Extended-spectrum beta-lactamase-producing Klebsiella pneumoniae and risk factors associated with high total bacterial count in bulk tank milk from dairy farms in Colombia.

The objective of the study was to evaluate the frequency and genetic characteristics of ESBL-producing Escherichia coli and Klebsiella spp. and the risk factors associated with a high total bacterial count in bulk tank milk samples of dairy farms in three municipalities of the Antioquia Department, Colombia. Fifteen samples were positive for E. coli and Klebsiella spp. Subsequent analysis of the 16 S rRNA gene sequences confirmed these isolates included E. coli (n = 3), K. oxytoca (n = 11), and K. pneumoniae (n = 1). None of the isolates was positive for ESBL identification by phenotypic methods, but the only the isolate of K. pneumoniae was positive for the blaSHV61 gene by sequence analysis. The antibiotic susceptibility evaluation for all Klebsiella spp. isolates identified resistance to fosfomycin (50%; 6/12) and ampicillin (100%; 12/12). While most of the herds maintain adequate hygienic quality, specific risk factors such as having more than 60 milking cows, frequent changes in milkers, milking in paddocks, and using a chlorinated product for pre-dipping have been identified as associated with a high total bacterial count > 100,000 CFU/mL in bulk tank milk. However, certain variables including the milker being the owner of the animals and the proper washing and disinfection of the milking machine contribute to maintain a high level of hygiene and quality in the raw milk stored in the tanks. In conclusion, the frequency of ESBL producers was relatively low, with only K. pneumoniae testing positive for the blaSHV ESBL type. The presence of these bacteria in milk tanks represents a potential risk to public health for consumers of raw milk and its derivatives.

Staphylococcus aureus, Escherichia coli, and Klebsiella spp. prevalence in bulk tank milk of Colombian herds and associated milking practices.

Background and Aim: Bovine mastitis is one of the most costly and prevalent diseases in dairy herds, which can be prevented and controlled through proper milking practices, diagnosis, and elimination of chronic animals, among others. Contagious pathogens such as Staphylococcus aureus and environmental pathogens such as Escherichia coli and Klebsiella spp. can affect cows and milk for human consumption, generating a public health risk. This study aimed to estimate the prevalence of herds with somatic cell count (SCC) ≥200,000 cells/mL, S. aureus, E. coli, and Klebsiella spp., in bulk tank milk (BTM) and its associated risk factors in Colombian dairy cow herds. Materials and Methods: A cross-sectional probabilistic study was carried out in 150 dairy herds located in the north of the Antioquia province. A single visit per herd was conducted, during which three BTM samples were aseptically collected. General data and milking practices were collected through an epidemiological survey applied in each herd. Results: The prevalence of S. aureus, E. coli, and Klebsiella spp. were 14% (21/150), 2% (3/150), and 8% (12/150), respectively. Moreover, 95% of the herds presented an SCC of ≥200,000 cells/mL. Practices such as in-paddock milking, change of milker during the last month, use of disposable gloves, and hand disinfection were associated with increased prevalence of S. aureus, whereas proper dipping was a protective factor. Proper washing and disinfection of the milking machine, use of chlorinated disinfectants for hand hygiene, and use of disposable gloves decreased the prevalence of E. coli and Klebsiella spp. Bulk tank SCC increased in herds with 30-60 milking cows, herds with >60 milking cows, and herds with a change of milker during the last month. Hand disinfection and dipping decreased the SCC. Conclusion: Staphylococcus aureus, E. coli, and Klebsiella spp. were prevalent in BTM from dairy cow herds. The risk of S. aureus isolation was higher in herds with an in-paddock milking system. The risk of E. coli and Klebsiella spp. isolation were higher in herds with >60 milking cows, with a change of milker during the last month. Processes such as avoiding changing the milker and greater control in medium and large herds could improve the SCC in BTM.

Anisakidae parasites in frozen fish fillets intended for human consumption. / Parásitos anisákidos en filetes de pescado congelado destinados a consumo humano

INTRODUCTION: Anisakiasis is a human parasitic disease caused by the consumption of raw fish or shellfish containing larvae of the Anisakidae family. It is currently considered an emerging disease of public health interest. OBJECTIVE: To identify the presence of larvae of the Anisakidae family in samples of frozen raw fish fillets intended for human consumption in markets in Medellín and its metropolitan area in Antioquia, Colombia. MATERIALS AND METHODS: A cross-sectional study was carried out, in which larvae of the Anisakidae family were detected and identified in frozen raw fish fillets from three representative markets in Medellín and its metropolitan area. A total of 384 ready for consumption fillets were analyzed (197 sawfish, 137 salmon, 37 tuna, and 13 hake), using the pressing and ultraviolet light method. Taxonomic keys were used to identify the collected parasites and to establish its genus. Conventional PCR and Sanger sequencing was performed to determine the species. RESULTS: Four larvae were found in 4 of the 384 (1.04%) fillets (CI95% 1.04 ± 1.01%). The species of fish in which the larvae were found was sawfish (Scomberomorus spp.) and the genus and species of the larvae was established as Anisakis pegreffii. CONCLUSIONS: According to the study, the presence of Anisakis parasites in frozen raw fish fillets in the influence area is evident.

Introducción. La anisakiasis es una infección producida por parásitos de la familia Anisakidae, transmitida a los humanos por el consumo de pescado o mariscos crudos. En la actualidad, se considera una enfermedad emergente de interés en salud pública. Objetivo. Identificar la presencia de larvas de la familia Anisakidae en muestras de filetes de pescado crudo congelado destinados a consumo humano, en mercados de Medellín y su área metropolitana en Antioquia (Colombia). Materiales y métodos. Se realizó un estudio transversal, en el cual se buscó la presencia de larvas de la familia Anisakidae en filetes de pescado crudo congelado de tres mercados representativos de Medellín y su área metropolitana. Se analizaron 384 filetes listos para el consumo (197 sierras, 137 salmones, 37 atunes y 13 merluzas). Cada filete fue analizado mediante el método de prensado y luz ultravioleta. Los parásitos recolectados se identificaron a partir de claves taxonómicas para establecer el género, así como PCR convencional y posterior secuenciación Sanger, para determinar la especie. Resultados. Se encontraron 4 larvas en 4 de los 384 filetes (1,04 %) (IC95% 1,04 ± 1,01 %). Las larvas encontradas fueron identificadas como Anisakis pegreffi y el tipo de pescado en el cual se encontraron fue la sierra (Scomberomorus spp.). Conclusiones. De acuerdo con el estudio realizado, se evidencia la presencia de parásitos anisákidos en filetes de pescado crudo congelado en el área de influencia.

Paratuberculosis control strategies in dairy cattle: A systematic review.

Background: Mycobacterium avium subsp. paratuberculosis is the causative agent of paratuberculosis (PTB), incurable enterocolitis, affecting domestic and wild ruminants. Economic losses, impacts on animal health and welfare, and public health concerns justify its herd-level control. Aim: To systematically collect information to answer: What are the control and eradication strategies of PTB in dairy cattle worldwide? Methods: The search procedure was carried out on October 2nd, 2019, and updated on August 3rd, 2021, using OVID®, SciELO, and Redalyc databases, and the registers from the International Colloquium on Paratuberculosis (1991-2018). The inclusion criteria considered articles published in English, Portuguese, and Spanish and in peer-reviewed journals. The exclusion criteria included irrelevant topics, species other-than bovines, and not original articles. Definitive studies were obtained through the consensus of the authors on eligibility and quality. Data extraction was performed, considering bibliographic information, control and outcome strategies, follow-up time, and results. Results: Twenty-six relevant studies were found, reporting the use of three grouped control strategies: hygiene and management strategy (HMS), test-and-cull strategy (TCS), and vaccination strategy (VS). The HMS was the most common one (20/26), followed by TCS (17/26) and VS (7/26). Combined control strategies such as TCS-HMS (12/26), TCS-VS (1/26), and HMS-VS (1/26) were also described, and the consideration of the three control strategies (TCS-HMS-VS) was reported in two articles. The HMS included practices such as neonates/juvenile livestock hygiene, biosecurity, prevention of infection introduction into the herd, and environmental management. Within HMS, the most frequent practices were to remove calves from their dams as soon as possible after birth and to keep the minimal exposure of calves and heifers to adult cattle. As limitations, within the HMS, it is considered that some strategies cannot be included due to lack of compliance, or the application of the same strategy among one study and another may have a different degree of interpretation; publication bias was not controlled since the results of the control programs in endemic countries may be not available. Conclusion: The main PTB control strategies in dairy cattle worldwide are HMS, TCS, and VS. The use of one or several combined strategies has been found to succeed in controlling the disease at the herd-level.

Mycobacterium avium subsp. paratuberculosis (MAP) molecular diversity in cattle, sheep, and goats from Latin America and the Caribbean: a systematic review.

This study aimed to systematically collect and appraise the scientific evidence to answer the research question: What MAP genotypes have been isolated from cattle, sheep, and goats in Latin America and the Caribbean? An electronic search was conducted on three platforms (i.e., OVID®, Web of Science®, SciELO) as well as on the proceedings of the International Colloquium on Paratuberculosis. Inclusion and exclusion criteria were defined a priori and conserved through the systematic process and only articles published in peer-reviewed journals were considered. A total of 26 articles met the definitive inclusion criteria. All were published in English, in 15 different journals, and between 1989 and 2020. The relevant articles reported the use of six different genotyping techniques (i.e., polymerase chain reaction-restriction endonuclease analysis, restriction fragment length polymorphism, type-specific-PCR, mycobacterial interspersed repetitive units-variable number of tandem repeats, multi-locus short sequence repeat, single nucleotide polymorphism) in isolates from seven countries. Genotypes found so far in the region using typing techniques were mainly C type. MIRU-VNTR mostly reported INMV 1, INMV 2, and INMV 11 subtypes, among others. MLSSR reported genotypes from four different countries, reporting nine different subtypes of which 7g-10g-4ggt was the most common for loci 1, 2, and 8, respectively. Regardless the high diversity of techniques used so far to genotype Latin American and Caribbean MAP isolates, the original question of this systematic review has been answered. In addition, a relative genetic similarity between MAP strains recovered from cattle, goats, and sheep unrelatedly of the matrix and geographic origin was identified.

Tipificación molecular de Mycobacterium avium subsp. paratuberculosis en lecherías de Antioquia, Colombia

RESUMEN Objetivo. Determinar la diversidad molecular de Mycobacterium avium subsp. paratuberculosis (MAP) en muestras ambientales de hatos lecheros colombianos. Materiales y métodos. Las muestras ambientales de 25 hatos lecheros positivos a MAP por IS900-qPCR se cultivaron por duplicado en medio de yema de huevo de Herrold con micobactina J para obtener aislamientos. Las colonias sospechosas fueron confirmadas para MAP por IS900-qPCR. El ADN positivo se subtipó utilizando técnicas de unidades micobacterialess repetitivas intercaladas - número variable de repeticiones en tándem (MIRU-VNTR) y técnicas de repeticiones de multilocus de secuencia corta (MLSSR) para analizar las diferencias genéticas entre los aislamientos. Resultados. El subtipado reveló dos genotipos diferentes por MIRU-VNTR (INMV 2 e INMV 36). La técnica de MLSSR se realizó para aumentar el poder discriminatorio de lo obtenido por MIRU-VNTR, pero no se observaron diferencias entre los aislamientos recuperados. Conclusiones. El presente estudio representa un enfoque importante para el conocimiento del estatus epidemiológico de MAP en la población de estudio.

ABSTRACT Objective. To determine Mycobacterium avium subsp. paratuberculosis (MAP) molecular diversity in environmental samples from Colombian dairy herds. Materials and methods. Environmental samples from 25 IS900-qPCR MAP-positive dairy herds were cultured by duplicate in Herrold's egg yolk medium with mycobactin J to obtain isolates. Suspicious colonies were confirmed by MAP-IS900-qPCR. Positive DNA was sub-typed using mycobacterial interspersed repetitive units-variable number of tandem repeat (MIRU-VNTR) and multilocus short sequence repeats (MLSSR) techniques to analyze the genetic differences between the isolates. Results. Sub-typing revealed two different genotypes by MIRU-VNTR (INMV 2 and INMV 36). MLSSR technique was carried out to increase the discriminatory power from what was obtained by MIRU-VNTR, but no differences were observed among the recovered isolates. Conclusions. The present study represents an important approach to the knowledge on MAP epidemiological status in the study population.

Detección de anticuerpos anti-Mycobacterium avium subsp. paratuberculosis en bovinos de hatos lecheros de trópico bajo en los departamentos de Sucre y Córdoba, Colombia / Detection of anti-Mycobacterium aviumsubsp. paratuberculosis antibodies in cows of low-tropic dairy herds in the Provinces of Sucre and Córdoba, Colombia

RESUMEN Objetivo. Reportar la frecuencia de la seropositividad a Mycobacterium avium subsp. paratuberculosis (MAP) y explorar los factores asociados al resultado positivo, tanto a nivel de hato como de animal. Materiales y métodos. Se llevó a cabo un estudio transversal en 204 vacas lecheras resultado del cruce de diferentes razas, en cinco hatos de zonas de trópico bajo ubicadas en tres municipios de los departamentos de Sucre y Córdoba (Colombia) en 2018. Los animales fueron seleccionados aleatoriamente y se colectaron muestras de sangre de cada uno. Se utilizó un kit comercial de ELISA para analizar los sueros. La información con respecto a los factores relacionados con los hatos y los animales se recopiló mediante cuestionarios. Se calcularon las estadísticas descriptivas para todas las variables (i.e. tamaño del hato, presencia de otros rumiantes en co-pastoreo con el ganado bovino en los últimos 2 años, edad de los animales, paridad y días desde el último parto), y se exploró la asociación entre dichas variables y el resultado a ELISA (p<0.05). Resultados. El 17.2% (35/204; IC 95%: 12.0-22.3%) de las vacas resultaron positivas a MAP mediante la prueba de ELISA, y los cinco hatos tenían animales seropositivos. La variable a nivel de hato presencia de otros rumiantes en co-pastoreo con el ganado bovino en los últimos 2 años y las variables a nivel de animal edad y paridad se encontraron asociados con los resultados positivos de ELISA. Conclusiones. El presente estudio encontró que el 17.2% de las vacas y el 100% de los hatos fueron positivos a MAP mediante la prueba de ELISA. Adicionalmente se identificaron variables asociadas que pueden ser de interés tanto para los productores como para los veterinarios de animales de producción, y orientar su enfoque para el manejo de la enfermedad.

ABSTRACT Objective. To report the frequency of seropositivity against Mycobacterium avium subsp. paratuberculosis (MAP) and to explore the factors associated with the positive outcome, both at herd and animal level. Materials and methods. A cross-sectional study was carried out on 204 dairy cows, result of the crossing of different breeds, from five low-tropic dairy herds, located in three municipalities of the Provinces of Sucre and Córdoba (Colombia) in 2018. The animals were randomly selected, and blood samples were collected from each one. A commercial ELISA kit was used to analyze the sera. Information regarding factors related to herds and animals was collected through questionnaires. Descriptive statistics were calculated for all variables (i.e. herd size, presence of other ruminants co-grazing with cattle in the last 2 years, age of the animals, parity, and days since the last birth), and the association between these variables and the result to ELISA (p<0.05) was explored. Results. The 17.2% (35/204; 95% CI: 12.0-22.3%) of the cows were positive for MAP by the ELISA test, and the five herds had seropositive animals. The herd-level variable presence of other ruminants in co-grazing with cattle in the last 2 years and the animal-level variables age and parity were associated with the positive ELISA results. Conclusions. The present study found that 17.2% of the cows and 100% of the herds were MAP positive by the ELISA test. Additionally, associated variables were identified and may be of interest to both producers and veterinarians of production animals, guiding their approach to disease management.

Detección de anticuerpos anti-Mycobacterium avium subsp. paratuberculosis (MAP) en venados silvestres en Colombia / Detection of anti-Mycobacterium aviumsubsp. paratuberculosis antibodies in wild deer in Colombia

RESUMEN Objetivo. Determinar la presencia de anticuerpos anti-Mycobacterium avium subsp. paratuberculosis (MAP) en venados de cola blanca (Odocoileus virginianus) y venados de páramo (Mazama rufína), capturados en las regiones de la Orinoquía y el Caribe en Colombia. Materiales y métodos. Un total de 44 muestras de suero sanguíneo de venados de las especies M. rufína y O. virginianus fueron colectadas en condiciones de campo entre 2014 y 2016. Se utilizó un kit comercial de ELISA para la detección de anticuerpos anti-MAP. Un animal se consideró positivo a ELISA cuando la relación muestra-positivo (S/P%) fue > 0.4, según lo recomendado por el fabricante. Resultados. El 50% (22/44) de los animales muestreados resultaron positivos, lo que corresponde a 10 hembras y 12 machos. Un 81.8% (18/22) y un 77.3% (17/22) de estos animales seropositivos fueron capturados en la región de la Orinoquía y fueron reportados como adultos, respectivamente. Conclusiones. No se sabe cómo o cuándo se introdujo MAP en la población de ciervos colombianos en las regiones de estudio. La hipótesis más plausible para explicar la presencia de anticuerpos anti-MAP en estas poblaciones silvestres es la transmisión por contacto con el ganado bovino infectado, ya que en ambas regiones estas especies comparten pasturas. Este es el primer estudio en explorar la infección por MAP en animales silvestres en Colombia. Estos hallazgos respaldan la necesidad de realizar más estudios utilizando técnicas de diagnóstico directo, y aproximaciones investigativas que permitan la definición de vínculos en la dinámica de la infección entre mamíferos silvestres y domésticos en Colombia.

ABSTRACT Objective. To determine the presence of anti-Mycobacterium avium subsp. paratuberculosis (MAP) antibodies in white-tailed deer (Odocoileus virginianus) and red brocket deer (Mazama rufína), captured in the Orinoquía and Caribbean regions in Colombia. Materials and methods. A total of 44 blood serum samples from deer of species M. rufina and O. virginianus were collected under field conditions between 2014 and 2016. An ELISA commercial kit was used to detect anti-MAP antibodies. An animal was considered ELISA-positive at a sample-to-positive ratio (S/P%) of > 0.4, as recommended by the manufacturer. Results. The 50% (22/44) of the animals were positive, corresponding to 10 females and 12 males. An 81.8% (18/22) and 77.3% (17/22) of these seropositive animals were captured in the Orinoquía region and were reported as adults, respectively. Conclusions. It is not known how or when MAP was introduced in the Colombian deer population in the study regions. The most plausible hypothesis to explain the presence of antibodies against MAP in these wild populations is transmission by contact with infected bovine cattle since, in both regions, these species share pastures. This is the first study to explore MAP infection in wild animals in Colombia. These findings support the need for further studies using different direct diagnostic techniques and research approaches that allow the definition of links in the infection dynamics between wild and domestic mammals in Colombia.

Ehrlichia canis TRP36 diversity in naturally infected-dogs from an urban area of Colombia.

Ehrlichia canis is the etiologic agent of a highly prevalent tick-borne disease, canine monocytic ehrlichiosis (CME). Four defined E. canis genotypes based on the trp36 gene sequences have been reported, three of them identified in North or South America. The diversity of E. canis has been investigated using genetic and serologic approaches based on distinct 36 kDa tandem repeat protein (trp36) gene sequences that have been reported. The main objectives of this study were to determine the prevalence of E. canis infection in dogs from Medellín, Colombia by PCR and determine the E. canis diversity using molecular and serologic approaches. Blood was collected from dogs (n = 300) with clinical signs of CME for PCR detection of E. canis 16S rRNA, dsb and trp36 DNA. Phylogenetic analysis of trp36 gene sequences was performed using MEGA. A serological evaluation was performed using immunofluorescence microscopy and ELISA with species-specific peptides from E. canis TRP19 and TRP36 (3 genotypes) and E. chaffeensis (TRP32). E. canis DNA (16S rRNA and/or dsb) was detected in 18 % (53/300) of dogs by PCR amplification. The trp36 gene was amplified and sequenced from 35/53 16S rRNA/dsb PCR positive samples revealing three genotypes: United States (US; n = 21), Costa Rica (CR; n = 11), and Brazil (BR; n = 3). Most dogs (33/35) with detectable trp36 DNA had anti-E. canis TRP19 and TRP36 peptide antibodies that corresponded to the genotype detected by PCR. Dogs that had antibodies to the TRP19 peptide (82/300; 38 %), also had antibodies to one or more genotype-specific TRP36 peptides. Based on TRP36 serology, the dogs exhibited highest frequency of infection with the US genogroup (US = 26), followed by the CR genogroup (CR = 19) and the BR genogroup (BR = 11). Notably, 26/53 trp36 PCR positive dogs had detectable antibodies to multiple E. canis genotypes (US/BR/CR = 8, BR/CR = 7, US/CR = 6 and US/BR = 5) suggesting coinfection or multiple sequential infections with different genotypes. Colombian dogs did not have antibodies to E. chaffeensis as determined by a TRP32 species-specific ELISA. Our results demonstrate the presence of three previously defined genotypes in North and South America in Colombian dogs (US, BR, CR). These results also demonstrate that TRP19 and TRP36 serology can provide valuable information regarding E. canis exposure and the potential genotype(s) involved in infection.

Prevalence of Mycobacterium avium subsp. paratuberculosis infection in dairy herds in Northern Antioquia (Colombia) and associated risk factors using environmental sampling.

This cross-sectional study aimed to determine Mycobacterium avium subsp. paratuberculosis (MAP) herd-level prevalence using a quantitative real-time PCR method (qPCR), performed on environmental samples. Secondly, the study aimed to explore herd-level risk factors associated with the presence of MAP in dairy herds with in-paddock milking facilities of the Northern region of the Province of Antioquia (Colombia). Study herds (n = 292) located in 61 different districts from six municipalities were randomly selected amongst 7794 dairies registered in the foot-and-mouth disease vaccination records from 2015. The sampling strategy considered a proportional allocation, both at municipality and district level. Participant herds were visited once between June and October 2016 to collect one composite environmental sample and to complete a risk assessment questionnaire. Each composite environmental sample contained material from six different sites of concentration of adult cattle and/or high traffic areas (e.g. areas surrounding waterers and feeders, areas surrounding the current mobile milking-unit places). Identification of MAP was achieved using a duplex qPCR (Bactotype MAP PCR Kit®, Qiagen). A herd was considered as MAP infected if the environmental sample was positive in the qPCR. Information about the general characteristics of the herd, management practices, and knowledge about the disease was collected using the risk-assessment questionnaire. The information on risk factors was analyzed using a multivariable logistic regression model. The apparent herd-level prevalence was 4.1% (12/292; 95% CI: 1.8-6.4). Herds with a history of mixed farming of cattle with other ruminants had higher odds of being MAP infected than herds without (OR = 3.9; 95% CI: 1.2-13.2). Our study demonstrates the MAP prevalence in dairy herds from Antioquia, Colombia and the possible relationship between MAP environmental positivity with the history of mixed farming of cattle with other susceptible ruminants.

Mycobacterium avium subsp. paratuberculosis in Colombia (1924-2016): A review / Mycobacterium avium subsp. paratuberculosis en Colombia (1924-2016): Revisión de literatura / Mycobacterium avium subsp. paratuberculosis na Colômbia (1924-2016): Revisão de literatura

Abstract Mycobacterium avium subsp. paratuberculosis (MAP) is an acid-fast, gram-positive bacillus. MAP is the causal agent of paratuberculosis (PTB) or Johne's disease, an infectious disease affecting domestic ruminants and some wild species. Its importance as a potentially zoonotic agent due to its relation to Crohn's disease (CD) in humans is still under debate and investigation. The aim of the present systematic review is to summarize original studies on MAP carried out in Colombia since 1924, as well as to establish strengths, weaknesses, and future research opportunities in the country with emphasis on diagnosis and epidemiology. The initial search for existing publications reporting original studies on MAP, PTB, and the relationship between MAP and CD was carried out in the available databases and national libraries. After compilation of the available studies (n = 20), the relevant data was extracted (year, province of report, species studied, diagnostic tests used, study design, summary of results, and authors). Recommendations for future research opportunities on MAP in Colombia are made.

Resumen Mycobacterium avium subsp. paratuberculosis (MAP) es un bacilo ácido resistente, gram-positivo. MAP es el agente causal de la paratuberculosis (PTB) o enfermedad de Johne, una enfermedad infecciosa que afecta rumiantes domésticos y algunas especies salvajes. Su importancia como agente zoonótico, debido a su relación con la enfermedad de Crohn (CD) en humanos, está aún en debate y bajo investigación. El objetivo de la presente revisión es exponer los estudios originales sobre MAP llevados a cabo en Colombia desde 1924, así como establecer sus fortalezas, debilidades y oportunidades de investigación futura, con énfasis en los puntos de vista diagnóstico y epidemiológico. La búsqueda inicial de las publicaciones existentes sobre estudios originales realizados acerca de MAP, PTB y la relación MAP y CD fue realizada en las bases de datos disponibles y en bibliotecas nacionales. Luego de la compilación de los estudios disponibles (n = 20), los datos relevantes fueron extraídos (año, provincia de reporte, especie estudiada, prueba diagnóstica usada, diseño del estudio, resumen de resultados y autores). Se hacen recomendaciones para futuras investigaciónes de MAP en Colombia.

Resumo Mycobacterium avium subsp. paratuberculosis (MAP) é um ácido forte, bacilo gram-positivo. O MAP é o agente causador da paratuberculosis (PTB) ou doença de Johne, uma doença infecciosa que afecta os ruminantes domésticos e algumas espécies selvagens. Sua importância como um agente zoonótico por causa de sua relação com a doença de Crohn (CD) em humanos ainda está em discussão e sob investigação. O objetivo desta revisão é apresentar a estudos mapa original realizado na Colômbia desde 1924, e estabelecer pontos fortes, pontos fracos e oportunidades para futuras pesquisas no país com ênfase no diagnóstico e epidemiológico pontos de vista. A busca inicial da literatura sobre estudos originais sobre MAP, PTB e a relação MAP e CD foi feita nas bases de dados disponíveis e bibliotecas nacionais. Após a compilação dos estudos disponíveis (n = 20), os dados relevantes foram extraídos (ano, relatório província, espécies estudadas, teste de diagnóstico utilizado, desenho do estudo, os resultados resumo e autores). Foram feitas recomendações sobre futuras oportunidades de pesquisa em MAP em Colômbia.

Tasa de incidencia de mastitis clínica y susceptibilidad antibiótica de patógenos productores de mastitis en ganado lechero del norte de Antioquia, Colombia* / Incidence rate of clinical mastitis and antibiotic susceptibility of mastitis-producing pathogens in dairy cattle from Northern Antioquia, Colombia / Taxa de incidência de mastite clínica e susceptibilidade antibiótica de patógenos produtores de mastite em gado leiteiro do norte de Antioquia, Colômbia

Resumen Introducción: la mastitis se define como la inflamación de la glándula mamaria causante de grandes pérdidas económicas en hatos lecheros, lo que amerita su investigación. Objetivo: determinar la tasa de incidencia de mastitis clínica TIMC, la etiología asociada y la sensibilidad antibiótica de patógenos aislados de casos de mastitis en ganado lechero. Materiales y métodos: se siguieron 37 hatos por 12 meses para el diagnóstico de mastitis cílínica (MC). A las muestras de leche de los casos de MC se les realizó cultivo bacteriológico y a estas, así como a los aislamientos de casos de mastitis subclínica de un estudio previo, se les efectuó prueba de susceptibilidad antibiótica. Para el análisis de la información se utilizó estadística descriptiva estándar. Resultados: TIMC fue de 13,8 casos por 100 vacas-año a riesgo. Se aislaron 188 patógenos totales de casos de MC, entre los cuales el Streptococcus agalactiae, el Streptococcus pyogenes y el Corynebacterium spp. fueron los más frecuentes, con 29,8; 11,7 y 5,9 %, respectivamente. Se halló una alta sensibilidad de las bacterias contagiosas para los antibióticos cloxacilina y cefoperazone. Conclusiones: en el presente estudio se encontró una TIMC de 13,8 casos por 100 vacas-año a riesgo. Los patógenos más prevalentes identificados en casos de MC fueron contagiosos. Se encontró una alta sensibilidad de las bacterias contagiosas para la mayoría de los antibióticos β-lactámicos.

Abstract Introduction: Mastitis is defined as the inflammation of the mammary gland causing great economic losses in dairy herds, which merits investigation. Objective: To determine the incidence rate of clinical mastitis IRCM, associated etiology, and antibiotic sensitivity of pathogens isolated from cases of mastitis in dairy cattle. Materials and methods: 37 herds were followed during 12 months for clinical mastitis (CM) diagnosis. Milk samples from CM cases were bacteriologically cultured, which underwent, along with isolates from subclinical mastitis cases from a previous study, antibiotic susceptibility testing. For data analysis, standard descriptive statistics were used. Results: The IRCM was 13.8 cases per 100 cows-year at risk. A total of 188 pathogens of CM cases were isolated, among which Streptococcus agalactiae, Streptococcus pyogenes, and Corynebacterium spp. were the most frequent, with 29.8%, 11.7%, and 5.9%, respectively. A high susceptibility of contagious bacteria was found for cloxacillin and cefoperazone. Conclusions: The present study found an IRCM of 13.8 cases per 100 cows-year at risk. The most prevalent pathogens identified in cases of CM were contagious. A high sensitivity of contagious bacteria was found for most β-lactam antibiotics.

Resumo Introdução: a mastite se define como a inflamação da glândula mamária causante de grandes perdas econômicas em rebanhos de gado leiteiro, o que justifica a sua pesquisa. Objetivo: determinar a taxa de incidências de mastite clínica TIMC, a etiologia associada e a sensibilidade antibiótica de patógenos isolados de casos de mastite em gado leiteiro. Materiais e métodos: seguiram-se 37 rebanhos bovinos durante 12 meses para o diagnóstico de mastitis clínica (MC). Com as amostras de leite dos casos de MC realizou-se cultivo bacteriológico e a estas, assim como aos isolamentos de casos de mastite subclínica de um estudo prévio, efetuou-se prova de susceptibilidade antibiótica. Para a análise da informação se utilizou estatística descritiva padrão. Resultados: a TIMC foi de 13,8 casos por 100 vacas-ano a risco. Foram isolados 188 patógenos totais de casos de MC, entre os quais o Streptococcus agalactiae, o Streptococcus pyogenes e o Corynebacterium spp. foram os mais frequentes, com 29,8; 11,7 e 5,9 %, respectivamente. Constatou-se uma alta sensibilidade das bactérias contagiosas para os antibióticos cloxacilina e cefoperazone. Conclusões: neste estudo evidenciou-se uma TIMC de 13,8 casos por 100 vacas-ano a risco. Os patógenos mais prevalentes identificados em casos de MC foram contagiosos. Encontrou-se uma alta sensibilidade das bactérias contagiosas para a maioria dos antibióticos β-lactâmicos.

Fecal culture and two fecal-PCR methods for the diagnosis of Mycobacterium avium subsp. paratuberculosis in a seropositive herd / Cultivo fecal y dos métodos de PCR en materia fecal para el diagnóstico de Mycobacterium avium subsp. paratuberculosis en un hato seropositivo / Cultivo fecal e dois métodos de PCR em matéria fecal para o diagnóstico de Mycobacterium avium subsp. paratuberculosis em um rebanho soropositivo

Abstract Background: paratuberculosis is a slow-developing infectious disease, characterized by chronic granulomatous enterocolitis. This disease has a variable incubation period from 6 months to over 15 years, and is caused by Mycobacterium avium subsp. paratuberculosis (MAP). Its detection by direct and indirect diagnostic techniques has been of special interest. Objective: to report the diagnosis and detection of MAP using several diagnostic tests in a herd of the Northern region of Antioquia, Colombia. Methods: serum samples from the study herd were analyzed, using a commercial ELISA (enzyme-linked immunosorbent assay) kit. Fecal samples were cultured by duplicate using Herrold´s egg yolk medium (HEYM), and analyzed by an end- point IS900-specific nested PCR protocol, and a commercial F57-real-time PCR kit. Results: eight out of 27 serum samples in the study herd resulted ELISA-positive. None of fecal samples resulted positive to HEYM culture by duplicate and none were found to be positive by F57-real-time PCR. Seven of the 27 fecal samples were found to be positive by end-point IS900-specific nested PCR. Agreement was found between ELISA and end-point IS900-specific nested PCR in one of the animals. Conclusion: the present study gives information about the agreement between direct and indirect MAP-detection techniques, using different matrixes from animals under the same husbandry conditions.

Resumen Antecedentes: la paratuberculosis es una enfermedad infecciosa de desarrollo lento, caracterizada por una enterocolitis granulomatosa crónica. Esta enfermedad tiene un periodo de incubación que varía entre los 6 meses hasta por más de 15 años, y es causada por Mycobacterium avium subsp. paratuberculosis (MAP). Su detección por técnicas diagnósticas directas e indirectas ha sido de interés especial. Objetivo: reportar el diagnóstico y detección de MAP utilizando varias técnicas diagnósticas en un hato de la región norte de Antioquia, Colombia. Métodos: se analizaron las muestras de suero del hato de estudio utilizando un kit comercial de ELISA (enzyme-linked immunosorbent assay). Las muestras de materia fecal fueron cultivadas por duplicado en Herrold´s egg yolk medium (HEYM), y analizadas mediante un protocolo de PCR anidado específico de IS900 y un kit comercial de PCR en tiempo real para F57. Resultados: ocho de las 27 muestras de suero resultaron positivas por ELISA. Ninguna de las muestras de materia fecal resultó positiva al cultivo en HEYM por duplicado ni por PCR en tiempo real para F57. Siete de las 27 muestras de materia fecal resultaron positivas a PCR anidado específico de IS900. Se encontró concordancia entre el resultado de ELISA y de PCR anidado específico de IS900 en uno de los animales. Conclusión: el presente estudio brinda información acerca de la concordancia entre técnicas directas e indirectas de detección de MAP, utilizando diferentes matrices a partir de animales bajo las mismas condiciones de manejo.

Resumo Antecedentes: a paratuberculosis é uma doença infecciosa de evolução lenta, caracterizada por uma enterocolite granulomatosa crônica. Esta doença tem um período de incubação que varia de 6 meses a 15 anos e é causada pelo Mycobacterium avium subsp. paratuberculosis (MAP). Sua detecção por técnicas de diagnóstico diretos e indiretos tem sido de especial interesse. Objetivo: reportar o diagnóstico e a detecção de MAP utilizando várias técnicas de diagnóstico em um rebanho na região norte de Antióquia, Colômbia. Métodos: foram analisadas amostras de soro do rebanho utilizando um kit comercial de ELISA (enzyme- linked immunosorbent assay). As amostras de fezes foram cultivadas em duplicado em Herrold´s egg yolk medium (HEYM) e analisadas utilizando um protocolo de PCR aninhada específico de IS900 e um kit de PCR em tempo real comercial para F57. Resultados: oito das 27 amostras de soro foram positivas para ELISA. Nenhuma das amostras testadas na cultura de fezes HEYM duplicado foram positivas ou na PCR em tempo real para F57. Sete das 27 amostras de fezes foram positivas na PCR aninhada específica para IS900. Foi encontrada concordância entre o resultado de ELISA e PCR aninhada específica para IS900 em um animal. Conclusão: este estudo fornece informações sobre a correlação entre técnicas de detecção direta e indireta do MAP, utilizando diferentes matrizes de animais sob as mesmas condições de condução.

Factors associated with Mycobacterium avium subsp. paratuberculosis in dairy cows from Northern Antioquia, Colombia / Factores asociados con el estatus serológico de Mycobacterium avium subsp. paratuberculosis en vacas lecheras del norte de Antioquia, Colombia / Fatores associados ao estado sorológico de Mycobacterium avium subsp. paratuberculosis em soro de gado leiteira no norte de Antioquia, Colombia

Summary Background: Johne´s disease (JD) is a severe enteritis that affects ruminants and has been diagnosed in cattle and sheep in Colombia. However, epidemiological information on the disease in this country is scarce. Objective: to identify factors associated with the JD serum enzyme-linked immunosorbent assay (ELISA) status of dairy cows. Methods: a cross-sectional study was carried out in 307 asymptomatic adult Holstein dairy cows from 14 herds in nine districts of Belmira and San Pedro de los Milagros municipalities during November, 2007. From 19 to 25 cows, ≥ 2 years of age were randomly selected and blood sampled from every herd. A commercial ELISA kit was used to analyze sera. Information regarding cow related factors (age, farm-born, parity, and daily milk yield) and herd management practices (i.e. herd size, herd average milk production, current presence of symptomatic animals, cattle purchase, own animals grazing in foreign pastures, feeding of calves before weaning, manure spread on pastures, and sighting of birds in feed storing areas) was collected using questionnaires. Descriptive statistics were computed for all variables and a multivariable logistic regression model was constructed (p<0.05). Results: ten percent (31/307; 95% CI: 7.0-14.0%) of the animals were positive by ELISA. In 70% (10/14) of the herds, ELISA detected at least one positive animal. Cow and herd factors "parity" and "feeding of calves before weaning" showed weak and strong associations with ELISA positive results, respectively. The odds ratio (OR) for JD seropositivity increased 20% (OR = 1.20; 95% CI: 0.98-1.47; p = 0.067) in cows with > 1 parity. The OR was 0.74 times lower (OR = 0.26; 95% CI: 0.096-0.70; p<0.01) in herds feeding calves with pooled colostrum from several cows, compared to herds feeding calves with colostrum from their own dams. Conclusion: JD seroprevalence was 10 and 70% at animal and herd-level, respectively. Cow and herd factors "parity" and "feeding of calves before weaning" showed weak and strong association with positive results, respectively.

Resumen Antecedentes: la enfermedad de Johne (EJ) es una enteritis severa que afecta rumiantes, y ha sido diagnosticada en bovinos y ovejas en Colombia. Sin embargo, la información epidemiológica de la enfermedad en el país es escaza. Objetivo: identificar los factores asociados con el estatus serológico de la EJ mediante ensayo de inmunoabsorción ligado a enzimas (ELISA) en vacas lecheras. Métodos: se realizó un estudio transversal en 307 vacas lecheras adultas asintomáticas de 14 hatos en 9 veredas en los municipios de Belmira y San Pedro de los Milagros, en noviembre de 2007. De cada hato se seleccionaron aleatoriamente 19-25 vacas, mayores de 2 años de edad y se tomaron muestras de sangre. Se utilizó un kit comercial de ELISA para analizar el suero. Se recolectó información mediante cuestionarios sobre características individuales de las vacas y prácticas de manejo de los hatos. Se realizó un estudio estadístico descriptivo para todas las variables de interés y se construyó un modelo de regresión logística multivariada, considerando un nivel de significancia de p<0,05. Resultados: el 10% de los animales fue positivo a ELISA (31/307; 95% CI: 7,0-14%). En el 70% (10/14) de los hatos al menos un animal fue detectado como positivo por ELISA. Los factores a nivel individual y a nivel hato "número de partos" y "tipo de alimentación de terneras antes del destete" mostraron una asociación débil y fuerte con los resultados positivos de ELISA, respectivamente. El odds ratio (OR) para la seropositividad a EJ aumentó en un 20% (OR = 1,20; 95% CI: 0,98-1,47; p = 0,067) en vacas con más de un parto, mientras que fue 0,74 veces menor (OR = 0,26; 95% CI: 0,096-0,70; p<0,01) en hatos que alimentan sus terneras con mezcla de calostro proveniente de varias vacas, en comparación con aquellos hatos que solo utilizan el calostro de la propia madre. Conclusión: la seroprevalencia de EJ fue del 10 y del 70% a nivel individual y a nivel hato, respectivamente. Los factores a nivel individual y a nivel hato "número de partos" y "tipo de alimentación de terneras antes del destete" mostraron una asociación débil y fuerte con los resultados positivos de ELISA, respectivamente.

Resumo Antecedentes: a doença de Johne (DJ) é uma enterite grave que afeta o ruminantes, e tem sido diagnosticada em bovinos e ovinos na Colômbia. No entanto, a informação epidemiológica da doença no país é escassa. Objetivo: identificar os fatores associados ao estado sorológico de DJ em vacas leiteiras por ensaio imunossorvente ligado a enzima (ELISA). Métodos: estudo transversal foi realizado em 307 vacas leiteiras adultos assintomáticos em 9 de 14 aldeias nos municípios de Belmira y San Pedro de los Milagros, em novembro de 2007. Em cada rebanho foram selecionados aleatoriamente 19-25 vacas com mais de 2 anos de idade e amostras de sangue foram tomadas. Um kit de ELISA comercial é utilizado para testar soros. As informações foram coletadas por meio de questionários sobre as características individuais de animais e práticas de gestão do rebanho. A análise estatística descritiva foi realizada para todas as variáveis de interesse e modelo de regressão logística multivariada foi construído, considerando um nível de significância de p<0,05. Resultados: dez por cento dos animais foram positivos no ELISA (31/307; 95% CI: 7,0-14%). No 70% (10/14) os rebanhos pelo menos, um animal foi detectado como positivo por ELISA. Os fatores a nível individual como a nível do rebanho "paridade" e "tipo de alimentação bezerros antes do desmame" mostraram uma associação fraca e forte com os resultados positivos de ELISA, respectivamente. O odds ratio (OR) para DJ soropositividade aumentou em um 20% (OR = 1,20; 95% CI: 0,98-1,47; p = 0,067) em vacas com mais de um nascimento, enquanto era 0,74 vezes menor (OR = 0,26; 95% CI: 0,096-0,70; p<0,01) em rebanhos que alimentam seus filhotes com a mistura de colostro de várias vacas, em comparação com apenas dados de uso que o colostro da mãe. Conclusão: DJ soroprevalência foi de 10 e 70% a nível individual como a nível do rebanho, respectivamente. Os fatores a nível individual e do rebanho "paridade" e "tipo de alimentação bezerros antes do desmame" mostrou fraca associação e forte com os resultados positivos de ELISA, respectivamente.