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1.
Exp Parasitol ; 212: 107872, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32165145

RESUMO

Theileriosis is a widespread and economically important disease of small ruminants in Pakistan. Ruminants are the intermediate hosts in the lifecycle of Theileria spp., with ticks of the family Ixodidae being the definitive hosts. To better understand the distribution and prevalence of theileriosis in Pakistan, a molecular survey was performed in small ruminants from the Lower Dir district of the Khyber Pakhtunkhwa province. A total of 200 healthy sheep and goats were screened from Maidan, Samar Bagh and Munda districts of district Dir Lower, Pakistan during December (2017) to April (2018). DNA samples were screened through nested PCR using universal primers. The amplified 492-498 bp amplicon was subjected to RLB analysis which was based on the hypervariable of the 18S rRNA gene to test for the presence of genotypes of Theileria in blood samples. A phylogeny was constructed to determine the species of Theileria genotypes. Nested PCR results indicated 53.5% prevalence of one or more Theileria genotypes in the blood of the host animal. From RLB assay, 27 animals (13.5%) showed infection with only a single species of Theileria while 80 animals (40%) showed coinfection by multiple Theileria spp. Based on the 18S rRNA phylogeny, the unknown genotype is of the species Theileria luwenshuni and is closely related to Chinese isolates. The present finding is the first report on molecular diagnosis of Theileria luwenshuni in small ruminants in Pakistan.


Assuntos
Doenças das Cabras/parasitologia , Doenças dos Ovinos/parasitologia , Theileria/isolamento & purificação , Theileriose/parasitologia , Distribuição por Idade , Análise de Variância , Animais , Coinfecção/epidemiologia , Coinfecção/parasitologia , DNA de Protozoário/química , DNA de Protozoário/genética , DNA de Protozoário/isolamento & purificação , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Ribossômico/isolamento & purificação , Feminino , Genótipo , Doenças das Cabras/epidemiologia , Cabras , Hibridização Genética , Funções Verossimilhança , Modelos Lineares , Masculino , Análise Multivariada , Sondas de Oligonucleotídeos , Paquistão/epidemiologia , Filogenia , Reação em Cadeia da Polimerase , Prevalência , RNA Ribossômico 18S/genética , Fatores de Risco , Análise de Sequência de DNA , Distribuição por Sexo , Ovinos , Doenças dos Ovinos/epidemiologia , Theileria/classificação , Theileria/genética , Theileriose/epidemiologia
2.
Epidemiol Infect ; 148: e49, 2020 02 14.
Artigo em Inglês | MEDLINE | ID: mdl-32054545

RESUMO

A new fast-growing mycobacterium, designated strain QGD101T, was isolated from the sputum of an 84-year-old man suspected of tuberculosis in Wuhan Medical Treatment Center, Hubei, China. This strain was a gram-staining-negative, aerobic, non-spore-forming and catalase-positive bacterium, which was further identified as the NTM by PNB and TCH tests. The moxifloxacin and levofloxacin exhibited strong suppressing function against QGD101T with MIC values of 0.06 and 0.125 µg/ml after drug susceptibility testing of six main antimicrobial agents on mycobacteria. Based on the sequence analysis of 16S rRNA, rpoB, hsp65 and 16S-23S rRNA internal transcribed spacer, the strain QGD101T could not be identified to a species level. Mycobacterium moriokaense ATCC43059T that shared the highest 16S rRNA gene sequence similarity (98%) with strain QGD101T was actually different in genomes average nucleotide identity (78.74%). In addition, the major cellular fatty acids of QGD101T were determined as C18:1ω9c, C16:0 and C18:2ω6c. The DNA G + C content was 64.9% measured by high performance liquid chromatography. Therefore, the phenotypic and genotypic characterisation of this strain led us to the conclusion that it represents a novel species of mycobacteria, for which the name Mycobacterium hubeiense sp. nov. (type strain QGD101T = CCTCCAA 2017003T = KCTC39927T) was proposed. Thus, the results of this study are very significant for the clinical diagnosis of tuberculosis and future personalised medicine.


Assuntos
Infecções por Mycobacterium não Tuberculosas/microbiologia , Mycobacterium/classificação , Mycobacterium/isolamento & purificação , Escarro/microbiologia , Idoso de 80 Anos ou mais , Antituberculosos/farmacologia , Proteínas de Bactérias/genética , Composição de Bases , Chaperonina 60/genética , China , Análise por Conglomerados , Citosol/química , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , RNA Polimerases Dirigidas por DNA/genética , Ácidos Graxos/análise , Humanos , Levofloxacino/farmacologia , Masculino , Testes de Sensibilidade Microbiana , Moxifloxacina/farmacologia , Mycobacterium/genética , Mycobacterium/fisiologia , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
3.
J Parasitol ; 106(1): 133-156, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-32048913

RESUMO

Four new species of Paraorygmatobothrium are described from carcharhinid shark species from the Gulf of Mexico and the U.S. coast of the northwestern Atlantic Ocean. Paraorygmatobothrium bullardi n. sp. is described from Carcharhinus brevipinna as its type host, and from Carcharhinus acronotus and Carcharhinus limbatus. Paraorygmatobothrium campbelli n. sp. is described from Rhizoprionodon terraenovae. Paraorygmatobothrium deburonae n. sp. is described from Carcharhinus isodon as its type host, as well as from C. brevipinna, C. limbatus, and R. terraenovae. Paraorygmatobothrium mattisi n. sp. is described from R. terraenovae as its type host, and from C. brevipinna and C. limbatus. These 4 species differ from other species of Paraorygmatobothrium and from each other in bothridial microthrix shape, and in various combinations of features such as the total length of the worm, apical sucker size, bothridial musculature, proglottid number, testes number, and genital pore position. Difficulty in differentiating these 4 species from one another, and from other species of Paraorygmatobothrium, points to limitations in the use of morphology alone to recognize species diversity in this genus. A phylogenetic analysis of Paraorygmatobothrium and related genera, based on partial (D1-D3) 28S rDNA sequence data, confirms that the genus is not monophyletic. Genetic distances of COI revealed that sequence divergence between species of Paraorygmatobothrium is generally at least an order of magnitude larger than COI divergence within a species. Patterns of host use for 3 of the new species are consistent with previous findings that point to a lack of host specificity in Paraorygmatobothrium.


Assuntos
Cestoides/classificação , Infecções por Cestoides/veterinária , Doenças dos Peixes/parasitologia , Tubarões/parasitologia , Animais , Oceano Atlântico , Cestoides/genética , Cestoides/fisiologia , Cestoides/ultraestrutura , Infecções por Cestoides/parasitologia , DNA Ribossômico/química , Complexo IV da Cadeia de Transporte de Elétrons/genética , Golfo do México , Especificidade de Hospedeiro , Intestinos/parasitologia , Microscopia Eletrônica de Varredura/veterinária , Filogenia , RNA Ribossômico 28S/genética , Tubarões/classificação
4.
J Parasitol ; 106(1): 157-166, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-32053468

RESUMO

This study provides additional taxonomic features based on scanning electron microscopy (SEM) and molecular data for Paracamallanus cyathopharynx ( Baylis, 1923 ) (Nematoda: Camallanidae). Parasites were collected from the posterior end of the intestine of cultured freshwater Sharptooth catfish Clarias gariepinus (Burchell, 1822) from Kibos fish farm, Kisumu County, Kenya, from December 2017 to March 2018. Additional taxonomic features recorded for P. cyathopharynx include the occurrence of 4-5 equal length digitate processes on the caudal end of the female, 4 processes (2 smaller and 2 larger) on the male caudal end, and a description of the shape of the distal tip of the right spicule. The study provides SEM images of the exposed sclerotized buccal capsule. This gives more information on the tridents, the sclerotized plate that extends laterally from the buccal capsule, and the narrow isthmus separating the anterior buccal capsule from the posterior. The prevalence, intensity, mean intensity, and mean abundance was 52.91%, 2-38, 12.37 and 6.54, respectively. 18S rDNA fragments were amplified, sequenced, and compared to other camallanid taxa, and 18S data confirmed the identity of the newly obtained sequences (MN396556) as that of P. cyathopharynx, being identical to sequence DQ813445 from Tanzania. This represents the first geographical record of P. cyathopharynx in Kenya.


Assuntos
Peixes-Gato/parasitologia , Doenças dos Peixes/parasitologia , Infecções por Spirurida/veterinária , Spirurina/genética , Spirurina/ultraestrutura , Animais , DNA Ribossômico/química , Feminino , Doenças dos Peixes/epidemiologia , Pesqueiros , Água Doce , Intestinos/parasitologia , Quênia/epidemiologia , Masculino , Microscopia Eletrônica de Varredura/veterinária , Prevalência , RNA Ribossômico 18S/genética , Alinhamento de Sequência/veterinária , Análise de Sequência de DNA/veterinária , Infecções por Spirurida/epidemiologia , Infecções por Spirurida/parasitologia
5.
Gene ; 737: 144433, 2020 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-32014563

RESUMO

The Carassius auratus (crucian carp) complex of the Dongting water system exhibits coexistence of diploid and triploid forms. As reported, triploid C. auratus is autotriploid origin. Ribosomal DNA (rDNA) with evolutionary conservation is widely used to study polyploidization. Here, we investigated genomic and transcribed rDNA sequences (18S and 5S) in diploid (2nCC, 2n = 100) and triploid (3nCC, 3n = 150) C. auratus. The results showed that the genetic traits and expression of 18S and 5S rDNA from 2nCC individuals were identified in 3nCC individuals. Moreover, pseudogenization of rDNA (18S and 5S) sequences were also observed in both 2nCC and 3nCC individuals, but expression of these variants was not detected. Based on the transcribed rDNA consensus sequence between 2nCC and 3nCC individuals, the functional secondary structures of 18S rRNA (expansion segments, ES6S) and 5S rRNA were predicted. These data demonstrated that complex evolutionary dynamics existed in the rDNA family of C. auratus. The evolutionary conservation of rDNA revealed that autotriploidization could not induce the divergence in Carassius taxa of the Dongting water system. These observations will expand our knowledge of the evolutionary dynamics of the rDNA family in vertebrates.


Assuntos
DNA Ribossômico/genética , Carpa Dourada/genética , RNA Ribossômico 18S/genética , RNA Ribossômico 5S/genética , Triploidia , Animais , Sequência de Bases , DNA Ribossômico/química , Evolução Molecular , Carpa Dourada/classificação , Conformação de Ácido Nucleico , Filogenia , RNA Ribossômico 18S/química , RNA Ribossômico 5S/química , Alinhamento de Sequência
6.
J Parasitol ; 106(1): 71-81, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31995717

RESUMO

An unusual coccidian parasite was described previously from the prostate of a male Antechinus flavipes (family: Dasyuridae; common name: yellow-footed antechinus). Morphometrics and a partial nuclear 18S small subunit rDNA (18S rDNA) sequence were used to assign this parasite to the genus Eimeria; it was named Eimeria taggarti. We generated full nuclear 18S rDNA and mitochondrial genome sequences from this parasite and used the newly completed 18S rDNA and mitochondrial cytochrome c oxidase subunit I (COI) sequences to perform a more in-depth phylogenetic analysis. The parasite clustered closely with Choleoeimeria spp. and Acroeimeria spp. infecting herptiles in a well-supported clade that was the sister lineage to the Eimeriidae sensu stricto. The mitochondrial genome of this parasite contained 2 inverted segments compared to mitochondrial genomes from parasites in the Eimeriidae sensu stricto (i.e., Stieda body-possessing coccidia with 4 dizoic sporocysts); this mitochondrial genome arrangement was shared with the only Choleoeimeria species for which sequence data were available publicly. Examination of histological preparations and TEM images uncovered bivalvate sporocysts and otherwise confirmed previously described morphological features of the parasite. Based on our phylogenetic analyses and histological observations, we propose the generic reclassification of E. taggarti to Choleoeimeria taggarti n. comb.


Assuntos
Coccidiose/veterinária , Eimeriidae/genética , Genoma Mitocondrial/genética , Marsupiais/parasitologia , Próstata/parasitologia , Animais , Coccidiose/parasitologia , DNA de Protozoário/química , DNA Ribossômico/química , Eimeriidae/classificação , Eimeriidae/isolamento & purificação , Eimeriidae/ultraestrutura , Complexo IV da Cadeia de Transporte de Elétrons/genética , Masculino , Anotação de Sequência Molecular , Oocistos/ultraestrutura , Filogenia , Reação em Cadeia da Polimerase , RNA de Protozoário/genética , RNA Ribossômico 18S/genética , Alinhamento de Sequência
7.
Parasit Vectors ; 13(1): 33, 2020 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-31959216

RESUMO

BACKGROUND: Triatomines are natural vectors of Chagas disease and are mainly prevalent in the Americas. In China, previous data from decades ago showed that there were two species of triatomine bugs, Triatoma rubrofasciata and T. sinica. However, the distribution, genetic characteristics and public health implications of triatomines in China are still relatively unknown. In order to gain knowledge on the distribution, genetic characteristics and public health implications of the triatomines in Guangxi, China, an entomological-epidemiological study and genetic research was conducted. METHODS: Different methods were used to elucidate the distribution of triatomines in Guangxi including consultations with county-level Center for Disease Prevention and Control staff and village doctors, the distribution of educational material on triatomines though the internet and social media apps such as Wechat and QQ, and conducting manual inspections and light trapping to collect triatomines. The morphological characteristics of the collected triatomines were identified under light microscopy. The mitochondrial 16S rRNA, cytochrome b (cytb) genes and nuclear 28S rRNA gene were amplified, sequenced and used in phylogenetic analyses. RESULTS: A total of 305 triatomines were captured from 54 different sites in 13 cities in Guangxi. All collected bugs were identified as T. rubrofasciata based on morphology. Most triatomine collection sites were around or inside houses. Four triatomines bite cases were observed during the investigation indicating that triatomine bites are common, the bites can cause serious anaphylaxis and skin papules and urticaria, suggesting a systemic skin response. The 16S rRNA, 28S rRNA and cytb sequence analyses of T. rubrofasciata from Guangxi and other countries showed that T. rubrofasciata sequences from different regions exhibit a high similarity, with no geographical differences. The phylogenetic tree based on the 16S rRNA and cytb genes showed that T. rubrofasciata sequences from different regions and continents were in the same cluster, indicating no differentiation among different geographical populations. CONCLUSIONS: Our study showed that T. rubrofasciata is widely distributed in Guangxi and that people are commonly bitten by this insect in some regions. This highlights the need to enhance surveillance for and control of T. rubrofasciata and to strengthen the monitoring of imported Trypanosoma cruzi in China. The 16S rRNA, 28S rRNA and cytb sequence analyses of T. rubrofasciata from different regions and continents suggested that T. rubrofasciata populations exhibit high similarity, and the clustering in the phylogenetic analyses indicates that T. rubrofasciata has a close ancestor originating in the Americas.


Assuntos
Doença de Chagas/transmissão , Mordeduras e Picadas de Insetos/epidemiologia , Insetos Vetores/fisiologia , Triatoma/fisiologia , Adulto , Animais , Doença de Chagas/epidemiologia , China/epidemiologia , Citocromos b/genética , DNA/química , DNA/isolamento & purificação , DNA Ribossômico/química , DNA Ribossômico/isolamento & purificação , Feminino , Habitação , Humanos , Mordeduras e Picadas de Insetos/parasitologia , Mordeduras e Picadas de Insetos/patologia , Insetos Vetores/classificação , Insetos Vetores/genética , Insetos Vetores/parasitologia , Masculino , Pessoa de Meia-Idade , Filogenia , RNA Ribossômico 16S/genética , RNA Ribossômico 28S/genética , Alinhamento de Sequência , Triatoma/classificação , Triatoma/genética , Triatoma/parasitologia
8.
Epidemiol Infect ; 148: e6, 2020 01 14.
Artigo em Inglês | MEDLINE | ID: mdl-31933451

RESUMO

Cervids represent a mammal group which plays an important role in the maintenance of ecological balance. Recent studies have highlighted the role of these species as reservoirs for several arthropods-borne pathogens. Globally, hemotropic mycoplasmas (haemoplasmas) are emerging or remerging bacteria that attach to red blood cells of several mammals species causing hemolytic anaemia. Therefore, the aim of this study was to investigate the occurrence and assess the phylogenetic positioning of Mycoplasma ovis in free-ranging deer from Brazil. Using a polymerase chain reaction targeting the 16S rRNA region, 18 (40%) out of 45 sampled deer were positive to M. ovis. Among the nine sequences analysed, four distinct genotypes were identified. The sequences detected in the present study were closely related to sequences previously identified in deer from Brazil and the USA. On the other hand, the Neighbour-Net network analysis showed that the human-associated M. ovis genotypes were related to genotypes detected in sheep and goats. The present study shows, for the first time, the occurrence of M. ovis in Mazama gouazoubira and Mazama bororo deer species, expanding the knowledge on the hosts harbouring this haemoplasma species. Once several deer species have your population in decline, additional studies are needed to evaluate the pathogenicity of M. ovis among deer populations around the world and assess its potential as reservoir hosts to human infections.


Assuntos
Doenças dos Animais/epidemiologia , Doenças dos Animais/microbiologia , Cervos/microbiologia , Variação Genética , Infecções por Mycoplasma/veterinária , Mycoplasma/classificação , Mycoplasma/isolamento & purificação , Animais , Brasil , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Genótipo , Mycoplasma/genética , Infecções por Mycoplasma/epidemiologia , Infecções por Mycoplasma/microbiologia , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
9.
Mol Phylogenet Evol ; 145: 106727, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-31899222

RESUMO

Trichophoreae is a nearly cosmopolitan Cyperaceae tribe that contains ~17 species displaying striking variation in size, inflorescence complexity, and perianth morphology. Although morphologically distinct, the status of its three genera (Cypringlea, Oreobolopsis and Trichophorum) is controversial because recent phylogenetic studies have suggested they might not be reciprocally monophyletic. However, previous analyses have shown conflicting topologies and consistently poor support due to an initial rapid diversification of the tribe. We analysed restriction-site associated DNA sequencing (RADseq) data from nearly all species of the clade, combined with five Sanger-based markers (matK, ndhF, rps16, ETS-1f, ITS) sampled extensively within species. This approach allowed us to resolve deep and shallow relationships within Trichophoreae for the first time, despite an anomaly zone spanning several successive short branches that produced considerable gene tree incongruence. Analyses reveal a primary phylogenetic split of the tribe into two clades roughly corresponding to an East Asian-North American disjunction that dates back to the mid-Miocene, with both clades comprised of a mixture of reduced unispicate and larger taxa with highly compound inflorescences. Morphological characters traditionally used in the circumscription of Trichophoreae genera are shown to be homoplasious. Several of these characters correlate best with climatic conditions, with the most reduced species occurring in open habitats at high latitudes and altitudes. Close relatives with highly compound inflorescences are found in temperate or subtropical forest understories. Cypringlea and Oreobolopsis are deeply nested within Trichophorum, and we merge all three genera into a more broadly circumscribed Trichophorum. We also show that Scirpus filipes is another previously unrecognized East Asian species of Trichophorum with highly compound inflorescences.


Assuntos
Cyperaceae/classificação , Teorema de Bayes , Biodiversidade , Cyperaceae/anatomia & histologia , Cyperaceae/genética , DNA de Plantas/química , DNA de Plantas/metabolismo , DNA Ribossômico/química , DNA Ribossômico/metabolismo , Funções Verossimilhança , Filogenia , Filogeografia , Plastídeos/genética , Análise de Sequência de DNA
10.
Appl Microbiol Biotechnol ; 104(2): 765-774, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31776608

RESUMO

25-hydroxyvitamin D3 has attracted considerable attention due to its great medical value and huge market demand in animal husbandry. Microbial production of 25-hydroxyvitamin D3 has been recognized as an alternative superior to traditional chemical synthesis. In this study, a Gram-positive bacteria zju 4-2 (CCTCC M 2019385) was isolated from the soil using vitamin D3 as the sole carbon source and was identified as Bacillus cereus according to its physiological characteristics and 16S rRNA analysis, which also showed a relatively high capacity for 25-hydroxyvitamin D3 production. Through systematic optimization of different catalytic conditions, the optimal solvent system of vitamin D3, vitamin D3 addition time and concentration, temperature, and pH were shown to be propylene glycol/ethanol (v/v = 9:1), early stationary phase, 2 g/L, 37 °C, and pH 7.2, respectively. With these optimal conditions, 796 mg/L of 25-hydroxyvitamin D3 was achieved after 48 h bioconversion with zju 4-2 at the shake flask level. Finally, up to 830 mg/L 25-hydroxyvitamin D3 with a yield of 41.5% was obtained in a 5 L fermentation tank. Our developed biotransformation process with this newly isolated strain provides a platform to produce 25-hydroxyvitamin D3 efficiently at industrialization scale.


Assuntos
Bacillus cereus/isolamento & purificação , Bacillus cereus/metabolismo , Calcifediol/metabolismo , Hormônios e Agentes Reguladores de Cálcio/metabolismo , Colecalciferol/metabolismo , Bacillus cereus/classificação , Bacillus cereus/genética , Biotransformação , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Fermentação , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Microbiologia do Solo , Temperatura
11.
Bull Environ Contam Toxicol ; 104(2): 200-205, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31781814

RESUMO

Phenoxy acid 2,4-D (2,4-dichlorophenoxy acid) is one of the most commonly-used herbicide in agriculture. Biodegradation of 2,4-D can be stimulated by structurally-related plant secondary metabolites such as ferulic acid (FA). The aim of this study is to: (1) assess the potential of indigenous soil bacteria to degrade 2,4-D in the presence of FA by PCR analysis of functional tfdA genes, (2) to determine the influence of 2,4-D and FA on samples ecotoxicity using Phytotoxkit® and Microtox® biotests. The detection of tfdA genes varied depending on the enrichment of samples with FA. FA suppressed detection of the tfdA genes, 100 µM 2,4-D induced higher detection of studied amplicons, while 500 µM 2,4-D delayed their detection. The ecotoxicity response was specific and differed between plants (PE% Lepidium sativum > Sinapis alba > Sorghum saccharatum) and bacteria (PE% up to 99% for Vibrio fischeri). Our findings confirm that 2,4-D and FA had a toxic influence on the used organisms.


Assuntos
Ácido 2,4-Diclorofenoxiacético/análise , Biodegradação Ambiental , Ácidos Cumáricos/análise , Herbicidas/análise , Poluentes do Solo/análise , Ácido 2,4-Diclorofenoxiacético/metabolismo , Aliivibrio fischeri/metabolismo , Ácidos Cumáricos/metabolismo , DNA Ribossômico/química , Genes Bacterianos , Herbicidas/metabolismo , Lepidium sativum/metabolismo , Extratos Vegetais , Sinapis/metabolismo , Solo , Microbiologia do Solo , Poluentes do Solo/metabolismo , Sorghum/metabolismo
12.
Vet Parasitol ; 276: 108965, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31726324

RESUMO

Tritrichomonas foetus isolates from feline and bovine origin has been previously shown to carry a certain degree of genetic heterogeneity. Here, novel candidate molecular markers were developed by means of multilocus sequence typing of the gap2 gene (encoding for T. foetus glyceraldehyde-3-phosphate dehydrogenase), ITS region, the TR7/TR8 variable-length repeat and microsatellite genotyping. These markers were used to characterize T. foetus field isolates from bulls and domestic cats and to compare phylogenetically with the following ATCC isolates: T. foetus isolated from cattle and pig (syn. Tritrichomonas suis), Tritrichomonas mobilensis, Tetratrichomonas gallinarum and Pentatrichomonas hominis. Among them, TFMS10 and TFMS7 were found to be the most polymorphic markers. Moreover, an 809 bp fragment of the gap2 gene was successfully amplified from all the trichomonads included in this study and the sequence analysis revealed differences between T. foetus porcine and feline genotypes and T. mobilensis in comparison to the bovine T. foetus ATCC isolate. The TR7/TR8 repeat pattern was not reproducible, being only consistent the fragments of approximately 110 and 217 bp. Sequence analysis of the latter revealed the existence of 3 SNPs resulting in 98.6 % homology between bovine and feline isolates. A search for similar sequences was carried out to develop a Restriction Length Fragment Polymorphism analysis. A 503 bp region, named TF1, revealed the existence of two BbvI restriction enzyme sites that were able to generate different length fragments for T. foetus feline and bovine isolates. Finally, the neighbour-joining analyses showed that T. foetus porcine genotype clusters together with bovine genotype, whereas T. mobilensis and the feline genotype form a separate cluster.


Assuntos
Doenças do Gato/parasitologia , Doenças dos Bovinos/parasitologia , Marcadores Genéticos , Infecções Protozoárias em Animais/parasitologia , Tritrichomonas foetus/genética , Animais , Sequência de Bases , Gatos , Bovinos , Sequência Consenso , DNA Ribossômico/química , Gliceraldeído-3-Fosfato Desidrogenase (Fosforiladora)/genética , Masculino , Repetições de Microssatélites/genética , Repetições Minissatélites , Dados de Sequência Molecular , Tipagem de Sequências Multilocus/veterinária , Filogenia , Polimorfismo de Fragmento de Restrição , Polimorfismo de Nucleotídeo Único , Sequências Repetitivas de Ácido Nucleico/genética , Alinhamento de Sequência/veterinária , Tritrichomonas foetus/classificação
13.
Genes Dev ; 33(23-24): 1688-1701, 2019 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-31727772

RESUMO

Human nucleolar organizer regions (NORs), containing ribosomal gene (rDNA) arrays, are located on the p-arms of acrocentric chromosomes (HSA13-15, 21, and 22). Absence of these p-arms from genome references has hampered research on nucleolar formation. Previously, we assembled a distal junction (DJ) DNA sequence contig that abuts rDNA arrays on their telomeric side, revealing that it is shared among the acrocentrics and impacts nucleolar organization. To facilitate inclusion into genome references, we describe sequencing the DJ from all acrocentrics, including three versions of HSA21, ∼3 Mb of novel sequence. This was achieved by exploiting monochromosomal somatic cell hybrids containing single human acrocentric chromosomes with NORs that retain functional potential. Analyses revealed remarkable DJ sequence and functional conservation among human acrocentrics. Exploring chimpanzee acrocentrics, we show that "DJ-like" sequences and abutting rDNA arrays are inverted as a unit in comparison to humans. Thus, rDNA arrays and linked DJs represent a conserved functional locus. We provide direct evidence for exchanges between heterologous human acrocentric p-arms, and uncover extensive structural variation between chromosomes and among individuals. These findings lead us to revaluate the molecular definition of NORs, identify novel genomic structural variation, and provide a rationale for the distinctive chromosomal organization of NORs.


Assuntos
Cromossomos/química , Cromossomos/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Região Organizadora do Nucléolo/química , Região Organizadora do Nucléolo/genética , Animais , Sequência de Bases , Linhagem Celular , Sequência Conservada/genética , Estruturas Genéticas/genética , Variação Genética , Humanos , Células Híbridas , Camundongos , Pan troglodytes/genética
14.
Parasit Vectors ; 12(1): 543, 2019 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-31730024

RESUMO

BACKGROUND: Cryptosporidium spp. and Giardia duodenalis are major intestinal pathogens that can cause diarrheal diseases in humans, especially children. Enterocytozoon bieneusi is another parasite which can cause gastrointestinal tract disorders, with diarrhea being the main clinical symptom. However, few genetic studies of these parasites in pediatric inpatients in China have been published. METHODS: To assess the genetic characteristics and epidemiological status of these parasites, a total of 2284 fecal samples were collected from children in the pediatric departments of three hospitals in Zhengzhou, central China, and screened for these protozoans with PCR, based on the small subunit ribosomal RNA (SSU rRNA) genes of Cryptosporidium spp. and G. duodenalis and the internal transcribed spacer (ITS) of E. bieneusi. RESULTS: Six (0.26%), 14 (0.61%), and 27 (1.18%) of the samples were positive for Cryptosporidium spp., G. duodenalis and E. bieneusi, respectively. Of the 12 successfully sequenced G. duodenalis isolates, four were identified as assemblage A and eight as assemblage B. In subtype and multilocus genotype (MLG) analyses, C. parvum IIdA19G1 (n = 4) and two novel G. duodenalis MLGs belonging to subassemblage AII (n = 3) and BIV (n = 5) were successfully identified. The E. bieneusi isolates included genotypes D (n = 17), J (n = 2), PigEBITS7 (n = 1), BEB6 (n = 1), and CM8 (n = 1). This is the first report of C. parvum subtype IIdA19G1 in HIV-negative children and E. bieneusi genotype CM8 in humans. CONCLUSIONS: The dominance of zoonotic C. parvum subtype IIdA19G1 indicates that this parasite is turning into zoonotic origin from human-to-human transmission. The phylogenetic analysis also revealed the zoonotic origins and anthroponotic transmission potential of G. duodenalis and E. bieneusi, suggesting more efforts must be made to minimize the threat these pathogens pose to public health.


Assuntos
Cryptosporidium/classificação , Cryptosporidium/genética , Enterocytozoon/classificação , Enterocytozoon/genética , Fezes/parasitologia , Giardia lamblia/classificação , Giardia lamblia/genética , Adolescente , Criança , Criança Hospitalizada , Pré-Escolar , China , Análise por Conglomerados , Criptosporidiose/parasitologia , Cryptosporidium/isolamento & purificação , DNA de Protozoário/química , DNA de Protozoário/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Transmissão de Doença Infecciosa , Enterocytozoon/isolamento & purificação , Feminino , Variação Genética , Genótipo , Giardia lamblia/isolamento & purificação , Giardíase/parasitologia , Hospitais Pediátricos , Humanos , Lactente , Masculino , Microsporidiose/parasitologia , Filogenia , RNA Ribossômico 18S/genética , Análise de Sequência de DNA
15.
J Parasitol ; 105(6): 878-881, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31730392

RESUMO

The microsporidium Nosema sp. SE is a pathogen that infects the beet armyworm Spodoptera exigua. The complete sequence of its 4,302-base pair (bp) ribosomal ribonucleic acid (rRNA) gene region was obtained by polymerase chain reaction amplification and sequencing. The rRNA organization of Nosema sp. SE was 5'-large subunit (LSU) rRNA-internal transcribed spacer-small subunit (SSU) rRNA-intergenic spacer-5S-3', which corresponded to the pattern of Nosema bombycis. Phylogenetic analysis based on LSU rRNA and SSU rRNA both indicated that the parasite had a close relationship with other true Nosema species, confirming that Nosema sp. SE belongs to true Nosema group of the genus Nosema.


Assuntos
Beta vulgaris/parasitologia , Nosema/genética , Spodoptera/microbiologia , Animais , DNA Fúngico/isolamento & purificação , DNA Ribossômico/química , DNA Ribossômico/genética , Estágios do Ciclo de Vida , Nosema/classificação , Nosema/crescimento & desenvolvimento , Nosema/ultraestrutura , Filogenia , RNA Ribossômico/química , Alinhamento de Sequência
16.
J Parasitol ; 105(5): 821-826, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31670613

RESUMO

Codonocephalus is a monotypic genus of diplostomid digeneans and is the only genus in the sub-family Codonocephalinae. The type-species Codonocephalus urniger has an unusual progenetic metacercaria that uses frogs as intermediate hosts and can use snakes as paratenic hosts. Adult C. urniger parasitize ardeid wading birds in the Palearctic. Despite the broad distribution of Codonocephalus, no DNA sequence data are currently available for the genus. In this study, we generated sequence data for nuclear ribosomal and mitochondrial DNA from progenetic metacercaria of the type-species C. urniger from marsh frog, Pelophylax ridibundus, collected in Ukraine. We used partial sequences of the nuclear ribosomal 28S gene to examine for the first time the phylogenetic position of Codonocephalus among the Diplostomoidea.


Assuntos
Filogenia , Ranidae/parasitologia , Trematódeos/classificação , Infecções por Trematódeos/veterinária , Animais , Teorema de Bayes , DNA de Helmintos/química , DNA de Helmintos/isolamento & purificação , DNA Ribossômico/química , Complexo IV da Cadeia de Transporte de Elétrons/química , Complexo IV da Cadeia de Transporte de Elétrons/genética , Estágios do Ciclo de Vida , Metacercárias/classificação , Metacercárias/genética , Microscopia Eletrônica de Varredura/veterinária , Mitocôndrias/enzimologia , RNA de Helmintos/genética , RNA Nuclear/genética , RNA Ribossômico 28S/genética , Trematódeos/genética , Trematódeos/ultraestrutura , Infecções por Trematódeos/parasitologia
17.
Parasit Vectors ; 12(1): 515, 2019 Nov 04.
Artigo em Inglês | MEDLINE | ID: mdl-31685003

RESUMO

BACKGROUND: Strongyloidiasis is a health problem in Vietnam, but appropriate information is still limited. The aim of this study was to determine the prevalence, geographical distribution, epidemiological aspects, symptoms and other health indicators of Strongyloides stercoralis infections in patients from 27 provinces of northern Vietnam attending the Hanoi Medical University Hospital during 2016 and 2017. METHODS: Blood samples of 2000 patients were analyzed for S. stercoralis infection with an IgG ELISA test. Seroprevalence was analyzed by gender, age group, locality of origin (rural or urban areas) and symptoms. Stools from the seropositive patients were examined for the detection of worms which were subsequently used for species identification by morphology and rDNA ITS1 sequencing. RESULTS: A seroprevalence of 20% was detected, showing an increasing prevalence from young to older age groups but without significant gender difference. Seroprevalence was higher in rural areas than in urban areas, both in general and individually in all provinces without exception, and lower in the mountainous areas than in the large valley lowlands. The follow-up of the 400 patients showed eosinophilia in 100% of cases, diarrhoea in 64.5%, digestion difficulties in 58.0%, stomachache in 45.5%, stomach and duodenal ulcers in 44.5%, itching in 28.0% and fever in 9.5%. The prevalence of symptoms and signs were also higher in older age groups than in younger age groups. Worms were detected in stools of 10.5% of the patients. Sequencing of a 501-bp nuclear ribosomal DNA ITS1 fragment allowed for the verification of infection by Strongyloides stercoralis. CONCLUSIONS: To our knowledge, this study is the largest survey of human strongyloidiasis in Vietnam so far and the first molecular identification of this nematode species in this country. Long-term chronicity may probably be usual in infected subjects, mainly in the older age groups.


Assuntos
Strongyloides stercoralis/genética , Estrongiloidíase/epidemiologia , Adolescente , Adulto , Distribuição por Idade , Idoso , Animais , Anticorpos Anti-Helmínticos/sangue , DNA de Helmintos/análise , DNA de Helmintos/química , DNA Ribossômico/análise , DNA Ribossômico/química , Diarreia/epidemiologia , Diarreia/parasitologia , Eosinofilia/epidemiologia , Eosinofilia/parasitologia , Feminino , Geografia , Humanos , Imunoglobulina G/sangue , Enteropatias Parasitárias/epidemiologia , Enteropatias Parasitárias/parasitologia , Masculino , Pessoa de Meia-Idade , Prevalência , População Rural , Estudos Soroepidemiológicos , Strongyloides stercoralis/imunologia , Strongyloides stercoralis/isolamento & purificação , Estrongiloidíase/parasitologia , Vietnã/epidemiologia , Adulto Jovem
18.
Parasit Vectors ; 12(1): 506, 2019 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-31666114

RESUMO

BACKGROUND: Chagas disease (CD) is caused by the protozoan parasite Trypanosoma cruzi and is transmitted by triatomine insects. Clinical manifestations vary according to the phase of the disease. Cutaneous manifestations are usually observed in the acute phase (chagoma and Romaña's sign) or after reactivation of the chronic phase by immunosuppression; however, a disseminated infection in the acute phase without immunosuppression has not been reported for CD. Here, we report an unusual case of disseminated cutaneous infection during the acute phase of CD in a Mexican woman. METHODS: Evaluation of the patient included a complete clinical history, a physical exam, and an exhaustive evaluation by laboratory tests, including ELISA, Western blot and PCR. RESULTS: Skin biopsies of a 50-year-old female revealed intracellular parasites affecting the lower extremities with lymphangitic spread in both legs. The PCR tests evaluated biopsy samples obtained from the lesions and blood samples, which showed a positive diagnosis for T. cruzi. Partial sequencing of the small subunit ribosomal DNA correlated with the genetic variant DTU II; however, serological tests were negative. CONCLUSIONS: We present a case of CD with disseminated skin lesions that was detected by PCR and showed negative serological results. In Mexico, an endemic CD area, there are no records of this type of manifestation, which demonstrates the ability of the parasite to initiate and maintain infections in atypical tissues .


Assuntos
Doença de Chagas/diagnóstico , Dermatopatias Parasitárias/diagnóstico , Trypanosoma cruzi/imunologia , Doença Aguda , Anticorpos Antiprotozoários/sangue , Antígenos de Protozoários/imunologia , Western Blotting , DNA de Protozoário/isolamento & purificação , DNA Ribossômico/química , Diagnóstico Diferencial , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Perna (Membro)/parasitologia , Perna (Membro)/patologia , Sistema Linfático/parasitologia , México , Pessoa de Meia-Idade , Filogenia , Reação em Cadeia da Polimerase , Proteínas de Protozoários/imunologia , Alinhamento de Sequência , Pele/parasitologia , Pele/patologia , Trypanosoma cruzi/classificação , Trypanosoma cruzi/genética , Trypanosoma cruzi/isolamento & purificação
19.
Int J Parasitol ; 49(12): 945-961, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31628939

RESUMO

We explore the growing issue of cryptic speciation in the Digenea through study of museum material and newly collected specimens consistent with the enenterid genus Koseiria from five species of the Kyphosidae and Chaetodontoplus meredithi Kuiter (Pomacanthidae) collected in the Indo-Pacific. We use an integrated approach, employing traditional morphometrics, principal components analysis (PCA), and molecular data (ITS2 and 28S rDNA). Our results support recombination of Koseiria allanwilliamsi Bray & Cribb, 2002 as Proenenterum allanwilliamsi (Bray & Cribb, 2002) n. comb. and transfer of Koseiria huxleyi Bray & Cribb, 2001 to a new genus as Enenterageitus huxleyi (Bray & Cribb, 2002) n. comb. Molecular data indicate the presence of four further species consistent with Koseiria, one from Western Australia (sequence data only) and three from eastern Australia. All three eastern Australian species are morphologically consistent with Koseiria xishaensis Gu & Shen, 1983, but distinct from all other previously described species. Although K. xishaensis has been reported from Australia, we conclude that the similarity of the present forms to the original description of K. xishaensis means records of this species from Japan, Palau and Australia are unreliable. Because the eastern Australian forms cannot be reliably ascribed to K. xishaensis, we describe Koseiria argalea n. sp., Koseiria laiphopharophora n. sp., and Koseiria pyknophora n. sp., following application of PCAs and iterative refinement of species concepts and type series. These analyses did not allow convincing identification hypotheses for all specimens examined. In this genus, both morphological and molecular data, together with reliable host identifications, are essential for species recognition, and thus we refrain from attempting to name samples lacking molecular data. The issues presented by these taxa encapsulate those of trematodes in the region as a whole. Many records require dramatically improved supporting data, leading to substantial uncertainly in the identification of this fauna.


Assuntos
Peixes/parasitologia , Filogenia , Trematódeos/anatomia & histologia , Trematódeos/classificação , Animais , Biometria , Análise por Conglomerados , DNA de Helmintos/química , DNA de Helmintos/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Microscopia , Oceano Pacífico , RNA Ribossômico 28S/genética , Análise de Sequência de DNA , Trematódeos/genética , Trematódeos/isolamento & purificação
20.
Mycologia ; 111(6): 998-1027, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31613712

RESUMO

The Pleuroascaceae (Leotiomycetes) is introduced for Phialophora hyalina (section Catenulatae) and its closest relatives based on analyses of DNA sequences of five gene regions and the comparison of cultural and micromorphological characters. The family is resolved as a strongly supported clade that encompasses Pleuroascus and the new anamorph genera Entimomentora and Venustampulla. The latter includes V. parva, a species placed formerly in Scopulariopsis, and V. echinocandica, which is established for the echinocandin-producing isolate BP-5553. Entimomentora includes E. hyalina, a species based on the ex-type strain of Ph. hyalina. Additional isolates identified as Ph. hyalina are distantly related to the Pleuroacaceae and include Psychrophila antarctica (Arachnopezizaceae) and Cryonesomyces dreyfussii, the sole member of the new genus Cryonesomyces (incertae sedis). Isolates identified or deposited as Ph. alba are also not closely related; they include a species for which we propose the name Neobulgaria koningiana (Gelatinodiscaceae) and a second psychrophilic species that we describe as Psychrophila lagodekhiensis. Of the 13 isolates assessed for in vitro antifungal activity, only V. echinocandica inhibited the growth of Candida albicans.


Assuntos
Técnicas Microbiológicas , Microscopia , Phialophora/classificação , Phialophora/genética , Filogenia , Antifúngicos/análise , Antifúngicos/farmacologia , Cromatografia Líquida de Alta Pressão , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Genes de RNAr , Espectrometria de Massas , Testes de Sensibilidade Microbiana , Phialophora/crescimento & desenvolvimento , RNA Polimerase II/genética , RNA Fúngico/genética , RNA Ribossômico 18S/genética , RNA Ribossômico 28S/genética , Análise de Sequência de DNA
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