Molecular Detection and Genotyping of Cryptosporidium spp. Isolates from Bats in Colombia.

PURPOSE: Cryptosporidiosis is a zoonotic infectious disease caused by the protozoan parasite Cryptosporidium spp., frequently found in several animal species, including bats. Several Cryptosporidium genotypes have been described in bats worldwide, suggesting that bats are infected by host-specific Cryptosporidium spp. To date, there are no published reports about Cryptosporidium spp. in bats from Colombia. Therefore, this study aimed to determine the presence and molecular diversity of Cryptosporidium spp. in Colombian bats. METHODS: A total of 63 gut samples from three bat species served for molecular detection of Cryptosporidium spp. 18S rDNA gene by qPCR. The sequenced amplicons were used in subsequent phylogenetic analyses to identify them as species or genotypes. RESULTS: Cryptosporidium spp. qPCR detection occurred in 9.5% (6/63) of bat intestines, and four sequences represented two new genotypes, called Cryptosporidium bat genotypes XIX and XX, were identified. CONCLUSIONS: This study describes the detection of two novel Cryptosporidium bat genotypes, in two species of bats from a region of Colombia, requiring further studies to determine the relationhip between Cryptosporidium and bats in Colombia.

Isolation and identification of Salmonella spp. in raw milk from dairy herds in Colombia / Isolamento e identificação de Salmonella spp. no leite cru de rebanhos leiteiros na Colômbia

Salmonellosis is a foodborne disease (FBD) that affects public health and can cause death in people. Many outbreaks of Salmonellosis have been reported due to the contamination of raw milk and dairy products with the pathogen. To determine the prevalence of Salmonella spp. in milk samples from four dairy herds in the Sabana of Bogotá in 2017, 112 milk samples were taken directly from the mammary gland during milking. All milk samples were cultured and tested to isolate and identify Salmonella spp. using microbiological and molecular methods. Salmonella spp. prevalence of milk samples was found to be 20.5% (n=23). The main Salmonella serovars isolated were S. Newport (60.87%), S.Typhimurium (17.4%), S. Virchow, S. Bredeney, and S. Anatum (4.3% each one of the serovars). However, it was not possible to determine the Salmonella serotype in two isolates. The prevalence of Salmonella spp. in milk has not been studied extensively in Colombia. The 20.5% in the prevalence might be due to fact that the sample was taken directly from the mammary gland allowing a better chance of isolation by avoiding the dilutional effect of mixed milk from different cows in the buckets. This also suggests that the infection of the udder could have occurred by hematogenous dissemination or by milking machine contamination. This study highlights the need to implement measures to prevent contamination and reduce the problem in the herds, which will result in milk and dairy products with high standards of innocuity and quality and decrease the risk of foodborne illness(AU)

A salmonelose é uma doença transmitida por alimentos que afeta a saúde pública e pode causar a morte de pessoas. Muitos surtos de salmonelose têm sido relatados devido à contaminação de leite cru e produtos lácteos com o patógeno. Para determinar a prevalência de Salmonella spp. em amostras de leite de quatro rebanhos leiteiros na Sabana de Bogotá em 2017, cento e doze amostras de leite foram colhidas diretamente da glândula mamária durante a ordenha. Todas as amostras de leite foram cultivadas para isolar e identificar Salmonella spp. usando métodos microbiológicos e moleculares. A prevalência de Salmonella spp. nas amostras de leite foi de 20,5% (n = 23). Os principais sorovares de Salmonellaidentificados foram S. Newport (60,87%), S. Typhimurium (17,4%), S. Virchow, S. Bredeney e S. Anatum (4,3% cada um dos sorovares). No entanto, não foram determinados os sorovares de dois isolados. A prevalência de Salmonella spp. no leite ainda não foi extensivamente estudada na Colômbia. Os 20,5% na prevalência podem ser devidos ao fato de a amostra ter sido colhida diretamente da glândula mamária, permitindo uma melhor chance de isolamento, evitando o efeito de diluição do leite misto de diferentes vacas nos baldes, o que pode indicar infecção do úbere pela disseminação hematogênica ou por contaminação da ordenhadeira. Este estudo destaca a necessidade da implementação de medidas destinadas a prevenir a contaminação e reduzir o problema nos rebanhos, resultando em leite e produtos lácteos com altos padrões de inocuidade e qualidade, diminuindo o risco de doenças de origem alimentar.(AU)

Identification of Malassezia species as part of normal skin and ear canal microbiota in horses / Identificación de especies de Malassezia como microbiota normal en piel y canal auditivo externo de equinos / Identificação de espécies de Malassezia como microbiota normal em pele e canal auditivo externo de cavalos

Abstract Background: The yeasts of the genus Malassezia are considered part of the normal skin microbiota in humans and animals. In horses, several species of the genus Malassezia have been reported in different areas of the skin and ear canal. Objective: Isolate, characterize and identify the different species belonging to the genus Malassezia isolated from the ear canal and skin of equine patients with no dermatological lesions that were referred to the large animal clinic of veterinary teaching hospital at the National University of Colombia. Methods: 22 horses were evaluated and sampled. Eighty-two samples were obtained by swabbing either the ear canals (left and right), skin areas of prepuce, mammary gland and inguinal region. The samples were examined by cytological evaluation and were cultured on modified Dixon's agar and phenotypic and molecular identification were performed for yeast colonies. Results: Fourteen yeast isolates were obtained from the 82 samples. Biochemical identification determined that 50% (n=7) were Malassezia spp., 35.7% (n=5) were identified as Candida spp. and 14.3% (n=2) as Cryptococcus spp.. Using molecular tests, the Malassezia species were M. slooffiae (28.6%) and M.nana (57.1%); only one isolate was classified as Trichosporo asahii. Conclusion: M.nana and M. slooffiae were identified as part of the normal ear canal and skin microbiota in the evaluated horses. The observed prevalence of Malassezia spp. was 18.2% (n=4/22) in this study sample.

Resumen Antecedentes: Las levaduras del género Malassezia hacen parte de la microbiota normal cutánea de humanos y animales. En equinos se han reportado diferentes especies de Malassezia aisladas de varias regiones de piel y canal auditivo externo. Objetivo: Aislar, caracterizar e identificar las especies del género Malassezia spp. a partir de canal auditivo externo y piel de equinos sin lesiones dermatológicas, remitidos a la Clínica de Grandes Animales de la Facultad de Medicina Veterinaria y de Zootecnia de la Universidad Nacional de Colombia. Metodología: Se evaluaron 22 equinos, a partir de los cuales se obtuvieron 82 muestras entre hisopados de canal auditivo externo (izquierdo y derecho) y diferentes regiones de piel (prepucio, glándula mamaria e ingle). Las muestras fueron procesadas mediante examen directo y cultivo en agar Dixon modificado. A partir de los aislamientos en los que se observaron colonias morfológicamente compatibles con Malassezia spp. se realizó la identificación fenotípica y molecular. Resultados: De las 82 muestras procesadas se obtuvieron 14 aislamientos de levaduras, de las cuales mediante identificación bioquímica el 50% (n=7) correspondió a Malassezia spp., el 35,7% (n=5) a Candida spp., y el 14,3% (n=2) a Cryptococcus spp. Luego mediante pruebas moleculares se identificaron las especies del género Malassezia como: M. slooffiae (28,6%) y M . nana (57,1%); y un aislamiento correspondió a Trichosporon asahii. Conclusión: Se logró identificar las especies M.nana y M. slooffiae como microbiota normal de la piel y el canal auditivo en los equinos evaluados. La prevalencia de Malassezia spp. para la población evaluada fue de 18,2% (n=4/22).

Resumo Antecedentes: As leveduras do gênero Malassezia fazem parte da microbiota cutânea normal de humanos e animais. Em cavalos, diferentes espécies de Malassezia isoladas de várias regiões da pele e do canal auditivo externo foram reproduzidas. Objetivo: Isolar, caracterizar e identificar as espécies do gênero Malassezia spp. do canal auditivo externo e pele eqüinos sem lesões cutâneas, referiu-se à Clínica de Grandes Animais da Faculdade de Medicina Veterinária e Zootecnia da Universidade Nacional da Colômbia. Métodos: 22 equinos foram avaliadas a partir dos quais 82 amostras a partir de esfregaços do canal auditivo externo (esquerda e direita) e diferentes regiões da pele (prepúcio, glândula mamaria e virilha) foram obtidos. As amostras foram processadas por exame direto e cultura em ágar Dixon modificado. Dos isolados nos quais as colônias foram observadas morfologicamente compatíveis com Malassezia spp. identificação fenotípica e molecular foi realizada. Resultados: Das 82 amostras processadas 14 isolados de levedura, que foram obtidos por identificação bioquímica de 50% (n=7) correspondia a Malassezia spp., 35,7% (n=5) a Candida spp., e 14,3% (n=2) para Cryptococcus spp.. Em seguida, usando o teste molecular espécie Malassezia foram identificadas como M. slooffiae (28,6%) e M . nana (57,1%); e um isolamento correspondia a Trichosporon asahii. Conclusão: As espécies M.nana e M. slooffiae foram identificadas como microbiota de pele normal e do canal auditivo nos equídeos avaliados. A prevalência de Malassezia spp. para a população avaliada foi 18,2% (n=4/22).

Acid base disorders in hospitalized cattle / Distúrbios ácido-base em bovinos hospitalizados

The objectives of this retrospective study were to determine the main acid-base and electrolytes disorders in hospitalized cattle, using both Henderson-Hasselbalch and the physicochemical approach and to compare their diagnostic and therapeutic utility. A total of 31 medical records were reviewed of bovines admitted to the Large Animal Hospital at Universidad Nacional de Colombia, that met the inclusion criteria of the measurement of blood gases, blood electrolytes and plasma protein on admission before providing any treatment. Using the Henderson-Hasselbalch approach, acid base abnormalities were found in 83.3% of the patients, compared to 93.5% using the physicochemical approach. The principal acid-base disorders found were strong ion acidosis (61.29%) and weak acid acidosis (38.7%); strong ion gap (SIG) acidosis was found in 73.68% of cases showing strong ion acidosis. These results highlight the importance of the diagnosis of acid-base disorders in sick cattle for proper recognition of pathophysiological phenomena and its understanding to guide treatment decisions.(AU)

Os objetivos deste estudo retrospectivo foram determinar os principais distúrbios ácido-base e eletrolíticos em bovinos internados, utilizando a abordagem Henderson-Hasselbalch e a abordagem físico-química comparando a utilidade diagnóstica e terapêutica de tais procedimentos. Foram revisados 31 prontuários de bovinos admitidos no Hospital de Animais de Grande Porte da Universidad Nacional de Colombia, que preencheram os critérios de inclusão da medição de gases sanguíneos, eletrólitos sanguíneos e proteínas plasmáticas na admissão antes de fornecer qualquer tratamento. Com o emprego da abordagem de Henderson-Hasselbalch, as anormalidades da base ácida foram encontradas em 83,3% dos pacientes, enquanto com a abordagem físico-química o valor obtido foi de 93,5%. Os principais distúrbios ácido-base encontrados foram forte acidose iônica (61,29%) e acidose ácida fraca (38,7%). A acidose com Ion Gap Forte (SIG) foi encontrada em 73,68% dos casos com forte acidose iônica. Estes resultados destacam a importância do diagnóstico de distúrbios ácido-base em bovinos doentes para o reconhecimento adequado dos fenômenos fisiopatológicos e sua compreensão para orientar as decisões de tratamento.(AU)

Genotyping of Malassezia pachydermatis disclosed genetic variation in isolates from dogs in Colombia / Genotipagem de Malassezia pachydermatis revelou variação genética em isolados de cães na Colômbia

Malassezia pachydermatis is a lipophilic and lipid-dependent yeast mostly isolated from animals' skin; hence, it is regarded as a zoophilic species causing otitis externa in dogs. Aspects associated with its epidemiology and pathogenicity is a matter of interest. This study aimed to conduct a molecular characterization of 43 isolates of M. pachydermatis obtained from dogs with otitis externa. For this purpose, the 5.8S internal transcribed spacer 2 (ITS2) and D1/D2 26S rRNA regions were amplified, sequenced and analyzed using restriction fragment length polymorphism (RFLP) with AluI, CfoI, and BstF5I endonucleases. Phylogenetic analyses revealed that these isolates grouped with the sequence types I, IV and V, previously proposed for M. pachydermatis. Interestingly, we found a new polymorphic RFLP pattern using BstF5I, these isolates were associated with the sequence types IV and V, nevertheless an association between polymorphic RFLP patterns, and fosfolipase activity or canine population data was not observed. These findings underline the genetic diversity of M. pachydermatis and provide new insights about the epidemiology of this species in the analyzed population.(AU)

Malassezia pachydermatis é uma levedura lipofílica e dependente de lipídios, principalmente da pele de animais. Sendo, por essa razão, considerada uma espécie zoofílica e causadora de otite externa em cães. Neste sentido, aspectos associados à sua epidemiologia e patogenicidade constituem um tema de interesse científico. O objetivo deste estudo foi realizar a caracterização molecular de 43 isolados de M. pachydermatis obtidos a partir de cães com otite externa. Para esta propósito, foram amplificadas, sequenciadas e analisadas com enzimas de restrição as regiões do gene 5.8S, do espaçador interno transcrito 2 (ITS2) e D1/D2 do 26S do rRNA pelo método RFLP, com as endonucleases AluI, CfOI e BstF5I. Análises filogenéticas revelaram que os isolados se agruparam com as sequências tipo I, IV e V de M. pachydermatis como já descrito anteriormente. De maneira interessante, se observou um novo RFLP polimórfico utilizando BstF5I. Os isolados que mostraram esse padrão foram associados com os padrões IV e V. No entanto, não foi observada associação entre padrões polimórficos de RFLP e atividade de fosfolipase ou dados da população canina. Estes resultados demonstram a diversidade genética de M. pachydermatis e fornecem novas perspectivas sobre a epidemiologia destas espécies na população analisada.(AU)

Salmonella spp. prevalence, antimicrobial resistance and risk factor determination in Colombian swine farms / Salmonella spp. prevalência, resistência antimicrobiana e determinação de fatores de risco em granjas suínas colombianas

To determine Salmonella spp. prevalence/seroprevalence, antimicrobial resistance patterns and risk factor identification associated with its presence in Colombian swine farms. 504 samples (Faeces, swabs and environment samples) were obtained from 21 farms distributed in four geographical regions in Colombia. Salmonella spp. microbiological and molecular detection were determined by two Salmonella spp. MDS3M™ and MALDI-TOF MS assays, respectively. In addition, for serological evaluation 231 serum samples were analyzed employing ELISA Salmonella Pigtype®-Salmonella Ab (QUIAGEN®). Additionally, 41 isolates were tested for antimicrobial susceptibility using broth microdilution technique (Panel B1016-180 Beckman Coulter NC72®) and verified with WHONET 2016 software. Risk factors were assessed from a survey and analyzed for statistical significance by U Mann-Whitney test. An 8.9% prevalence (n=45) and 38.1% (n=88) seroprevalence were determined. All isolates presented 100% antimicrobial susceptibility against amikacin. However, resistance against penicillin, tetracycline, cefuroxime and trimethoprim/sulfamethoxazole was present in more than 50% of evaluated strains. Risk factors associated with Salmonella spp. presence were surface water use, rough-surfaced on floors, presence of hoppers as feeders and worker's boots. Bacteria were present in animals and environmental samples from evaluated farms. Animal contact and/or exposure with the microorganism were also evident in obtained serological response. Bacteria presence depended on management practices and infrastructure, likewise antibiotic use, supplemented in the diet may have induced an increase in Salmonella spp. antimicrobial resistance.(AU)

Para determinar Salmonellaspp. prevalência/soroprevalência, padrões de resistência antimicrobiana e identificação de fatores de risco associados à sua presença em granjas suínas colombianas. Foram obtidas 504 amostras (fezes, zaragatoas e amostras do ambiente) de 21 fazendas distribuídas em quatro regiões geográficas da Colômbia. Salmonella spp., a detecção microbiológica e molecular foi determinada por 2 Salmonella spp. Ensaios MDS3M™ e MALDI-TOF MS, respectivamente. Além disso, para avaliação sorológica, foram analisadas 231 amostras de soro empregando ELISA Salmonella Pigtype® - Salmonella Ab (QUIAGEN®). Além disso, 41 isolados foram testados quanto à suscetibilidade antimicrobiana usando a técnica de microdiluição em caldo (Painel B1016-180 Beckman Coulter NC72®) e verificados com o software WHONET 2016. Os fatores de risco foram avaliados em uma pesquisa e analisados quanto à significância estatística pelo teste U Mann-Whitney. Foram determinadas prevalências de 8,9% (n=45) e 38,1% (n=88). Todos os isolados apresentaram 100% de suscetibilidade antimicrobiana à amicacina. No entanto, resistência à penicilina, tetraciclina, cefuroxima e trimetoprim/sulfametoxazol estava presente em mais de 50% das cepas avaliadas. Fatores de risco associados à Salmonella spp., presença de uso de água de superfície, superfície áspera no chão, presença de tremonhas como alimentadores e botas de trabalho. Bactérias estavam presentes em animais e amostras ambientais de fazendas avaliadas. O contato animal e/ou a exposição ao microrganismo também foram evidentes na resposta sorológica obtida. A presença de bactérias dependia de práticas de manejo e infraestrutura, assim como o uso de antibióticos suplementados na dieta pode ter induzido um aumento de Salmonella spp. resistência antimicrobiana.(AU)

Estimación de la prevalencia de Salmonella spp. en pechugas de pollo para consumo humano provenientes de cuatro localidades de Bogotá- Colombia / Prevalence of Salmonella spp. in chicken breasts for human consumption in four districts in Bogotá - Colombia

Objetivo: Determinar la prevalencia de Salmonella spp. en muestras de pechugas de pollo para consumo humano en diferentes localidades de Bogotá. Materiales y métodos: Se obtuvieron 72 muestras en tiendas y plazas de mercado de 4 localidades de la ciudad. Las muestras fueron procesadas por métodos microbiológicos y moleculares estandarizados para la detección e identificación de Salmonella spp. Resultados: La prevalencia estimada para Salmonella spp. fue del 29.2% (n=21), de las cuales el 52.4% (n=11) fueron obtenidas a partir de muestras de tiendas y el 47.6% (n=10) de las muestras de plazas de mercado. Los serovares identificados con mayor frecuencia fueron Salmonella enterica group IIIb, S. Bredeney y S. Virchow. Las localidades con mayor presencia del patógeno fueron Usaquén y Fontibón. Discusión: Estudios realizados en Colombia reportan una prevalencia del 27% para Salmonella spp. en muestras de carcasas completas, lo que coincide con lo observado en este estudio, esto podría asociarse con inadecuadas condiciones de manejo y/o almacenamiento del producto. Conclusión: Se determinó la presencia de Salmonella spp. en las muestras de pechugas de pollo evaluadas, lo que implica un riesgo potencial para la salud pública, por lo que es necesario ampliar este tipo de estudios para conocer la situación real a nivel nacional frente a este patógeno.

Objective: To determine the prevalence of Salmonella spp. in chicken breasts for human consumption in four different districs in Bogotá. Materials and Methods: Seventy two samples were collected from different convinience stores and market places in the city. All the samples were processed using standarized microbiological and molecular methods to isolate and identify Salmonella spp. Results: The determined prevalence of Salmonella spp. was 29.2% (n=21), out of vwhich 52.4% (n=11) were obtained from convinience stores and 47.6% (n=10) from market places. The identified serovars were Salmonella enterica group IIIb, S. Bredeney and S. Virchow. The districs with highest prevalence of the pathogen were Usaqén and Fontibón Discussion: Previous studies done in Colombia have reported prevalence of Salmonella spp. around 27% in chicken carcasses, this is very similar to the prevalence observed in this study; these findings could be due to inadequate handling and storage practices of the product. Conclusion: The presence of Salmonella spp. was determined in the studied samples; this finding indicates the potential public health risk. These results demand further studies in countrywide base to determine the actual situation of this pathogen.

Evaluación de la capacidad patogénica de Fusarium spp. en modelos vegetal y murino / Evaluation of Fusarium spp. pathogenicity in plant and murine models

El género Fusarium es ampliamente conocido por su capacidad fitopatógena, típicamente asociada al marchitamiento vascular. Sin embargo, se ha reportado como un patógeno oportunista en pacientes inmunocompetentes e inmunocomprometidos, por lo que puede ser considerado como un microorganismo de interés en estudios de patogenicidad en diferentes hospederos. Este trabajo evaluó la capacidad patogénica de aislamientos de Fusarium spp. de diferentes orígenes en hospederos vegetales y en un hospedero animal (modelo murino). Doce aislamientos de Fusarium spp. de origen vegetal, animal superficial, humano superficial y humano sistémico fueron inoculados en plantas de tomate, gulupa y clavel, y en ratones BALB/c, inmunocompetentes e inmunosuprimidos. Las pruebas de patogenicidad en plantas no mostraron todos los síntomas asociados al marchitamiento vascular en los tres modelos vegetales, pero la colonización y la necrosis de los haces vasculares observada en todos los casos, independientemente de la especie de Fusarium y el origen del aislamiento, demostró el potencial infeccioso de Fusarium spp. en las diferentes especies de plantas. Por otro lado, las pruebas de patogenicidad en el modelo murino evidenciaron alteraciones del comportamiento. Asimismo, se observó en el modelo murino que todos los aislamientos infectaron y colonizaron diferentes órganos, independientemente de su origen, de la especie o del estado inmunitario del hospedero, pero solamente cinco (de diferente origen y correspondientes a diferentes especies) generaron mortalidad. En contraste, la prueba de inoculación superficial no evidenció lesiones ni colonización. Los resultados observados indican el potencial papel patogénico de los aislamientos de Fusarium spp. en los diferentes tipos de hospederos. Sin embargo, es necesario profundizar en estudios de factores de patogenicidad que expliquen la capacidad de este género para colonizar múltiples hospederos.

The genus Fusarium is widely recognized for its phytopathogenic capacity. However, it has been reported as an opportunistic pathogen in immunocompetent and immunocompromised patients. Thus, it can be considered a microorganism of interest in pathogenicity studies on different hosts. Therefore, this work evaluated the pathogenicity of Fusarium spp. isolates from different origins in plants and animals (murine hosts). Twelve isolates of Fusarium spp. from plants, animal superficial mycoses, and human superficial and systemic mycoses were inoculated in tomato, passion fruit and carnation plants, and in immunocompetent and immunosuppressed BALB/c mice. Pathogenicity tests in plants did not show all the symptoms associated with vascular wilt in the three plant models; however, colonization and necrosis of the vascular bundles, regardless of the species and origin of the isolates, showed the infective potential of Fusarium spp. in different plant species. Moreover, the pathogenicity tests in the murine model revealed behavioral changes. It was noteworthy that only five isolates (different origin and species) caused mortality. Additionally, it was observed that all isolates infected and colonized different organs, regardless of the species and origin of the isolates or host immune status. In contrast, the superficial inoculation test showed no evidence of epidermal injury or colonization. The observed results in plant and murine models suggest the pathogenic potential of Fusarium spp. isolates in different types of hosts. However, further studies on pathogenicity are needed to confirm the multihost capacity of this genus.

[Evaluation of Fusarium spp. pathogenicity in plant and murine models]. / Evaluación de la capacidad patogénica de Fusarium spp. en modelos vegetal y murino.

The genus Fusarium is widely recognized for its phytopathogenic capacity. However, it has been reported as an opportunistic pathogen in immunocompetent and immunocompromised patients. Thus, it can be considered a microorganism of interest in pathogenicity studies on different hosts. Therefore, this work evaluated the pathogenicity of Fusarium spp. isolates from different origins in plants and animals (murine hosts). Twelve isolates of Fusarium spp. from plants, animal superficial mycoses, and human superficial and systemic mycoses were inoculated in tomato, passion fruit and carnation plants, and in immunocompetent and immunosuppressed BALB/c mice. Pathogenicity tests in plants did not show all the symptoms associated with vascular wilt in the three plant models; however, colonization and necrosis of the vascular bundles, regardless of the species and origin of the isolates, showed the infective potential of Fusarium spp. in different plant species. Moreover, the pathogenicity tests in the murine model revealed behavioral changes. It was noteworthy that only five isolates (different origin and species) caused mortality. Additionally, it was observed that all isolates infected and colonized different organs, regardless of the species and origin of the isolates or host immune status. In contrast, the superficial inoculation test showed no evidence of epidermal injury or colonization. The observed results in plant and murine models suggest the pathogenic potential of Fusarium spp. isolates in different types of hosts. However, further studies on pathogenicity are needed to confirm the multihost capacity of this genus.

Detección e identificación de Salmonella spp. en huevos para consumo humano, provenientes de diferentes localidades de Bogotá, Colombia, 2015 / Salmonella spp. isolation and identification in eggs for human consumption from different urban areas in Bogotá, Colombia, 2015

Objetivo: Determinar la prevalencia de Salmonella spp. en muestras de huevos para consumo humano en localidades de Bogotá. Materiales y métodos: Se obtuvieron 96 muestras en tiendas y plazas de mercado de 4 localidades de la ciudad. Se procesó de forma separada la cáscara y el contenido interno mediante métodos microbiológicos y moleculares para aislamiento e identificación Salmonella spp. Resultados: Se determinó una prevalencia total de Salmonella spp. de 9,4% (n=9), de ésta el 55% (n = 5) provenían del contenido interno y 44% (n = 4) de cáscara, sin embargo no se logró tipificar el serovar de Salmonella enterica presente. Las localidades con mayor presencia del patógeno fueron Usaquén y Fontibón. Discusión: Estudios realizados en Colombia evidencian bajas prevalencias de Salmonella spp. (0 -1,74%) en muestras de huevos, sin embargo los hallazgos de este estudio evidencian una mayor recuperación, lo que podría asociarse con inadecuadas condiciones de manejo y/o almacenamiento del producto. Conclusión: Se estableció la presencia de Salmonella spp. en las muestras de huevo evaluadas, lo que implica un riesgo potencial para la salud pública, por lo que es necesario ampliar este tipo de estudios para conocer la situación real a nivel nacional frente a este patógeno.

Objective: To determine the prevalence of Salmonella spp presence in eggs for human consumption in urban areas in Bogota. Materials and methods: 96 samples were collected from convenience stores and markets in 4 urban areas in the city. The eggshells were separated from the egg's internal content and both were processed separately using microbiological and molecular techniques to isolate and identify Salmonella spp. strains. Results: A Salmonella spp. prevalence of 9.4% (n=9) was found. Salmonella spp was isolated from the egg's internal content in 55% (n=5) of samples and 44% (n=4) from the eggshells. The Salmonella enterica serovar could not be identified. The pathogen was more frequently isolate in samples from Usaquén and Fontibón urban areas. Discussion: Studies of Salmonella spp. prevalence in eggs done in Colombia have shown it to be low (0-1.74%); However, this study determined a higher prevalence. These results suggest that inadequate handling/storage conditions could have been associated with them. Conclusion: Salmonella spp. was isolated from the egg samples from 4 different urban areas in Bogotá. These findings suggest the existence of a public health risk; therefore, there is a need to perform wider and more complete studies to determine the actual situation of Salmonella ssp. egg contamination in the country.

Actividad antifúngica in vitro de azoles y anfotericina B frente a Malassezia furfur por el método de microdilución M27-A3 del CLSI y Etest® / In vitro antifungal activity of azoles and amphotericin B against Malassezia furfur by the CLSI M27-A3 microdilution and Etest® methods

Antecedentes. Malassezia furfur es una levadura comensal de la piel del ser humano que ha sido asociada con la presencia de algunas entidades dermatológicas e infecciones sistémicas oportunistas. Por su condición dependiente de lípidos, los métodos de referencia establecidos para las levaduras por el Clinical and Laboratory Standards Institute (CLSI) para evaluar la sensibilidad antifúngica no son aplicables. Objetivos. Evaluar la sensibilidad in vitro de aislamientos de M. furfur asociados a procesos patológicos en el ser humano frente a antifúngicos de uso clínico. Métodos. Se evaluó el perfil de sensibilidad a la anfotericina B, el itraconazol, el ketoconazol y el voriconazol de 20 aislamientos de M. furfur mediante el método de microdilución en caldo (CLSI M27-A3) y Etest®. Resultados. El itraconazol y el voriconazol presentaron la mayor actividad antifúngica frente a los aislamientos evaluados. El acuerdo esencial entre los dos métodos usados para evaluar la actividad antifúngica de los azoles estuvo en el 60-85%, y el acuerdo categórico en el 70-80%; para la anfotericina B tanto el acuerdo esencial como el categórico fueron del 10%. Conclusiones. De acuerdo con los dos métodos evaluados los azoles fueron los compuestos que presentaron la mayor actividad antifúngica frente a M. furfur; sin embargo, es necesario realizar más estudios que permitan afirmar que Etest® es un método confiable para ser implementado en la rutina del laboratorio clínico (AU)

Background. Malassezia furfur is a human skin commensal yeast that can cause skin and opportunistic systemic infections. Given its lipid dependant status, the reference methods established by the Clinical and Laboratory Standards Institute (CLSI) to evaluate antifungal susceptibility in yeasts are not applicable. Aims. To evaluate the in vitro susceptibility of M. furfur isolates from infections in humans to antifungals of clinical use. Methods. The susceptibility profile to amphotericin B, itraconazole, ketoconazole and voriconazole of 20 isolates of M. furfur, using the broth microdilution method (CLSI M27-A3) and Etest®, was evaluated. Results. Itraconazole and voriconazole had the highest antifungal activity against the isolates tested. The essential agreement between the two methods for azoles antifungal activity was in the region of 60-85% and the categorical agreement was around 70-80%, while the essential and categorical agreement for amphotericin B was 10%. Conclusions. The azoles were the compounds that showed the highest antifungal activity against M. furfur, as determined by the two techniques used; however more studies need to be performed to support that Etest® is a reliable method before its implementation as a routine clinical laboratory test (AU)

[In vitro antifungal activity of azoles and amphotericin B against Malassezia furfur by the CLSI M27-A3 microdilution and Etest® methods]. / Actividad antifúngica in vitro de azoles y anfotericina B frente a Malassezia furfur por el método de microdilución M27-A3 del CLSI y Etest®.

BACKGROUND: Malassezia furfur is a human skin commensal yeast that can cause skin and opportunistic systemic infections. Given its lipid dependant status, the reference methods established by the Clinical and Laboratory Standards Institute (CLSI) to evaluate antifungal susceptibility in yeasts are not applicable. AIMS: To evaluate the in vitro susceptibility of M. furfur isolates from infections in humans to antifungals of clinical use. METHODS: The susceptibility profile to amphotericin B, itraconazole, ketoconazole and voriconazole of 20 isolates of M. furfur, using the broth microdilution method (CLSI M27-A3) and Etest®, was evaluated. RESULTS: Itraconazole and voriconazole had the highest antifungal activity against the isolates tested. The essential agreement between the two methods for azoles antifungal activity was in the region of 60-85% and the categorical agreement was around 70-80%, while the essential and categorical agreement for amphotericin B was 10%. CONCLUSIONS: The azoles were the compounds that showed the highest antifungal activity against M. furfur, as determined by the two techniques used; however more studies need to be performed to support that Etest® is a reliable method before its implementation as a routine clinical laboratory test.

Phenotypic characterization of canine Malassezia spp., isolates / Caracterización fenotípica de aislamientos de Malassezia spp., de origen canino

ABSTRACT Objective . To characterize and identify yeasts of the genus Malassezia by phenotypic features. Materials and methods. First, the macroscopic and microscopic morphological characteristics were described. In addition we performed biochemical and physiological assays as Tweens and Cremophor, including more. Results. Our results evidenced of 105 isolates obtained from dogs diagnosed with external otitis, it was possible to identify two distinct species from 46 isolates within the Malassezia genus: 36.19% (n=38) were identified as M. pachydermatis and 7.62% (n=8) as M. furfur. According to phenotypic patterns the remaining 56.19% (n=59) were reported as Malassezia spp., possibly corresponding to M. furfur and/or M. pachydermatis. Conclusions. Results emphasize the necessity to characterize according to species. It is not feasible to define Malassezia by species based on morphological, biochemical, and physiological findings. Therefore, molecular genotyping should be performed to identify markers allowing a more precise isolate identification. This would broaden our epidemiological knowledge regarding different species involved in canine otitis pathologies.

RESUMEN Objetivo . Caracterizar e identificar levaduras del género Malassezia, mediante características fenotípicas. Materiales y métodos . Inicialmente se describieron las características morfológicas macroscópicas y microscópicas, adicionalmente se realizaron pruebas bioquímicas y fisiológicas como Tween y Cremophor, entre otras. Resultados . De 105 aislamientos de caninos diagnosticados previamente con otitis, 46 fueron caracterizados hasta especie, así: El 36.19% (n=38) correspondió a M. pachydermatis, el 7.2% (n=8) a M. furfur; y 56.19% (n=59) restante fueron reportados como Malassezia spp., debido a los patrones fenotípicos atípicos que presentaron, y que podrían corresponder a variantes de M. furfur y/o M. pachydermatis. Conclusión. Estos resultados enfatizan la necesidad de hacer una caracterización a nivel de especie y/o genotipos mediante marcadores moleculares que permitan una identificación más precisa de los aislamientos. Con el presente estudio, se contribuye al conocimiento de las diferentes especies involucradas en patologías óticas en caninos.

Concordance between otic cytology and culture in diagnosis of external otitis canine by Malassezia spp / Concordancia entre citología ótica y cultivo en el diagnóstico de otitis externa canina por Malassezia spp

Objective. To determine the correlation between microbiological culture and otic cytology for diagnoses of external otitis by Malassezia in dogs. Materials and Methods. 158 ear swabs of dogs with clinical diagnosis of external otitis were analyzed by cytology, mycological culture and metabolic tests. Results. Were obtained a positive results by cytology of 62% and 75.3% by culture. The 31.1% of isolates were identified as M. pachydermatis, 12.6% as M. furfur and 56.3% were classified as Malassezia spp., because was not possible to define the species. We found a positive concordance between cytology and culture for Malassezia spp., of 0.76 with a kappa index of 0.448 (95% CI 0.30 to 0.60) which represents a moderate strength of concordance between the two techniques, without regard the identified species. Conclusions. The use of a diagnostic test is not enough to establish the participation of Malassezia spp., as a causal disease agent.

Objetivo. Determinar la concordancia entre el cultivo microbiológico y la citología ótica para el diagnóstico de otitis externa causada por Malassezia spp. en caninos. Materiales y Métodos. Se analizaron 158 muestras de hisopados de caninos con diagnóstico clínico de otitis externa, todas las muestras fueron analizadas mediante citología, cultivo micológico y pruebas metabólicas. Resultados. Se obtuvo una positividad mediante citología del 62% y por cultivo del 75.3%. El 31.1% de los aislamientos fueron identificados como M. pachydermatis, el 12.6% como M. furfur y un 56.3% se clasificó como Malassezia spp., dado que bioquímicamente no fue posible hallar su especie. Se determinó una concordancia observada entre técnicas positivas para Malassezia spp. de 0.76 con índice Kappa de 0.448 IC 95% (0.30 - 0.60) lo que representa una fuerza de concordancia moderada entre las dos técnicas, sin tener en cuenta la especie identificada. Conclusiones. El uso de una prueba diagnóstica no es suficiente para establecer la participación de Malassezia pachydermatis como agente causal de enfermedad.

Determination of presence of Tritrichomonas foetus in uterine lavages from cows with reproductive problems.

The aim of this study was to determine the presence of Tritrichomonas foetus in two dairy herds on the Altiplano Cundiboyacense. Twenty-one low-volume uterine lavages from cows with a history of reproductive problems in two dairy herds located in the municipality of Sibaté (Cundinamarca) and Ventaquemada (Boyacá) were evaluated. In the first herd, 10 cows were sampled and in the second, 11 cows, based on three inclusion criteria. The uterine lavages were obtained through infusion of physiological saline solution into the uterine body. The samples were centrifuged and seeded in Tritrichomonas basal medium for 10-15 days at 37 ºC. The protozoa were evaluated on the day of sampling and 10 and 15 days after incubation by means of direct viewing under a dark-field microscope. Positive samples were stained with Wright and Lugol to identify the morphological characteristics. This study showed that T. foetus was present in 61.8% of the animals sampled. The determination that T. foetus was present in 61.8% of the samples analyzed is a significant finding given that in the herds evaluated, this agent had not previously been diagnosed.

Determination of presence of Tritrichomonas foetus in uterine lavages from cows with reproductive problems / Determinação da presença de Tritrichomonas foetus em lavados uterinos de vacas com problemas reprodutivos

The aim of this study was to determine the presence of Tritrichomonasfoetus in two dairy herds on the Altiplano Cundiboyacense. Twenty-one low-volume uterine lavages from cows with a history of reproductive problems in two dairy herds located in the municipality of Sibaté (Cundinamarca) and Ventaquemada (Boyacá) were evaluated. In the first herd, 10 cows were sampled and in the second, 11 cows, based on three inclusion criteria. The uterine lavages were obtained through infusion of physiological saline solution into the uterine body. The samples were centrifuged and seeded in Tritrichomonas basal medium for 10-15 days at 37 ºC. The protozoa were evaluated on the day of sampling and 10 and 15 days after incubation by means of direct viewing under a dark-field microscope. Positive samples were stained with Wright and Lugol to identify the morphological characteristics. This study showed that T. foetus was present in 61.8% of the animals sampled. The determination that T. foetus was present in 61.8% of the samples analyzed is a significant finding given that in the herds evaluated, this agent had not previously been diagnosed.

O objetivo do presente estudo, foi determinar a presença de Tritrichomonas foetus em dois rebanhos leiteiros no Altiplano Cundiboyacense. De vacas com um histórico de problemas reprodutivos, foram avaliados 21 lavados uterinos de pequeno volume, em dois rebanhos leiteiros localizados nos municípios de Sibaté (Cundinamarca) e Ventaquemada (Boyacá). No primeiro rebanho, 10 vacas foram amostradas e no segundo 11, baseado em três critérios de inclusão. Os lavados uterinos foram obtidos mediante infusão de solução salina fisiológica no corpo do útero. As amostras foram centrifugadas e semeadas em meio de cultura básico para Tritrichomonas por 10-15 dias a 37 ºC. Os protozoários foram avaliados no dia da colheita de amostras e 10 e 15 dias após incubação por meio de visualização direta sob um microscópio de campo escuro. As amostras positivas foram coradas com Wright e Lugol para identificar as características morfológicas. Este estudo mostrou que T. foetus estava presente em 61,8% dos animais amostrados. A determinação de que T. foetus estava presente em 61,8% das amostras analisadas é um achado significativo, dado que nos rebanhos avaliados, esse agente não tinha sido previamente diagnosticado.

Análisis retrospectivo (2009-2010) de las alteraciones dermatológicas, óticas y oftalmológicas con diagnóstico clínico presuntivo de micosis en caninos y felinos / Análise retrospectiva (2009-2010) das alterações dermatológicas, óticas e oftalmológicas com diagnóstico clínico presuntivo de micose em cães e gatos / A retrospective analysis (2009-2010) of the dermatological, ear and ophtalmological alterations with presumptive clinical diagnosis of mycosis in canines and felines

Objetivo. Determinar la prevalencia de las alteraciones dermatológicas, óticas y oftalmológicas clínicamente presuntivas de ser causadas por hongos y analizar demográficamente su presentación. Materiales y métodos. Se realizó un estudio retrospectivo en una clínica para pequeños animales en Bogotá; se revisaron historias clínicas de pacientes que consultaron por alteraciones dermatológicas, óticas y oftalmológicas durante 2009 y 2010. Resultados. En el período evaluado la prevalencia de las alteraciones dermatológicas en caninos fue de 7,8%, las óticas de 4,2% y las oftalmológicas de 3,3%. Dentro de las alteraciones dermatológicas, la sospecha etiológica principal por diagnóstico clínico fueron los dermatofitos con 86,9%, en las alteraciones óticas fue Malassezia con 86,5% y para las oftalmológicas fue Candida con 83,3%. El análisis estadístico de variables no mostró asociación con los síntomas ni con los agentes etiológicos diagnosticados clínicamente; sin embargo, la evaluación categorizada evidenció asociaciones entre razas y edades con las etiologías por diagnóstico clínico. El diagnóstico de las etiologías micóticas se estableció en la mayoría de los casos por hallazgos clínicos y con menor frecuencia por raspado de piel y citología ótica; en ninguno de los casos se realizó cultivo micológico. La población felina no presentó alteraciones dermatológicas, óticas y oftalmológicas clínicamente compatibles con etiología fúngica. Conclusión. En caninos, la prevalencia de las alteraciones dermatológicas clínicamente compatibles con dermatofitos fue de 86,9%, los procesos óticos asociados con Malassezia de 86,5% y los oftalmológicos sospechosos de Candida con 83,3%. Los diagnósticos definitivos se establecieron por hallazgos clínicos sin la realización de metodologías diagnósticas estándar como el cultivo...

A retrospective analysis (2009-2010) of the dermatological, ear and ophtalmological alterations with presumptive clinical diagnosis of mycosis in canines and felines. Objective. To determine the prevalence of dermatological, ear and ophthalmological alterations clinically presumptive to be caused by fungi and to analyze their occurrence demographically. Materials and methods. We carried out a retrospective study in a clinic in Bogota for small animals. We reviewed the medical histories of patients who requested medical appointments due to dermatological, ear and ophtalmological alterations in 2009 and 2010. Results. In the assessed period, the prevalence of dermatological alterations in dogs was of 7.8%, 4.2% of ear alterations and of 3.3% of ophtalmological alterations. The main etiological suspects through clinical diagnosis were: dermatophytes with an incidence of 86.9% among dermatological alterations, Malassezia with an 86.5% of incidence in ear alterations, and Candida with 83.3% of incidence in ophtalmological alterations. Statistical analysis of variables showed no association with symptoms or clinically diagnosed etiological agents, but the categorized evaluation showed associations of races and ages with the etiologies through clinical diagnosis. The diagnosis of fungal etiologies was accomplished in most cases by clinical findings and less frequently by skin scrapings and ear cytology; in none of the cases a mycological culture was done. The feline population showed no dermatological, ear or ophtalmological alterations compatible with a fungal etiology. Conclusion. In canines, the prevalence of dermatological alterations clinically compatible with dermatophytes was of 86.9%; ear alterations associated with Malassezia were of 86.5%; and ophtalmological alterations Candida-suspected were 83.3%. Final diagnoses were accomplished with the help of clinical findings without following standard diagnostic methodologies such as cultivation...

Análise retrospectiva (2009-2010) das alterações dermatológicas, óticas e oftalmológicas com diagnóstico clínico presuntivo de micose em cães e gatos. Objetivo. Determinar a prevalência das alterações dérmicas, óticas e oftálmicas clinicamente presuntivo de serem causadas por fungos e analisar demograficamente sua apresentação. Materiais e métodos. Foi realizado um estudo retrospectivo numa clínica para pequenos animais em Bogotá; foram revisadas as histórias clínicas de pacientes com alterações dérmicas, óticas e oftalmicas em 2009 e 2010. Resultados. Durante o período de avaliação a prevalência das alterações dérmicas em cães foi de 7,8%, as óticas de 4,2% e as oftálmicas de 3,3%. Dentro das alterações dérmicas, a principal suspeita etiológica por diagnóstico clínico foram os dermatófitos com 86,9%, nas alterações óticas foi Malassezia com 86,5% e para as oftálmicas foi Candida com 83,3%. A análise estatística das variáveis não apresentou associação com os sintomas ou com os agentes etiológicos diagnosticados clínicamente; embora, a avaliação categorizada evidenciou associações entre raças e idades com as etiologias por diagnóstico clínico. O diagnóstico das etiologias micóticas foi estabelecido na maioria dos casos pelos achados clínicos e, menos freqüentemente por raspados de pele e citologia ótica, em nenhum dos casos foi realizada uma cultura micótica. A população felina não apresentou alterações dérmicas, óticas e oftálmicas clinicamente compatíveis com etiologia fúngica. Conclusão. Em cães, a prevalência das alterações dérmicas clinicamente compatíveis com dermatófitos foi de 86,9%, os processos óticos associados com Malassezia foi de 86,5% e os oftálmicos suspeitos de Candida foi de 83,3%. Os diagnósticos definitivos foram estabelecidos pelos achados clínicos sem a realização de métodos diagnósticos padroes, como o cultivo...