The aim of the present study was to evaluate the relative expression levels of cytokines IL-10 and TGF-β in the intestinal mucosa of clinically healthy alpacas from 2 to 47 days old. The animals were classified in three age groups (six animals per group): 2-8 days old (group 1), 10-21days old (group 2) and 26-47-days old (group 3), regardless sex or breed. The midportion of jejunum of each animal was collected; total RNA was extracted and then analyzed by real-time RT-PCR technique using IL-10 and TGF-β specific primers. The relative mRNA expression analysis was done using the comparative 2-ΔΔCt method. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as housekeeping gene and jejunum of three 11-month-old fetuses were used as calibrators. The results showed that IL-10 mRNA expression levels were 7.21±1.02 (group 1), 13.53±1.26 (group 2), and 18.77±1.48 (group 3) times as ...

El objetivo del trabajo fue evaluar los niveles de expresión relativa de los genes de las citoquinas IL-10 y TGF-β en la mucosa intestinal de alpacas de 2 a 47 días de edad, clínicamente sanas. Se formaron tres grupos etarios (seis crías por grupo) conformados por alpacas de 2 a 8 días (grupo 1), 10 a 21 días (grupo 2) y 26 a 47 días (grupo 3), sin distinción de sexo o raza. Se tomó la porción media del yeyuno de cada animal, se extrajo el ARN total y se empleó la técnica RT-PCR en tiempo-real con cebadores específicos para las citoquinas en estudio. La expresión relativa de ARNm de IL10 y TGF-β fue determinada por el método comparativo 2-ΔΔCt usando como calibrador el yeyuno de tres fetos de 11 meses de gestación y como gen endógeno el gliceraldehído-3-fosfato deshidrogenasa (GAPDH). La expresión promedio de ARNm de IL-10 fue de 7.21±1.02 (grupo 1), 13.53±1.26 (...

The aim of this study was to compare histological lesions of rainbow trout (Oncorhynchus mikyss) affected by the bacterial cold water disease (BCDW) with asymptomatic trout skin samples from fish farms in Lake Titicaca (Puno, Peru) and identify the presence of Flavobacterium psychrophilum by conventional PCR. Thirteen trout with skin lesions and seven without lesions, with sizes greater than 25 cm and average weight of 350 g were used. Conventional PCR was performed for the identification of Flavobacterium psychrophilum, using primers directed to the 16S rRNA region. Molecularly, both groups of animals were positive for the presence of F. psycrophilum. It can be concluded that this pathogen is present in the southern part of the country.

The aim of this study was to compare histological lesions of rainbow trout (Oncorhynchus mikyss) affected by the bacterial cold water disease (BCDW) with asymptomatic trout skin samples from fish farms in Lake Titicaca (Puno, Peru) and identify the presence of Flavobacterium psychrophilum by conventional PCR. Thirteen trout with skin lesions and seven without lesions, with sizes greater than 25 cm and average weight of 350 g were used. Conventional PCR was performed for the identification of Flavobacterium psychrophilum, using primers directed to the 16S rRNA region. Molecularly, both groups of animals were positive for the presence of F. psycrophilum. It can be concluded that this pathogen is present in the southern part of the country.

The aim of this study was to determine the in vitro toxic effect of the protein extract of macrocysts of Sarcocystis aucheniae on circulating rabbit leukocytes (cytotoxicity). Five concentrations of the protein extract (0.5, 1, 50, 500, 1000 ng/100 μl) were exposed to a population of 500 000 leukocystes/ml for 1 and 12 h in a culture medium at 37 °C. The cell viability was evaluated by trypan blue exclusion and the enzyme degranulation of leukocytes (myeloperoxidase and glucosaminidase ) by polyacrylamide gel electrophoresis and transfered to nitrocellulose membranes (Western blotting). The cell count at 1 h of incubation showed high mortality (57.7 to 58.1%), whereas at 12 h varied from 65.2 to 99.4%, and without statistical differences between the mean of leukocytes that survive in each treatment. Mortality in the control group (saline) was 5 and 57% at 1 and 12 h of incubation respe...

En el presente trabajo se buscó determinar el efecto tóxico in vitro del extracto proteico de macroquistes de Sarcocystis aucheniae sobre leucocitos circulantes de conejo (citotoxicidad). Se trabajó con cinco concentraciones de extracto proteico (0.5, 1, 50, 500, 1000 ng/100 ìl) en una población de 500 000 leucocitos/ml incubados por 1 y 12 horas en un medio de cultivo a 37 ºC. Se determinó la viabilidad celular por exclusión con azul de tripán y la degranulación de enzimas leucocitarias (mieloperoxidasa y glucosaminidasa) mediante electroforesis en geles de poliacrilamida y transferencia a membranas de nitrocelulosa (Western blotting). El recuento de células a 1 h de incubación demostró una alta mortalidad (57.7 a 58.1%), siendo a las 12 h entre 65.2 y 99.4%, y sin diferencia estadística entre las medias del número de leucocitos que sobreviven a cada tratamiento. La morta...

The aim of this study was to determine and compare the expression levels of IgA genes and associated cytokines IL-5, IL-6 and TGF-β in the intestinal mucosa of clinically healthy baby alpaca, that were orally vaccinated with a clostridial antigen. Ten unvaccinated baby alpacas (control) and 15 vaccinated (two doses with a seven-day interval) were used. Sampling was done seven days after the second dose. Likewise, two age subgroups (15-28 and 35-56 days of age at the time of sampling) were considered. Intestinal segments of 2 cm in length were taken from the middle portion of the jejunum and stored at -196 °C. Total RNA extraction was performed and the complementary DNA (cDNA) was synthesized by reverse transcription. Subsequently, real-time PCR was run using specific primers for IL-5, IL-6, TGF-β and exon 3 of IgA. Relative expression evaluation was performed using the 2-ΔΔÄCt meth...

The aim of this study was to determine and compare the expression levels of IgA genes and associated cytokines IL-5, IL-6 and TGF-β in the intestinal mucosa of clinically healthy baby alpaca, that were orally vaccinated with a clostridial antigen. Ten unvaccinated baby alpacas (control) and 15 vaccinated (two doses with a seven-day interval) were used. Sampling was done seven days after the second dose. Likewise, two age subgroups (15-28 and 35-56 days of age at the time of sampling) were considered. Intestinal segments of 2 cm in length were taken from the middle portion of the jejunum and stored at -196 °C. Total RNA extraction was performed and the complementary DNA (cDNA) was synthesized by reverse transcription. Subsequently, real-time PCR was run using specific primers for IL-5, IL-6, TGF-β and exon 3 of IgA. Relative expression evaluation was performed using the 2-ΔΔÄCt meth...