The aim of this study was to determine the presence of genomic ?-and ? -defensins (bovine enteric beta defensin – ?-def alp – and Defa 8 respectively) and their levels of expression as messenger RNA in samples of intestinal mucosa of newborn alpacas. Samples of jejunum of 30 animals from 0 to 45 days of age were collected. Genomic DNA and mRNA were extracted for PCR analysis and real-time RT-PCR respectively. All samples amplified a single DNA segment (between 800 and 900 pb) by PCR which corresponds to the alpha defensin 8 (defa 8), while 90% (27/30) amplified a segment expected 300-350 pb for ?-def alp and two additional bands of 800-900 pb evidenced by agarose gel electrophoresis. The detection of mRNAs by real time RT-PCR was performed based on Cycle threshold (Ct) analysis and Melting temperature (Tm) of the amplified products. The Cts for defa 8 products were found between 25.1...

El objetivo del presente estudio fue determinar la presencia genómica de ? - y ?- defensinas (Defa 8 y??-def alp) y sus niveles de expresión como ARN mensajeros, en muestras de mucosa intestinal de crías alpaca. Se colectaron muestras de yeyuno y sangre de 30 animales de 0 a 45 días de edad. Se extrajo el ADN genómico y los ARN mensajeros para el análisis por PCR y RT-PCR tiempo real, respectivamente. La prueba de PCR convencional determinó que todas las muestras amplificaron un segmento genómico de entre 800 y 900 pb correspondiente a una Defa 8, y el 90% (27/30) de las muestras amplificó un segmento esperado de 300 a 350 pb para ?-def alp y dos bandas adicionales de 800 a 900 pb, evidenciados en la electroforesis en gel de agarosa. La detección de ARN mensajeros por RT-PCR tiempo real se realizó en base al análisis de las curvas de amplificación (Ct) y curvas de disociaci�...

In 2013, clinically suggestive cases of PEDv were reported in pig farms in Lima and Arequipa, reaching up to 100% mortality in piglets and negative results to other viral agents such as classical swine fever (CPPv) and transmissible gastroenteritis (TGEv). For this reason, the aim of this study was to isolate and molecularly detect PEDv strains in Lima pig farms. A total of 37 stool samples and intestinal contents of piglets between 2 and 21 days of age with clinical signs suggestive of PEDv from pig farms of the department of Lima, Peru were collected. Results showed that 94.3% (35/37) were positive by the immunochromatography (IHC) test; 97.1% (34/35) of them were confirmed by RT-PCR real-time that amplified a 101-bp segment of the ORF3 gene of the PEDv. The control and positive samples showed a threshold cycle (Ct) between 10 and 21 cycles with a melting temperature (Tm) of 77.7 °C. ...

The aim of this study was to determine the seroprevalence of Bovine Viral Diarrheavirus (BVDV) in grazing cattle without history of vaccination, in the province of SanPablo, Cajamarca, Peru. It was used 385 samples from the serum bank, collected in 2004.Samples were stratified in four age groups (2 to <6, 6 to <12, 12 to <24, and >24 months)and by sex. The detection of antibodies against BVDV was done by the viral neutralizationtest. The 27.0 ± 4.4% (104/385) of samples had antibodies against BVDV, and withoutstatistical difference due to age or sex; however, 71.2 ± 8.7% (47/66) of animals between 12 and 24 months of age showed antibody titres between 128 and >256. It was concludedthat the BVDV is present with a low seroprevalence in the cattle population of San Pabloprovince, Cajamarca.

El objetivo del presente estudio fue determinar la seroprevalencia del virus de la Diarrea Viral Bovina (VDVB) en bovinos criollos de crianza extensiva, sin historia de vacunación, en la provincia de San Pablo, Cajamarca. Se emplearon 385 muestras del banco de sueros, colectadas en el 2004. Las muestras se estratificaron en cuatro grupos etarios (2 a <6, 6 a <12, 12 a <24 y >24 meses) y por sexo. La detección de anticuerpos contra el VDVB se hizo mediante la prueba de neutralización viral. El 27.1 ± 4.4% (104/385) de los bovinos presentó anticuerpos contra el VDVB indistintamente del grupo etario o sexo; sin embargo, el 71.2 ± 8.7% (47/66) de los animales entre 12 y 24 meses de edad presentaron títulos de anticuerpos entre 128 a >256. Se concluye que el VDVB está presente con una prevalencia baja en la población de bovinos de la provincia de San Pablo.

En 2013 se observaron cuadros clínicos sugerentes a PEDv en granjas porcinas de Lima y Arequipa llegando hasta el 100% de mortalidad en lechones y con resultados negativos a otros agentes virales como peste porcina clásica (PPCv) y gastroenteritis transmisible (TGEv). Por esta razón, el objetivo del trabajo fue aislar y detectar molecularmente cepas del PEDv en granjas porcinas de Lima. Se colectaron 37 muestras de heces y contenido intestinal de lechones entre 2 y 21 días de edad con cuadros clínicos sugerentes a PEDV procedentes granjas porcinas del departamento de Lima, Perú. El 94.3% (35/37) resultaron positivos al test de inmunocromatografía (IHC). El 97.1% (34/35) de estos fueron confirmados por RT-PCR en tiempo real que amplifica un segmento de 101 pb del gen ORF3 del PEDv. El control y las muestras positivas mostraron un ciclo umbral (Ct) entre 10 y 21 ciclos con una tempe...

The aim of this study was to isolate and genotyping the Porcine Respiratory and Reproductive Syndrome Virus (PRRSV) strain in pig farms in Lima and Arequipa, Peru. Seropositive pig farms to PRRSV were identified by ELISA test. Blood samples were collected from weaned pigs from positive pig farms in Lima (A=44, B=20; C=16) and Arequipa (D=32, E=92). The serum samples (n=204) were processed in 51 pools of 4 samples each for virus isolation using Porcine Alveolar Macrophage (PAM) cell line. The genome of the virus isolated was identified by Reverse Transcription-nested Polimerase Chain Reaction (RT-nPCR). The complementary DNA (cDNA) of genotype 1 and 2 of PRRSV vaccine strains were used as positive controls and cDNA of equine viral arteritis virus, classical swine fever virus and PAM cells as negative controls in the RT-nPCR. Three blind passages in PAM cell line with each of the 51 pool w...

The aim of this research was to determine the expression of proinflammatory cytokines from alpaca leukocytes through the antigenic challenge with macrocyst extract of Sarcocystis aucheniae at various doses and exposure times. Leukocytes in a concentration of 500 000 cells/ml were exposed to concentrations of 0.5, 1, 50, 500 and 1000 ng of macrocyst extract ofS. aucheniae and incubated for 1, 12 and 24 h. The total messenger RNA (mRNA) was extracted for each treatment using Trizol and used to perform real-time RT-PCR with specific primers for cytokine TNF-α and interleukins IL-1α, IL-1β and IL-6. The generated mRNA levels of IL-1α and TNF-α at 1 h were detectable and higher in comparison to the calibrator (leukocytes not exposed to the extract). IL-1β increased at the 1 ng/ml concentration at 24 h showing a negative kinetic expression when compared with the untreated control group. ...

The aim of this study was to isolate and genotyping the Porcine Respiratory and Reproductive Syndrome Virus (PRRSV) strain in pig farms in Lima and Arequipa, Peru. Seropositive pig farms to PRRSV were identified by ELISA test. Blood samples were collected from weaned pigs from positive pig farms in Lima (A=44, B=20; C=16) and Arequipa (D=32, E=92). The serum samples (n=204) were processed in 51 pools of 4 samples each for virus isolation using Porcine Alveolar Macrophage (PAM) cell line. The genome of the virus isolated was identified by Reverse Transcription-nested Polimerase Chain Reaction (RT-nPCR). The complementary DNA (cDNA) of genotype 1 and 2 of PRRSV vaccine strains were used as positive controls and cDNA of equine viral arteritis virus, classical swine fever virus and PAM cells as negative controls in the RT-nPCR. Three blind passages in PAM cell line with each of the 51 pool w...

The aim of this research was to determine the expression of proinflammatory cytokines from alpaca leukocytes through the antigenic challenge with macrocyst extract of Sarcocystis aucheniae at various doses and exposure times. Leukocytes in a concentration of 500 000 cells/ml were exposed to concentrations of 0.5, 1, 50, 500 and 1000 ng of macrocyst extract ofS. aucheniae and incubated for 1, 12 and 24 h. The total messenger RNA (mRNA) was extracted for each treatment using Trizol and used to perform real-time RT-PCR with specific primers for cytokine TNF-α and interleukins IL-1α, IL-1β and IL-6. The generated mRNA levels of IL-1α and TNF-α at 1 h were detectable and higher in comparison to the calibrator (leukocytes not exposed to the extract). IL-1β increased at the 1 ng/ml concentration at 24 h showing a negative kinetic expression when compared with the untreated control group. ...

The aim of the study was to determine the presence of antibodies against New Jersey (NJ) and Indiana subtype 1 (IND-1) Vesicular Stomatitis Virus (VSV) in free-living white-lipped peccaries (Tayassu pecari) in three localities of Madre de Dios, Peru. The presence of antibodies against VSV by virus neutralization test was determined in 88 serum samples of adult male and females in apparent good health condition. Results showed that 53.4% (47/88) and 18.2% (16/88) of samples were positive to antibodies against serotypes IND-1 and NJ respectively, whereas 55.3 and 12.6% of the serum samples had neutralizing antibodies titers equal or greater than 1:32 against serotype IND-1 and NJ respectively. There was a no significant association between seropositivity of VSV and source of samples.

El objetivo del presente estudio fue determinar la presencia de anticuerpos neutralizantes contra los serotipos New Jersey (NJ) e Indiana subtipo 1 (IND-1) del virus Estomatitis Vesicular (VEV) en huanganas (Tayassu pecari) de vida libre de las localidades de Boca de Manu (n=30), Concesión para la Conservación Los Amigos (n=10) y La Reserva Nacional Tambopata/Parque Nacional Bahuaja Sonene (n=48) en el departamento de Madre de Dios. La presencia de anticuerpos contra el VEV fue determinado mediante la prueba de neutralización viral en las 88 muestras de suero de huanganas machos y hembras adultos de apariencia normal. El 53.4% (47/88) y 18.2% (16/88) de las muestras fue positiva a anticuerpos contra los serotipos IND-1 y NJ, respectivamente, en tanto que el 29.5 y el 2.3% de las muestras tuvieron anticuerpos neutralizantes igual o mayor a 1:32 contra los serotipos IND-1 y NJ, respecti...

The aim of this study was to evaluate the expression of proinflammatory cytokines of alpaca circulating leukocytes when confronting total extract of Clostridum perfringens (structural components and toxins). Whole blood was collected from the jugular vein of 5 female and 5 male adult alpacas. The leukocytes were separated using ammonium chloride to lyse erythrocytes followed by centrifugation and then cultured at a concentration of 500 000 cells/ml of MEM (Minimal Essential Medium) in culture plates of 12 wells. Simultaneously were challenged with total extract of C. perfringens grown in thioglycolate broth at concentrations of 400, 80, 16, 0.8 and 0.2 μg/ml and then quantifying by RT-PCR the expression of cytokines TNF∝, IL-1∝, IL-6 and IL-1ß at 1, 12 and 24 h of exposure. The results showed that low doses of clostridial extracts (0.2 to 0.8 mg/ml) are capable of inducing expressi...

The aim of this study was to evaluate the expression of proinflammatory cytokines of alpaca circulating leukocytes when confronting total extract of Clostridum perfringens (structural components and toxins). Whole blood was collected from the jugular vein of 5 female and 5 male adult alpacas. The leukocytes were separated using ammonium chloride to lyse erythrocytes followed by centrifugation and then cultured at a concentration of 500 000 cells/ml of MEM (Minimal Essential Medium) in culture plates of 12 wells. Simultaneously were challenged with total extract of C. perfringens grown in thioglycolate broth at concentrations of 400, 80, 16, 0.8 and 0.2 μg/ml and then quantifying by RT-PCR the expression of cytokines TNF∝, IL-1∝, IL-6 and IL-1ß at 1, 12 and 24 h of exposure. The results showed that low doses of clostridial extracts (0.2 to 0.8 mg/ml) are capable of inducing expressi...

Peru has 13% of the Amazonian tropical forest which contains over 460 species of mammals, including the white lipped peccary or huangana (Tayassu peccari). This species is of great ecological and commercial importance and a major source of protein for the local inhabitants; however, information about the sanitary situation of this species is scarce. The aim of this study was to determine antibodies against bluetongue virus (BTV) and other Orbivirus in free-living and healthy white lipped peccaries from the Madre de Dios region, Peru. One hundred and six serum samples were evaluated to determine antibodies against BTV by competitive ELISA and other Orbivirus by immunodiffusion agar gel test. Results showed that 7.5% (8/106) of the samples had antibodies against BTV and 29.2% (31/106) against BTV/other Orbivirus. Antibodies against BTV and other Orbivirus were detected in all three localit...

This study aimed to determine the presence ofAnaplasma platys in domestic dogs of Metropolitan Lima with clinic signs consistent with anaplasmosis by identifying inclusion bodies in platelets and through the Hemi-Nested PCR technique using peripheral blood samples. For this purpose, 144 samples were collected between January to December 2012. The results showed that 29.2% (42/144) were positive (thrombocytopenic individuals and presence ofinclusion bodies in platelets) and 12.5% (18/144) were suspicious (nonthrombocytopenic individuals and presence of inclusion bodies in platelets) by hematologic identification. Moreover, 1.4% (2/144) resulted positive to the Hemi-Nested PCR test. These results confirm the presence ofA. platys in domestic dogs in the country

The aim of this study was to determine the seroprevalence of antibodies against Equine Herpes Virus type 1 (EHV-1) and Equine Herpes Virus type 4 (EHV-4), which causes rhinopneumonitis in horses. Blood samples (n=825) from Peruvian horses older than six months of age, both sexes, identified as racehorses, Peruvian Paso, equitation and criollo, in apparent healthy conditions, were collected for detection of neutralizing antibodies against EHV-1/EHV-4 by virus neutralization test. The 48.9 ± 5.3% (403/825) of the samples had antibodies against EHV-1/EHV-4. Antibody titers ranged from 2 to >256, where 58.5% ranged from 2 to 8, 29.5% from 16 to 64, and 11.9% from 128 to >256. The sex and region of the country did not represent a risk for presenting antibodies against EHV-1/EHV-4.

The aim of this study was to compare the relative expression levels of the gene from exon 1 of the IgA in the intestinal epithelium of baby alpacas untreated and treated orally with Clostridium perfringens antigens and all-trans retinoic acid (ATRA). Thirty two animals were sampled: 14 treated (6 of 1 day old and 8 between 7-14 days old) and 18 untreated (10 of 1 day old and 8 between 7-14 days old). The animals were slaughtered after one week of the treatment and 2 cm of the jejunum was collected and kept at -196 °C. Total RNA was extracted from each sample using the TRIzol® method and complementary DNA (cDNA) was synthesized. PCR and RT-PCR Real Time were conducted using specific oligonucleotides designed for the detection of exon 1 of the region Fc IgA in the alpaca. The quantification of the relative expression by normalization with GAPDH gene mRNA and the relative expression calcu...

El Perú posee el 13% de los bosques tropicales amazónicos con más de 460 especies de mamíferos, incluyendo el pecarí labiado o huangana (Tayassu pecari). Esta especie es de gran importancia ecológica y comercial, así como una importante fuente de proteína para la población amazónica; sin embargo, las informaciones sobre su estado sanitario son escasas. El objetivo del estudio fue determinar anticuerpos contra el virus de lengua azul (VLA) y otros Orbivirus en huanganas de vida libre de apariencia normal de tres localidades del departamento de Madre de Dios. Se evaluaron 106 muestras de suero para determinar los anticuerpos contra el VLA mediante un kit de ELISA de competición y contra otros Orbivirus mediante la prueba de inmunodifusión en gel de agar. El 7.5% (8/106) presentaron anticuerpos específicos contra el VLA y el 29.2% (31/106) contra VLA/otro Orbivirus. Anticuerpos...

This study aimed to determine the presence ofAnaplasma platys in domestic dogs of Metropolitan Lima with clinic signs consistent with anaplasmosis by identifying inclusion bodies in platelets and through the Hemi-Nested PCR technique using peripheral blood samples. For this purpose, 144 samples were collected between January to December 2012. The results showed that 29.2% (42/144) were positive (thrombocytopenic individuals and presence ofinclusion bodies in platelets) and 12.5% (18/144) were suspicious (nonthrombocytopenic individuals and presence of inclusion bodies in platelets) by hematologic identification. Moreover, 1.4% (2/144) resulted positive to the Hemi-Nested PCR test. These results confirm the presence ofA. platys in domestic dogs in the country