/classificação , Repetições de Microssatélites/genética , Filogenia , Análise de Componente Principal
7.
Parasitol Res ; 118(8): 2399-2408, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31222390

RESUMO

A new Isospora (Apicomplexa: Eimeriidae) species is described from an Australian raven (Corvus coronoides) in Western Australia. Sporulated oocysts (n = 21) are ovoid, 21.2 (18.4-23.9) µm in length and 18.8 (16.9-20.6) µm in width, with a shape index of 1.13. The bi-layered oocyst wall is smooth and colourless, 1.2 µm thick. A polar granule and oocyst residuum is present, but the micropyle is absent. The sporocysts are ovoid-shaped, 16.3 (13.7-18.9) × 10.7 (8.4-12.9) µm, with a shape index (length/width) of 1.52. Stieda and substieda bodies are present, the Stieda body being small and hemidome-shaped and the substieda being indistinct. Each sporocyst with four vermiform sporozoites arranged head to tail. The sporozoites are crescent-shaped, 9.0 (8.9-9.2) × 2.7 (2.3-3.0) µm, with a shape index (length/width) of 3.33. The sporocyst residuum is present. The isolated oocysts had different morphological characteristics when compared with all known Isospora spp. The coccidian parasite was analysed at the 18S and 28S ribosomal RNA and the mitochondrial cytochrome oxidase (COI) loci. At the 18S locus, I. coronoideae n. sp. exhibited 98.9% similarity to I. neochmiae from a captive-bred red-browed finch (KT224380) and Isospora sp. from domestic pigeons (Columba livia domestica) (AB757860), 98.5% similarity to I. gryphoni (AF080613) from an American goldfinch and 98.3% similarity to I. manorinae (KT224379) from a yellow-throated miner. At the 28S locus, it exhibited 95.4% and 94.8% similarity to I. manorinae (KT224381) and I. anthochaerae (KF766053), respectively. At the COI locus, it exhibited 99.8% and 99.7% similarity to I. butcherae (KY801687) and I. neochmiae (KT224378), respectively. Based on morphological and molecular data, this isolate is a new species of Isospora, which is named Isospora coronoideae n. sp. after its host, the Australian raven (Corvus coronoides) (Passeriformes: Corvidae) (Linnaeus, 1758).


Assuntos
Doenças das Aves/parasitologia , Corvos/parasitologia , Isospora/isolamento & purificação , Isosporíase/veterinária , Animais , Austrália , Columbidae/parasitologia , Complexo IV da Cadeia de Transporte de Elétrons/genética , Isospora/classificação , Isospora/genética , Isosporíase/parasitologia , Oocistos/classificação , Oocistos/genética , Oocistos/isolamento & purificação , Filogenia , Proteínas de Protozoários/genética , RNA Ribossômico 18S/genética , RNA Ribossômico 28S/genética , Austrália Ocidental
8.
Biol Bull ; 236(3): 199-206, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-31167090

RESUMO

The barnacle Balanus glandula is a broadly distributed species in the temperate northeastern Pacific that is notable for a robust genetic cline between about 36° and 40° N latitude. Prior work established the evolutionary origins of this pattern and proposed that it is maintained by environmental selection. In recent years, "climate velocity" studies in marine habitats have shown dramatic distributional shifts for many species as they track their preferred temperature range in a warming ocean. We re-sampled B. glandula across its entire geographic range to determine whether there has been any shift in this genetic distribution, a development signaling that temperature or other climate factors are maintaining this genetic cline. Additionally, we asked whether the spatially distributed mitochondrial lineages also vary in reproductive output with latitude, using location as a proxy for temperature and other coastal environmental factors. Here we show that although the distribution of the genetic cline has not appreciably changed, there is a notable association of decreased reproductive output at lower latitudes of the distribution in the "northern" lineage of B. glandula.


Assuntos
Distribuição Animal/fisiologia , Thoracica/genética , Animais , Mudança Climática , DNA Mitocondrial/genética , Ecossistema , Complexo IV da Cadeia de Transporte de Elétrons/genética , Genética Populacional , Oceano Pacífico , Reprodução/fisiologia , Temperatura Ambiente , Thoracica/fisiologia
9.
Dokl Biochem Biophys ; 485(1): 119-122, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31201629

RESUMO

Samples from 11 populations of the Arctic char of the North-European part of Russia belonging to the anadromous and resident forms and two samples from Lake Sobach'e (Taimyr) were studied. The nucleotide sequence of the mitochondrial COI gene was determined in 60 individuals. In the majority of populations, the same COI haplotype was found. In some populations of the resident chars, haplotypes differing from the widespread haplotype in a single nucleotide substitution were found. The obtained genetic data give no reason to distinguish the resident form of the Arctic char from lakes of Karelia and the Kola Peninsula as an independent species, Salvelinus lepechini. The adaptation of the Arctic char to the unstable environmental conditions is ensured primarily by its phenotypic plasticity.


Assuntos
DNA Mitocondrial/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Proteínas de Peixes/genética , Proteínas Mitocondriais/genética , Filogenia , Truta/genética , Animais , Europa (Continente)
10.
J Chem Ecol ; 45(5-6): 429-439, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31152352

RESUMO

Animal classification is primarily based on morphological characters, even though these may not be the first to diverge during speciation. In many cases, closely related taxa are actually difficult to distinguish based on morphological characters alone, especially when there is no substantial niche separation. As a consequence, the diversity of certain groups is likely to be underestimated. Lepidoptera -moths and butterflies- represent the largest group of herbivorous insects. The extensive diversification in the group is generally assumed to have its origin in the spectacular radiation of flowering plants and the resulting abundance of ecological niches. However, speciation can also occur without strong ecological divergence. For example, reproductive isolation can evolve as the result of divergence in mate preference and the associated pheromone communication system. We combined pheromone trapping and genetic analysis to elucidate the evolutionary relationships within a complex of primitive moth species (Lepidoptera: Eriocraniidae). Mitochondrial and nuclear DNA markers provided evidence that Eriocrania semipurpurella, as currently defined by morphological characters, includes three cryptic species in Northern and Western Europe. Male moths of these cryptic species, as well as of the closely related E. sangii, exhibited relative specificity in terms of their attraction to specific ratios of two major pheromone components, (2S,6Z)-nonen-2-ol and (2R,6Z)-nonen-2-ol. Our data suggest strong assortative mating in these species in the absence of apparent niche separation, indicating that Eriocrania moths may represent an example of non-ecological speciation. Finally, our study argues in favour of combining pheromone investigations and DNA barcoding as powerful tools for identifying and delimitating species boundaries.


Assuntos
Mariposas/genética , Atrativos Sexuais/química , Animais , DNA/isolamento & purificação , DNA/metabolismo , Complexo IV da Cadeia de Transporte de Elétrons/classificação , Complexo IV da Cadeia de Transporte de Elétrons/genética , Feminino , Variação Genética , Gliceraldeído 3-Fosfato Desidrogenase (NADP+)/classificação , Gliceraldeído 3-Fosfato Desidrogenase (NADP+)/genética , Masculino , Mitocôndrias/genética , Filogenia , Atrativos Sexuais/metabolismo
11.
Korean J Parasitol ; 57(2): 153-159, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31104407

RESUMO

Echinococcus granulosus is an important zoonotic parasite globally causing cystic echinococcosis (CE) in humans and animals. In this study, prevalence of CE and variation of cox1 gene sequence were analyzed with isolates E. granulosus collected from different areas in northern Xinjiang, China. The survey showed that 3.5% of sheep and 4.1% of cattle were infected with CE. Fragment of cox1 was amplified from all the positive sheep and cattle samples by PCR. In addition, 26 positive samples across the 4 areas were included. The isolates were all E. granulosus sensu stricto (s.s.) containing 15 haplotypes (Hap1-15), and clustered into 2 genotypes, G1 (90.1%, 91/101) and G3 (9.9%, 10/101). Hap1 was the most common haplotype (48.5%, 49/101). Hap9 were found in humans samples, indicating that sheep and cattle reservoir human CE. It is indicate that E. granulosus may impact on control of CE in livestock and humans in the region.


Assuntos
Doenças dos Bovinos/epidemiologia , Equinococose/epidemiologia , Equinococose/veterinária , Echinococcus granulosus/genética , Echinococcus granulosus/isolamento & purificação , Doenças dos Ovinos/epidemiologia , Animais , Bovinos , Doenças dos Bovinos/parasitologia , China/epidemiologia , Análise por Conglomerados , Equinococose/parasitologia , Echinococcus granulosus/classificação , Complexo IV da Cadeia de Transporte de Elétrons/genética , Genótipo , Humanos , Epidemiologia Molecular , Prevalência , Ovinos , Doenças dos Ovinos/parasitologia
12.
Mol Med Rep ; 19(6): 5464-5472, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31059062

RESUMO

We previously reported that the collapse of ATP production via mitochondrial damage causes ATPase dysfunction, resulting in the onset or progression of lens opacification in cataracts in model rats. In the present study, it was investigated whether the mRNA expression levels of the three subtypes of mitochondrial cytochrome c oxidase (MTCO)1, 2 and 3 and ATP content change with the type and severity of cataracts in human lens. Samples of lens epithelium were collected from Japanese patients during cataract surgery, and the type and severity of the cataracts (grade) were determined according to the WHO classification [cortical (COR), nuclear (NUC), posterior subcapsular (PSC) opacification]. The MTCO1­3 mRNA expression levels in patients with grade­1 COR, NUC and PSC opacification were significantly enhanced compared with those of normal patients. The enhanced MTCO1­3 mRNA levels subsequently decreased in patients with COR, and the MTCO1­3 mRNA levels and ATP levels in patients with grade­3 COR were similar to those in normal patients. However, the mRNA expression levels of MTCO3 in patients with grade 3­NUC opacification and MTCO1­3 in patients with grade­3 PSC opacification, along with the ATP content, were significantly lower than in patients without cataracts. In conclusion, it was revealed that ATP production in lens epithelium is enhanced in early­stage cataracts (grade­1) in Japanese patients with COR, NUC and PSC opacification. In addition, in severe cataracts (grade­3), ATP production and content are strongly decreased in Japanese patients with PSC opacification. ATP depletion in human lens epithelium with PSC opacification may promote lens opacification by ATPase dysfunction.


Assuntos
Catarata/patologia , Complexo IV da Cadeia de Transporte de Elétrons/genética , Cristalino/metabolismo , Mitocôndrias/metabolismo , RNA Mensageiro/metabolismo , Trifosfato de Adenosina/metabolismo , Idoso , Catarata/metabolismo , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Epitélio/metabolismo , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Índice de Gravidade de Doença
13.
Zoolog Sci ; 36(1): 82-94, 2019 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-31116542

RESUMO

To elucidate the genetic population structure of two coastal weevils, Scepticus griseus and S. tigrinus, we conducted molecular phylogenetic analyses of the mitochondrial DNA cytochrome c oxidase subunit I (COI) region (1308 bp) and cytochrome c oxidase subunit II (COII) region (584 bp). A total of 650 individuals (S. griseus, 444 individuals; S. tigrinus, 206 individuals) were obtained from 64 sites. The haplotype networks of both species showed three major lineages with roughly regional distribution. However, the two species show quite different genetic structures; S. griseus has a complicated structure while that of S. tigrinus is simple. We hypothesize that the genetic structure of each of these two weevil species reflects climatic oscillations during the Pleistocene, and the differences in genetic structure between S. griseus and S. tigrinus may represent a unique evolutionary history scenario in each species.


Assuntos
Gorgulhos/classificação , Gorgulhos/genética , Animais , Evolução Biológica , Mudança Climática , DNA Mitocondrial/genética , Ecossistema , Complexo IV da Cadeia de Transporte de Elétrons/genética , Japão , Filogenia , Análise de Sequência de DNA
14.
J Food Sci ; 84(6): 1256-1265, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31120564

RESUMO

DNA barcoding possesses advantages of high resolution, high sensitivity, and capability in capturing as much identity information as possible. However, highly varying sources of food materials and a complicated supply chain bring about challenge to the application of barcoding methods. In this study, different barcode systems were compared to establish a robust method for tracing animal species in food. Experiments on food samples from mammal, poultry, and fish proved that a mini barcode system targeting a 192 bp COI gene fragment was able to accurately identify both raw and highly processed animal food. In order to distinguish species in a mixed food sample, cloning technique was used by which as low as 10% target animal ingredient could be detected. Testing of marketed food products verified the capability of the mini barcoding method in identifying illegally claimed product.


Assuntos
Código de Barras de DNA Taxonômico/métodos , Complexo IV da Cadeia de Transporte de Elétrons/genética , Peixes/genética , Contaminação de Alimentos/análise , Mamíferos/genética , Carne/análise , Aves Domésticas/genética , Sequência de Aminoácidos , Animais , Análise Discriminante , Alinhamento de Sequência
15.
Hum Genet ; 138(7): 749-756, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31079202

RESUMO

Cytochrome c oxidase 20 (COX20)/FAM36A encodes a conserved protein that is important for the assembly of COX, complex IV of the mitochondrial respiratory chain. A homozygous mutation (p.Thr52Pro) in COX20 gene has been previously described to cause muscle hypotonia and ataxia. In this study, we describe two patients from a non-consanguineous family exhibiting autosomal recessive sensory-dominant axonal neuropathy and static encephalopathy. The whole-exome sequencing analysis revealed that both patients harbored compound heterozygous mutations (p.Lys14Arg and p.Trp74Cys) of COX20 gene. The pathogenicity of the variants was further supported by morphological alternations of mitochondria observed in sural nerve and decreased COX20 protein level of peripheral blood leucocytes derived from the patients. In conclusion, COX20 might be considered as a candidate gene for the complex inherited disease. This observation broadens the clinical and genetic spectrum of COX20-related disease. However, due to the limitation of a single-family study, additional cases and studies are definitely needed to further confirm the association.


Assuntos
Encefalopatias/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Deformidades do Pé/genética , Neuropatia Axonal Gigante/genética , Mutação , Adolescente , Adulto , Encefalopatias/patologia , Feminino , Deformidades do Pé/patologia , Neuropatia Axonal Gigante/patologia , Humanos , Masculino , Adulto Jovem
16.
Mol Med Rep ; 19(6): 4852-4862, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31059068

RESUMO

Cerebrovascular disease (CVD) is one of the leading causes of mortality worldwide. The role of cytochrome c oxidase subunit 6B1 (COX6B1) in the central nervous system remains unclear. The present study aimed to analyze the role of COX6B1 in rat hippocampal neurons extracted from fetal rats. The subcellular localization of the neuron­specific marker microtubule­associated protein 2 was detected by immunofluorescence assay. Cell viability was assessed using a cell counting kit, and the levels of apoptosis and cytosolic Ca2+ were analyzed by flow cytometry. The expression levels of the molecular factors downstream to COX6B1 were determined using reverse transcription­quantitative polymerase chain reaction and western blotting. Reoxygenation following oxygen­glucose deprivation (OGD) decreased cell viability and the expression levels of COX6B1 in a time­dependent manner, and 60 min of reoxygenation was identified as the optimal time period for establishing an ischemia/reperfusion (I/R) model. Overexpression of COX6B1 was demonstrated to reverse the viability of hippocampal neurons following I/R treatment. Specifically, COX6B1 overexpression decreased the cytosolic concentration of Ca2+ and suppressed neuronal apoptosis, which were increased following I/R treatment. Furthermore, overexpression of COX6B1 increased the protein expression levels of apoptosis regulator BCL­2 and mitochondrial cytochrome c (cyt c), and decreased the protein expression levels of apoptosis regulator BCL2­associated X and cytosolic cyt c in I/R model cells. Collectively, the present study results suggested that COX6B1 overexpression may reverse I/R­induced neuronal damage by increasing the viability of neurons, by decreasing the cytosolic levels of Ca2+ and by suppressing apoptosis. These results may facilitate the development of novel strategies for the prevention and treatment of CVD.


Assuntos
Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Complexo IV da Cadeia de Transporte de Elétrons/farmacologia , Neurônios/metabolismo , Traumatismo por Reperfusão/prevenção & controle , Lobo Temporal/metabolismo , Animais , Apoptose/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Transtornos Cerebrovasculares/prevenção & controle , Citocromos c/metabolismo , Complexo IV da Cadeia de Transporte de Elétrons/genética , Feminino , Glucose/metabolismo , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Neurônios/patologia , Oxigênio/metabolismo , Substâncias Protetoras/metabolismo , Substâncias Protetoras/farmacologia , Ratos , Ratos Sprague-Dawley , Lobo Temporal/patologia , Proteína bcl-X/metabolismo
17.
BMC Evol Biol ; 19(1): 105, 2019 05 17.
Artigo em Inglês | MEDLINE | ID: mdl-31101005

RESUMO

BACKGROUND: The Pleistocene cyclic sea-level fluctuations are thought to have markedly affected the distribution and genetic architecture of Atlanto-Mediterranean biota. Despite the acknowledged key role played by these historical events in shaping population genetic structure of marine species, little is still known about the processes involved in shaping the spatial distribution of genetic variation within intertidal species. We intended in this study to reconstruct the phylogeography of a common and widely distributed coastal species across the East Atlantic and Mediterranean Sea (the warty crab Eriphia verrucosa), aiming to unravel potential microevolutionary processes likely involved in shaping its genetic polymorphism. For this purpose, a total of 155 specimens of E. verrucosa from 35 locations across the entire distribution range were analyzed by comparing a 453 basepairs region of the mitochondrial gene cytochrome oxidase subunit 1 (Cox1). RESULTS: Our results unveiled the prevalence of high genetic connectivity among East Atlantic and Mediterranean populations, with noticeable genetic distinctiveness of the peripheral population from the Azores. Spatio-temporal patterns of genetic diversification and demographic history allowed retrieving genetic imprints of late Pleistocene vicariant event across the Gibraltar Strait followed by subsequent postglacial expansion events for both the East Atlantic and Mediterranean regions. Integrative evidences from the outcomes of comparison of regional genetic diversification, as well as evolutionary and biogeographic histories reconstructions, support the existence of potential glacial refugia for E. verrucosa in the East Atlantic and western Mediterranean. Our results also revealed low levels of genetic variability along with recent demographic and spatial expansion events for eastern Mediterranean warty crabs, suggesting that the eastern areas within the distribution range of the species might have been recently colonized from putative glacial refugia. CONCLUSIONS: These findings provide new insights into the phylogeography and evolutionary history of a common but poorly studied Atlanto-Mediterranean decapod species. Specifically, they contribute to the understanding of the impact of historical processes on shaping contemporary population genetic structure and diversity in intertidal marine species.


Assuntos
Braquiúros/genética , Camada de Gelo , Filogenia , Filogeografia , Refúgio , Animais , Sequência de Bases , Teorema de Bayes , Complexo IV da Cadeia de Transporte de Elétrons/genética , Variação Genética , Gibraltar , Haplótipos/genética , Mar Mediterrâneo , Fatores de Tempo
18.
Ann Parasitol ; 65(1): 87-97, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31127891

RESUMO

One of the fundamental aspects in understanding the biology, diversity and epidemiology of a parasite lies in its proper identification. In the present study, morphological and molecular characterization of Clinostomum metacercariae recovered from an ornamental fish, Trichogaster fasciata, was carried out in order to ascertain its identity. To serve the purpose, scanning electron micrographs and gene sequences for two commonly used molecular markers, i.e., nuclear ribosomal internal transcribed spacer 2 (rDNA-ITS2) and mitochondrial cytochrome c oxidase subunit 1 (mtCO1) were obtained. The sequences were further used for generating similarity index matrix as well as inferring phylogenies. Light and electron microscopic observations on metacercariae of the parasite revealed that it belongs to the genus Clinostomum. Identification of the same up to the level of species was made possible through sequence and phylogenetic analyses. The ITS2 sequence analyses of our species (KX758630) showed similarity to unidentified Clinostomum sp. reported from Nigeria (KY865625) and China (KP110579), and C. tilapiae recorded from South Africa (KX034048) and Nigeria (KY649353). However, the CO1 gene analyses suggested it to be highly identical to C. philippinense and the same was also corroborated in the phylogenetic analysis. Thus, morphological and molecular characterization revealed that the recovered metacercariae belong to the species C. philippinense. Additionally, a brief description of secondary structures of ITS2 of various species of Clinostomum has also been discussed.


Assuntos
Peixes , Metacercárias , Trematódeos , Infecções por Trematódeos , Animais , DNA Espaçador Ribossômico/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Peixes/parasitologia , Metacercárias/classificação , Metacercárias/genética , Metacercárias/ultraestrutura , Filogenia , Trematódeos/classificação , Trematódeos/genética , Trematódeos/ultraestrutura , Infecções por Trematódeos/parasitologia , Infecções por Trematódeos/veterinária
19.
Parasit Vectors ; 12(1): 213, 2019 May 07.
Artigo em Inglês | MEDLINE | ID: mdl-31064400

RESUMO

BACKGROUND: The poultry red mite (PRM), Dermanyssus gallinae, is one of the most economically deleterious ectoparasites affecting egg-laying hens worldwide. It may be possible to control D. gallinae populations by manipulating lighting regimes within poultry units. However, no studies have clearly shown the effects of darkness on the population growth rate of D. gallinae. METHODS: The effect of darkness on the population growth rate of D. gallinae was investigated, together with the first description of the molecular identity of the mite from China. Mite variables under two lighting regimens (1:23 h L:D and 12:12 h L:D) were compared, including number of mites and eggs, survival and feeding rates, engorgement, oviposition, hatchability and the life-cycle of D. gallinae. RESULTS: The results showed that the number of mites (13,763 ± 956) and eggs (5424 ± 317) in the rearing system with prolonged darkness of 1:23 h L:D at 4th week were 2.4- and 3.6-fold higher than those under a conventional lighting regimen of 12:12 h L:D, respectively. The feeding rates of mites under prolonged darkness ranged from 36.7 ± 1.1% to 52.0 ± 7.0%, which were significantly higher than those under conventional lighting regimen (ranging from 22.6 ± 1.9% to 37.3 ± 1.6%). The mean weight of engorged females (0.26 ± 0.01 mg) and the mean number of eggs per female (on average 5.87 ± 0.36) under prolonged darkness were significantly higher than those under conventional lighting regimen (0.22 ± 0.01 mg and 3.62 ± 0.31, respectively). However, the survival rate ranging from 98.07 ± 0.10% to 98.93 ± 0.19%, hatchability of 97.93 ± 0.01% and the life-cycle of D. gallinae (9 days) was not affected by the lighting period. CONCLUSIONS: Our findings demonstrated that prolonged darkness significantly promoted the proliferation levels of D. gallinae, resulting in increased number of mites and eggs in the rearing system. The promoted population growth of D. gallinae was found to be related to the increased feeding rate, engorgement level and oviposition level of mites under prolonged darkness. The egg hatchability, the survival rates and the duration of life-cycle of D. gallinae were not affected by the light regimes.


Assuntos
Escuridão , Ácaros/efeitos da radiação , Animais , Galinhas , DNA Intergênico , Complexo IV da Cadeia de Transporte de Elétrons/genética , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Comportamento Alimentar/efeitos da radiação , Feminino , Estágios do Ciclo de Vida , Luz , Infestações por Ácaros/veterinária , Ácaros/genética , Ácaros/crescimento & desenvolvimento , Oviposição/efeitos da radiação , Fotoperíodo , Crescimento Demográfico , Doenças das Aves Domésticas/parasitologia , Reprodução/efeitos da radiação , Fatores de Tempo
20.
Mem Inst Oswaldo Cruz ; 114: e180595, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31116227

RESUMO

The genetic information of ancient Paragonimus westermani, the oriental lung fluke infecting over 20 million people worldwide, has not been thoroughly investigated thus far. We analysed genetic markers (COI and ITS2) of P. westermani from coprolite specimens (n = 6) obtained from 15th to 18th century Korean mummies. Our results indicated that all P. westermani sequences were generally distinct from the other species of the genus Paragonimus. The sequences were clustered into three groups: Group I for East Asia; Group II for South and Southeast Asia; and Group III for India and Sri Lanka. In this study, we found that ancient P. westermani sequences in Korea belong to Group I, adding invaluable information to the existing knowledge of Paragonimus paleogenetics.


Assuntos
DNA de Helmintos/genética , DNA Espaçador Ribossômico/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Fezes/parasitologia , Múmias/parasitologia , Paragonimus westermani/isolamento & purificação , Animais , Ásia , Humanos , Paleodontologia , Paragonimus westermani/genética , Contagem de Ovos de Parasitas , Filogenia
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