Actividad antimicótica de = la Monensina sobre Malassezia in vitro aislada de caninos con otitis externa

<= o:p>

Antifungal activity of monensin on Ma= lassezia in vitro isolated from canines with otitis external

= ¹

Andrea Fernanda Cevallos Minchala

https://orcid.org/0000-0002-9425-463= 9

Maestría en Medici= na Veterinaria, Universidad Católica de Cuenca, Ecuador.
afcevallosm70@est.ucacue.edu.ec

= ²

Manuel Esteban Maldonado Cornejo

https://orcid.org/0000-0002-= 1507-2280

Maestría en Medici= na Veterinaria, Universidad Católica de Cuenca, Ecuador.
mmaldonadoc@ucacue.edu.ec

= ³

Pablo Giovanny Rubio Arias

https://orcid.org/0000-0002-9185-482= 3

Maestría en Medici= na Veterinaria, Universidad Católica de Cuenca, Ecuador.
prubioa@ucacue.edu.ec

=

Artículo de Investigación Científica y Tecnológica

Enviado: 11/05/2023

Revisado: 26/06/2023

Aceptado: 06/07/2023

Publicado:28/07/2023

DOI: https://doi.org/10.33262/ap.v5i3.380

<= span lang=3DEN-US style=3D'font-size:12.0pt;line-height:115%;font-family:"Time= s New Roman",serif; mso-fareast-font-family:Calibri;color:blue;mso-ansi-language:EN-US'>=

Cítese:

Cevallos Minchala, A. F., Maldonado Cornejo, M. E., & Rubio Arias, P. G. (2023= ). Actividad antimicótica de la Monensina sobre Malassezia in vitro aislada = de caninos con otitis externa. AlfaPublicaciones, 5(3), 103–115. https://d= oi.org/10.33262/ap.v5i3.380

ALFA PUBLICACIONES, es una Revista Multidisciplinar, Trimestral, que se public= ará en soporte electrónico tiene como misión contribuir a la &nbs= p; formación de profesionales competentes con visión humanística y crítica q= ue sean capaces de exponer sus resultados investigativos y científicos en la misma medida que se promueva mediante su intervención cambios positivos e= n la sociedad. https://alfapubli= caciones.com

La revista es editada por la Editorial Ciencia Digital (Editorial de prestigio registrada en la Cámara Ecuatoria= na de Libro con No de Afiliación 663) www.celibro.org.ec

Esta revista está protegida bajo una licencia Creative Common= s Attribution Non Commerci= al No Derivatives 4.0 International. Copia de la licencia: http://creativecommons.org/licenses/by-nc-nd/4.0/

Palabras claves:

Mico= sis; Ionoforos; Infecciones; Control; Caninos.

Resumen

Objetivo. Evaluar la actividad antimicótica de la monensina sobre = Malassezia in vitro aislada de caninos con otitis externa (OE), y determinar su susceptibilidad a monensina que serán cultivados in vitr= o. Metodología. Esta investigac= ión fue realizada sobre 30 paciente caninos con otitis externa clínica que fu= eron atendidos en Fundación Arca y en clínicas veterinarias particulares. Los mismos que presentaron signos clínicos como: dolor, eritema del pabellón auricular, prurito, sacudidas de la cabeza e inclinación cefálica. Para la identificación del agente etiológico (Malassezi pachidermatis), mediante citología con tinción = diff quick para ser eva= luadas bajo microscopio con su forma de zapato dando positivo a presencia de Malassezia para ser cultivadas in vitro y de esta manera ser evaluada conjuntamente la actividad antimicrobiana de monensina = que se utilizó el tratamiento= de estudio en cajas Petri con Monensina con cuat= ro diferentes concentraciones 1%, 5%, 10% y 20%. Resultado. Obteniend= o como resultados que los 30 pacientes caninos dieron positivos a malassezia, pero su actividad antimicrobia= na de monensina no obtuvo ningún resultado favorable fren= te a otros agentes antimicóticos de uso comercial como el ketoconazol, fluconazol y griseofulvi= na, siendo el ketoconazol el medicamento con mayor actividad antimicótica frente a malassezia. C= onclusión. Que la = Malassezia desempeńa un gran rol en esta enfermedad, por lo que varios tratamien= tos se han investigado e implementado, sin embargo, con la evolución de la especie, nuevas resistencias se han originado dando como resultado infecciones crónicas persistente. La Monensina no actúa contra este microorganismo con las concentraciones manejadas en el presente estudio, por lo tanto, se necesita nuevos estudios que analicen componentes farmacológicos, entre los que se han planteado diversos aceit= es naturales con el fin de evitar efecto tóxicos y adversos para los canes. = Área de= estudio general: Medicina Veterinaria. Área de estudio Especifica: Dermatología.

Keywords:= Mycosis; Ionophores; Infections; Contr= ol; Canines.

Abstract

Objective. Evaluate the antifungal activity of monensin on Malassezia in Vitro, isolated from canines= with external otitis (EO), and delimitate their susceptibility to monensin, wich will be = cultured in vitro. Methodology. This research was conducted on 30 canine patients, suffering from clinical external otitis,= and treated at ¨Fundación Ar= ca¨ and private veterinary clinics. = These canine patients exhibited clinical signs such as pain, erythema of pinna, pruritus, head shacking and head tilting. Malassezi<= /span> pachidermatis was identified as the etiologic= al agent through cytology with diff-quick staining, and samples were evaluat= ed under a microscope to confirm the presence of Malass= ezia for in vitro culturing. Results. The results showed that all 30 canine patients tested positive for = Malassezia, but the antimicrobial activity of monensin did not yield any favorable results compar= ed to other commercially used antifungal agents like ketoconazole, fluconazole,= and griseofulvin. Ketoconazole exhibited the high= est antifungal activity against Malassezia. Conclusion. Malassezia develops a significant role in this disease, and various treatments have been investigated and implemented form this fungus. However, with the evolution of the species,= new resistances have emerged, resulting in persistent chronic infections. Monensin does not act against this microorganism at= the concentrations used in this study; therefore, new studies are needed to analyze pharmacological components, including various natural oils, with = the aim of avoiding toxic and adverse effects for the canines.

Introducción

La otitis externa es una enfermedad inflamatoria del conducto auditivo externo= y dependiendo su duración puede ser aguda o crónica, esta última es persisten= te y dura 3 meses o más (Bajwa, 2019). Esta inflamación está acompańada de constante prurito y signos de malestar = en los animales. Además, puede desencadenar hiperplasia glandular, dilatación glandular, hiperplasia epitelial e hiperqueratosis (Bajwa, 2019; Bond et al., 2020). <= /span>

Entre los microorganismos que más comúnmente se encuentran en los oídos de los ca= nes infectados se encuentra Staphylococcus spp., pseudomonas, enterococ= os, estreptococos, y Corynebacterium spp (Bajwa, 2019). Por otro lado, la levadura Malassezia también repre= senta otro agente patógeno en la otitis externa canina. El género Malassezia comprende un grupo de levaduras lipofílicas que ha evolucionado como un comensal oportunista que puede volverse patógeno y habita tanto en humanos = como en animales de sangre caliente (Ashbee, 2007; Bond et al., 2020; Gaitanis et al., 2012; Guillot & Bond, 2020). Cabe mencion= ar que el género comprende actualmente 18 especies (Takashima & Sugita, 2022), y recientemente se le ha asigna= do su propia clase, Malasseziomycetes. Su relevancia como patógeno en la veterinaria canina se dio desde el ańo 1955 = con precisamente el contexto de otitis externa canina por Gustafson. En 1961 Fraiser también los identificó en oídos de perros saludables y enfermos. Se ha propuesto que Malassezia <= !--[if supportFields]>ADDIN paperpile_citation <clusterId>U318I386X976B769</clusterId><m= etadata><citation><id>2c75cbf6-264c-432f-bc6d-bb239d282fc8&l= t;/id></citation></metadata><data>eJyNWO1u2zgWfRXCPzoz= gO3ow7akLopdx3bbtPnwJmm6i2JQUBJls6FFDSnZ9RZ9gH2U+b9v0Bfbcyk7cd1g0GAGlSjyfpx= 77iHpD186H2Xeed4JsmiYpcWoF4wGWW8QBkUvzUZ5L02DMMmDOCiyuNPt5E310S65jxXxwAvzJM= oizxNZHntD3x+NsiIajJIo9eJ0OMoC3ys4VklrG4ElPp4bozrPP3SWdV09PznJP/dzLfvaLE58r= +/j72Sdi74fxF6CyTTLYtpms+mXWSr7pVr1S7nsL/T6pGrSlchPQj8ZRoE3OJiuSyVLoWRquNn2= N1KJbT/TqxO4esLN70hLSwrviQjgREm7FMDoS2ely3pJYOHDVnBDj17gdb66FMuPXNVkJhwNe6E= /ooh4UUgleS11iS9TUXFTr0RZM12wCYKUGVfsJpOizIRlvMzZjTBriZcuu9ZbfLwTtTCyRCJsop= USC9Flr/nmHjHxnJ3zEu+X2tRLdrFdrSx9rAspVI4nYepCm9wupcGs8XnCottxl71722c/E8o4X= 3O85OxC5DKT5OkmW2qtaMlBXI+f35VyLYyV9ZamzEVZ2q2CEcm7LEw8D24VTG7ZTW2EqLtsvpSK= 50JVS5oyR3B+4nsDWLoZ99m3/2ZaCVY6/Die1t/+rL/9SW6REct/GSvkBzOnUs+//Q8T9gOzMYv= CGP62JV9tM9ibzyYYLtdwccGl1aV9mHymRC8XvZeGsu2y9t9+C8ofjWB3h14RYcROdaPEmpuc5c= DqpURdLMsbNuHI1k2acyNRjGjoeeGjyXEpV4TyvSwdwjNu2E0lMslBshrzL4RKdWNKcYjv4ww2Q= cHIfOSxU8Wz+xRzQRSOar9SomTvOfAH3bvsTma1Ng54f+ih+o2tDYxwqr1ZZdyI3lrUbH77b3Z+= O4VJZ1w4j63FG/Jg2QSluu+yfzZClBblw5eBH0RHJg/pdBD7nNdLnUqt9AJBXcqlLg9ZsqM0LUI= NjbBIPhMPNATY8WjE3vKVkKUGjs0nafkGSb3lJV+4p2AY9Lw4RopveMXLv4hlZ/WYpmPwj8ucOu= Th6UY36KiHBLsMdRweZky9ndJLRi1/Op68fXV99e5y+pzdLgVbiLKx4Jni1or/SM6kZRCgCpxAO= 8EnZwujm4oelaw06K9kxqApYAGrl7xmS1SSibVWa6zgllniDGwgL8tV26C6qkDgpgQzsDpraqiB= huOKQEcMtnUFokrR5rriq9V+dSohDX12dfpmNrk9u5vdtLHr9JPIagBE8+slIs911jg48VxrVhm= 9Bkj4hoZ8RJdiQyjw40LDV8MkCIWi1kiBaiLRJRsJYLPGGGewRAeuXHsz/EcWRbbjikuiWm7tnj= y55ItSW2or9ABvK0y+KiPWeHZGihaoHEBzSzpmrUbzUATO80FNdmjTbL1oIcl4DUQuZrevr6Y3b= Hw5ZRfj29n12fi8xYaDFdqgGBDptiycvWoABu034Do2Hfbrq/lvzhiikmKDaZSWkkCC1w1ki0Mi= FE+hVuCDNgTpFXo1BV6B58d95+nVnNICaOQGKlNjDR4PzLTmnacVJBTvLTtyB6clPK1QKCVFoCu= Z2dbye20UtH2Hyw60gy4hdeC1w5z9+n58N/3tcXuYwDBoBYo9Zk1eZV0LsSdGzhb4SILXMsG12m= JZs4KSJUJhIpp7F88DuzacvghLKoSka1ZvdMug/QbAVtg0ZLlwvD6gXn4QMqUlHNOccwR4ZGLbE= oxttMEmqqu/Mdn6dig+FoeibRGt99wkNNhGpFCO9jtnFWLQrqWt/MzcAQF5Tdp1YDvoz3BgMnv+= 1W1tXWGMdqiq7Y4utiJ4iZGZNuhsx1pEr92qQlL4e7Tg4+pycv7u5uzqsmXq5Pzs8mwyPmdnF/O= r69vx5WTmSIsAlsguhXojXiNW3Ny7BMVnyKXdEeC+1BsQjKS4fKJJKEAEIQ0ztCO77cttZbtG67= b9nTWKG+RjJVXftbN795PE+4U669FwWzNZyx870MW7l77C6BV1QOuHQtWphX7shSYD5MZxbEuhS= ywXnyGIrdIDOySG5MEayh6yTpx9kBLyDaFET6lDQlWk6/LhEEDUB4XRVCgkaqqU67ZF6eScotgZ= JBnmFejNs6U4mJ1DoJSuRI4jAMNZoN3xXFfCNBmwW4S8wnLSTV4RfxEdVaWkJFIgWRStLncZmOJ= OBEB2s0RZ0TXAp0JCyLLgtPG3ztGN0EUcB/MdVpBdEgTavoC9okP56Q/62lb7QWIR6U/W7a90wr= bbyVP6Q530tADRAZwvhKWDdtyL6ERdaIUQMPIB5/ZPdFbiqmgULhWduycVAWv2N5bIC/2Qh4Ufp= EHGw0EcjJJwFPI8Gsaen3tpyLM4Ee2NhQ7sXjJMeoNBEtJ277Qffr90kGUN+CmsawpJGkungGtt= LV61ydyVysNfD/+HvQB/vWQYJy5+iqymC0XnVAM5sgB06907LhPfOZheHXiYouuwxVwd2blAXVE= PN3Vnqh06NvbmwNYbnuFwa49MoV5K6Zq9eTS1Gzq2NTs0NsOxomwQ2psnAMD5bOANcCWKh8cAgC= ToK9R/duDwYfQHLL6DAnvNkblrSAP4NH001Y4c23l7d2DoLW4pAsp8dwwqJ1tu6h5T/oSt7wjQy= Cfr70ejQW8UJP86coFDrD6sP70f25+fHjiYE8PZ6ZGZ16gPc/N2dmig8xX9kWGjyk+357K8x/Du= zm1xh4NM4w6tF0q4i/Fu6O9L9UKUz5R58axsVi9C7xkZsC9+7rKOc5crgjuXtFfneluRxMznH8f= Xt2eT85mLMRVq179k/l5sd9Snu3QP5+mHvm57uv+gCCRbEERBF+8oiiez6WAyjodT359NxhPv5e= RlMPJjfzoOByNMxQ6CvRJzQ9/5LRcNxATvosR7RgJH+PmjKA48bxTHfW8wHCVDhL5yFTz4aQEn/= fZXAK/b+UhhX+RDikKkBe7hXhYOszgUaTIoiiwKRJQM0yIXHomPu9K0v7IMI+6JPMh7eYJUvTSN= eknsFT2Rj5IgG0ZJMSL1wSY/+wyPU15z+t1B2pe4G6ElOs8LsEKQPp2V5+3vG53ntWnc0LnOCLP= dFAfUbIXjDBzzVGb+wA//saABqvkeykvuIBqnkk2Wna9ff/8/kY0V1g=3D=3D</data> \* MERGEFORMAT(Bond et al., 2020), actúa en un modo de transición entre un ser comensal y un patógeno que se ve influenciado por la piel del huésped y el ambiente externo. Se ha propuesto= que la colonización en la piel canina ocurre desde su nacimiento debido al tras= paso desde sus madres. Es así que se vio en un estudio que en el 40% de muestras obtenida de labios, uńas y oídos en 22 cachorros presentaba Malassezia (Bond et al., 2020). Malassezia pachydermatis es la especie más común en perros y diferentes estudios han mostrado su conexi= ón con la otitis externa canina. M. pachydermatis ha sido reportada como lipofílica pero no dependiente de los mismos debido a que contrario a otras especies de Malassezia puede utilizar fracciones de lípido que se encuentran en la peptona del agar dextrosa Sabouraud (Bond et al., 2020).

Entre los principales tratamientos que se utilizan se encuentran los azoles, entre ellos los imidazol= es como clotrimazole, climbaz= ole y miconazole son usado de manera tópica usualme= nte utilizados en cremas y shampoos (Bond et al., 2020).

La Monensina es un ionóforo bioactivo natural liposoluble producido por Streptomyc= es spp (Aowicki & Huczyński, 2013). Su actividad antimicrobiana está mediada por su cap= acidad de intercambiar iones Na+ y K+ a través de la membrana celular, alterando así los gradientes iónicos y la fisiología celu= lar. Está aprobado por la Food and Drug Administration como antibiótico veterinario para tratar la coccidiosis (Aowicki & Huczyński, 2013; Potter et al., 1= 984).

La Monensina es un antibiótico que se utiliza como = coccidiostático y agente promotor del crecimiento en = la práctica veterinaria. Debido a sus propiedades antibacterianas y coccidiostáticas, se la usa en la avicultura industri= al, debido a que tiene un efecto positivo sobre el crecimiento de las aves de corral al reducir la proliferación de parásitos, eliminando así la atenuaci= ón sobre los animales. Para la prevención de la coccidios= is en aves de corral se utiliza la formulación Mondolar, que contiene 10% o 20% de Monensina sódica. Est= udios posteriores han demostrado que la Monensina tam= bién puede mejorar el metabolismo de los alimentos en los rumiantes, lo que garantiza su mejor aprovechamiento y, en consecuencia, conduce a un crecimi= ento más rápido del ganado. La estimulación del crecimiento se asocia en este ca= so a cambios favorables de la microflora intestinal bovina y al aumento de las cantidades de proteína digerida asimilable. La rumenes= ina, que contiene un 6,6% de Monensina, se utiliza c= omo promotor no hormonal del crecimiento de los animales, es por ello que se utiliza principalmente en la producción de carne y leche debido a que reduce significativamente los costos de alimentación (Goodrich et al., 1984; Potter et al., 1984; Rajendra= n et al., 2018).

Además= de las aplicaciones veterinarias, la Monensina pre= senta un amplio espectro de actividad contra patógenos oportunistas del ser human= o, como bacterias, virus, hongos y parásitos, tanto en cepas sensibles a los fármacos como en cepas resistentes. Este ionóforo puede matar selectivamente patógenos con un efecto tóxico insignificante en células de mamíferos (Aowicki & Huczyński, 2013).

Difere= ntes estudios han investigado el éxito de la Monensina como tratamiento en contra de microorganismos. Se ha encontrado que tienen efecto en bacterias gram-positivo. Sin embargo,= no han demostrado tener efecto contra bacterias gram-negativo y hongos (Chan et al., 2018). Adicionalmente, se ha visto que los protozoar= ios Eimeria acervulina y Eimeria tenella causantes= de la coccidiosis llegan a desarrollar mecanismos de resist= encia contra la Monensina (Bafun= do & Jeffers, 1990).

En cua= nto al uso de la Monensina en contra de enfermedades zoonóticas, ha sido utilizada en el tratamiento de toxoplasmosis bovina, la tripanosomiasis en el ganado y la coccidiosis e= n las aves de corral, lo que demuestra un gran potencial en el tratamiento de otr= as enfermedades animales (Buxton et al., 1988; Chapman et al., 2010; Steverding et al., 2016).

La morbilidad, la mortalidad y los costos de salud asociados con las infeccion= es por hongos son altos y la emergencia de nuevos patógenos está aumentando el= número de personas susceptibles a estas enfermedades. Entre ellos, se encuentra la= Malassezia que es parte de la microflora normal, pero= bajo ciertas condiciones representa una amenaza patogénica. La Malassezia está principalmente asociada con enfermedades de la piel como: pitiriasis <= span class=3DSpellE>versicolor, foliculitis, dermatitis seborreica, caspa, dermatitis atópica, psoriasis, onicomicosis y dermatosis acantolítica transitoria. Adicionalmente, para las personas inmunocomprimidas e infantes prematuros pueden causar graves infecciones sistémicas cuando ingresan a unidades de cuidado intensi= vo (Dixon et al., 1996; Pedrosa et al., 2018).

Metodología

Se realizó un análisis prospectivo= en la Fundación Arca y en clínicas veterinarias particulares en Cuenca en pacientes caninos indistintamente de edad, se= xo, raza, lugar de vivienda, alimentación, tipo de orejas en los que se ha observado infección del oído y que a pesar de llevar un tiempo prolongado de tratamiento (especificar que tratamiento) no han exhibido muestras de mejor= ía. Se seleccionaron 30 pacientes caninos que cumplieron los siguientes criteri= os de inclusión: presentación de otitis externa, positividad bajo prueba de tinción para infección por Malassezia y no haber recibido tratamiento 5 días previos a la toma de muestra para cultivo. De e= stos canes, se procedió a tomar muestras de sus oídos por raspado con la implementación de un hisopo esterilizado que luego de la toma fue almacenad= o en medio de crecimiento caldo tripticasa soya. Siguiente, se procedió a sembrar cada muestra en dos cajas Petri de agar saburoe : una de experimentación y la otra de control. Estos platos se incubaron en u= na temperatura de 37 °C por dos semanas. Pasado este tiempo se procedió a apli= car a cada grupo el agente antimicótico correspondiente. Es así como, en la pri= mera caja Petri, de investigación, se procedió a colocar en cada cuarto de caja Petri, papel filtro (cuanto media) sumergido en Monens= ina con cuatro diferentes concentraciones: 1%, 5%, 10% y 20% respectivamente, c= on el afán de observar si la Monensina daba halos = de crecimiento contra Malassezia. Estas concentraciones se obtuvieron al mezclar 100 mililitros de agua destilada c= on 1gramo, 5 gramos, 10 gramos y 20 gramos de Monensina, respectivamente. Se utilizó agua destilada cómo solvente debido a que la mi= sma no causa efectos adversos en los canes, y se utilizó la concentración máxim= a de 20% debido a que se observó sobrenadante al mezclar el agua destilada y la = Monensina. Por otro lado, la segunda caja Petri sirve= , de control, sirve para comprobar que la siembra microbiana haya sido correcta y adecuada pues en esta se colocaron 3 antimicóticos: Fl= uconazole, Ketoconazole y Griseofulvi= na conocidos en los que de hecho se esperaba encontrar el halo de crecimiento = de la Malassezia. Finalmente, después de 24 horas = se procedió a observar la formación o ausencia de halos.

Se observaron los datos, se midier= on los halos de inhibición, compararon con el testigo y sus intervalos de acci= ón (IC 95%), para de esta forma poder describir la actividad antimicótica de l= a Monensina sobre Malassezia in vitro aislada de caninos con otitis externa.

Resultados y discusión

La Malassezi= a es sensible a Fluco= nazole y moderadamente sensible al Ketoconazole. Por e= l contrario la actividad otros agentes como la Monensina sobre la misma muestra es nula según se evi= dencia en la Figura 1. Actividad Antimicótica de los Agentes Micóticos.

Figura 1

Actividad Antimicótica de los Agentes Micóticos

Al com= parar los halos de inhibición de las concentraciones de 5%, 10%, 20%, Fluconazole y Ketoconazole de acuerdo con la tabla 1. se puede establecer que los estudios en azoles específicos en Fluconazol= e y Ketoconazole, ya han comprobado el efecto de = estos en Malassezia (Miranda et al. 2007), donde se ha observado un efecto cruzado entre agentes micoticos que disminuye el efecto de estos (Jesus et al. = 2011), comprobándose una mayor actividad de Fluconazole, frente a otros agentes para su control (Rojas et al. 2022; Chebil et al. 2022). Por otro lado se busca potencializ= ar su efecto junto a otros productos como Cinnamaldehido, Thymol y Carvacrol, que h= an demostrado actividad micotica media y que en co= njunto a los Azoles, llegan a potencializarse significativamente (Schlemmer et al. 2019).

Tabla 1

Intervalo Inhibición frente a Fluconazole, Ketaconazole y Mobensina<= o:p>

Fluconazole

Ketoconazole

Monensina

Rango (GM) (mg/ml)

Rango (GM) (mg/ml)

5%

10%

20%

Estudio In Vitro

0,5-4,0

0,1-2,0

0

0

0

Mirande et al. (2007)

0,125-16,0

0,03-0,25

Jesus et al. (2011)

0,01-4,0

0,25-2

Schlemmer et al. (2019)

1,0-6,4

Chebil et al. (2022)

0,125-8,0

0,06-8,0

= ;

El uso continuo de antimicrobianos trae consigo algunas complicaciones, entre ella= s la resistencia microbiana. Es así que el uso excesivo de antibióticos en anima= les ha provocado el desarrollo de resistencias bacterianas y la generalización = de bacterias resistentes en el mundo (Christaki et al., 2020; Huemer et al., 2020; Morrison & Zembower, 2020). Las bacterias resistentes a los antibióticos (ARB) y los genes resistentes a los antibióticos (ARG) en animales destinados a la alimentaci= ón se consideran actualmente contaminantes emergentes, que suponen una grave amenaza para la salud pública en todo el mundo (Xiong et al., 2018). Es por ello por lo que se debe buscar alternativas.=

En la práctica clínica, la mayoría de los casos de OE aguda se tratan con product= os tópicos polivalentes para el oído que incluyen un glucocorticoide (utilizado para controlar la inflamación aguda leve) y agentes antimicrobianos utiliza= dos para tratar infecciones concurrentes. Varios detergentes, como el ácido etilendiaminotetraacético (TrizE= DTA) y la clorhexidina, se utilizan en gran medida para limpiar el conducto audi= tivo externo, eliminar los desechos y la producción excesiva de cerumen y para interrumpir la biopelícula. Los antibióticos, especialmente las fluoroquinolonas, los aminoglucósidos y las polimixinas, deben utilizarse s= olo después de la identificación de los agentes etiológicos y solo después de comprobar la integridad de la membrana timpánica. Los tratamientos contra <= span class=3DSpellE>Malassezia incluyen antifúngicos= como los derivados azólicos (tiabendazol, clotrimazol, miconazol e itraconazol), nistatina y terbin= afina. Las principales causas de fracaso terapéutico son el manejo incorrecto de la medicación del paciente, y la falta de identificación de las afecciones primarias y la progresión a otitis media obligan a cambiar a terapia sistém= ica. La resistencia a los antimicrobianos ha surgido a nivel mundial como una gr= ave amenaza para la salud humana y animal. Publicaciones recientes apoyan= las observaciones anteriores de que la mayoría de las levaduras Malassezia de tipo salvaje siguen siendo susceptibles a los fármacos azólicos de uso común, aunque la eficacia del fluconazol es más variable (Bajwa, 2019; Gotthelf, 2004; Rosychuk, 1994)<= !--[if supportFields]><= ![endif]-->.

Es por= ello por lo que constantemente se busca implementar nuevos tratamientos más eficaces= y específicos. Es así que se tomó como alternativa el uso de monencina para el tratamiento de otitis externa en perros debido a que diferentes estudios han seńalado su seguridad en uso veterinario (Vercelli et al., 2021). Se ha comprobado que es seguro tanto para aves como pollos, gallinas, pavos y ganado vacuno. Sin embargo, como se pudo observar= en el estudio presente la Monensina no actúa sobre= Malassezia en concentraciones de 1%, 5% ni 20 %.

Una re= ciente revisión basada en la evidencia sobre el tratamiento de la dermatitis canina por Malassezia informó de una "fuerte" evidencia para el uso de un champú con Miconazol al 2% y Clorhexidina al 2%, utilizado dos veces por semana. Se dispuso de evidencia "moderada" para un champú de Clorhexidina al 3%. Para los casos caninos en los que la terapia tópica es ineficaz o poco práctica, había evidencia "moderada" para el uso de Ketoconazol a 5-10 mg/kg por vía oral u= na o dos veces al día; e Itraconazol a 5 mg/kg por vía oral una vez al día o dos días consecutivos por semana. Recientemente, se evaluaron los efectos clíni= cos y citológicos de una aplicación una vez al día de una formulación en spray = sin enjuague que contiene zinc, etil lauroil arginato, laureth-9, urea, pantenol, glicerina y butilenglicol en un estudio aleator= izado, ciego y controlado de 18 perros con pododermatitis crónica asociada a levaduras de Malassezia. En comparación con el tratamiento con placebo del pie contralateral, la reducc= ión de los recuentos de levaduras del pie tratado activamente se asoció con una reducción de las puntuaciones clínicas (Bond et al., 2020; Guillot & Bond, 2020)<= !--[if supportFields]><= ![endif]-->.

Entre = las diferentes posibilidades de tratamiento también sobresale el uso del efecto placebo, que acorde a recientes investigaciones está conectado a mecanismos psicológicos complejos que se originan en el cerebro como la expectativa y = el condicionamiento. Así, se ha visto este efecto en el tratamiento de Parkins= on en el que a través del efecto placebo se produce dopamina que reduce el deterioro motor (Enck et al., 2008; Pollo & Benedetti, 2009).

La resistencia antifúngica constituye un problema creciente debido al escaso número de posibles tratamientos y el desarrollo de mecanismos de resistenci= a de estos. Es así como se ha visto que la resistencia a fl= uconazole de non-Candida albicans se ha incrementado (Wiederhold, 2017).<= /span>

Conclusiones

·         La otitis externa representa un gran problem= a en la veterinaria canina desde hace décadas, es por ello que diversos estudios= se han centrado en esta temática y así se ha visto que la Malassezia desempeńa un gran rol en esta enfermedad, por lo que varios tratamiento= s se han investigado e implementado, sin embargo con la evolución de la especie, nuevas resistencias se han origado dando como resultado infecciones crónicas persistente, es así que en este trabajo se q= uiso revisar el desempeńo de Monensina como un posib= le tratamiento. Sin embargo, se evidenció, que la Monensi= na no actúa contra este microorganismo con las concentraciones manejadas en el presente estudio. Esto demostró que se necesitan nuevos estudios que analic= en nuevos componentes farmacológicos, entre los que se han planteado diversos aceites naturales con el fin de evitar efecto tóxicos y adversos para los canes.

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= ¹	Andrea Fernanda Cevallos Minchala	https://orcid.org/0000-0002-9425-463= 9
	Maestría en Medici= na Veterinaria, Universidad Católica de Cuenca, Ecuador. afcevallosm70@est.ucacue.edu.ec
= ²	Manuel Esteban Maldonado Cornejo	https://orcid.org/0000-0002-= 1507-2280
	Maestría en Medici= na Veterinaria, Universidad Católica de Cuenca, Ecuador. mmaldonadoc@ucacue.edu.ec
= ³	Pablo Giovanny Rubio Arias	https://orcid.org/0000-0002-9185-482= 3
	Maestría en Medici= na Veterinaria, Universidad Católica de Cuenca, Ecuador. prubioa@ucacue.edu.ec

=	Artículo de Investigación Científica y Tecnológica Enviado: 11/05/2023 Revisado: 26/06/2023 Aceptado: 06/07/2023 Publicado:28/07/2023 DOI: https://doi.org/10.33262/ap.v5i3.380
<= span lang=3DEN-US style=3D'font-size:12.0pt;line-height:115%;font-family:"Time= s New Roman",serif; mso-fareast-font-family:Calibri;color:blue;mso-ansi-language:EN-US'>=
Cítese:		Cevallos Minchala, A. F., Maldonado Cornejo, M. E., & Rubio Arias, P. G. (2023= ). Actividad antimicótica de la Monensina sobre Malassezia in vitro aislada = de caninos con otitis externa. AlfaPublicaciones, 5(3), 103–115. https://d= oi.org/10.33262/ap.v5i3.380
		ALFA PUBLICACIONES, es una Revista Multidisciplinar, Trimestral, que se public= ará en soporte electrónico tiene como misión contribuir a la &nbs= p; formación de profesionales competentes con visión humanística y crítica q= ue sean capaces de exponer sus resultados investigativos y científicos en la misma medida que se promueva mediante su intervención cambios positivos e= n la sociedad. https://alfapubli= caciones.com La revista es editada por la Editorial Ciencia Digital (Editorial de prestigio registrada en la Cámara Ecuatoria= na de Libro con No de Afiliación 663) www.celibro.org.ec
		Esta revista está protegida bajo una licencia Creative Common= s Attribution Non Commerci= al No Derivatives 4.0 International. Copia de la licencia: http://creativecommons.org/licenses/by-nc-nd/4.0/
Palabras claves: Mico= sis; Ionoforos; Infecciones; Control; Caninos.		Resumen Objetivo. Evaluar la actividad antimicótica de la monensina sobre = Malassezia in vitro aislada de caninos con otitis externa (OE), y determinar su susceptibilidad a monensina que serán cultivados in vitr= o. Metodología. Esta investigac= ión fue realizada sobre 30 paciente caninos con otitis externa clínica que fu= eron atendidos en Fundación Arca y en clínicas veterinarias particulares. Los mismos que presentaron signos clínicos como: dolor, eritema del pabellón auricular, prurito, sacudidas de la cabeza e inclinación cefálica. Para la identificación del agente etiológico (Malassezi pachidermatis), mediante citología con tinción = diff quick para ser eva= luadas bajo microscopio con su forma de zapato dando positivo a presencia de Malassezia para ser cultivadas in vitro y de esta manera ser evaluada conjuntamente la actividad antimicrobiana de monensina = que se utilizó el tratamiento= de estudio en cajas Petri con Monensina con cuat= ro diferentes concentraciones 1%, 5%, 10% y 20%. Resultado. Obteniend= o como resultados que los 30 pacientes caninos dieron positivos a malassezia, pero su actividad antimicrobia= na de monensina no obtuvo ningún resultado favorable fren= te a otros agentes antimicóticos de uso comercial como el ketoconazol, fluconazol y griseofulvi= na, siendo el ketoconazol el medicamento con mayor actividad antimicótica frente a malassezia. C= onclusión. Que la = Malassezia desempeńa un gran rol en esta enfermedad, por lo que varios tratamien= tos se han investigado e implementado, sin embargo, con la evolución de la especie, nuevas resistencias se han originado dando como resultado infecciones crónicas persistente. La Monensina no actúa contra este microorganismo con las concentraciones manejadas en el presente estudio, por lo tanto, se necesita nuevos estudios que analicen componentes farmacológicos, entre los que se han planteado diversos aceit= es naturales con el fin de evitar efecto tóxicos y adversos para los canes. = Área de= estudio general: Medicina Veterinaria. Área de estudio Especifica: Dermatología.
Keywords:= Mycosis; Ionophores; Infections; Contr= ol; Canines.		Abstract Objective. Evaluate the antifungal activity of monensin on Malassezia in Vitro, isolated from canines= with external otitis (EO), and delimitate their susceptibility to monensin, wich will be = cultured in vitro. Methodology. This research was conducted on 30 canine patients, suffering from clinical external otitis,= and treated at ¨Fundación Ar= ca¨ and private veterinary clinics. = These canine patients exhibited clinical signs such as pain, erythema of pinna, pruritus, head shacking and head tilting. Malassezi<= /span> pachidermatis was identified as the etiologic= al agent through cytology with diff-quick staining, and samples were evaluat= ed under a microscope to confirm the presence of Malass= ezia for in vitro culturing. Results. The results showed that all 30 canine patients tested positive for = Malassezia, but the antimicrobial activity of monensin did not yield any favorable results compar= ed to other commercially used antifungal agents like ketoconazole, fluconazole,= and griseofulvin. Ketoconazole exhibited the high= est antifungal activity against Malassezia. Conclusion. Malassezia develops a significant role in this disease, and various treatments have been investigated and implemented form this fungus. However, with the evolution of the species,= new resistances have emerged, resulting in persistent chronic infections. Monensin does not act against this microorganism at= the concentrations used in this study; therefore, new studies are needed to analyze pharmacological components, including various natural oils, with = the aim of avoiding toxic and adverse effects for the canines.

	Fluconazole	Ketoconazole	Monensina
	Rango (GM) (mg/ml)	Rango (GM) (mg/ml)	5%	10%	20%
Estudio In Vitro	0,5-4,0	0,1-2,0	0	0	0
Mirande et al. (2007)	0,125-16,0	0,03-0,25
Jesus et al. (2011)	0,01-4,0	0,25-2
Schlemmer et al. (2019)	1,0-6,4
Chebil et al. (2022)	0,125-8,0	0,06-8,0