Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 4.474
Filtrar
1.
J Helminthol ; 97: e98, 2023 Dec 14.
Artigo em Inglês | MEDLINE | ID: mdl-38095098

RESUMO

Scaphanocephalus is a small trematode genus belonging to the family Opistorchiidae. The genus currently contains only three species associated with marine fish as intermediate hosts and fish-eating birds as definitive hosts. Here, specimens of Scaphanocephalus were collected from the Osprey, Pandion haliaetus, and the White mullet, Mugil curema in the Yucatán Peninsula, Mexico. We report for the first-time DNA sequences of adult specimens of Scaphanocephalus, particularly S. expansus, as well as a sequence of a different species sampled as metacercaria. Morphological comparisons of Scaphanocephalus expansus confirmed the identity of the adult specimens, with minor morphological variations; Scanning electron photomicrographs were included, and the species was re-described. Phylogenetic analysis based on 28S rDNA sequences showed that Scaphanocephalus is monophyletic within Opisthorchiidae and consists of three independent lineages. Sequences of adults are identical to those of S. expansus. Instead, the sequence of the metacercaria sampled from the mesentery of Mugil curema nested with specimens reported as Scaphanocephalus sp. from a labrid fish in the Mediterranean Sea, herein named it as Scaphanocephalus sp. 2.


Assuntos
Falconiformes , Doenças dos Peixes , Heterophyidae , Smegmamorpha , Trematódeos , Infecções por Trematódeos , Animais , México , Filogenia , DNA de Helmintos/genética , Heterophyidae/genética , Peixes , Metacercárias , Infecções por Trematódeos/veterinária
2.
Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi ; 35(5): 492-496, 2023 Dec 01.
Artigo em Chinês | MEDLINE | ID: mdl-38148538

RESUMO

OBJECTIVE: To investigate the prevalence of Echinococcus infections in wild carnivores in Serthar County, Sichuan Province, so as to provide insights into echinococcosis control in local areas. METHODS: Stool samples were collected from wild carnivores in Serthar County, Sichuan Province in May 2021, and the host sources of stool samples and Echinococcus infections were identified using PCR assays. The prevalence of E. multilocularis, E. granulosus and E. shiquicus infections was estimated in different hosts. RESULTS: A total of 583 stool samples were collected from wild carnivores, including 147 stool samples from fox, 154 from wolf, 227 from wild dogs and 11 from lynx. The overall prevalence of E. multilocularis, E. granulosus and E. shiquicus infections was 5.68%, 0.19% and 14.20% in canine stool samples, and no E. granulosus infection was detected in fox stool samples, while the prevalence of E. multilocularis and E. shiquicus infections was 0.68% and 47.62% in fox stool samples (χ2 = 88.41, P < 0.001). No E. granulosus or E. shiquicus infection was detected in wolf stool samples, and the prevalence of E. multilocularis infection was 10.39% in wolf stool samples. The prevalence of E. multilocularis, E. granulosus and E. shiquicus infections was 5.73%, 0.44% and 2.20% in canine stool samples (χ2 = 12.13, P < 0.01). In addition, the prevalence of E. multilocularis infections was significantly higher in wolf stool samples than in canine and fox stool samples (χ2 = 13.23, P < 0.01), and the prevalence of E. shiquicus infections was significantly higher in fox stool samples than in canine and wolf stool samples (χ2 = 187.01, P < 0.001). No Echinococcus infection was identified in 11 lynx stool samples. CONCLUSIONS: The prevalence of Echinococcus infections is high in wild canines in Serthar County, Sichuan Province. Wolf, wild dog and fox all participate in the wild life cycle of E. multilocularis in Serthar County, and wolf and wild dogs may play a more important role.


Assuntos
Carnívoros , Equinococose , Animais , Cães/microbiologia , China/epidemiologia , DNA de Helmintos/genética , Equinococose/epidemiologia , Equinococose/veterinária , Fezes , Raposas/microbiologia , Lynx/microbiologia , Prevalência , Lobos/microbiologia , Carnívoros/microbiologia
3.
Sci Rep ; 13(1): 19420, 2023 11 08.
Artigo em Inglês | MEDLINE | ID: mdl-37940673

RESUMO

Lymphatic filariasis is a mosquito borne disease which leads to abnormal painful enlarged body parts, severe disability and social stigma. We screened Wuchereria bancrofti in Matayos constituency in Busia County. Blood samples were collected from 23 villages selected purposively based on clinical case reports. Finger prick and/or venous blood sampling and mosquito collections was carried out. Antigenaemia and filarial DNA prevalence were determined. Infection rates on mosquito pools were estimated and SPSS version 26 was used for descriptive statistics analysis. A total of 262 participants were recruited, 73.3% (n = 192) of the participants had no symptoms, 14.1% (n = 5.3) had swollen legs, 5.3% (n = 14) had painful legs and 3.8% (n = 10) with scrotal swellings. Average antigenemia prevalence was 35.9% (n = 94) and DNA prevalence was at 8.0% (n = 21). A total of 1305 mosquitoes were collected and pooled into 2-20 mosquitoes of the same species and from the same village. Two pools out of 78 were positive for filarial DNA with a minimum infection rate of 0.15%. From this study, antigenaemia and infected mosquitoes are an indication of active transmission. The clinical signs are evidence that filarial infections have been in circulation for over 10 years. The global climate change phenomenon currently happening has been shown to adversely affect the transmission of vector borne diseases and is likely to increase lymphatic filariasis transmission in the area. This study therefore recommends further screening before Mass Drug Administration, morbidity management and enhanced mosquito control Programmes are recommended in the study area.


Assuntos
Culex , Filariose Linfática , Animais , Humanos , Wuchereria bancrofti , Quênia , Culex/genética , DNA de Helmintos/genética
4.
Res Vet Sci ; 165: 105049, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37856947

RESUMO

Haemonchus contortus is the most pathogenic and economically restrictive gastrointestinal nematode in the small ruminant industry globally. Morbidity, poor cross-bodily state, and mortality of sheep in Lesotho suggest the presence of H. contortus. The present study investigated the morphological, molecular, and population genetics of H. contortus third-stage larvae infecting sheep in four ecological zones (EZ) of Lesotho. Coprocultures were prepared for larval morphological identification and PCR determination. Larvae were identified morphologically as 100% H. contortus. The Second Internal Transcribed Spacer (ITS-2) gene of the ribosomal DNA of H. contortus isolates in the present study revealed nucleotide homology ranging from 97 to 100% when compared with selected GenBank reference sequences. Pairwise evolutionary divergence among H. contortus isolates was low, with 0.01318 recorded as the highest in the present study. Five haplotypes resulted from 14 Lesotho sequences. Haplotype diversity and nucleotide diversity were 0.76923 and 0.00590, respectively. Genetic differentiation among isolates was low but not statistically significant. An analysis of molecular variance revealed that most molecular variation was distributed within topographic populations at 94.79% (FST = 0.05206, p > 0.05) and 5.21% among populations. There was high gene flow and no definite population genetic structure among Lesotho isolates.


Assuntos
Hemoncose , Haemonchus , Doenças dos Ovinos , Animais , Ovinos/genética , Haemonchus/genética , Lesoto , Variação Genética , DNA Espaçador Ribossômico/genética , DNA Espaçador Ribossômico/química , Hemoncose/veterinária , DNA de Helmintos/genética , Carneiro Doméstico/genética , Genética Populacional , Ruminantes , Doenças dos Ovinos/genética , Doenças dos Ovinos/epidemiologia , Nucleotídeos
5.
Parasitol Res ; 122(11): 2467-2476, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-37642769

RESUMO

The aim of this study was to characterize the Tunisian Fasciola spp. flukes by morphometric and molecular analyses. Flukes were collected from livers of sheep slaughtered in Sejnane slaughterhouses (Bizerte gouvernorate, Northwest Tunisia) between January and March 2021.Five morphometric parameters were determined for all the liver flukes, as follows: (i) total body length (BL), (ii) distance between ventral sucker and the tail (VS-T), (iii) distance between oral sucker and ventral sucker (OS-VS), (iv) abdomen diameter (AD), (v) tail diameter (TD) and the body length to width ratio (BL/BW). Molecular identification of the fluke specimens was carried out by polymerase chain reaction, restriction fragment polymorphism (PCR-RFLP) of a 680 bp sequence of the internal transcribes spacer 1 (ITS1) gene and by amplification, sequencing, and phylogenetic analysis of a 500 bp sequence of the ITS2 gene. Morphometric measurements showed that the mean of the total body length of the adult flukes was 21.1 ± 2.7 mm with minimum and maximum lengths of 13 and 31 mm, respectively. The PCR-RFLP analysis revealed a single profile consisting of three bands of approximately 370, 100, and 60 bp. Fasciola sequences described in the present study (GenBank numbers: OQ457027 and OQ457028) showed 99.58-100% identity to Fasciola hepatica. In conclusion, the results of this study show that molecular and phylogenetic analyses confirm the presence of a single species of F. hepatica in the Sejnane region Northwest of Tunisia. However, further studies are needed to identify the occurrence of Fasciola species in other Tunisian regions.


Assuntos
Doenças dos Bovinos , Fasciola hepatica , Fasciola , Fasciolíase , Ovinos/genética , Animais , Bovinos , Fasciola/genética , Filogenia , Tunísia/epidemiologia , Fasciolíase/epidemiologia , Fasciolíase/veterinária , Fasciola hepatica/genética , Doenças dos Bovinos/epidemiologia , DNA de Helmintos/genética
6.
Infect Dis Poverty ; 12(1): 71, 2023 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-37550723

RESUMO

BACKGROUND: Schistosomiasis remains a public health issue and the need for accurate and affordable diagnostics is crucial in the elimination of the disease. While molecular diagnostics are highly effective, they are expensive, with the main costs been associated with DNA extraction. The DNA dipstick is a rapid, affordable and simple purification method that allows DNA to be extracted from diagnostic samples within 30 s. We aimed to optimise the DNA dipstick method for samples from mice and egg-spiked human samples. METHODS: Urine, blood and faeces were collected from mice exposed to Schistosoma japonicum infection at weekly intervals from Day 0 to Day 42. Urine and faecal samples were also collected from volunteer, uninfected humans and spiked with S. japonicum eggs. All samples were subject to several optimisation procedures and DNA extracted with the DNA dipstick. Amplification of the target DNA was carried out using LAMP and visualised using agarose gel electrophoresis and flocculation. RESULTS: The DNA dipstick successfully identified S. japonicum from infected mice and human clinical samples spiked with cracked eggs or genomic DNA from S. japonicum. Amplification was observed from week 4 post infection in infected mice. For human samples, amplification was observed in sieved faecal samples, filtered urine samples heated at 95 °C for 30 min, and sera samples heated at 95 °C for 30 min. CONCLUSIONS: The DNA dipstick combined with LAMP has huge potential in providing cost-effective, simple and accurate detection of schistosomiasis infection in endemic regions. This will allow for rapid treatment, tracking outbreaks-such as occur after typhoons, leading to better health outcomes and contributing to control and eventual elimination of schistosomiasis.


Assuntos
Schistosoma japonicum , Esquistossomose Japônica , Esquistossomose , Humanos , Camundongos , Animais , Técnicas de Amplificação de Ácido Nucleico/métodos , Esquistossomose Japônica/diagnóstico , Esquistossomose Japônica/tratamento farmacológico , Esquistossomose/diagnóstico , DNA de Helmintos , Sensibilidade e Especificidade
7.
J Helminthol ; 97: e37, 2023 Apr 18.
Artigo em Inglês | MEDLINE | ID: mdl-37070361

RESUMO

Ancylostoma caninum is the most common nematode parasite of dogs in the United States. The present study aimed to describe the molecular epidemiology of A. caninum isolates from the central and eastern states of the United States using the partial mitochondrial cytochrome oxidase (cox1) gene and to compare them with those reported globally. We isolated eggs from faecal samples of dogs and characterized each isolate based on cox1 sequences. A total of 60 samples originating from Kansas, Iowa, New York, Florida and Massachusetts were included. 25 haplotypes were identified in the United States dataset with high haplotype diversity (0.904). Sequence data were compared to sequences from other world regions available in GenBank. Global haplotype analysis demonstrated 35 haplotypes with a haplotype diversity of 0.931. Phylogenetic and network analysis provide evidence for the existence of moderate geographical structuring of A. caninum haplotypes. Our results provide an updated summary of A. caninum haplotypes and data for neutral genetic markers with utility for tracking hookworm populations. Sequences have been deposited in GenBank (ON980650-ON980674). Further studies of isolates from other regions are essential to understand the genetic diversity of this parasite.


Assuntos
Doenças do Cão , Parasitos , Estados Unidos/epidemiologia , Animais , Cães , Ancylostoma/genética , Parasitos/genética , Filogenia , DNA de Helmintos/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Doenças do Cão/epidemiologia , Doenças do Cão/parasitologia , Florida
8.
J Helminthol ; 97: e35, 2023 Apr 18.
Artigo em Inglês | MEDLINE | ID: mdl-37070390

RESUMO

Members of the genus Strigea Abildgaard, 1790 are endoparasites of birds distributed worldwide. Adults of an undescribed species of the genus Strigea were collected from the intestines of two hawk species (Rupornis magnirostris and Accipiter coperii). Other species identified as Parastrigea macrobursa that were described in Argentina were also recovered from two hawk species (Buteogallus urubitinga and Buteogallus anthracinus) in three localities along the coasts of Mexico. Specimens of the two species were sequenced for three molecular markers, the internal transcribed spacers locus (ITS1-5.8S rDNA- ITS2) and the domains D1-D3 from the large subunit from nuclear ribosomal DNA and the cytochrome c oxidase subunit 1 from mitochondrial DNA. The newly sequenced specimens were aligned with other strigeids sequences downloaded from GenBank. Maximum likelihood and Bayesian analyses inferred with each molecular marker revealed that our specimens of Strigea sp. formed an independent lineage, which is recognized herein as a new species, Strigea magnirostris n. sp., representing the first species in Mexico and the 16th in the Neotropical region. Morphologically, the new species is distinguished from other congeneric species from the Americas by having an oral sucker with several papillae around it, well-developed pseudosuckers (118-248 µm), a tegument covered with tiny spines, a larger cone genital (193-361 × 296-637) and a larger copulatory bursa (247-531 × 468-784). Our phylogenetic analyses revealed that P. macrobursa is not closely related to other members of the genus Parastrigea and is nested within Strigea, suggesting that P. macrobursa should be transferred to Strigea to form Strigea macrobursa n. comb., expanding its distribution range from Mexico to Argentina. Finally, the analyses also revealed that the taxonomy and systematics of Strigea should be re-evaluated, combining morphological and molecular characteristics.


Assuntos
Aves Predatórias , Trematódeos , Animais , Filogenia , Teorema de Bayes , Aves , DNA Ribossômico/genética , México , DNA de Helmintos/genética
9.
Int J Mol Sci ; 24(1)2023 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-36614336

RESUMO

Loop-mediated isothermal amplification (LAMP) is the most popular technology for point-of-care testing applications due its rapid, sensitive and specific detection with simple instrumentation compared to PCR-based methods. Many systems for reading the results of LAMP amplifications exist, including real-time fluorescence detection using fluorophore-labelled probes attached to oligonucleotide sequences complementary to the target nucleic acid. This methodology allows the simultaneous detection of multiple targets (multiplexing) in one LAMP assay. A method for multiplexing LAMP is the amplification by release of quenching (DARQ) technique by using a 5'-quencher modified LAMP primer annealed to 3'-fluorophore-labelled acting as detection oligonucleotide. The main application of multiplex LAMP is the rapid and accurate diagnosis of infectious diseases, allowing differentiation of co-infecting pathogens in a single reaction. Schistosomiasis, caused among other species by Schistosoma mansoni and strongyloidiasis, caused by Strongyloides stercoralis, are the most common helminth-parasite infections worldwide with overlapping distribution areas and high possibility of coinfections in the human population. It would be of great interest to develop a duplex LAMP to detect both pathogens in the same reaction. In this study, we investigate the use of our two previously developed and well-stablished LAMP assays for S. mansoni and Strongyloides spp. DNA detection in a new duplex real-time eight-primer system based on a modified DARQ probe method that can be performed in a portable isothermal fluorimeter with minimal laboratory resources. We also applied a strategy to stabilize the duplexed DARQ-LAMP mixtures at room temperature for use as ready-to-use formats facilitating analysis in field settings as point-of-care diagnostics for schistosomiasis and strongyloidiasis.


Assuntos
Esquistossomose , Strongyloides stercoralis , Estrongiloidíase , Animais , Humanos , Schistosoma mansoni/genética , Sistemas Automatizados de Assistência Junto ao Leito , DNA de Helmintos/genética , Técnicas de Amplificação de Ácido Nucleico/métodos , Strongyloides stercoralis/genética , Oligonucleotídeos , Corantes Fluorescentes , Sensibilidade e Especificidade
10.
Vet Res Commun ; 47(2): 947-952, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-36333528

RESUMO

Echinococcosis, caused by tapeworms of the genus Echinococcus, is a zoonotic parasitic disease. Various Echinococcus spp. are endemic and distributed in the Qinghai Province of China. Currently, few studies on the prevalence of Echinococcus spp. in the wild foxes are available. Hence, the aim of the study was to evaluate the prevalence of Echinococcus spp. in wild foxes in highly endemic areas of Qinghai Province, China. A total of 600 wild canid fecal samples were collected from Yushu, Qilian and Guinan in the study region, and 521 samples were successfully molecularly identified as wild foxes (Tibetan fox: 448, red foxes: 70, corsac fox: 3). Among the wild foxes, 5.57% (29/521) tested positive for Echinococcus spp. The prevalence rates of Echinococcus spp. in wild foxes in the Yushu, Qilian and Guinan areas were 2.51%, 15.22% and 0.96%, respectively. Furthermore, sequencing analysis indicated that E. multilocularis was the most prevalent species, occurring in 4.03% (21/521) of the wild foxes. Compared to E. granulosus occurring in 0.58% (3/521) of the foxes, E. shiquicus occurred in 1.54% (8/521), and E. shiquicus was first reported with 2.17% (3/138) prevalence in the Qilian area, indicating its transmission range is expanding. The current results provide useful epidemiological data for understanding and monitoring the dissemination of Echinococcus spp. by wild foxes in Qinghai Province, China.


Assuntos
Echinococcus , Animais , Echinococcus/genética , Raposas , Prevalência , DNA de Helmintos , China/epidemiologia , Zoonoses
11.
J Microbiol Methods ; 204: 106661, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36565938

RESUMO

Although several protocols were developed to extract DNA for soil-transmitted helminthiasis diagnostic, amplifying these extracts remains challenging due to DNA polymerase inhibitors. This study aimed to assess a DNA extraction method for efficient detection of soil-transmitted helminth species by determining stool mass and the type of DNA polymerase that can be used for this extraction method. For this study, 141 stool samples harbouring soil-transmitted eggs and 50 samples without egg were obtained from school-aged children of Makenene in the Centre region of Cameroon. DNA was extracted from 10, 20, 40 and 80 mg of stool using commercial kit and/or cetyltrimethylammonium bromide (CTAB)-based method. The amount of stool for molecular diagnostic of soil-transmitted helminthiasis was determined by amplifying Ascaris lumbricoides DNA. The performances of three DNA polymerases and CTAB-based method were assessed by amplifying DNA of different soil-transmitted helminth species. For this study, 94 stools with A. lumbricoides eggs, 39 with Trichuris trichuria and 15 with hookworm were analyzed. DNA of A. lumbricoides, T. trichuria, Necator americanus and Ancylostoma duodenale were detected in 97.9% of extracts from stools harbouring soil-transmitted helminth eggs. Soil-transmitted helminth DNAs were significantly (X2 = 17.66; df = 3; p ã€ˆ00001) more amplified in extracts from 10 and 20 mg than those from 40 and 80 mg. The amplification rate with "Q5 high fidelity DNA polymerase" was significantly (X2 = 30.54; df = 2; p < 0.00001) higher than that of other DNA polymerases. Multiplex-PCR confirmed co-infections of A. lumbricoides with either T. trichuria or N. americanus. The extraction cost for the CTAB-based method was $1.45. This method appearedis reliable and 3 times cost effective than commercial kit. Its combination with the "Q5 high fidelity DNA polymerase" may improve soil-transmitted helminthiasis diagnostic.


Assuntos
Helmintíase , Helmintos , Criança , Animais , Humanos , Cetrimônio , DNA de Helmintos , Solo , Helmintíase/diagnóstico , Fezes , Prevalência
12.
Parasit Vectors ; 15(1): 379, 2022 Oct 20.
Artigo em Inglês | MEDLINE | ID: mdl-36266710

RESUMO

BACKGROUND: Multiplex polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (RFLP) for nuclear phosphoenolpyruvate carboxykinase (pepck) and polymerase delta (pold), respectively, have been used to differentiate Fasciola hepatica, F. gigantica, and hybrid Fasciola flukes. However, discrimination errors have been reported in both methods. This study aimed to develop a multiplex PCR based on a novel nuclear marker, the fatty acid binding protein type I (FABP) type I gene. METHODS: Nucleotide sequence variations of FABP type I were analyzed using DNA samples of F. hepatica, F. gigantica, and hybrid Fasciola flukes obtained from 11 countries in Europe, Latin America, Africa, and Asia. A common forward primer for F. hepatica and F. gigantica and two specific reverse primers for F. hepatica and F. gigantica were designed for multiplex PCR. RESULTS: Specific fragments of F. hepatica (290 bp) and F. gigantica (190 bp) were successfully amplified using multiplex PCR. However, the hybrid flukes contained fragments of both species. The multiplex PCR for FABP type I could precisely discriminate the 1312 Fasciola samples used in this study. Notably, no discrimination errors were observed with this novel method. CONCLUSIONS: Multiplex PCR for FABP type I can be used as a species discrimination marker in place of pepck and pold. The robustness of the species-specific primer should be continuously examined using a larger number of Fasciola flukes worldwide in the future since nucleotide substitutions in the primer regions may cause amplification errors.


Assuntos
Fasciola , Fasciolíase , Animais , Fasciola/genética , Marcadores Genéticos , Proteínas de Ligação a Ácido Graxo/genética , Fosfoenolpiruvato , DNA de Helmintos/genética , Fosfoenolpiruvato Carboxiquinase (ATP)/genética , Nucleotídeos
13.
Infect Genet Evol ; 105: 105373, 2022 11.
Artigo em Inglês | MEDLINE | ID: mdl-36202207

RESUMO

Fasciola gigantica and hybrid Fasciola flukes, responsible for the disease fasciolosis, are found in Southeast Asian countries. In the present study, we performed molecular species identification of Fasciola flukes distributed in Terengganu, Malaysia using multiplex PCR for phosphoenolpyruvate carboxykinase (pepck) and PCR-restriction fragment length polymorphism (RFLP) for DNA polymerase delta (pold). Simultaneously, phylogenetic analysis based on mitochondrial NADH dehydrogenase subunit 1 (nad1) was performed for the first time on Malaysian Fasciola flukes to infer the dispersal direction among neighboring countries. A total of 40 flukes used in this study were identified as F. gigantica. Eight nad1 haplotypes were identified in the F. gigantica population of Terengganu. Median-joining network analysis revealed that the Malaysian population was related to those obtained from bordering countries such as Thailand and Indonesia. However, genetic differentiation was detected using population genetics analyses. Nevertheless, the nucleotide diversity (π) value suggested that F. gigantica with the predominant haplotypes was introduced into Malaysia from Thailand and Indonesia. The dispersal direction suggested by population genetics in the present study may not be fully reliable since Fasciola flukes were collected from a single location in one state of Malaysia. Further studies analyzing more samples from many locations are required to validate the dispersal direction proposed herein.


Assuntos
Distribuição Animal , DNA de Helmintos , Fasciola , Animais , Sudeste Asiático , DNA de Helmintos/genética , DNA Mitocondrial/genética , Fasciola/genética , Malásia , NADH Desidrogenase/genética , Filogenia , Filogeografia/métodos
14.
Infect Genet Evol ; 104: 105359, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-36070805

RESUMO

Fasciola gigantica is a major pathogen that causes fasciolosis in Africa. A recent study in Uganda demonstrated that Fasciola flukes were present in 65.7% of slaughtered cattle. However, molecular identification of Fasciola species has not yet been performed in the country. In the present study, 292 Fasciola flukes were collected from Kampala and Gulu, Uganda. The samples were identified as F. gigantica using a multiplex polymerase chain reaction (PCR) assay for phosphoenolpyruvate carboxykinase (pepck) and a PCR-restriction fragment length polymorphism (RFLP) assay for DNA polymerase delta (pold). A significant genetic difference between F. gigantica obtained from cattle slaughtered at Kampala and Gulu was observed by analyzing the mitochondrial markers NADH dehydrogenase subunit 1 (nad1) and cytochrome C oxidase subunit 1 (cox1). Fasciola collected from Gulu had a more diversified population than that collected from Kampala, probably because of differences in livestock management systems. One of the possible reasons for this observation is that cattle slaughtered in Gulu were reared under an extensive communal grazing system, which is suitable for maintaining parasite diversity, whereas cattle slaughtered in Kampala mainly originated from fenced/closed farms, which limits parasite diversity. However, the cause of the difference between these two locations was not clearly defined by the results of this study. The F. gigantica population from Uganda was related to that obtained from Zambia. A star-like phylogeny was detected in a median-joining network analysis, which indicated rapid population expansion and suggested that the F. gigantica populations from both countries are maintained by domestic ruminants in eastern Africa. Interestingly, the F. gigantica population from Uganda was not related to those from Egypt and Nigeria. The results of the present study suggest that F. gigantica populations in African countries are indigenous to each country or region.


Assuntos
Doenças dos Bovinos , Fasciola , Fasciolíase , Animais , Bovinos , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/parasitologia , DNA Polimerase III/genética , DNA de Helmintos/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Fasciola/genética , Fasciolíase/epidemiologia , Fasciolíase/parasitologia , Fasciolíase/veterinária , Haplótipos , Estrutura Molecular , NADH Desidrogenase/genética , Fosfoenolpiruvato , Filogenia , Ruminantes , Uganda/epidemiologia
15.
J Helminthol ; 96: e59, 2022 Aug 04.
Artigo em Inglês | MEDLINE | ID: mdl-35924599

RESUMO

Gastropod-associated nematodes have been previously studied and documented worldwide, with some species forming host-specific association as obligate parasites of molluscs while others form intermediate and temporary association. Philippinella moellendorffi from Imelda, Zamboanga Sibugay, Philippines, were collected, washed and maintained in the laboratory until death. Cadavers were placed on nutrient agar to allow nematode proliferation. Nematode pure culture was obtained using one gravid female for propagation. Morphology and molecular analyses (18S ribosomal DNA (rDNA) and D2-D3 expansion segments of 28S rDNA) were employed as diagnostic tools in identifying the nematode species isolated from P. moellendorffi. The newly isolated nematode was identified as Caenorhabditis brenneri, thus designated as C. brenneri strain IZSP from the Philippines. This is the first record of C. brenneri isolated from the terrestrial slug P. moellendorffi.


Assuntos
Caenorhabditis , Gastrópodes , Nematoides , Rabditídios , Animais , Cadáver , Caenorhabditis/genética , DNA de Helmintos/genética , DNA Ribossômico/genética , Feminino , Gastrópodes/parasitologia , Filipinas , Filogenia , Rabditídios/anatomia & histologia
16.
BMC Vet Res ; 18(1): 283, 2022 Jul 21.
Artigo em Inglês | MEDLINE | ID: mdl-35864506

RESUMO

BACKGROUND: Dicrocoelium dendriticum is a broadly distributed zoonotic helminth, which is mainly reported from domesticated and wild ruminants. There is little data covering the molecular features of this trematode; therefore, current study aimed to molecularly analyze D. dendriticum in livestock. METHODS: Totally, 23 samples of D. dendriticum were collected from cattle, sheep, and goat from Ilam, Lorestan, and Khuzestan, three west and south-west provinces of Iran from February to August 2018. After genomic DNA extraction, the internal transcribed spacer (ITS) 2 fragment was amplified and sequenced in samples. To investigate genetic variations through the ITS 2 fragment of obtained D. dendriticum, phylogenetic tree and network analysis were employed. RESULTS: All 23 samples were successfully amplified and sequenced. Phylogenetic tree showed that our samples were clearly grouped in a clade together with reference sequences. There was no grouping based on either geographical regions or hosts. Network analysis confirmed the phylogenetic findings and showed the presence of nine distinct haplotypes, while our samples together most of sequences, which were previously submitted to the GenBank, were grouped in the Hap1. CONCLUSIONS: Our findings indicated that although ITS 2 fragment discriminate D. dendriticum, this fragment is not suitable to study intra-species genetic variations. Therefore, exploring and describing new genetic markers could be more appropriate to provide new data about the genetic distribution of this trematode.


Assuntos
Doenças dos Bovinos , Dicrocoelium , Doenças das Cabras , Doenças dos Ovinos , Animais , Bovinos , DNA de Helmintos/genética , Dicrocoelium/genética , Cabras/genética , Irã (Geográfico) , Filogenia , Ovinos/genética
17.
J Helminthol ; 96: e54, 2022 Jul 27.
Artigo em Inglês | MEDLINE | ID: mdl-35894440

RESUMO

We obtained new data on the complete mitochondrial DNA (mtDNA) and the ribosomal operon of the trematode Carassotrema koreanum (Digenea: Haploporata: Haploporidae), an intestinal parasite of Carassius auratus, using next-generation sequencing. The mtDNA of C. koreanum contained 13,965 bp, including 12 protein-coding genes, two ribosomal genes, 22 transport RNA (tRNA) genes and a non-coding region. The ribosomal operon of C. koreanum was 10,644 bp in length, including ETS1 (1449 bp), 18S ribosomal RNA (rRNA) gene (1988 bp), ITS1 ribosomal DNA (rDNA) (558 bp), 5.8S rRNA gene (157 bp), ITS2 rDNA (274 bp), 28S rRNA gene (4152 bp) and ETS2 (2066 bp). Phylogenetic analysis based on mtDNA protein-coding regions showed that C. koreanum was closely related to Parasaccocoelium mugili, a species from the same suborder Haploporata. Bayesian phylogenetic tree topology was the most reliable and confirmed the validity of the Haploporata. The results of sequence cluster analysis based on codon usage bias demonstrated some agreement with the results of the phylogenetic analysis. In particular, Schistosoma spp. were differentiated from the other members of Digenea and the members of Pronocephalata were localized within the same cluster. Carassotrema koreanum and P. mugili fell within different clusters. The grouping of C. koreanum and P. mugili within the same cluster was obtained on the basis of frequencies of 13 specified codons, of which three codon pairs were degenerate. A similarity was found between two haploporid species and two Dicrocoelium spp. in the presence of TTG start codon of the mitochondrial nad5 gene. Our results confirmed the taxonomical status of the Haploporata identified in the previous studies and revealed some characteristic features of the codon usage in its representatives.


Assuntos
Genoma Mitocondrial , Trematódeos , Animais , Teorema de Bayes , DNA de Helmintos/genética , DNA Mitocondrial/genética , DNA Ribossômico/genética , Sequenciamento de Nucleotídeos em Larga Escala , Óperon , Filogenia , RNA Ribossômico , Análise de Sequência de DNA/métodos , Trematódeos/genética
18.
Sci Rep ; 12(1): 9947, 2022 06 15.
Artigo em Inglês | MEDLINE | ID: mdl-35705676

RESUMO

Next-generation sequencing technologies have accelerated the pace of helminth DNA metabarcoding research, enabling species detection in bulk community samples. However, finding suitable genetic markers with robust species-level resolution and primers targeting a broad species range among parasitic helminths are some of the challenges faced. This study aimed to demonstrate the potential use of the mitochondrial 12S and 16S rRNA genes for parasitic helminth (nematodes, trematodes, cestodes) DNA metabarcoding. To demonstrate the robustness of the 12S and 16S rRNA genes for DNA metabarcoding, we determined the proportion of species successfully recovered using mock helminth communities without environment matrix and mock helminth communities artificially spiked with environmental matrices. The environmental matrices are human fecal material, garden soil, tissue, and pond water. Our results revealed the robustness of the mitochondrial rRNA genes, through the high sensitivity of the 12S rRNA gene, and the effectiveness of the 12S and 16S primers targeting platyhelminths. With the mitochondrial rRNA genes, a broad range of parasitc helminths were successfully detected to the species level. The potential of the mitochondrial rRNA genes for helminth DNA metabarcoding was demonstrated, providing a valuable gateway for future helminth DNA metabarcoding applications like helminth detection and biodiversity studies.


Assuntos
Código de Barras de DNA Taxonômico , Helmintos , Animais , Código de Barras de DNA Taxonômico/métodos , Primers do DNA/genética , DNA de Helmintos/genética , Genes de RNAr , Helmintos/genética , Humanos , RNA Ribossômico 16S/genética
19.
Parasitol Res ; 121(7): 1921-1935, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35488923

RESUMO

Patagifer Dietz, 1909 is a small genus of echinostomatids, with 12 recognized species, mostly parasitising threskiornithid birds, distributed worldwide. In the current research, adult specimens of the type species, Patagifer bilobus (Rudolphi, 1819) Dietz, 1909 from the white faced ibis (Plegadis chihi) and white ibis (Eudocimus albus) were re-described, providing new metrical data for the number of head collar spines. Those specimens were recorded from eight localities in Mexico and compared morphologically with specimens previously identified as Patagifer lamothei. A total of 19 specimens identified as P. bilobus including two hologenophores were sequenced with three molecular markers: domains D1-D3 of the large subunit (LSU), the internal transcribed spacer (ITS1, ITS2) plus 5.8S from the nuclear rDNA, and nicotinamide adenine dinucleotide dehydrogenase subunit 1 (nad1) from mitochondrial DNA. The new sequences were aligned with other sequences of Patagifer spp., downloaded from GenBank. Phylogenetic trees inferred from each data set, placed all the specimens in a clade, confirming that the isolates belonged to the same species. The morphological examination of specimens previously identified as P. lamothei by Ortega-Olivares MP, Hernández-Mena DI, Pérez-Ponce de León G, García-Varela M (2011) Helminths of the white ibis, Eudocimus albus (Aves Therskiornithidae) in Mexico. (Zootaxa 3088, 15-26. 10.11646/zootaxa.3088.1.2) and in combination with molecular data confirms that those specimens should be reassigned to P. bilobus. In addition, this is the first study in P. bilobus using an integrative taxonomy approach.


Assuntos
Echinostomatidae , Trematódeos , Animais , Aves/parasitologia , DNA de Helmintos/genética , DNA Ribossômico/genética , Echinostomatidae/genética , México , Filogenia
20.
J Parasitol ; 108(1): 70-78, 2022 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-35167699

RESUMO

Although Clonorchis sinensis is a parasite that still infects many people in East Asia, its genetics remain largely unknown. We conducted ancient DNA analysis of C. sinensis eggs obtained from a Joseon period mummy newly discovered in South Korea. Clonorchis sinensis DNA was amplified for internal transcribed spacer 1, cytochrome c oxidase subunit 1, and NADH dehydrogenase subunit 2 and 5 genes. The results of BLAST/NCBI showed that the consensus sequences were 98.24 to 100% identical to the modern and ancient C. sinensis sequences reported from Korea, China, Japan, and other Asian countries. Our report helps to fill in the genetic profile of ancient C. sinensis strains that infected East Asian people hundreds of years ago.


Assuntos
Clonorquíase/história , Clonorchis sinensis/genética , Múmias/parasitologia , Animais , Clonorquíase/parasitologia , Clonorchis sinensis/classificação , DNA de Helmintos/química , DNA de Helmintos/genética , História Antiga , Óvulo , Filogenia , República da Coreia
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...