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In this study, the frequency of canines infected with Leishmania spp. in an area endemic to leishmaniasis in humans was determined. A descriptive pilot study was conducted between the months of October and December 2020 on dogs from Rota, a community in the municipality of León, which included 45 specimens from the peridomestic area. Different variables from each specimen were monitored, such as age, sex, breed, body condition, and clinical characteristics, as well as information on the owners and cases of human leishmaniasis presented in less than 5 years. Blood samples were collected from the cephalic vein and peripheral blood was separated. A complete blood count (CBC) was performed using venous blood samples with ethylene diamine tetraacetic acid (EDTA), as well as a conventional PCR was applied for the detection of Leishmania spp. Amastigotes were found in 22% of venous or peripheral blood samples, whereas a high prevalence of 28.89% (95% CI: 14.53-43.24) was found by PCR. Only 1/12 of positive dogs in PCR presented dry exfoliative dermatitis, therefore, there was no significant difference (p ≥ 0.05), the age and sex of the dogs were also not factors associated with infection (p ≥ 0.05). This study reports for the first time the molecular detection of Leishmania in dogs in an endemic area of leishmaniasis in humans in Nicaragua. The high frequency of dogs infected with Leishmania suggests that they play an important role in the transmission cycle of human leishmaniasis.
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Various subspecies of Apis mellifera L. were introduced to Central America since colonization 500 years ago. Hybridization increased with the entrance of the Africanized bee in Nicaragua in 1984. Rustic beekeeping activities and numerous feral swarms define the genetic pattern, reflected in phenotypic heterogeneity and remarkable differences in the behaviour of the bee colonies, especially the nest defence. Due to these facts, the question emerge about the degree of Africanization of honeybee colonies in Nicaragua. In this study, we identified Africanized honeybee colonies based on the single character "mean forewing length" and we corroborated our results by determining mitotypes using mtDNA analysis. Morphometric and genetic approaches were realized in three different geographical zones of Nicaragua and related to beehive characteristics and management. Worker bee samples were taken from the inside of 146 hives from 26 apiaries. Abdominal colour as phenotypic character was the first examination, followed by measurement of 1460 right forewings to determine corresponding probability of Africanization. More than 60% of the beehives showed phenotypic heterogeneity and mean forewing length of 8.74 mm (SD 0.16 mm) indicated a high degree of Africanization. Those results provided a selection of 96 worker bees to perform PCR of two worker bees per hive. For mitochondrial DNA analysis 14 samples from sentinel apiaries were added. Three from 61 beehives presented bees with different mtDNA. Throughout, three mitotypes of the African (A) lineage were detected; one mitotype is still unidentified. Mitotype A1 A. mellifera iberiensis was represented by 88 bees and mitotype A4 A. mellifera scutellata by 21 bees. Phylogenetic analysis confirmed the PCR findings. No associations were found between mitotypes, forewing length, beehive characteristics and management. A high degree of Africanization in A. mellifera colonies represented by two predominating mitotypes from the A lineage, prevail in Neotropical Nicaragua, with mitotype A4 predominating at higher altitudes.
Assuntos
DNA Mitocondrial , Genética Populacional , Animais , Abelhas/genética , DNA Mitocondrial/genética , Hibridização Genética , Nicarágua , FilogeniaRESUMO
BACKGROUND: The black spiny-tailed iguana (Ctenosaura similis) is an endemic animal in Mesoamerica, whose meat is consumed by the local population. OBJECTIVES: Because the black spiny-tailed iguana may be potential reservoirs of pathogens, this study aimed to isolate and characterise Salmonella spp. in their meat commercialised in markets of the city of León, Nicaragua. METHODS: Thirteen specimens were analysed for the isolation of Salmonella spp., as well as their antimicrobial resistance patterns, including the presence of genes encoding extended-spectrum ß-lactamases. RESULTS: Salmonella spp. isolates were found in eight out of 13 samples, with S. enterica serovar Enteritidis being found in six out of eight samples. Moreover, eight Salmonella spp. isolates were resistant to amoxicillin plus clavulanic acid and cephalexin, but sensitive to other tested antibiotics. The blaSHV gene was detected in seven out of eight Salmonella spp. isolates, followed by the blaTEM (two out of eight) and blaCXT-M (one out of eight) genes. CONCLUSIONS: These findings represent an important contribution to the implementation of appropriate strategies to prevent foodborne diseases.
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Iguanas , Animais , Antibacterianos/farmacologia , Carne , Nicarágua/epidemiologia , Salmonella/genéticaRESUMO
In Central America, outbreaks of trypanosomiasis by Trypanosoma vivax have been recorded only in cattle. This is the first report of an outbreak of trypanosomiasis by T. vivax in 30 Pelibuey sheep (2 to 7 years old, male and female) from Nicaragua, which occurred in 2009. Clinical signs included fever, apathy, pale mucous membranes, weakness, progressive weight loss, and sudden death. Infection by T. vivax was detected in 22 (73.3%) sheep by blood smear analysis and/or PCR. Trypanosomes were morphologically identified in 11 (36.7%) blood smear samples, whereas 17 (85%) of the 20 samples subjected to PCR were positive for T. vivax. Eighteen (81.8%) of the 22 infected sheep presented a packed red cell volume (PCV) lower than 25%. Upon diagnosis, the treated animals were clinically recovered and no parasites could be observed in subsequent examinations. Tabanids were potential mechanical vectors of T. vivax in the farm. This is the first report of T. vivax in Nicaragua and for the first time this haemoparasite is recorded in sheep in Central America.
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Doenças dos Bovinos , Doenças dos Ovinos , Trypanosoma , Tripanossomíase , Animais , Bovinos , Doenças dos Bovinos/parasitologia , Feminino , Masculino , Nicarágua/epidemiologia , Ovinos , Doenças dos Ovinos/epidemiologia , Doenças dos Ovinos/parasitologia , Trypanosoma vivax/genética , Tripanossomíase/veterináriaRESUMO
The ectoparasite Varroa (Acari: Varroidae) is considered to be the main pest of honey bees (Apis mellifera L.) in Nicaragua. The aim of this study was to determine morphotypes and mitochondrial haplotypes of the Varroa mites, related to infestation levels in A. mellifera hives in Nicaragua in a cross-sectional study (2013-2016). Samples were taken from 34 sentinel apiaries in five geographical zones; from 713 Varroa females collected during the study, 153 were selected for measurement of various morphometric characters for further classification into morphotypes. The mitochondrial haplotype was assigned to one of the two (Korean or Japanese), using the restriction by SacI of the PCR product of a fragment of the gene CO-I. Morphometric measurement and sequencing revealed the affiliation to the species Varroa destructor with a mean (± SD) body width of 1699.1 ± 60.2 µm and a body length of 1161.1 ± 34.9 µm. Body characters were significantly different among the 34 apiaries. Varroa destructor samples were classified into four morphotypes, with no significant differences in the geographical zones. As GAGCTC SacI enzyme cleavage sequences were not observed, all PCR products were identified as V. destructor Korean haplotype. The CO-I gene nucleotide sequences from two samples V. destructor showed both 100% similarity with the Korean haplotype and 99.8% similarity to the Japanese haplotype. Although the V. destructor mite was identified as a Korean haplotype, host-parasite association in 2 decades has led into a balance without entering into severe losses in the Nicaraguan apiculture.
Assuntos
Varroidae , Animais , Criação de Abelhas , Abelhas , Estudos Transversais , Feminino , Haplótipos , NicaráguaRESUMO
Introduction: Molecular biology diagnostic methods such as real-time PCR should be used in Nicaragua to improve the diagnosis of leptospirosis in humans and animals. Objective: To evaluate three qPCR methods for pathogenic Leptospira detection in domestic animals. Materials and methods: Real-time PCR primers were designed for the amplification of specific regions from the Lip 32 gene of Leptospira in SYBER Green (SYBER Green-A) and TaqMan, as well in SYBER Green-B as previously published. The sequences of 12 strains obtained from the database of the National Center for Biotechnology Information (NCBI) were aligned to select probes and primers. The analytical sensitivity was determined by calculating the detectable genomic equivalent while 18 pathogenic references strains and 28 negative controls were used to evaluate the sensitivity and specificity of each one of the three sets in 129 urine samples of domestic animals. Results: The detection limit of four genomic equivalents per reaction was obtained from SYBR Green-A. The specificities were 94.4% (95% CI: 81.1-100.0) for TaqMan, 77.8% (95% CI: 55.8-99.8) for SYBR Green-A, while for SYBR Green-B it was 61.1% (95% CI: 35.8-86.4). In the three tests, we obtained a specificity of 100% (95% CI: 98.2-100.0). In the field samples, 26.4% were positive with SYBR Green-A and 6.1% with SYBR Green-B. Conclusion: SYBR Green-A presented the lowest detection limit while the three techniques under evaluation showed high specificity while TaqMan was the most sensitive.
Introducción. En Nicaragua es necesario estandarizar pruebas moleculares como la PCR en tiempo real (quantitative Polymerase Chain Reaction, qPCR) que mejoren el diagnóstico de leptospirosis en humanos y animales. Objetivo. Evaluar tres qPCR para la detección de leptospiras patógenas en animales domésticos de Nicaragua. Materiales y métodos. Se diseñaron cebadores para la amplificación del gen LipL32 en SYBR Green (SYBR Green-A) y TaqMan, y en otros descritos previamente (SYBR Green-B). Las secuencias de 12 cepas obtenidas de la base de datos del National Center for Biotechnology Information (NCBI) se alinearon para la búsqueda de sondas y cebadores. La sensibilidad analítica se determinó calculando el equivalente genómico detectable, se utilizaron 18 cepas de referencia para la sensibilidad diagnóstica y 28 controles negativos para la especificidad. Los métodos se aplicaron en 129 muestras de orina de animales domésticos. Resultados. En SYBR Green-A se obtuvo un límite de detección de cuatro equivalentes genómicos; en TaqMan, la sensibilidad fue del 94,4 % (IC95% 81,1-100,0). Con SYBR Green-A, se obtuvo una sensibilidad del 77,8 % (IC95% 55,8-99,8), en tanto que con SYBR Green-B fue del 61,1 % (IC95% 35,8-86,4). En las tres pruebas se logró una especificidad del 100 % (IC95% 98,2-100,0). El 26,4 % de las muestras de animales domésticos fueron positivas con SYBR Green-A y el 6,2 % con SYBR Green-B. Conclusiones. El SYBR Green-A presentó un límite de detección bajo, en tanto que las tres técnicas evaluadas mostraron alta especificidad, en tanto que la TaqMan tuvo la mayor sensibilidad.
Assuntos
Animais Domésticos/microbiologia , Proteínas da Membrana Bacteriana Externa/genética , Genes Bacterianos , Leptospira/genética , Lipoproteínas/genética , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Animais , Animais Domésticos/urina , Bovinos , Sondas de DNA/genética , Cães , Amplificação de Genes , Cavalos , Leptospira/isolamento & purificação , Nicarágua , Técnicas de Amplificação de Ácido Nucleico/veterinária , Reação em Cadeia da Polimerase em Tempo Real/métodos , Sensibilidade e Especificidade , Ovinos , Sus scrofaRESUMO
Resumen: Introducción. En Nicaragua es necesario estandarizar pruebas moleculares como la PCR en tiempo real (quantitative Polymerase Chain Reaction, qPCR) que mejoren el diagnóstico de leptospirosis en humanos y animales. Objetivo. Evaluar tres qPCR para la detección de leptospiras patógenas en animales domésticos de Nicaragua. Materiales y métodos. Se diseñaron cebadores para la amplificación del gen LipL32 en SYBR Green (SYBR Green-A) y TaqMan, y en otros descritos previamente (SYBR Green-B). Las secuencias de 12 cepas obtenidas de la base de datos del National Center for Biotechnology Information (NCBI) se alinearon para la búsqueda de sondas y cebadores. La sensibilidad analítica se determinó calculando el equivalente genómico detectable, se utilizaron 18 cepas de referencia para la sensibilidad diagnóstica y 28 controles negativos para la especificidad. Los métodos se aplicaron en 129 muestras de orina de animales domésticos. Resultados. En SYBR Green-A se obtuvo un límite de detección de cuatro equivalentes genómicos; en TaqMan, la sensibilidad fue del 94,4 % (IC95% 81,1-100,0). Con SYBR Green-A, se obtuvo una sensibilidad del 77,8 % (IC95% 55,8-99,8), en tanto que con SYBR Green-B fue del 61,1 % (IC95% 35,8-86,4). En las tres pruebas se logró una especificidad del 100 % (IC95% 98,2-100,0). El 26,4 % de las muestras de animales domésticos fueron positivas con SYBR Green-A y el 6,2 % con SYBR Green-B. Conclusiones. El SYBR Green-A presentó un límite de detección bajo, en tanto que las tres técnicas evaluadas mostraron alta especificidad, en tanto que la TaqMan tuvo la mayor sensibilidad.
Abstract: Introduction: Molecular biology diagnostic methods such as real-time PCR should be used in Nicaragua to improve the diagnosis of leptospirosis in humans and animals. Objective: To evaluate three qPCR methods for pathogenic Leptospira detection in domestic animals. Materials and methods: Real-time PCR primers were designed for the amplification of specific regions from the Lip 32 gene of Leptospira in SYBER Green (SYBER Green-A) and TaqMan, as well in SYBER Green-B as previously published. The sequences of 12 strains obtained from the database of the National Center for Biotechnology Information (NCBI) were aligned to select probes and primers. The analytical sensitivity was determined by calculating the detectable genomic equivalent while 18 pathogenic references strains and 28 negative controls were used to evaluate the sensitivity and specificity of each one of the three sets in 129 urine samples of domestic animals. Results: The detection limit of four genomic equivalents per reaction was obtained from SYBR Green-A. The specificities were 94.4% (95% CI: 81.1-100.0) for TaqMan, 77.8% (95% CI: 55.8-99.8) for SYBR Green-A, while for SYBR Green-B it was 61.1% (95% CI: 35.886.4). In the three tests, we obtained a specificity of 100% (95% CI: 98.2-100.0). In the field samples, 26.4% were positive with SYBR Green-A and 6.1% with SYBR Green-B. Conclusion: SYBR Green-A presented the lowest detection limit while the three techniques under evaluation showed high specificity while TaqMan was the most sensitive.
Assuntos
Leptospirose/diagnóstico , Animais Domésticos , Reação em Cadeia da Polimerase , Leptospira , NicaráguaRESUMO
In Nicaragua, there are ideal environmental conditions for leptospirosis. The objective of this investigation was to detect pathogenic and saprophytic leptospires in water and soil samples from leptospirosis-endemic areas in Nicaragua. Seventy-eight water and 42 soil samples were collected from houses and rivers close to confirmed human cases. Leptospira spp was isolated in Ellinghausen-McCullough-Johnson-Harris (EMJH) culture medium with 5-fluororacil and positive samples were analyzed through PCR for the LipL32 gene, specific for pathogenic leptospires (P1 clade). There were 73 positive cultures from 120 samples, however only six of these (5% of all collected samples) were confirmed to be pathogenic, based on the presence of the LipL32 gene (P1 clade). Of these six pathogenic isolates, four were from Leon and two from Chinandega. Four pathogenic isolates were obtained from water and two from soil. This study proved the contamination of water and soil with pathogenic leptospires, which represents a potential risk for public health.
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RESUMEN Objetivo. Identificar los genotipos de parvovirus canino-circulantes en cachorros en dos municipios de Nicaragua. Materiales y métodos. Se recolectaron muestras por hisopado rectal de 45 cachorros con y sin antecedentes de vacunación, menores de 6 meses de edad, con y sin sintomatología compatible con parvovirosis. Las muestras y dos de las vacunas que se comercializan en Nicaragua (vacuna nº1 y vacuna nº2) fueron analizadas por Reacción en Cadena de la Polimerasa (PCR) convencional para un producto de ≈630 pb del gen VP2. Además, el producto directo del PCR se secuenciaron en sentido reverso cuatro muestras de campo elegidas aleatoriamente y las dos cepas vacunales. Resultados. El 28.9% (13/45) de las muestras analizadas fueron positivas en PCR. No se encontraron diferencias significativas en la detección por PCR del fragmento de VP2, respecto al estado de vacunación de los animales (p≥0.05). Las cuatro muestras de campo secuenciadas fueron identificadas como genotipo CPV-2C y las dos cepas vacunales se identificaron como genotipo CPV- 2A. Conclusiones. la inferencia evolutiva de las secuencias alineadas de cepas vacunales mostró alta divergencia evolutiva respecto a las cepas de campo, este hallazgo lleva a replantear el tema sobre la eficacia de las vacunas analizadas en este trabajo y que son aplicadas en Nicaragua.
ABSTRACT Objective. To identify genotypes of canine parvovirus circulating in puppies in two municipalities of Nicaragua. Materials and methods. Rectal swab samples were collected from 45 puppies (less than 6 months of age) with or without a vaccination history, showing or not symptomatology compatible with parvovirosis. The samples and two of the vaccines that are marketed in Nicaragua (vaccine nº1 and vaccine nº2) were analyzed by conventional Polymerase Chain Reaction (PCR) to a product of ≈630 bp of the VP2 gene. In addition, four randomly chosen field samples and both vaccine strains were sequenced in reverse sense. Results. 28.9% (13/45) of the analyzed samples were positive by PCR, for the fragment of CPV VP2 gene. No significative difference (p≥0.05) was seen in PCR detection between dogs with or without vaccination history. The four sequenced field samples were identified as CPV-2C genotype while both vaccine strains were identified as CPV-2A genotype. Conclusions. The aligned sequences showed high evolutionary divergence of filed strains with respect to vaccines strains, leading us to rethink the efficacy of the analyzed vaccines which are nowadays commercially available in Nicaragua.
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Animais , Cães , Parvovirus CaninoRESUMO
Phenotypic variability in prion diseases, such as scrapie, is associated to the existence of prion strains, which are different pathogenic prion protein (PrPSc) conformations with distinct pathobiological properties. To faithfully study scrapie strain variability in natural sheep isolates, transgenic mice expressing sheep cellular prion protein (PrPC) are used. In this study, we used two of such models to bioassay 20 scrapie isolates from the Spain-France-Andorra transboundary territory. Animals were intracerebrally inoculated and survival periods, proteinase K-resistant PrP (PrPres) banding patterns, lesion profiles and PrPSc distribution were studied. Inocula showed a remarkable homogeneity on banding patterns, all of them but one showing 19-kDa PrPres. However, a number of isolates caused accumulation of 21-kDa PrPres in TgShp XI. A different subgroup of isolates caused long survival periods and presence of 21-kDa PrPres in Tg338 mice. It seemed that one major 19-kDa prion isoform and two distinct 21-kDa variants coexisted in source inocula, and that they could be separated by bioassay in each transgenic model. The reason why each model favours a specific component of the mixture is unknown, although PrPC expression level may play a role. Our results indicate that coinfection with more than one substrain is more frequent than infection with a single component.
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Proteínas Priônicas/metabolismo , Scrapie/etiologia , Scrapie/patologia , Animais , Encéfalo/metabolismo , Bovinos , Modelos Animais de Doenças , França , Camundongos Transgênicos , Proteínas PrPSc/metabolismo , Proteínas PrPSc/patogenicidade , Scrapie/metabolismo , Scrapie/prevenção & controle , Ovinos , EspanhaRESUMO
Leptospirosis is one of the most important zoonoses in tropical countries, including Nicaragua, where it is considered endemic. The aim of this study was to determine the frequency of Leptospira spp in rodents captured from peridomestic sites in leptospirosis endemic regions of Nicaragua. Using live traps, 191 rodents were captured in 2012 and 2013 between April and December. Kidney samples were collected and processed for Leptospira detection from 166 animals by direct culture and isolation. The isolates were tested by PCR for LipL32 and lfb1-F genes specific to pathogenic Leptospira species. The trapping success over all sites was 20.2%, with higher rates of success in rainy season (p < 0.05). Leptospira spp were detected in 22.3% of rodents by direct culture methods. Significant differences (p < 0.01) were found in the frequencies of Leptospira positive rodents per month as well as per region. Of the isolated Leptospira spp, 37.5% were positive for pathogenic species by PCR. The frequency of Leptospira positive rodents by isolation could be used as a predictive indicator for the risk of human leptospirosis in Nicaragua.
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Leptospira , Leptospirose , Animais , Leptospirose/epidemiologia , Leptospirose/veterinária , Nicarágua/epidemiologia , Roedores , Zoonoses/epidemiologiaRESUMO
OBJECTIVE: The objective of this study was to determine the epidemiological characteristics of leptospirosis in pets and in humans in peridomestic settings in Nicaragua between 2014 and 2016. METHODS: The samples were taken in areas where cases were confirmed in humans using non-probabilistic sampling in 10 of the country's 17 departments. This included 112 urine samples from pets, 129 water samples, and 69 soil samples in order to isolate leptospires in Ellinghausen-McCullough-Johnson-Harris (EMJH) medium. Furthermore, the microscopic agglutination test (MAT) was applied to 263 samples of animal serum, and 88 isolates were analyzed using PCR. RESULTS: In 32.6% (101/310) of the samples, spirochetes were isolated: 23.2% (26/112) in the pet urine, 47.3% (61/129) in water samples, and 20.3% (14/69) in soil samples. Isolation analysis showed significant differences (p<0.05) between departments for the different types of samples, and isolation was more frequent in water than in soil (OR = 3.49; CI95%: 1.56-7.80). In total, 14.1% (37/263) of the animals were reactors in the microscopic agglutination test. The most frequent serogroup was Icterohaemorrhagiae (40%). PCR analysis to identify pathogenic species of leptospires resulted in 10.2% (9/88) positive isolations. CONCLUSIONS: This research demonstrates that pets and environment conditions play an important role in the emergence of outbreaks of leptospirosis, and confirms the endemic behavior of the disease in Nicaragua.
OBJETIVO: Descrever as características epidemiológicas da leptospirose em animais domésticos e em casos de leptospirose humana em áreas peridomiciliares na Nicarágua entre 2014 e 2016. MÉTODOS: As amostras foram coletadas por amostragem não probabilística em áreas com casos confirmados de leptospirose humana em 10 das 17 províncias do país. Foram analisadas 112 amostras de urina de animais domésticos, 129 amostras de água e 69 amostras de solo com o uso do meio de cultura padrão para o isolamento de leptospiras (Ellinghausen-McCullough-Johnson-Harris, EMJH). Além disso, foi realizado o teste de aglutinação microscópica em 263 amostras séricas de animais e 88 isolados foram analisados com a técnica de PCR. RESULTADOS: Em 32,6% (101/310) das amostras foram isoladas espiroquetas, sendo 23,2% (26/112) isoladas na urina de animais domésticos, 47,3% (61/129) nas amostras de água e 20,3% (14/69) nas amostras de solo. Houve diferença significativa (P < 0,05) entre as províncias no isolamento nos diferentes tipos de amostras analisadas, sendo o isolamento mais frequente nas amostras de água que de solo (OR = 3,49; IC95%: 1,567,80). Reatividade no teste de aglutinação microscópica foi observada em 14,1% (37/263) das amostras de animais. O sorogrupo mais frequentemente isolado foi Icterohaemorrhagiae (40%). A técnica de PCR demonstrou que 10,2% (9/88) dos isolados eram positivos para espécies patogênicas de leptospiras. CONCLUSÕES: Esta pesquisa demonstra que os animais domésticos e o entorno têm papel importante no surgimento de surtos de leptospirose e confirma o comportamento endêmico da doença na Nicarágua.
RESUMO
RESUMEN Objetivo El objetivo de este estudio fue conocer las características epidemiológicas de la leptospirosis en animales domésticos y en los casos de leptospirosis humana en áreas peridomésticas en Nicaragua entre 2014 y 2016. Métodos Las muestras se extrajeron en áreas donde se confirmaron casos en humanos utilizando un muestreo no probabilístico en 10 de los 17 departamentos del país. Se incluyeron 112 muestras de orina de animales domésticos, 129 muestras de agua y 69 de tierra para aislar leptospiras en medio Ellinghausen-McCullough-Johnson-Harris (EMJH). Además, se aplicó la prueba de microaglutinación (MAT) en 263 muestras de suero de animales y 88 aislados se analizaron mediante PCR. Resultados En 32,6% (101/310) de las muestras se aislaron espiroquetas, 23,2% (26/112) se aislaron en la orina de animales domésticos, 47,3% (61/129), en las muestras de agua y 20,3 % (14/69), en las de tierra. El análisis de aislamiento mostró diferencias significativas (P < 0,05) entre los departamentos para los diferentes tipos de muestras, y el aislamiento fue más frecuente en agua que en tierra (OR = 3,49; IC95%: 1,56-7,80). El 14,1% (37/263) de los animales fueron reactores en la prueba de microaglutinación. El serogrupo más frecuente fue Icterohaemorrhagiae (40%). En el análisis con la PCR para identificar leptospiras de las especies patógenas 10,2% (9/88) de los aislamientos fueron positivos. Conclusiones Esta investigación demuestra que los animales domésticos y el ambiente desempeñan un papel importante en la aparición de brotes de la leptospirosis y confirma el comportamiento endémico de la enfermedad en Nicaragua.
ABSTRACT Objective The objective of this study was to determine the epidemiological characteristics of leptospirosis in pets and in humans in peridomestic settings in Nicaragua between 2014 and 2016. Methods The samples were taken in areas where cases were confirmed in humans using non-probabilistic sampling in 10 of the country's 17 departments. This included 112 urine samples from pets, 129 water samples, and 69 soil samples in order to isolate leptospires in Ellinghausen-McCullough-Johnson-Harris (EMJH) medium. Furthermore, the microscopic agglutination test (MAT) was applied to 263 samples of animal serum, and 88 isolates were analyzed using PCR. Results In 32.6% (101/310) of the samples, spirochetes were isolated: 23.2% (26/112) in the pet urine, 47.3% (61/129) in water samples, and 20.3% (14/69) in soil samples. Isolation analysis showed significant differences (p<0.05) between departments for the different types of samples, and isolation was more frequent in water than in soil (OR = 3.49; CI95%: 1.56-7.80). In total, 14.1% (37/263) of the animals were reactors in the microscopic agglutination test. The most frequent serogroup was Icterohaemorrhagiae (40%). PCR analysis to identify pathogenic species of leptospires resulted in 10.2% (9/88) positive isolations. Conclusions This research demonstrates that pets and environment conditions play an important role in the emergence of outbreaks of leptospirosis, and confirms the endemic behavior of the disease in Nicaragua.
RESUMO Objetivo Descrever as características epidemiológicas da leptospirose em animais domésticos e em casos de leptospirose humana em áreas peridomiciliares na Nicarágua entre 2014 e 2016. Métodos As amostras foram coletadas por amostragem não probabilística em áreas com casos confirmados de leptospirose humana em 10 das 17 províncias do país. Foram analisadas 112 amostras de urina de animais domésticos, 129 amostras de água e 69 amostras de solo com o uso do meio de cultura padrão para o isolamento de leptospiras (Ellinghausen-McCullough-Johnson-Harris, EMJH). Além disso, foi realizado o teste de aglutinação microscópica em 263 amostras séricas de animais e 88 isolados foram analisados com a técnica de PCR. Resultados Em 32,6% (101/310) das amostras foram isoladas espiroquetas, sendo 23,2% (26/112) isoladas na urina de animais domésticos, 47,3% (61/129) nas amostras de água e 20,3% (14/69) nas amostras de solo. Houve diferença significativa (P < 0,05) entre as províncias no isolamento nos diferentes tipos de amostras analisadas, sendo o isolamento mais frequente nas amostras de água que de solo (OR = 3,49; IC95%: 1,56-7,80). Reatividade no teste de aglutinação microscópica foi observada em 14,1% (37/263) das amostras de animais. O sorogrupo mais frequentemente isolado foi Icterohaemorrhagiae (40%). A técnica de PCR demonstrou que 10,2% (9/88) dos isolados eram positivos para espécies patogênicas de leptospiras. Conclusões Esta pesquisa demonstra que os animais domésticos e o entorno têm papel importante no surgimento de surtos de leptospirose e confirma o comportamento endêmico da doença na Nicarágua.
Assuntos
Humanos , Zoonoses , Zoonoses/transmissão , Fatores Epidemiológicos , Leptospirose/prevenção & controle , NicaráguaRESUMO
Leptospirosis is one of the most extended zoonosis worldwide and humans become infected most commonly through contact with the urine of carrier animals, either directly or via contaminated water or soil. The aim in this study was to analyse the epidemiological behaviour of Leptospira spp., from domestic animals around the sites of human leptospirosis cases in Nicaragua, from 2007 through 2013. We report the results of a cross-sectional epidemiological study with a non-probability sampling of blood (n=3050) and urine (n=299) from Domestic Animals (DA) around the sites of human leptospirosis cases in Nicaragua. We analysed data obtained through Microscopic Agglutination Test (MAT), in-vitro culture, real time PCR and sequencing of lfb1 locus. Frequencies of 30.31% (95% CI: 28.66-31.95) and 15.38% (95% CI: 11.12-19.64) were obtained from serological test and from in-vitro culture, respectively. Although similar frequencies from serology test (P≥0.05) were found in DA species, in-vitro culture frequencies were significantly higher from bovine, equine and sheep (P<0.05) in comparison with swine and canine species. Ten serogroups of pathogenic Leptospira spp. were encountered, with the highest presence of Icterohaemorrhagiae serogroup 34.65% (95% CI: 29.35-39.94). We identified 7 samples homologous to L. interrogans species Pyrogenes serovar and 3 samples as L. noguchii Louisiana or Panama serovars by analysis of lfb1 sequences. We were able to establish a temporal and spatial correlation from DA and cumulative incidence of human cases. Therefore an effective epidemiological surveillance should be implemented with a specific control program toward DA in order to reduce human leptospirosis incidence.
Assuntos
Animais Domésticos/microbiologia , Leptospira/classificação , Leptospirose/epidemiologia , Testes de Aglutinação/veterinária , Animais , Bovinos/microbiologia , Estudos Transversais , Cães/microbiologia , Equidae , Cavalos/microbiologia , Humanos , Nicarágua/epidemiologia , Reação em Cadeia da Polimerase em Tempo Real , Sorogrupo , Ovinos/microbiologia , Suínos/microbiologia , ZoonosesRESUMO
Oropharyngeal and cloacal swabs from 117 captive psittacine birds presented at veterinary clinics (88) and from shelters/rescue centers of wildlife (29) were collected to determine the prevalence of C. psittaci in captive birds in Costa Rica. Samples were collected during 2009 from a total of 19 different species of parrots, with Ara macao (33), Amazona autumnalis (24), Amazona ochrocephala (21), and Ara ararauna (8) being the most representative species sampled. C. psittaci was detected in four (3.4%) birds using molecular detection (PCR). The positive samples belonged to birds presented at veterinary clinics; three of them were Ara macao and one Amazona ochrocephala. Three birds were adults; all positive birds showed no symptoms of illness and lived in homes with other birds, two in San José and two in Heredia. Sequencing was used to confirm the PCR positive results, showing that two samples of C. psittaci belonged to genotype A, representing the first report of the presence of this genotype in Costa Rica. The detection of this bacterium in captive psittacine birds shows that there is a potential risk for people living or having contact with them and that there is a possibility of infecting other birds.