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1.
BMC Health Serv Res ; 23(1): 1350, 2023 Dec 04.
Artículo en Inglés | MEDLINE | ID: mdl-38049849

RESUMEN

BACKGROUND: Discarding pharmaceuticals in the garbage or into the sewage system are still the most common methods in many countries. This study aims to investigate the guidance provided by pharmacists to customers on the disposal of unused and expired household medications in São Paulo State, Brazil. METHOD: The study population consisted of 630 pharmacists from the State of São Paulo, who work in community pharmacies. They answered an online questionnaire with questions composed in three blocks: demographic, work, and academic information on the pharmacist; guidance about the disposal of household medications; and knowledge regarding the reverse logistics of these medications. An invitation to participate in the questionnaire was made via WhatsApp, individually and collectively. Inferential statistics were performed using the chi-square test and were considered significant when p < 0.05%. RESULTS: Among the participating pharmacists, the majority were women under 60 years old,56 (8.89%) stated that they never orient the customer regarding the disposal of unused and expired household medications, while 574 (91,12%) indicated that they almost provide guidance. The frequency with which they provided guidance was influenced by the number of years since graduation (p = 0.0047), the time they had worked in pharmacies and drugstores (p = 0.0007), and whether or not they had a graduate degree (p = 0.0181). Regarding the disposal of medications, among the 643 responses provided by the pharmacists,516 (80.25%) indicated that they oriented customers to return them to a pharmacy. CONCLUSION: A small number of pharmacists always orient customers on the proper disposal that should be followed for unused and expired household medications, prioritizing their return to a pharmacy. In general, these pharmacists have longer periods of work experience and higher academic qualifications. Thus, it is important to increase knowledge through professional training and further education programs.


Asunto(s)
Servicios Farmacéuticos , Farmacéuticos , Humanos , Masculino , Femenino , Persona de Mediana Edad , Brasil , Encuestas y Cuestionarios , Muerte , Preparaciones Farmacéuticas , Conocimientos, Actitudes y Práctica en Salud
2.
Pathogens ; 12(12)2023 Nov 27.
Artículo en Inglés | MEDLINE | ID: mdl-38133278

RESUMEN

We investigated the zoonotic transmission of Cryptosporidium among the children (n = 188), dogs (n = 133), and cats (n = 55) living in 188 households. Fecal samples were examined using ELISA and confirmed via nested PCR. Coproantigens oocysts were detected in 3.7% of children, 8.3% of dogs, and 5.5% of cats. We found strong evidence of two cases of the zoonotic transmission of Cryptosporidium canis between children and dogs. Furthermore, four children and their respective pets (one dog and three cats) were infected with Cryptosporidium parvum, but we cannot exclude the hypotheses that the oocysts were transmitted from children to animals or that both hosts were infected by a shared source, such as contaminated water or food. The presence of an infected animal elevated the risk of zoonotic transmission by 129.7-fold (95% CI: 13.92-1209.68). Furthermore, sharing a bed with pets was identified as a risk factor for infection in children (OR: 9.9, 95% CI: 1.37-71.2). In conclusion, the zoonotic transmission of Cryptosporidium among children and pets cohabiting in the same household may be quite common, especially when infected animals lie or sleep on children's beds. These findings unequivocally highlight the public health concern surrounding C. canis.

3.
Pathogens ; 12(5)2023 May 13.
Artículo en Inglés | MEDLINE | ID: mdl-37242380

RESUMEN

Cockatiels (Nymphicus hollandicus) are among the most commonly sold psittacines pets. The aim of this study was to evaluate the occurrence of Cryptosporidium spp. in domestic N. hollandicus and identify risk factors for this infection. We collected fecal samples from 100 domestic cockatiels in the city of Araçatuba, São Paulo, Brazil. Feces from birds of both genders and older than two months were collected. Owners were asked to complete a questionnaire to identify how they handle and care for their birds. Based on nested PCR targeting the 18S rRNA gene, the prevalence of Cryptosporidium spp. in the cockatiels sampled was 9.00%, 6.00% based on Malachite green staining, 5.00% based on modified Kinyoun straining, and 7.00% when the Malachite green was combined with Kinyoun. Applying multivariate logistic regression to test the association between Cryptosporidium proventriculi positivity and potential predictors showed that gastrointestinal alterations was a significant predictor (p < 0.01). Amplicons from five samples were sequenced successfully and showed 100% similarity with C. proventriculi. In summary, this study demonstrates the occurrence of C. proventriculi in captive cockatiels.

4.
Vet Ital ; 58(2)2022 12 30.
Artículo en Inglés | MEDLINE | ID: mdl-36586119

RESUMEN

Intestinal parasites inhabit the intestinal tract of humans and animals, causing damages whose severity depends on several factors related to the parasite and the host. Immunocompromised individuals are more likely to develop severe forms of parasitic infestation. The diagnosis of the gastrointestinal parasitosis is mainly performed by the examination of the feces, which consists of the direct visualization and identification of the parasites eliminated through the feces. These tests are generally low sensitive and the microscope slides contain a large number of impurities, which can impair the result of the diagnosis. In order to improve the diagnostic accuracy, a new parasitological technique called Three Fecal Test (TF­Test) was developed. To further improve its diagnostic accuracy, few modifications of the original protocols have been made with the years. In this study the performance of these new techniques to detect gastrointestinal parasites in human and animal fecal samples was described and discussed in relation to the performance of other conventional coprological tests. It could be concluded that the TF­Test conventional and modified can be used for the diagnosis of several human and animal parasites, with satisfactory results.


Asunto(s)
Parasitosis Intestinales , Parásitos , Humanos , Animales , Parasitosis Intestinales/diagnóstico , Parasitosis Intestinales/veterinaria , Parasitosis Intestinales/parasitología , Heces/parasitología
5.
J. Health Biol. Sci. (Online) ; 10(1): 1-6, 01/jan./2022. tab, ilus
Artículo en Inglés | LILACS | ID: biblio-1411467

RESUMEN

Objective: to describe risk factors associated with SARS-CoV-2 infection. Methods: this is a retrospective descriptive cross-sectional study aimed at describing the epidemiological profile of laboratory and clinical diagnosis of unvaccinated patients seen at a basic health unit in Araçatuba ­ SP, infected with SARS-CoV-2 between June 2020 and January 2021.The results were analyzed through inferential and descriptive statistics. Additionally, Chi-square and Fisher exact tests were used (p<0.05). Results: of 313 patients, 128 were positive for COVID-19, with 68.75% diagnosed by RT-PCR and the others by immunochromatography. Women were 51.56% of those infected with adults corresponding to the main age group (76.56%), and 57% of patients had only a basic educational level concluded. A total of 88.26% of the patients progressed to cure without complications; eight patients died, most of whom were men and elderly. Of the variables analyzed for positive/negative outcomes, only "basic educational level" was significant for a positive result(p=0.0019). Conclusion: the deaths of infected patients may be associated with the existence of at least one comorbidity and advanced age of men.


Objetivo: descrever os fatores de risco associados com a infecção por SARS-CoV-2. Métodos: trata-se de um estudo descritivo retrospectivo e transversal, voltado a descrever o perfil epidemiológico de diagnóstico laboratorial e clínico, de pacientes não vacinados, atendidos em uma unidade básica de saúde de Araçatuba-SP, infectados por SARS-CoV-2, no período entre junho de 2020 e janeiro de 2021. Os resultados foram analisados por estatística inferencial e descritiva. Adicionalmente, foram aplicados os testes de Qui-quadrado e exato de Fisher (p<0.05). Resultados: dos 313 pacientes, 128 apresentaram resultado positivo para COVID-19, com 68,75% diagnosticados por RT-PCR e o restante por imunocromatografia. Mulheres foram 51,56% dos infectados, com adultos correspondendo à principal faixa etária (76,56%), 57% dos pacientes apresentavam apenas o nível educacional básico concluído. O quadro de 88.26%dos pacientes evoluiu para cura sem complicações;8 pacientes foram a óbito, sendo, em sua maioria, homens e idosos. Das variáveis analisadas para grau de dependência de resultado positivo/negativo, apenas "nível escolar básico" apresentou resultado significante para resultado positivo (p=0.0019). Conclusão: os óbitos dos pacientes infectados podem ser associados à existência de, pelo menos, uma comorbidade e à idade avançada de homens.


Asunto(s)
SARS-CoV-2 , COVID-19 , Comorbilidad , Diagnóstico Clínico , Incidencia , Estudios Transversales , Factores de Riesgo
6.
Exp Parasitol ; 239: 108303, 2022 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-35716768

RESUMEN

Countless research is carried out until new discoveries are transformed into products or services available to the population. This trajectory can be slower and more costly or even impossible when irreproducible data are obtained in the most diverse fields of science. Thus, quality management appears as an essential tool to guarantee the reliability of academic research results. In this work, we demonstrate the applied strategy to implement a Quality Management System (QMS) in a research laboratory in Veterinary Parasitology and we highlight the adaptable quality requirements in this scientific research environment. For this, the Plan-Do-Check-Act (PDCA) quality tool was used, and two internal audits were performed, one before and one after implementation. The audits reached 67 (41.36%) and 157 (96.91%) points before and after implementation, respectively, with a significant difference between the moments studied. Thus, we demonstrate that the adoption of QMS principles in research is feasible. The methodology applied in this work can be adopted by managers from other laboratories interested in the implementation of quality standards as a support in the reproducibility of research.


Asunto(s)
Laboratorios , Control de Calidad , Reproducibilidad de los Resultados
7.
Transbound Emerg Dis ; 69(5): e2302-e2311, 2022 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-35478434

RESUMEN

The Leishmania infantum (synonym, Leishmania chagasi) causes life-threatening infection, namely canine leishmaniosis (CanL), which is a chronic zoonosis prevalent in various countries and spread by the bite of the infected Lutzomyia female sandfly in South America. The objective of the study was to assess the effectiveness of a polymer matrix collar containing made up of 10% imidacloprid and 4.5% flumethrin for the prevention of canine leishmaniosis from the hyperendemic region falling under Araçatuba municipality (Brazil). The research included a total of 146 dogs chosen from 75 households. Test were initiated via physical examination; weighing and biological sample collection (blood, popliteal lymph node and conjunctival swab) of these dogs were done in March 2018 (Day 0; GA, control = 69, GB, treated = 77) to initiate laboratory tests. Post-inclusion, the animals were monitored on the 120th, 240th, 360th and 480th days, respectively. The usage of collars continued between 0 and 480 days before being substituted in second (D240) and fourth (D480) follow-up visits. On the whole, 25 dogs in GA (36.2%) and three in GB (3.9%) were found positive for L. infantum infection in a minimum of one diagnostic test used in the research. Therefore, the average collar effectiveness for protection from L. infantum infection was 89.2% (p < .01). In the last follow-up, the average incidence density rate for GA was 30.7%, whereas for GB, it was 2.9%. The imidacloprid/flumethrin collars evaluated in the research were found to be safe and extremely efficient for the prevention of L. infantum infection through Lutzomyia species among the large population of dogs in highly prone endemic regions. This is a dependable and efficient technique aimed at reducing the occurrence and propagation of this illness among the population of canines, which would eventually reduce the human-health-related hazards. In Brazil, Lutzomyia spp. is a leading vector of the infection; thus, the collar can be used to limit infection in dogs and humans.


Asunto(s)
Enfermedades de los Perros , Insecticidas , Leishmania infantum , Leishmaniasis Visceral , Leishmaniasis , Psychodidae , Animales , Brasil/epidemiología , Enfermedades de los Perros/epidemiología , Enfermedades de los Perros/prevención & control , Perros , Femenino , Humanos , Leishmaniasis/epidemiología , Leishmaniasis/prevención & control , Leishmaniasis/veterinaria , Leishmaniasis Visceral/epidemiología , Leishmaniasis Visceral/veterinaria , Neonicotinoides , Nitrocompuestos , Polímeros , Piretrinas
8.
Vet Res Commun ; 46(1): 49-58, 2022 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-34499298

RESUMEN

We analyzed Cryptosporidium spp. in fecal samples of wild cervids (Ozotoceros bezoarticus, Blastocerus dichotomus, Mazama nana, Mazama americana, and Mazama bororo) from many Brazilian regions, a fact unprecedented in the literature. Sniffer dogs were used to collect 936 fecal samples of cervids from 14 Brazilian localities. Cervids species were identified using polymerase chain reaction (PCR) performed from genomic DNA extracted from 563 fecal samples of Ozotoceros bezoarticus, Blastocerus dichotomus, Mazama nana, Mazama americana, and Mazama bororo. Cryptosporidium spp. oocyst screening was performed using malachite green negative staining. Nested PCR (nPCR) protocols targeting the 18S rRNA and GP60 genes followed by genetic sequencing were performed for Cryptosporidium spp. detection and Cryptosporidium parvum subtyping, respectively. Nested PCR targeting actin gene and genetic sequencing were performed in samples with non-identified Cryptosporidium species by 18S rRNA amplicon sequencing. The association between the occurrence of Cryptosporidium and the presence of bovines in the same locality was evaluated using Fisher's exact test. The positivity rates of diagnostic methods were compared by McNemar test and the Kappa correlation coefficient. The prevalence rates of Cryptosporidium spp. in cervids were 1.42% (8/563) and 0.36% (2/563) by nPCR and malachite green negative staining, respectively. C. parvum IIaA16G3R1 isolate was identified in three fecal samples from M. americana, two from M. nana and one from B. dichotomus. Cryptosporidium ryanae were found in one sample from B. dichotomus. We identified a new Cryptosporidium genotype, named Cryptosporidium deer genotype BR, from one M. americana fecal sample.


Asunto(s)
Criptosporidiosis , Cryptosporidium , Ciervos , Animales , Criptosporidiosis/epidemiología , Cryptosporidium/genética , Heces , Reacción en Cadena de la Polimerasa/veterinaria
9.
Parasitol Int ; 80: 102216, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-33137502

RESUMEN

This research had as objective to evaluate the occurrence and to characterize genetically the infections by Cryptosporidium in Mazama gouazoubira. By a non-invasive harvest methodology using trained sniffer dogs to locate fecal samples of cervids, 642 fecal samples were obtained from six Brazilian localities. The cervids species responsible for the excretion of each fecal sample were identified by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), using the mitochondrial cytochrome b target gene (cyst b) and the restriction enzymes Sspl, AflIII and BstN. From this identification, 437 fecal samples of M. gouazoubira were selected for research of Cryptosporidium spp. performed through negative staining with malachite green and polymerase chain reaction (nPCR), with the subunit of 18S rRNA gene, followed by sequencing the amplified products. In the samples that were diagnosed the presence of parasite species with zoonotic potential, genotyping was also performed using nPCR with the subunit of GP60 gene. Statistical analysis consisted of the Fisher exact test to verify the association of the presence of the enteroparasite in relation to the presence of cattle in each locality, and the McNemar tests and Kappa correlation coefficient used to compare the results obtained between the two diagnostic techniques. In the fecal samples of M. gouazoubira the occurrences of Cryptosporidium were diagnosed in 1.6% (7/437) and 1.1% (5/437), respectively, through nPCR and microscopy. Cryptosporidium. parvum was diagnosed in 100% (7/7) of the samples submitted to sequencing (18S gene). The IIaA16G3R1 subtype was diagnosed in five of the C. parvum samples submitted to genotyping (GP60 gene). This is the first world report of C. parvum in M. gouazoubira and subtype IIaA16G3R1 in cervids.


Asunto(s)
Criptosporidiosis/diagnóstico , Cryptosporidium parvum/aislamiento & purificación , Ciervos , Heces/parasitología , Animales , Brasil , Bovinos , Criptosporidiosis/parasitología , Reacción en Cadena de la Polimerasa/veterinaria , Polimorfismo de Longitud del Fragmento de Restricción , ARN de Helminto/análisis , ARN Ribosómico 18S/análisis
10.
Rev Soc Bras Med Trop ; 53: e20190535, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32491097

RESUMEN

Since the early 20th century, the detection of intestinal parasites has improved with the development of several techniques for parasitic structures recovery and identification, which differ in sensitivity, specificity, practicality, cost, and infrastructure demand. This study aims to review, in chronological order, the stool examination techniques and discuss their advantages, limitations, and perspectives, and to provide professionals and specialists in this field with data that lays a foundation for critical analysis on the use of such procedures. The concentration procedures that constitute the main techniques applied in routine research and in parasitological kits are a) spontaneous sedimentation; b) centrifugation-sedimentation with formalin-ethyl acetate; and c) flotation with zinc sulfate solution. While selecting a technique, one should consider the purpose of its application and the technical-operational, biological, and physicochemical factors inherent in the procedures used in stool processing, which may restrict its use. These intrinsic limitations may have undergone procedural changes driven by scientific and technological development and by development of alternative methods, which now contribute to the improvement of diagnostic accuracy.


Asunto(s)
Heces/parasitología , Parasitosis Intestinales/diagnóstico , Parasitología/historia , Manejo de Especímenes/historia , Animales , Historia del Siglo XX , Historia del Siglo XXI , Humanos , Parasitología/métodos , Sensibilidad y Especificidad , Manejo de Especímenes/métodos
11.
Rev. Soc. Bras. Med. Trop ; 53: e20190535, 2020. tab
Artículo en Inglés | LILACS, Coleciona SUS, Sec. Est. Saúde SP | ID: biblio-1136801

RESUMEN

Abstract Since the early 20th century, the detection of intestinal parasites has improved with the development of several techniques for parasitic structures recovery and identification, which differ in sensitivity, specificity, practicality, cost, and infrastructure demand. This study aims to review, in chronological order, the stool examination techniques and discuss their advantages, limitations, and perspectives, and to provide professionals and specialists in this field with data that lays a foundation for critical analysis on the use of such procedures. The concentration procedures that constitute the main techniques applied in routine research and in parasitological kits are a) spontaneous sedimentation; b) centrifugation-sedimentation with formalin-ethyl acetate; and c) flotation with zinc sulfate solution. While selecting a technique, one should consider the purpose of its application and the technical-operational, biological, and physicochemical factors inherent in the procedures used in stool processing, which may restrict its use. These intrinsic limitations may have undergone procedural changes driven by scientific and technological development and by development of alternative methods, which now contribute to the improvement of diagnostic accuracy.


Asunto(s)
Humanos , Animales , Historia del Siglo XX , Historia del Siglo XXI , Parasitología/historia , Manejo de Especímenes/historia , Heces/parasitología , Parasitosis Intestinales/diagnóstico , Parasitología/métodos , Manejo de Especímenes/métodos , Sensibilidad y Especificidad
12.
Food Waterborne Parasitol ; 16: e00062, 2019 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-32095632

RESUMEN

We performed molecular characterization of Giardia duodenalis in buffalo calves from the Southwest region of São Paulo State, Brazil. A total of 183 fecal samples of Murrah breed buffaloes up to six months of age were collected. We examined these samples by the polymerase chain reaction (PCR) targeting the small-subunit ribosomal RNA gene and positive samples were characterized using additional PCR assays targeting a portion of the beta-giardin, the glutamate dehydrogenase and the triose-phosphate isomerase genes. Based on the SSU rRNA nPCR, the presence of G. duodenalis was confirmed in 12 (6.56%) of fecal samples, of these, five, four and three samples were positive for the tpi, bg and gdh genes, respectively. Assemblage identification by sequencing was successful in 6 of 12 samples and sequence analysis showed 100% genetic similarity with G. duodenalis assemblage E. This observation represents the first detection of G. duodenalis assemblage E in buffaloes calves in Brazil.

13.
Rev Bras Parasitol Vet ; 27(1): 61-66, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-29641795

RESUMEN

This study used several diagnostic methods to examine the occurrence of and molecularly characterize Cryptosporidium spp. in captive canaries (Serinus canaria) in southern and southeastern Brazil. A total of 498 fecal samples were purified by centrifugal-flotation using Sheather's solution. Cryptosporidium spp. diagnosis was performed using three diagnostic methods: malachite green negative staining, nested PCR targeting the 18S rRNA gene, followed by sequencing the amplified fragments, and duplex real-time PCR targeting the 18S rRNA specific to detect Cryptosporidium galli and Cryptosporidium avian genotype III. The overall positivity for Cryptosporidium spp. (total samples positive in at least one protocol) from the microscopic analysis, nested PCR and duplex real-time PCR protocol results was 13.3% (66/498). The positivity rates were 2.0% (10/498) and 4.6% (23/498) for Cryptosporidium spp. by microscopy and nested PCR, respectively. Sequencing of 20 samples amplified by nested PCR identified C. galli (3.0%; 15/498), Cryptosporidium avian genotype I (0.8%; 4/498) and Cryptosporidium avium (0.2%; 1/498). Duplex real-time PCR revealed a positivity of 7.8% (39/498) for C. galli and 2.4% (12/498) for avian genotype III. Malachite green negative staining differed significantly from nested PCR in detecting Cryptosporidium spp. Duplex real-time PCR was more sensitive than nested PCR/sequencing for detecting gastric Cryptosporidium in canaries.


Asunto(s)
Canarios/parasitología , Criptosporidiosis/diagnóstico , Criptosporidiosis/parasitología , Cryptosporidium/aislamiento & purificación , Animales , Animales Domésticos , Brasil , Cryptosporidium/genética , ADN/análisis , Técnicas de Diagnóstico Molecular
14.
Prev Vet Med ; 151: 73-78, 2018 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-29496109

RESUMEN

The objective of this study was to determine the occurrence of Cryptosporidium spp. in domestic chickens raised in different chicken production systems in Brazil using three nested PCR protocols. The purification and concentration of oocysts present in 190 fecal samples from chickens raised in extensive, semi-intensive and intensive production systems were accomplished by centrifugal flotation in Sheather's solution and were followed by the extraction of genomic DNA. The detection and molecular characterization of Cryptosporidium species and genotypes were performed using three nested polymerase chain reaction (nested PCR) protocols targeting the 18S rRNA gene followed by sequencing of the amplified fragments. Subgenotyping of C. meleagridis was performed using a nested PCR reaction targeting the gp60 gene. Sample identified as Cryptosporidium sp. genetically similar to Cryptosporidium xiaoi and Cryptosporidium bovis by 18S rRNA gene sequencing were further analyzed by nested PCR targeting the actin gene and subsequent sequencing of the amplified fragment. Positive amplification for Cryptosporidium spp. was observed in 12.6% (24/190) of the samples, including C. baileyi (9.8%; 18/190), C. meleagridis (0.5%, 1/190), C. parvum (2.1%; 4/190) and Cryptosporidium sp. (0.5%; 1/190). Subgenotyping of C. meleagridis revealed the presence of the zoonotic subtype IIIgA23G3R1. Sequencing of the 18S rRNA gene and the actin gene fragments revealed a Cryptosporidium genotype in an extensive poultry system genetically related to C. xiaoi and C. bovis. There was no significant difference in the frequency of positive results obtained by the three nested PCR protocols (p > 0.05); additionally, the agreement obtained by Kappa index ranged from substantial (0.70) to almost perfect (0.9).


Asunto(s)
Pollos , Criptosporidiosis/epidemiología , Cryptosporidium/aislamiento & purificación , Enfermedades de las Aves de Corral/epidemiología , Actinas/genética , Crianza de Animales Domésticos/métodos , Animales , Proteínas Bacterianas/genética , Brasil/epidemiología , Criptosporidiosis/parasitología , Cryptosporidium/clasificación , ADN Bacteriano/genética , Femenino , Reacción en Cadena de la Polimerasa/veterinaria , Enfermedades de las Aves de Corral/parasitología , Prevalencia , ARN Ribosómico 18S/genética , Análisis de Secuencia de ADN/veterinaria
15.
Rev. bras. parasitol. vet ; 27(1): 60-65, Jan.-Mar. 2018. tab
Artículo en Inglés | LILACS | ID: biblio-899315

RESUMEN

Abstract This study used several diagnostic methods to examine the occurrence of and molecularly characterize Cryptosporidium spp. in captive canaries (Serinus canaria) in southern and southeastern Brazil. A total of 498 fecal samples were purified by centrifugal-flotation using Sheather's solution. Cryptosporidium spp. diagnosis was performed using three diagnostic methods: malachite green negative staining, nested PCR targeting the 18S rRNA gene, followed by sequencing the amplified fragments, and duplex real-time PCR targeting the 18S rRNA specific to detect Cryptosporidium galli and Cryptosporidium avian genotype III. The overall positivity for Cryptosporidium spp. (total samples positive in at least one protocol) from the microscopic analysis, nested PCR and duplex real-time PCR protocol results was 13.3% (66/498). The positivity rates were 2.0% (10/498) and 4.6% (23/498) for Cryptosporidium spp. by microscopy and nested PCR, respectively. Sequencing of 20 samples amplified by nested PCR identified C. galli (3.0%; 15/498), Cryptosporidium avian genotype I (0.8%; 4/498) and Cryptosporidium avium (0.2%; 1/498). Duplex real-time PCR revealed a positivity of 7.8% (39/498) for C. galli and 2.4% (12/498) for avian genotype III. Malachite green negative staining differed significantly from nested PCR in detecting Cryptosporidium spp. Duplex real-time PCR was more sensitive than nested PCR/sequencing for detecting gastric Cryptosporidium in canaries.


Resumo Este trabalho teve como objetivos determinar a ocorrência e realizar a caracterização molecular de Cryptosporidium spp. em 498 amostras fecais de canários (Serinus canaria) criados em cativeiro, utilizando três métodos de diagnóstico: análise microscópica pela coloração negativa com verde malaquita, nested PCR seguida de sequenciamento dos fragmentos amplificados e PCR duplex em tempo real específica para detecção de Cryptosporidium galli e Cryptosporidium genótipo III de aves. A positividade total para Cryptosporidium spp. (total de amostras positivas em pelo menos um método de diagnóstico) obtida pela análise microscópica, nested PCR e PCR duplex em tempo real foi de 13,3% (66/498). As taxas de positividade para Cryptosporidium spp. foram 2,0% (10/498) e 4,6% (23/498) por microscopia e nested PCR, respectivamente. O sequenciamento de 20 amostras amplificadas pela nested PCR identificou C. galli (3,0%; 15/498), Cryptosporidium genótipo I de aves (0,8%; 4/498) e Cryptosporidium avium (0,2%; 1/498). A PCR duplex em tempo real revelou positividade de 7,8% (39/498) para C. galli e 2,4% (12/498) para Cryptosporidium genótipo III de aves. A análise microscópica diferiu significativamente da nested PCR para detecção de Cryptosporidium spp. A PCR duplex em tempo real apresentou maior sensibilidade que a nested PCR/sequenciamento para detectar as espécies/genótipos gástricos de Cryptosporidium.


Asunto(s)
Animales , Canarios/parasitología , Criptosporidiosis/diagnóstico , Criptosporidiosis/parasitología , Cryptosporidium/aislamiento & purificación , Brasil , ADN/análisis , Cryptosporidium/genética , Técnicas de Diagnóstico Molecular , Animales Domésticos
16.
Exp Parasitol ; 184: 109-114, 2018 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-29247662

RESUMEN

The aim of this study was to evaluate the prevalence of and diagnostic methods for Cryptosporidium spp. in caged adult exotic parrots from Southern and Southeastern Brazil. Oocysts were purified from fecal samples from 463 psittacines by centrifugal-flotation in Sheather's sugar solution. Cryptosporidium spp. were detected by malachite green negative staining and nested PCR targeting the 18S rRNA gene. Cryptosporidium species were identified by sequencing nested PCR amplicons. Samples were also tested by duplex real-time PCR targeting the 18S rRNA gene of Cryptosporidium galli and Cryptosporidium avian genotype III. The prevalence rates of Cryptosporidium spp. determined by microscopy and nested PCR were 3.0% (14/463) and 5.0% (23/463), respectively. The nested PCR/sequencing identified avian genotype III (1.7%; 8/463), Cryptosporidium parvum (0.9%; 4/463) and Cryptosporidium canis (0.2%; 1/463). Duplex real-time PCR was positive for gastric Cryptosporidium in 9.5% (44/463) of the samples. Among them, 1.9% (9/463) were positive for C. galli, 5.8% (27/463) were positive for avian genotype III and 1.7% (8/463) showed mixed infections with C. galli and avian genotype III. With regards to the positive detection of Cryptosporidium spp., there was no statistically significant difference between nested PCR and microscopic analysis (p = .1237), and a fair agreement existed between them (Kappa = 0.242). A statistically significant difference (p < .0001) and fair agreement (Kappa = 0.317) were obtained between nested PCR/sequencing and duplex real-time PCR for the detection of gastric Cryptosporidium. We determined that nested PCR and duplex real-time PCR are the best options for the detection of Cryptosporidium spp. and gastric Cryptosporidium, respectively, and that avian genotype III is the most common Cryptosporidium genotype/species in psittacines.


Asunto(s)
Enfermedades de las Aves/diagnóstico , Criptosporidiosis/diagnóstico , Cryptosporidium/aislamiento & purificación , Loros/parasitología , Animales , Animales Domésticos , Enfermedades de las Aves/epidemiología , Enfermedades de las Aves/parasitología , Brasil/epidemiología , Clonación Molecular , Criptosporidiosis/epidemiología , Criptosporidiosis/parasitología , Cryptosporidium/clasificación , Cryptosporidium/genética , ADN Protozoario/química , Heces/parasitología , Reacción en Cadena de la Polimerasa/veterinaria , Prevalencia , ARN Ribosómico 18S/genética , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Análisis de Secuencia de ADN/veterinaria
17.
Rev. patol. trop ; 46(4): 321-330, dez. 2017. mapa, tab, graf
Artículo en Inglés | LILACS | ID: biblio-913724

RESUMEN

Intestinal parasites are among the major causative agents of diseases that affect animals and humans, especially children. In view of this, the current study evaluated the occurrence of these parasitic agents in 737 children in an urban region with excellent sanitation condition of the city of Pedreira, São Paulo, Brazil. Fecal samples from the children were processed with the use of a technique of high diagnostic efficiency (TF-Test®). The diagnosis of these samples resulted in the detection of 557 parasitic structures among eleven genera of parasites, and of 46.4% (342/737) infected children. Blastocystis spp. was found in 69.6% (238/342) of the positive samples and the monoparasitism was accompanied by symptoms in 44 children. Furthermore, 67.8% (232/342) of the infected children had close contact with pets, suggesting a possible zoonotic transmission. Lastly, this study allowed to perform health education to the children, aiming the reduction of new intestinal parasitic infections.


Asunto(s)
Humanos , Niño , Parásitos , Niño , Salud Pública , Diagnóstico , Infecciones , Intestinos/parasitología
18.
Vet Parasitol ; 243: 148-150, 2017 Aug 30.
Artículo en Inglés | MEDLINE | ID: mdl-28807284

RESUMEN

The carrier pigeon and the domestic pigeon are different breeds of the species Columba livia. Carrier pigeons are used for recreational activities such as bird contests and exhibitions. Due to the close contact with humans, these birds may potentially represent a public health risk, since they can host and disseminate zoonotic parasites, such as those belonging to the genus Cryptosporidium (phylum Apicomplexa). The purpose of this work was the detection by microscopic and molecular techniques of Cryptosporidium spp. oocysts in fecal samples of carrier pigeons, and subsequently to sequence the 18S ribosomal RNA marker of positive samples to identify the species. A total of 100 fecal samples were collected individually in two pigeon breeding facilities from Formiga and Araçatuba, cities located in Minas Gerais state and São Paulo state, Brazil, respectively. The age of the birds ranged from one to 12 years; 56 were females and 44 males. Fecal smears were stained with negative malachite green, whereas the molecular characterization was based on the sequence of a ∼800bp fragment of the 18S rRNA gene. Microscopic examination of fecal smears revealed 4% (4/100) oocyst positivity. On the other hand, 7% (7/100) of positivity were found using nested PCR. Three samples were 99% to 100% similar to Cryptosporidium parvum 18S rDNA type A (Genbank AH006572) and the other three samples had 99% to 100% similarity to C. parvum 18S rDNA type B (Genbank AF308600). To our knowledge, this is the first report of C. parvum oocysts in carrier pigeons.


Asunto(s)
Enfermedades de las Aves/parasitología , Columbidae , Criptosporidiosis/parasitología , Cryptosporidium parvum/aislamiento & purificación , Animales , Heces/parasitología , Femenino , Masculino , ARN Protozoario/genética , ARN Ribosómico 18S
19.
Vet Parasitol Reg Stud Reports ; 10: 154-156, 2017 12.
Artículo en Inglés | MEDLINE | ID: mdl-31014590

RESUMEN

Cryptosporidiosis in birds manifests as an acute or chronic disease of the respiratory or digestive tracts. The objective of our study was to perform the molecular characterization of Cryptosporidium spp. in wild psittacines kept in captivity at the Araçatuba Municipal Zoo, São Paulo, Brazil. A total of 47 fecal samples were collected from Amazona aestiva, Psittacara leucophthalma, and Ara ararauna. All samples were collected at the time the birds defecated. DNA extraction was performed using the ZR Fecal DNA MiniPrep™ kit (Zymo Research). Screening for Cryptosporidium spp. was accomplished using nested PCR targeting the 18S RNA gene and sequencing of amplified fragments. Positivity for Cryptosporidium spp. (10.64%; 5/47) was found in A. ararauna (4) and P. leucophthalma (1) samples. The amplified fragments were sequenced and showed 100% genetic similarity with Cryptosporidium baileyi.


Asunto(s)
Animales de Zoológico/parasitología , Enfermedades de las Aves/parasitología , Cryptosporidium/aislamiento & purificación , Loros/parasitología , Animales , Enfermedades de las Aves/epidemiología , Brasil/epidemiología , Cryptosporidium/clasificación , Heces/parasitología
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