The aim of this work consisted to study the clinical and hematological effects that included blood coagulation and antibody production of a group of three horses immunized for the production of monovalent serum against Bothrops alternatus venom, in order to correlate the results with the humoral immune response of the animals. The results showed that immunization of horses with the essayed protocol induce slight clinic alterations and discrete changes in some parameters of the hemogram. Results also showed variations in the neutralizing capacity of the serum on different venom components, particularly toxins that affect blood coagulation. Also, it was verified a direct relationship between the rate of white blood cells and serum immunoglobulin production. Furthermore, they also indicate that the three-month rest period may be considered as too long as it causes a marked decrease in antibody titles, losing the serum its neutralizing capacity to venom compounds. Nevertheless, after re-administration of venom horses provided a rapid reposition of anti-toxins against B. alternatus venom, tolerating high venom doses during this second period of immunization.

El objetivo del trabajo consistió en estudiar los efectos clínicos, hemáticos y coagulativos, así como la producción de anticuerpos en tres equinos inmunizados para la producción de suero antiofídico monovalente contra Bothrops alternatus y evaluar los resultados obtenidos correlacionando la respuesta inmune humoral con el estado de salud de los animales. Los resultados indicaron que la inmunización de los equinos con el protocolo utilizado, indujo alteraciones clínicas menores y cambios en ciertos parámetros del hemograma y pruebas coagulativas. Se registraron variaciones de la capacidad neutralizante sobre distintos componentes del veneno, en especial las toxinas que afectan la coagulación de la sangre. Asimismo, se constató una correlación directa entre las tasas séricas de leucocitos y la producción de inmunoglobulinas. Los resultados indicaron que el período de descanso de un trimestre resulta extenso y causa una franca disminución del título de anticuerpos, perdiendo el suero capacidad neutralizante de la toxicidad del veneno; no obstante, al reanudar las inoculaciones los equinos presentaron una rápida reposición de los anticuerpos anti-toxinas de B. alternatus, tolerando altas dosis de veneno en el segundo período de inmunización.

The aim of this work was to evaluate the pharmacological activities (citotoxicity, bactericidal, platelet aggregation) and the changes in the activity/stability of the isolated phospholipase A2 (PLA2) from Bothrops alternatus venom. In order to elucidate myotoxic and bactericidal activities, murine mioblast /miotubules (C2C12) and strains of Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922) were used, respectively. Also, normal mammary epithelial cell culture (NMuMG) and tumoral type (LM3) were tested for a possible role in oncological therapy. With the purpose of studying the inhibitory effect of the PLA2 on platelet aggregation, washed human platelet and thrombin as physiological inductor, were used. Finally, changes in the activity and stability of the isolated PLA2 at different temperatures (4-90ºC) by kinetic assays were recorded; while the enzyme was subjected to radial indirect hemolysis to evaluate the effect of different pH (2.5-10). The PLA2 presented dose-dependent inhibitory effect on thrombin-induced platelet aggregation. Also, it showed high structural stability when exposed to extreme conditions of temperature and pH during several hours, and catalytic activity remained almost unchanged. Nevertheless, this enzyme tested on different cells cultures did not induce cytotoxic (even at high doses) or bactericidal activities. These findings have a scientific value for toxicology since they expand the knowledge of the characterization (biological, biochemical and structural aspects) of one of the most abundant enzyme (PLA2 Ba SpII RP4) from the protein package of B. alternatus snake venom from the northeast of Argentina. Likewise, the confirmed safety and stability of this PLA2 strongly suggest that it could be proposed as an alternative immunogen for the antiserum production against Bothrops snake venom

El objetivo del trabajo fue evaluar actividades farmacológicas (citotóxica, bactericida, agregante plaquetaria) y factores que afectan la actividad/estabilidad de la fosfolipasa A2 (PLA2) aislada del veneno de Bothrops alternatus. Se utilizaron mioblastos/miotúbulos murinos (C2C12) y cepas bacterianas de Staphylococcus aureus (cepa ATCC 25923) y Escherichia coli (cepa ATCC 25922) para valorar respectivamente su acción miotóxica (in vitro) y bactericida, como así también líneas celulares epiteliales mamarias normales (NMuMG) y tumorales (LM3) para detectar su posible aplicación en terapia oncológica. Con el fin de evaluar la acción inhibitoria de la PLA2 aislada sobre la agregación plaquetaria se empleó plasma rico en plaquetas y trombina como inductor fisiológico. Por último, se registraron los cambios en la actividad y estabilidad de la PLA2 a diferentes temperaturas (4-90ºC) mediante ensayo cinético, en tanto que, por hemólisis radial indirecta se hizo lo mismo para evaluar efecto del pH (2,5-10). La PLA2 mostró una acción inhibitoria sobre la agregación plaquetaria dosis-dependiente. Presentó una alta estabilidad estructural al ser sometida a condiciones extremas de pH y temperatura durante varias horas, conservando la actividad catalítica prácticamente invariable. En cambio, cuando PLA2 fue evaluada sobre las diferentes líneas celulares, no produjo citotoxicidad (aún a dosis elevadas) ni efecto bactericida sobre las cepas utilizadas. Estos hallazgos amplían la caracterización de la enzima, una de las más abundantes del paquete proteico del veneno total de B. alternatus del nordeste argentino. A la vez, la inocuidad y estabilidad detectadas, permiten proponer a esta enzima como un inmunógeno alternativo en la producción de sueros antiofídicos.

Os efeitos terapêuticos da formulação em gotas óticas de própolis procedentes da Argentina foram avaliados no tratamento da otite externa canina. Quarenta e oito cães com sintomas de otite externa foram distribuídos aleatoriamente em ensaio clínico duplo-cego controlado para avaliar a eficácia da formulação tópica com a própolis contra um placebo tópico no tratamento da otite externa. A preparação de própolis e placebo foi administrada em ambos os canais da orelha externa, duas vezes por dia, durante 14 dias. Ao longo do estudo, os cães foram submetidos a exame físico e à análise microbiológica de exsudatos auriculares. Os mais frequentes microrganismos isolados em meios de cultura foram: Malassezia pachydermatis (54,2%), Staphylococcus aureus (43,8%), Staphylococcus coagulase-negativo (25,0%), Pseudomonas aeruginosa (20,8%), Candida albicans (18,8%), Proteus mirabilis (16,7%), Streptococcus spp.(16,7%), Enterococcus faecalis (12,5%), Escherichia coli (12,5%), Staphylococcus intermedius (6,3%), Klebsiella spp.(4,2%) e Candida glabrata (2,1%). A preparação de própolis apresentou atividade antimicrobiana contra a maioria dos microrganismos isolados de amostras do grupo de tratamento, sendo que os do grupo-controle não se recuperaram da doença infecciosa auricular, e não foram observados efeitos adversos à própolis. Isso permitiu aos pacientes tratados com própolis melhora significativa dos parâmetros clínicos. Essa nova formulação da própolis argentina para o ouvido apresenta potencial utilidade no tratamento tópico da otite externa em cães.

The therapeutic effects of Argentine propolis ear drop formulation on canine otitis externa were evaluated. Forty-eight dogs with symptoms of otitis externa were randomly assigned to double-blinded, controlled clinical trial to evaluate the efficacy of topical formulation with propolis versus a topical placebo in the treatment of otitis externa. The propolis preparation and placebo were administrated into both external ear canals, twice daily for 14 days. Throughout the study, clinical examination and microbiological analysis of dogs ear exudates were made. The most frequent microorganisms isolated in culture media were: Malassezia pachydermatis (54.2%), Staphylococcus aureus (43.8%), coagulase-negative Staphylococcus (25.0%), Pseudomonas aeruginosa (20.8%), Candida albicans (18.8%), Proteus mirabilis (16.7%), Streptococcus spp. (16.7%), Enteroccocus faecalis (12.5%), Escherichia coli (12.5%), Staphylococcus intermedius (6.3%), Klebsiella spp. (4.2%), andCandida glabrata (2.1%). Whereas the control group did not recover from the infectious ear disease, the propolis preparation exhibited antimicrobial activity against most of the microorganisms isolated from samples of the treated group. In addition, no propolis-adverse effects were observed. This allowed propolis-treated patients to show a significant improvement of the clinical parameters. Thus, this new Argentine propolis ear drop formulation may be used for topical treatment of otitis externa in dogs.