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1.
PLoS Negl Trop Dis ; 14(6): e0008392, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32542036

RESUMO

The canine hookworms Ancylostoma braziliense, Ancylostoma ceylanicum, Ancylostoma caninum and Uncinaria stenocephala are not only capable of producing morbidity and mortality in dogs but are also neglected tropical zoonoses. Each hookworm species differs considerably in its geographical distribution, life cycle, biology, pathogenic impacts on both canine and human hosts, zoonotic potential, and response to treatment with anthelminthics. Here we describe the development and validation of two Taq-Man based multiplex PCR assays capable of detecting and differentiating all four canine hookworm species in faeces of naturally infected dogs. The analytical sensitivity of both assays was assessed using 10-fold serial dilutions of synthetic gene block fragments containing individual sequence targets of each hookworm species. The sensitivity of the assays and ability to detect mixed species infections were compared to a conventional PCR-Restriction Fragment Length Polymorphism based-approach when applied to laboratory and field samples from endemic areas. The qPCRs detected at least one species of hookworms in 82.4% of PCR-RFLP-negative but microscopy-positive samples. The qPCRs detected an additional 68% mixed infections with different species of canine hookworms, and additional single species infection with A. caninum (47%), U. stenocephala (33%) and A. ceylanicum (0.02%) that were missed by PCR-RFLP. These multiplex qPCR assays will assist field based epidemiological surveillance studies towards an accurate and sensitive monitoring of canine hookworm infections in dogs, to inform their species-specific zoonotic risks to populations living in endemic areas, globally.


Assuntos
Ancylostomatoidea/genética , Ancylostomatoidea/isolamento & purificação , Doenças do Cão/diagnóstico , Infecções por Uncinaria/diagnóstico , Reação em Cadeia da Polimerase Multiplex/métodos , Reação em Cadeia da Polimerase Multiplex/veterinária , Ancylostoma/genética , Ancylostoma/isolamento & purificação , Ancylostomatoidea/classificação , Ancilostomíase/diagnóstico , Ancilostomíase/epidemiologia , Ancilostomíase/fisiopatologia , Animais , DNA de Helmintos/análise , DNA de Helmintos/genética , Doenças do Cão/epidemiologia , Doenças do Cão/fisiopatologia , Cães , Fezes/parasitologia , Infecções por Uncinaria/fisiopatologia , Humanos , Polimorfismo de Fragmento de Restrição , Sensibilidade e Especificidade , Zoonoses/diagnóstico , Zoonoses/epidemiologia , Zoonoses/fisiopatologia
2.
Parasitol Res ; 119(7): 2343-2346, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32435896

RESUMO

Marsupials of the genus Didelphis, such as black-eared opossums (Didelphis aurita), are common synanthropic animals in urban areas of Brazil. These marsupials are frequently parasitized by numerous helminth species, including ancylostomatid nematodes. This study aimed to report the occurrence of Ancylostoma caninum in black-eared opossums captured in an urban environment of Southeastern Brazil and discuss the potential impact of these findings for public health. From January to June 2019, we collected fecal samples from 49 restrained opossums and evaluated by a simple flotation method; Helminth eggs were observed at different magnifications and identified according to morphological and morphometric features. Genomic DNA was extracted from Ancylostomatidae eggs and screened by duplex PCR for Ancylostoma spp. and Necator americanus using primers that amplify a region of internal transcribed spacer 2 and the 28S ribosomal RNA (ITS2-28S rRNA). Ancylostoma spp. eggs were detected in 65.3% (32/49) of the animals. Sequence analysis revealed 100% homology with A. caninum sequences from GenBank. Our results demonstrate a new host-parasite interaction for A. caninum, suggesting that black-eared opossums may participate in the zoonotic cycle of this parasite in urban areas of Brazil.


Assuntos
Ancylostoma/isolamento & purificação , Ancilostomíase/veterinária , Didelphis/parasitologia , Ancylostoma/genética , Ancilostomíase/epidemiologia , Ancilostomíase/parasitologia , Animais , Brasil/epidemiologia , Cidades/epidemiologia , Fezes/parasitologia , Genoma Helmíntico/genética , Contagem de Ovos de Parasitas , Prevalência
3.
Am J Trop Med Hyg ; 103(1): 64-68, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32342850

RESUMO

Hookworm infections are classified as the most impactful of the human soil-transmitted helminth (STH) infections, causing a disease burden of ∼4 million disability-adjusted life years, with a global prevalence of 406-480 million infections. Until a decade ago, epidemiological surveys largely assumed Necator americanus and Ancylostoma duodenale as the relevant human hookworm species implicated as contributing to iron-deficiency anemia. This assumption was based on the indistinguishable morphology of the Ancylostoma spp. eggs in stool and the absence of awareness of a third zoonotic hookworm species, Ancylostoma ceylanicum. The expanded use of molecular diagnostic assays for differentiating hookworm species infections during STH surveys has now implicated A. ceylanicum, a predominant hookworm of dogs in Asia, as the second most common hookworm species infecting humans in Southeast Asia and the Pacific. Despite this, with the exception of sporadic case reports, there is a paucity of data available on the impact of this emerging zoonosis on human health at a population level. This situation also challenges the current paradigm, necessitating a One Health approach to hookworm control in populations in which this zoonosis is endemic. Here, we have summarized the available research studies and case reports on human A. ceylanicum infections in Southeast Asia and the Pacific after 2013 using a systematic review approach. We summarized eight research articles and five clinical case studies, highlighting the importance of future in-depth investigation of zoonotic A. ceylanicum infections using sensitive and cost-effective diagnostic tools.


Assuntos
Ancylostoma/patogenicidade , Ancilostomíase/epidemiologia , Doenças do Cão/epidemiologia , Doenças Negligenciadas/epidemiologia , Zoonoses/epidemiologia , Ancylostoma/isolamento & purificação , Ancilostomíase/diagnóstico , Ancilostomíase/parasitologia , Ancilostomíase/transmissão , Animais , Ásia Sudeste/epidemiologia , Doenças do Cão/diagnóstico , Doenças do Cão/parasitologia , Doenças do Cão/transmissão , Cães , Fezes/parasitologia , Humanos , Doenças Negligenciadas/diagnóstico , Doenças Negligenciadas/parasitologia , Saúde Única/legislação & jurisprudência , Ilhas do Pacífico/epidemiologia , Contagem de Ovos de Parasitas , Zoonoses/diagnóstico , Zoonoses/parasitologia , Zoonoses/transmissão
4.
Int J Parasitol ; 50(3): 217-225, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32135180

RESUMO

Polyparasitism, involving soil-transmitted helminths. and Schistosoma blood flukes, is common in low to middle income countries. These helminths impact on the gut environment and can cause changes to the gut microbiome composition. Here we examined the gut microbiome in individuals with polyparasitism from two human cohorts in the Philippines utilising DNA sequencing-based profiling. Multiple helminth species infections were high with 70.3% of study participants harbouring at least two parasite species, and 16% harbouring at least five species. Increased numbers of helminth co-infections, in particular with the gut-resident soil-transmitted helminths, were significantly associated with increased bacterial diversity; however no significant parasite-gut microbiome associations were evident for individuals infected only with Schistosoma japonicum. In general, a healthy gut is associated with high bacterial diversity, which in these human cohorts may be the result of helminth-mediated immune modulation, or due to changes in the gut environment caused by these parasitic helminths.


Assuntos
Coinfecção , Microbioma Gastrointestinal/genética , Helmintíase/epidemiologia , Helmintos/isolamento & purificação , Esquistossomose/epidemiologia , Adolescente , Adulto , Albendazol/uso terapêutico , Ancylostoma/isolamento & purificação , Ancylostomatoidea/isolamento & purificação , Animais , Ascaris/isolamento & purificação , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Criança , Pré-Escolar , Estudos de Coortes , Coinfecção/microbiologia , Coinfecção/parasitologia , Fezes/microbiologia , Fezes/parasitologia , Feminino , Genes Bacterianos , Helmintíase/tratamento farmacológico , Helmintos/genética , Interações Hospedeiro-Parasita , Humanos , Masculino , Metagenômica , Interações Microbianas , Pessoa de Meia-Idade , Patologia Molecular , Filipinas/epidemiologia , Schistosoma/isolamento & purificação , Esquistossomose/tratamento farmacológico , Solo/parasitologia , Trichuris/isolamento & purificação , Adulto Jovem
5.
Sci Rep ; 10(1): 3006, 2020 02 20.
Artigo em Inglês | MEDLINE | ID: mdl-32080267

RESUMO

Hookworms are intestinal parasites that cause major public health problems, especially in developing countries. To differentiate eggs from different hookworm species, it is necessary to use molecular methodologies, since the eggs are morphologically similar. Here, we performed the molecular identification of single hookworm eggs from six Brazilian states. Of the 634 eggs individually analyzed, 98.1% (622/634) represented Necator americanus, and surprisingly, 1.9% (12/634 eggs from the same patient) represented Ancylostoma caninum. DNA analysis of the A. caninum-positive stool sample revealed no contamination with animal feces. This is the first report of the presence of A. caninum eggs in human feces, which may have a direct implication for the epidemiology of hookworm infection caused by this species. This suggests the need for special attention regarding prophylaxis, as different reservoirs, previously not described, may have great relevance for the spread of A. caninum.


Assuntos
Ancylostoma/genética , Ancilostomíase/epidemiologia , DNA de Helmintos/genética , Necator americanus/genética , Necatoríase/epidemiologia , Ancylostoma/classificação , Ancylostoma/isolamento & purificação , Ancilostomíase/diagnóstico , Ancilostomíase/parasitologia , Animais , Brasil/epidemiologia , Fezes/parasitologia , Genótipo , Humanos , Intestinos/parasitologia , Necator americanus/classificação , Necator americanus/isolamento & purificação , Necatoríase/diagnóstico , Necatoríase/parasitologia
6.
Parasit Vectors ; 13(1): 6, 2020 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-31915050

RESUMO

BACKGROUND: Japan is one of the few countries believed to have eliminated soil-transmitted helminths (STHs). In 1949, the national prevalence of Ascaris lumbricoides was 62.9%, which decreased to 0.6% in 1973 due to improvements in infrastructure, socioeconomic status, and the implementation of national STH control measures. The Parasitosis Prevention Law ended in 1994 and population-level screening ceased in Japan; therefore, current transmission status of STH in Japan is not well characterized. Sporadic cases of STH infections continue to be reported, raising the possibility of a larger-scale recrudescence of STH infections. Given that traditional microscopic detection methods are not sensitive to low-intensity STH infections, we conducted targeted prevalence surveys using sensitive PCR-based assays to evaluate the current STH-transmission status and to describe epidemiological characteristics of areas of Japan believed to have achieved historical elimination of STHs. METHODS: Stool samples were collected from 682 preschool- and school-aged children from six localities of Japan with previously high prevalence of STH. Caregivers of participants completed a questionnaire to ascertain access to water, sanitation and hygiene (WASH), and potential exposures to environmental contamination. For fecal testing, multi-parallel real-time PCR assays were used to detect infections of Ascaris lumbricoides, Necator americanus, Ancylostoma duodenale and Trichuris trichiura. RESULTS: Among the 682 children, no positive samples were identified, and participants reported high standards of WASH. CONCLUSIONS: To our knowledge, this is the first STH-surveillance study in Japan to use sensitive molecular techniques for STH detection. The results suggest that recrudescence of STH infections has not occurred, and that declines in prevalence have been sustained in the sampled areas. These findings suggest that reductions in prevalence below the elimination thresholds, suggestive of transmission interruption, are possible. Additionally, this study provides circumstantial evidence that multi-parallel real-time PCR methods are applicable for evaluating elimination status in areas where STH prevalence is extremely low.


Assuntos
Ancylostoma/isolamento & purificação , Ascaris lumbricoides/isolamento & purificação , Necator americanus/isolamento & purificação , Trichuris/isolamento & purificação , Adolescente , Ancylostoma/genética , Ancilostomíase/parasitologia , Animais , Ascaríase/parasitologia , Ascaris lumbricoides/genética , Criança , Pré-Escolar , Fezes/parasitologia , Feminino , Helmintos , Humanos , Higiene , Japão , Masculino , Necator americanus/genética , Necatoríase/parasitologia , Solo/parasitologia , Inquéritos e Questionários , Tricuríase/parasitologia , Trichuris/genética
7.
Vet Parasitol Reg Stud Reports ; 18: 100324, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31796199

RESUMO

Monthly canine parasite prophylactic products prevent not only adult heartworm infection, but also patent infections with specific gastrointestinal parasites. While most monthly products control and treat certain hookworm and roundworm infections, fewer are labeled for whipworm (Trichuris vulpis). Therefore, we hypothesized that fecal samples collected from municipal dog parks will have a greater prevalence of whipworm eggs compared to hookworm and roundworm eggs. In this study, canine fecal samples were collected from municipal dog parks in three southeastern states, with up to 20 fecal samples were collected from each park. A total of 200 fecal samples were obtained from dog parks in Georgia, North Carolina, and South Carolina. All fecal samples were examined for the presence of gastrointestinal helminths by a simple centrifugal flotation using sheather's sugar flotation solution. Of the 200 samples collected, 27% were positive for gastrointestinal helminths by fecal flotation. Of these infected fecal samples, 8.5%, 17%, and 1.5% contained whipworm, hookworm, and roundworm, respectively. However, the majority of hookworm-positive samples were collected from one park, whereas whipworm and roundworm samples were collected from multiple parks. These results could indicate that dogs are at risk of infection by all three parasites at dog parks, and that preventive strategies may need to be tailored not only to the specific region, but to specific infected dog parks.


Assuntos
Ancilostomíase/veterinária , Doenças do Cão/transmissão , Toxocaríase/transmissão , Tricuríase/veterinária , Ancylostoma/isolamento & purificação , Ancilostomíase/parasitologia , Ancilostomíase/transmissão , Animais , Doenças do Cão/parasitologia , Cães , Georgia , North Carolina , South Carolina , Toxocara canis/isolamento & purificação , Toxocaríase/parasitologia , Tricuríase/parasitologia , Tricuríase/transmissão , Trichuris/isolamento & purificação
8.
J Parasitol ; 105(6): 904-912, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31805245

RESUMO

From the small intestines of both Ctenomys boliviensis and Ctenomys steinbachi collected from August 1984 through June 1990 from the eastern lowlands of the Department of Santa Cruz, Bolivia a total of 36 specimens of Ancylostoma were recovered. Morphological investigation and comparisons with known species described and reported from mammals in the Neotropical Region show that this is an undescribed species, herein described as new. These nematans were collected from individuals of C. steinbachi collected from near a locality called Caranda (northwest of Santa Cruz de la Sierra) and from C. boliviensis from near Santa Rosa de la Roca (northeast of Santa Cruz de la Sierra) and from cajuchis collected from 3 km west of Estación El Pailón, 30 km east of Santa Cruz de la Sierra. The new species of Ancylostoma differs from all other species of Ancylostoma known from the Neotropical Region in the presence of paired sub-terminal papillae on the dorsal ray of males.


Assuntos
Ancylostoma/classificação , Ancilostomíase/veterinária , Doenças dos Roedores/parasitologia , Ancylostoma/anatomia & histologia , Ancylostoma/isolamento & purificação , Ancilostomíase/diagnóstico , Ancilostomíase/parasitologia , Animais , Bolívia , Diagnóstico Diferencial , Feminino , Intestino Delgado/parasitologia , Masculino , Doenças dos Roedores/diagnóstico , Roedores/parasitologia
10.
PLoS Negl Trop Dis ; 13(10): e0007778, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31658264

RESUMO

BACKGROUND: A DNA extraction and preservation protocol that yields sufficient and qualitative DNA is pivotal for the success of any nucleic acid amplification test (NAAT), but it still poses a challenge for soil-transmitted helminths (STHs), including Ascaris lumbricoides, Trichuris trichiura and the two hookworms (Necator americanus and Ancylostoma duodenale). In the present study, we assessed the impact of different DNA extraction and preservativation protocols on STH-specific DNA amplification from stool. METHODOLOGY AND PRINCIPAL FINDINGS: In a first experiment, DNA was extracted from 37 stool samples with variable egg counts for T. trichiura and N. americanus applying two commercial kits, both with and without a prior bead beating step. The DNA concentration of T. trichiura and N. americanus was estimated by means of qPCR. The results showed clear differences in DNA concentration across both DNA extraction kits, which varied across both STHs. They also indicated that adding a bead beating step substantially improved DNA recovery, particularly when the FECs were high. In a second experiment, 20 stool samples with variable egg counts for A. lumbricoides, T. trichiura and N. americanus were preserved in either 96% ethanol, 5% potassium dichromate or RNAlater and were stored at 4°C for 65, 245 and 425 days. DNA was extracted using the DNeasy Blood & Tissue kit with a bead beating step. Stool samples preserved in ethanol proved to yield higher DNA concentrations as FEC increased, although stool samples appeared to be stable over time in all preservatives. CONCLUSIONS: The choice of DNA extraction kit significantly affects the outcome of NAATs. Given the clear benefit of bead beating and our validation of ethanol for (long-term) preservation, we recommend that these aspects of the protocol should be adopted by any stool sampling and DNA extraction protocol for downstream NAAT-based detection and quantification of STHs.


Assuntos
Fezes/parasitologia , Helmintíase/diagnóstico , Helmintos/isolamento & purificação , Técnicas de Diagnóstico Molecular/métodos , Preservação Biológica/métodos , Solo/parasitologia , Adolescente , Ancylostoma/genética , Ancylostoma/isolamento & purificação , Ancylostomatoidea/genética , Ancylostomatoidea/isolamento & purificação , Ancylostomatoidea/parasitologia , Animais , Ascaríase/diagnóstico , Ascaríase/parasitologia , Ascaris lumbricoides/genética , Ascaris lumbricoides/isolamento & purificação , Criança , Pré-Escolar , DNA/isolamento & purificação , Helmintíase/parasitologia , Helmintos/genética , Humanos , Necator americanus/isolamento & purificação , Necatoríase/diagnóstico , Necatoríase/patologia , Contagem de Ovos de Parasitas , Sensibilidade e Especificidade , Tricuríase/diagnóstico , Tricuríase/parasitologia , Trichuris/genética , Trichuris/isolamento & purificação
12.
Parasitol Int ; 73: 101974, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31421266

RESUMO

Hookworms are blood-sucking nematodes that infect dogs, cats, and humans, causing iron-deficiency anemia, abdominal pain, diarrhea, and skin inflammation. Amplification refractory mutation system (ARMS) is a modified technology based on allele-specific PCR, which is widely used in mutation detection and genotyping. However, no data about ARMS application in hookworm detection. This study aims to establish a multi-ARMS-qPCR method for the detection of three hookworm species from dogs and cats. A universal forward primer and three specific primers (ARMS-Cey, ARMS-Can, and ARMS-Tub) were designed based on the three ITS SNPs (ITS250, ITS78 and ITS153) of Ancylostoma ceylanicum, A. caninum, and A. tubaeforme, respectively. The results showed that the three designed ARMS primers generated specific melting curves for the three hookworms' standard plasmids. The melting temperature (Tm) values were 88.40 °C (A. ceylanicum), 83.15 °C (A. caninum), and 85.65 °C (A. tubaeforme), with good reproducibility of intra- and inter-assay. No amplification was observed with other intestinal parasites. The limit of detection using the established technique was 1, 2, and 104 egg per gram feces (EPG) for A. caninum, A. tubaeforme and A. ceylanicum, respectively. Using multi-ARMS-qPCR assay, 17 out of 50 fecal samples were positive for hookworms, including ten single and seven mixed infections, and single infections were quantified. In conclusion, the used multi-ARMS-qPCR method has the advantages of high efficiency, sensitivity, specificity, and quantitative analysis and can be used for the clinical detection, epidemiological investigation, and zoonotic risk assessment of canine and feline hookworms.


Assuntos
Ancylostoma/isolamento & purificação , Ancilostomíase/veterinária , Doenças do Gato/diagnóstico , Doenças do Cão/diagnóstico , Mutação , Técnicas de Amplificação de Ácido Nucleico/métodos , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Ancilostomíase/diagnóstico , Ancilostomíase/parasitologia , Animais , Doenças do Gato/parasitologia , Gatos , Doenças do Cão/parasitologia , Cães , Técnicas de Amplificação de Ácido Nucleico/veterinária , Reação em Cadeia da Polimerase em Tempo Real/métodos , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
13.
Infect Dis Poverty ; 8(1): 41, 2019 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-31262367

RESUMO

BACKGROUND: Baseline mapping of soil-transmitted helminth (STH) infections among school age children (SAC) in 2008-2009 found high or moderate prevalence in 13 of the 14 districts in Sierra Leone. Following these surveys, mass drug administration (MDA) of mebendazole/albendazole was conducted biannually at national level targeting pre-school children (PSC) aged 12-59 months and intermittently at sub-national level targeting SAC. In addition, MDA with ivermectin and albendazole for eliminating lymphatic filariasis (LF) has been conducted nationwide since 2010 targeting individuals over 5 years of age. Each MDA achieved high coverage, except in 2014 when all but one round of MDA for PSC was cancelled due to the Ebola emergency. The objective of the current study was to determine the prevalence and intensity of STH infections among SAC after a decade of these deworming campaigns. METHODS: Seventy-three schools in 14 districts were purposefully selected, including 39 schools from the baseline surveys, with approximately two sites from each of low, moderate and high prevalence categories at baseline per district. Fresh stool samples were collected from 3632 children aged 9-14 years (male 51%, female 49%) and examined using the Kato Katz technique. RESULTS: The prevalence of STH infections in Sierra Leone decreased in 2016 compared to 2008: Ascaris lumbricoides 4.4% (95% confidence interval [CI]: 3.7-5.1%) versus 6.6% (95% CI: 0-25%), Trichuris trichiura 0.7% (95% CI: 0.5-1.1%) versus 1.8% (95% CI: 0-30.2%), hookworm 14.9% (95% CI: 13.8-16.1) versus 38.5% (95% CI: 5.4-95.1%), and any STH 18.3% (95% CI:17.0-19.5%) versus 48.3% (CI: 5.4-96.3%), respectively. In 2016, no district had high hookworm prevalence and four districts had moderate prevalence, compared with eight and four districts respectively in 2008. In 2016, the arithmetic mean hookworm egg count in all children examined was light: 45.5 eggs per gram (EPG) of faeces, (95% CI:\ 35.96-55.07 EPG); three (0.08%) children had heavy infections and nine (0.25%) children had moderate infections. CONCLUSIONS: Sierra Leone has made considerable progress toward controlling STH as a public health problem among SAC. As LF MDA phases out (between 2017 and 2021), transition of deworming to other platforms and water and sanitation strategies need to be strengthened to maintain STH control and ultimately interrupt transmission.


Assuntos
Ancylostoma/isolamento & purificação , Ancilostomíase/epidemiologia , Ascaríase/epidemiologia , Ascaris lumbricoides/isolamento & purificação , Tricuríase/epidemiologia , Trichuris/isolamento & purificação , Adolescente , Ancilostomíase/prevenção & controle , Animais , Ascaríase/prevenção & controle , Quimioprevenção , Criança , Fezes/parasitologia , Feminino , Helmintíase/epidemiologia , Helmintíase/prevenção & controle , Humanos , Masculino , Prevalência , Instituições Acadêmicas , Serra Leoa/epidemiologia , Solo/parasitologia , Tricuríase/prevenção & controle
14.
Acta Derm Venereol ; 99(9): 805-808, 2019 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-31073620

RESUMO

Cutaneous larva migrans is a dermatitis, typically acquired in warm tropical or sub-tropical countries, caused by migration of the larvae of nematodes (hookworm; mainly Ancylostoma braziliense and, occasionally, Ancylostoma caninum or Uncinaria stenocephala), which are parasitic on animals such as cats and dogs, into the patient's skin. The larvae penetrate the skin after contact with infected soil and cause a typical creeping eruption. Patients with cutaneous larva migrans seen in Europe have usually acquired the disease following a stay in a tropical or sub-tropical area. How-ever, some cases of cutaneous larval migrans are acquired in Europe. We report here 5 autochthonous cases in France and give an overview of European autochthonous cases.


Assuntos
Ancylostoma/isolamento & purificação , Transmissão de Doença Infecciosa , Larva Migrans/parasitologia , Pele/parasitologia , Adulto , Ancylostoma/efeitos dos fármacos , Animais , Antinematódeos/uso terapêutico , Feminino , França , Humanos , Lactente , Ivermectina/uso terapêutico , Larva Migrans/tratamento farmacológico , Larva Migrans/transmissão , Masculino , Pessoa de Meia-Idade , Indução de Remissão , Pele/efeitos dos fármacos , Resultado do Tratamento , Adulto Jovem
15.
Vet Parasitol Reg Stud Reports ; 16: 100285, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-31027601

RESUMO

As laws change around the United States, wildlife that were once kept as companion animals are now often confiscated by local authorities. They are then euthanized unless a home is found for them at a sanctuary. Wolf sanctuaries are, therefore, becoming increasingly important for their conservation and management. However, little data is available on best practices for the health management of captive wolves, including data on parasitic diseases. Our objective was to assess the prevalence of parasites of captive wolves combining classical coprological techniques and immunoassays based on the detection of coproantigen of selected canid parasites. Fecal samples of 39 animals were collected upon observation of individual animals defecating. All samples were processed using the Fecal Dx® tests, a suite of coproantigen ELISAs for detection of ascarid, hookworm, whipworm, and Giardia (IDEXX Laboratories Inc.). Out of the 39 samples, 38 were processed using the double-centrifugation sugar flotation (DCSF) and 34 using a modification of the Baermann technique. Twenty-eight samples (71.8%) were positive for hookworm, and none positive for the other parasites tested using coproantigen ELISA. Ancylostoma sp. (26, 68.4%), Eucoleus boehmi (13, 34.2%), and Trichuris sp. (2; 5.3%), and Sarcocystis sp. (13, 34.2%) were detected using DCSF. No metastrongyloid lungworm larvae were found. The Cohen's kappa index (0.97) showed excellent agreement between the hookworm coproantigen ELISA and the DCSF using feces preserved in ethanol for a short period of time. This study provides a baseline on the parasites of captive wolves, and shows that recent innovative diagnostics in veterinary parasitology, developed and optimized for dogs, may be used for assessing the health of wolves.


Assuntos
Fezes/parasitologia , Helmintíase Animal/diagnóstico , Infecções Protozoárias em Animais/diagnóstico , Lobos/parasitologia , Ancylostoma/imunologia , Ancylostoma/isolamento & purificação , Ancylostomatoidea/imunologia , Ancylostomatoidea/isolamento & purificação , Animais , Antígenos de Helmintos/análise , Antígenos de Helmintos/isolamento & purificação , Antígenos de Protozoários/análise , Antígenos de Protozoários/isolamento & purificação , Centrifugação/métodos , Centrifugação/veterinária , Ensaio de Imunoadsorção Enzimática/veterinária , Helmintíase Animal/epidemiologia , Helmintíase Animal/parasitologia , Nematoides/imunologia , Nematoides/isolamento & purificação , Pennsylvania , Infecções Protozoárias em Animais/epidemiologia , Infecções Protozoárias em Animais/parasitologia , Sarcocystis/imunologia , Sarcocystis/isolamento & purificação , Sensibilidade e Especificidade , Trichuris/imunologia , Trichuris/isolamento & purificação , Estados Unidos
16.
Vet Parasitol Reg Stud Reports ; 16: 100270, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-31027603

RESUMO

The goal of this study was to assess the prevalence of gastrointestinal and respiratory parasites of shelter cats from northeast Georgia, thus promoting a more targeted approach in parasite diagnosis and treatment. Fecal samples of cats kept in a shelter located in Lavonia, northeastern Georgia, USA, were processed for the presence of parasites using double centrifugation sugar flotation (n = 103) and Baermann techniques (n = 98). Flotation revealed eggs of Toxocara cati (17.5%), Ancylostoma sp. (11.7%), Taeniidae (3.9%), Spirometra mansonoides (2.9%), Mesocestoides sp. (1%), Dipylidium caninum (1%), and Eucoleus aerophilus (1%), and oocysts of Cystoisospora felis (16.5%), and Cystoisospora rivolta (8.7%). Baermann diagnosed Aelurostrongylus abstrusus larvae in 5 cats (5.1%), while fecal flotation alone identified only 2 of these infections. Taeniidae eggs were identified to species-level by PCR and sequencing targeting the cytochrome oxidase c subunit 1 (cox1) of the mitochondrial DNA. All isolates belong to Hydatigera taeniaeformis sensu stricto, which is the first unequivocal report of the species in North America. Overall, 45.6% of the cats were infected with at least one parasite. This prevalence of infection is much higher than what is generally reported in client owned animals, highlighting the importance of using appropriate fecal diagnostic techniques to detect gastrointestinal and respiratory parasites on newly adopted cats. Correct diagnosis may direct appropriate treatment and control strategies, which would mitigate the risk of infection of other animals in household, and human exposure to zoonotic parasites.


Assuntos
Doenças do Gato/epidemiologia , Gastroenteropatias/veterinária , Doenças Parasitárias em Animais/epidemiologia , Doenças Respiratórias/veterinária , Distribuição por Idade , Ancylostoma/classificação , Ancylostoma/isolamento & purificação , Animais , Doenças do Gato/parasitologia , Gatos , Cestoides/classificação , Cestoides/isolamento & purificação , Fezes/parasitologia , Feminino , Gastroenteropatias/epidemiologia , Gastroenteropatias/parasitologia , Georgia/epidemiologia , Isospora/classificação , Isospora/isolamento & purificação , Funções Verossimilhança , Pneumopatias Parasitárias/epidemiologia , Pneumopatias Parasitárias/parasitologia , Pneumopatias Parasitárias/veterinária , Masculino , Mesocestoides/classificação , Mesocestoides/isolamento & purificação , Doenças Parasitárias em Animais/parasitologia , Filogenia , Prevalência , Doenças Respiratórias/epidemiologia , Doenças Respiratórias/parasitologia , Distribuição por Sexo , Spirometra/classificação , Spirometra/isolamento & purificação , Toxocara/classificação , Toxocara/isolamento & purificação
17.
Korean J Parasitol ; 57(1): 9-15, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30840793

RESUMO

Melting temperature shift (Tm-shift) is a new detection method that analyze the melting curve on real-time PCR thermocycler using SYBR Green I fluorescent dye. To establish a Tm-shift method for the detection of Ancylostoma ceylanicum and A. tubaeforme in cats, specific primers, with GC tail of unequal length attached to their 5 ́ end, were designed based on 2 SNP loci (ITS101 and ITS296) of the internal transcribed spacer 1 (ITS1) sequences. The standard curve of Tm-shift was established using the standard plasmids of A. ceylanicum (AceP) and A. tubaeforme (AtuP). The Tm-shift method stability, sensitivity, and accuracy were tested with reference to the standard curve, and clinical fecal samples were also examined. The results demonstrated that the 2 sets of primers based on the 2 SNPs could accurately distinguish between A. ceylanicum and A. tubaeforme. The coefficient of variation (CV) of Tm-values of AceP and AtuP was 0.07% and 0.06% in ITS101 and was 0.06% and 0.08% in ITS296, respectively. The minimum detectable DNA concentration was 5.22×10-6 and 5.28×10-6 ng/µl samples of AceP and AtuP, respectively. The accuracy of Tm-shift method reached 100% based on examination of 10 hookworm DNA samples with known species. In the clinical detection of hookworm in 69 stray cat fecal sample, the Tm-shift detection results were consistent with the microscopic examination and successfully differentiated between the 2-hookworm species. In conclusion, the developed method is a rapid, sensitive and accurate technique and can provide a promising tool for clinical detection and epidemiological investigation of cat-derived hookworms.


Assuntos
Ancylostoma/classificação , Ancylostoma/genética , Ancilostomíase/veterinária , Doenças do Gato/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Reação em Cadeia da Polimerase em Tempo Real/métodos , Temperatura de Transição , Ancylostoma/isolamento & purificação , Ancilostomíase/parasitologia , Animais , Doenças do Gato/parasitologia , Gatos , Primers do DNA/genética , DNA de Helmintos/genética , DNA Espaçador Ribossômico/genética , Fezes/parasitologia , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
18.
Int J Parasitol ; 49(5): 397-406, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30771359

RESUMO

Soil-transmitted nematodes infect over a billion people and place several billion more at risk of infection. Hookworm disease is the most significant of these soil-transmitted nematodes, with over 500 million people infected. Hookworm infection can result in debilitating and sometimes fatal iron-deficiency anemia, which is particularly devastating in children and pregnant women. Currently, hookworms and other soil-transmitted nematodes are controlled by administration of a single dose of a benzimidazole to targeted populations in endemic areas. While effective, people are quickly re-infected, necessitating frequent treatment. Widespread exposure to anthelmintic drugs can place significant selective pressure on parasitic nematodes to generate resistance, which has severely compromised benzimidazole anthelmintics for control of livestock nematodes in many areas of the world. Here we report, to our knowledge, the first naturally occurring multidrug-resistant strain of the canine hookworm Ancylostoma caninum. We reveal that this isolate is resistant to fenbendazole at the clinical dosage of 50 mg/kg for 3 days. Our data shows that this strain harbors a fixed, single base pair mutation at amino acid 167 of the ß-tubulin isotype 1 gene, and by using CRISPR/Cas9 we demonstrate that introduction of this mutation into the corresponding amino acid in the orthologous ß-tubulin gene of Caenorhabditis elegans confers a similar level of resistance to thiabendazole. We also show that the isolate is resistant to the macrocyclic lactone anthelmintic ivermectin. Understanding the mechanism of anthelmintic resistance is important for rational design of control strategies to maintain the usefulness of current drugs, and to monitor the emergence of resistance. The isolate we describe represents the first multidrug-resistant strain of A. caninum reported, and our data reveal a resistance marker that can emerge naturally in response to heavy anthelminthic treatment.


Assuntos
Ancylostoma/efeitos dos fármacos , Ancylostoma/isolamento & purificação , Doenças do Cão/parasitologia , Resistência a Medicamentos , Infecções por Uncinaria/veterinária , Ancylostoma/genética , Ancylostoma/crescimento & desenvolvimento , Animais , Anti-Helmínticos/farmacologia , Sequência de Bases , Cães , Feminino , Proteínas de Helminto/genética , Infecções por Uncinaria/parasitologia , Ivermectina/farmacologia , Masculino , Filogenia , Tiabendazol/farmacologia , Tubulina (Proteína)/genética
19.
Ann Agric Environ Med ; 25(3): 460-463, 2018 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-30260190

RESUMO

INTRODUCTION: While using recreational areas, people take with them accompanying animals (dogs, cats). These animals are the main source and reservoir of dangerous zoonoses, including parasitoses caused by genera of nematodes, also called roundworms, Toxocara spp., Trichuris spp. and Ancylostoma spp. Attention should also be paid to the possibility of contamination of recreational areas with the eggs of intestinal parasites by wild animals (e.g. red fox). MATERIAL AND METHODS: Sand was collected on beaches and leisure terrains located in recreational areas in Zwierzyniec, Rudka reservoir on the river Wieprz, 'Echo' reservoir, accessible to tourists to the Roztocze National Park, and the 'Florianiecki' reservoir, chich is inaccessible to tourists due to its location in the depths of the National Park. Material for the research was also collected from the 'Zemborzycki reservoir in Lublin, A total of 300 sand samples were collected. The samples, approximately 500 g each, were collected into plastic bags from the superficial layer of sand, at a depth up to 3 cm. For egg detection, flotation with the Wasylikowa metod and the modified Quinn method were used. In the Quinn metod, the flotation solution is NaCl saturated solution, in the Wasylikowa method, 5% solution of NaOH is used. RESULTS: The most contaminated grounds were areas by the water 'Zemborzycki' reservoir in Lublin. The eggs of intestinal parasites were isolated from 13 out of 24 samples examined. In sand collected from the beach by the 'Echo', 'Florianiecki' and 'Rudka' water resevoirs in Zwierzyniec, no eggs of intestinal parasites were detected. CONCLUSIONS: Such a dissemination of results may indicate a connection between the degree of occurrence of geohelmint eggs and the intensity of human movement, together with accompanying animals, in places that can be used for recreation.


Assuntos
Ancylostoma/isolamento & purificação , Óvulo/química , Solo/parasitologia , Toxocara/isolamento & purificação , Trichuris/isolamento & purificação , Ancylostoma/fisiologia , Animais , Monitoramento Ambiental , Fezes/parasitologia , Humanos , Contagem de Ovos de Parasitas , Polônia , Recreação , Toxocara/fisiologia , Trichuris/fisiologia
20.
Am J Trop Med Hyg ; 99(5): 1186-1193, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-30226132

RESUMO

Hookworm disease caused by Necator americanus, Ancylostoma duodenale, and Ancylostoma ceylanicum affects half a billion people worldwide. The prevalence and intensity of infection of individual hookworm species are vital for assessing morbidity and generating targeted intervention programs for their control. The present study aims to evaluate a multiplex real-time quantitative PCR (qPCR) assay to determine the prevalence and egg intensity of all three hookworm species and compare this with standard microscopy and published genus-based conventional and real-time multiplex qPCRs. Performance of the diagnostic assays was evaluated using DNA extracted from 192 fecal samples collected as part of a soil-transmitted helminth (STH) survey in northern Cambodia. The prevalence of hookworms as detected by the multiplex hookworm qPCR of 84/192 (43.8%) was significantly higher than that using microscopy of 49/192 (25.5%). The hookworm multiplex qPCR showed very good agreement for the detection of both N. americanus (Kappa 0.943) and Ancylostoma spp. (Kappa 0.936) with a multiplex STH qPCR. A strong and moderate quantitative correlation between cycle threshold and eggs per gram (EPG) feces was obtained for the hookworm qPCR for seeded DNA egg extracts (R 2 ≥ 0.9004) and naturally egg-infected individuals (R 2 = 0.6848), respectively. The newly developed hookworm quantitative multiplex qPCR has the potential for application in anthelmintic efficacy trials and for monitoring the success of mass deworming programs targeting individual species of anthroponotic and zoonotic hookworms.


Assuntos
Ancylostoma/isolamento & purificação , Fezes/parasitologia , Infecções por Uncinaria/diagnóstico , Reação em Cadeia da Polimerase Multiplex/métodos , Ancylostoma/genética , Ancilostomíase/diagnóstico , Ancilostomíase/epidemiologia , Animais , Anti-Helmínticos/uso terapêutico , Camboja/epidemiologia , Estudos Transversais , DNA de Helmintos/genética , Infecções por Uncinaria/epidemiologia , Humanos , Óvulo , Contagem de Ovos de Parasitas , Prevalência
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