Clinical rectal diarrheic swabs (n=27) and pathological intestinal contents (n=24) from 51 alpacas between 1 to 7 weeks of age were used to isolate and genotype Escherichia coli, and to test for antimicrobial sensibility. All the E. coli isolates, confirmed by the API system, were genotyped by Multiplex PCR to determine the presence of virulence genes: stx1 and stx2 (shigatoxin 1 and 2), eae (intimin), bfp (Bundle-Forming Pili), lt (heat-labile toxin), sta and stb (heat-stable toxin A and B), in enterohemorrhagic (EHEC), enteropathogenic (EPEC) or enterotoxigenic (ETEC) E. coli strains. Positive stx1 and stx2 strains were tested on Vero cells for verocytotoxicity, and all isolates were tested for antimicrobial sensibility to the eight most frequent used antibiotics in Andean livestock. Microbiological and molecular analysis revealed the presence of pathogenic E. coli strains in 19/27 (70...

Se analizaron 27 hisopos diarreicos rectales y 24 contenidos patológicos del intestino de 51 alpacas de 1 a 7 semanas de edad para aislar y genotificar Escherichia coli, y caracterizar su sensibilidad antimicrobiana. Los aislamientos de E. coli fueron genotipificados por PCR múltiple para detectar genes de virulencia: stx1 y stx2 (shigatoxina 1 y 2), eae (intimina) y bfp (Bundle-Forming Pili), lt (toxina termolábil), sta y stb (toxina termoestable A y B), presentes en E. coli enterohemorrágica (EHEC), enteropatógena (EPEC) y enterotoxigénica (ETEC). Paralelamente, cepas positivas a stx1 o stx2 fueron analizadas en células Vero para detectar efectos verocitotóxicos. Posteriormente, todas las cepas aisladas fueron evaluadas para determinar su sensibilidad antimicrobiana a ocho antibióticos frecuentemente utilizados en ganaderías alpaqueras altoandinas. Los análisis microbiológi...

Advances in research on the major causative agents of morbidity and mortality in newborn alpacas associated with enteric processes in southern Peru were reviewed. Microbiology and molecular analyses performed on intestinal samples from enterotoxemia fatalities confirmed the predominance of C. perfringens type A carrying only the gene coding for the major α exotoxin and identifiying for the first time the presence of the novel β2 toxin gene. In vitro studies have yielded three profiles for phospholipase activity (high, medium and low) with biological activity when high and medium strains were inoculated intraintestinally in mice and rabbits, but did not induce intestinal pathology in an alpaca cria. A detailed histopathological investigation has reveled that within necrotizing hemorrhagic enteritis Clostridium coexist with massive presence of Eimeria macusaniensis suggesting that primar...

Advances in research on the major causative agents of morbidity and mortality in newborn alpacas associated with enteric processes in southern Peru were reviewed. Microbiology and molecular analyses performed on intestinal samples from enterotoxemia fatalities confirmed the predominance of C. perfringens type A carrying only the gene coding for the major α exotoxin and identifiying for the first time the presence of the novel β2 toxin gene. In vitro studies have yielded three profiles for phospholipase activity (high, medium and low) with biological activity when high and medium strains were inoculated intraintestinally in mice and rabbits, but did not induce intestinal pathology in an alpaca cria. A detailed histopathological investigation has reveled that within necrotizing hemorrhagic enteritis Clostridium coexist with massive presence of Eimeria macusaniensis suggesting that primar...

The canine distemper (CD) is a viral infection caused by a Morbillivirus of the family Paramyxoviridae. It can generate digestive, respiratory, and nervous symptomatology, depending on the infecting strain. This study was designed to detect the CD virus through the RT-PCR technique and perform the analysis of the data collected to have a current view of the behavior of this virus in Lima, Peru. Whole blood samples from 52 unvaccinated canines with clinical signs compatible with canine distemper were collected between June 2012 and January 2015. The nucleoprotein of 287 bp was successfully amplified in 32.7% (17/52) of the samples analyzed. In addition, it was found a greater detection of the virus, but not significative, in individuals that manifested systemic clinical signs (respiratory and digestive signs vs nervous) and within the age range of 1.5-4 months.

El distemper canino (DC) es una infección vírica causada por un Morbilivirus de la familia Paramyxoviridae. Puede generar sintomatología digestiva, respiratoria y nerviosa, dependiendo de la cepa infectante. Este estudio estuvo destinado a detectar el virus del DC a través de la técnica de RT-PCR y analizar los datos recolectados para tener una visión actual del comportamiento de este virus en la zona de Lima, Perú. Se trabajó con muestras de sangre entera recolectadas entre junio de 2012 y enero de 2015 de 52 caninos no vacunados con signos clínicos compatibles con distemper canino. Se logró amplificar la nucleoproteína de 287 pb en el 32.7% (17/52) de las muestras analizadas, encontrando, además, una mayor detección del virus, aunque no significativa, en individuos que manifestaban signos clínicos sistémicos (signos respiratorios y digestivos vs nerviosos) y dentro del r...

The aim of this study was to evaluate the in vivo immunogenic capacity of a recombinant outer membrane protein of Pasteurella multocida called P6-like (rP6-like). The field trial was carried out in an alpaca production unit in Puno, Peru, and the processing of the samples was done in the Faculty of Veterinary Medicine of San Marcos University in Lima. Four groups of 5 animals were formed, and they were inoculated according to group with a) normal saline solution, b) bacterin, c) rP6-like, d) combination of bacterin + rP6-like. Blood samples were collected on days 0, 5, 7, 9, 12 and 15 post-inoculation. Single-point indirect ELISA tests were designed for the relative quantification of specific anti-rP6-like antibodies and for total anti-total P. multocida protein antibodies. The group inoculated with rP6-like showed significant elevation of antibodies from day 7 (p<0.05) until day 15 p...

The purpose of this study was to characterize strains of Pasteurella multocida isolated from lungs of 1-2 months old alpacas with signs of pneumonia, collected in 2014. The BOX-PCR technique was used to demonstrate genetic diversity among strains. Twenty-four strains of P. multocida were isolated from 46 animals through biochemical identification tests. The BOX-PCR analysis showed two amplified band patterns, where 22 strains were grouped in one cluster and two strains in another cluster. The present study showed genetic homogeneity in most of the strains, evidencing a common source of infection that affected the animals.

The objective of the study was the detection of canine parvovirus type 2 (CPV-2) in young dogs of Lima city with and without clinical symptoms compatible with parvovirus by the PCR technique using primers that can allow the amplification of a fragment of the gene coding for the protein VP2. Rectal swabs were collected from 78 dogs younger than one year old and without a history of previous vaccinations, of which 39 individuals had a clinical diagnosis of canine parvovirus and the other 39 were clinically healthy animals. For the extraction of viral DNA, the fast boiling method was used. Samples were boiled at 100 °C for 10 minutes and then centrifuged to extract the supernatant, which was used as a template for the PCR reaction. Specific primers that amplify a 1316 base pair fragment of the VP2 gene of the CPV-2 virus were used, using a commercial vaccine as a positive control. The viru...

The present study aimed to evaluate the immunogenic activity of three life attenuated strains of Salmonella Typhimurium isolated from guinea pig, by evaluating the excretion of faecal IgA in mice. The study was carried out with 28 four-week-old female mice of the Balb/c strain, distributed into four groups (seven mice per group): strain 1, strain 2, strain 3 and control group (PBS). Each mouse was inoculated with 109 CFU/100 µl of the attenuated strains using an orogastric probe. Fresh stool samples were collected at days 1, 9, 15 and 21 post-immunizations. The quantification of faecal IgA was carried out by an indirect in-house ELISA test. The results indicate that the mice inoculated with strains 1 and 2 presented a higher immunogenic activity (faecal IgA) compared to the Control group.

The present study aims to obtain data that facilitate the design of an animal model that replicates salmonellosis produced by the serovar Typhimurium under experimental conditions by a single dose of streptomycin 50 mg orally 24 hours prior to the challenge with three concentrations of a virulent strain of Salmonella Typhimurium. Female guinea pigs (n=58) were used, distributed in 6 treatment groups of 8 animals each (3 groups to which streptomycin was administered prior to the challenge and 3 groups that were only challenged) and 2 control groups of 5 animals each (1 group treated with streptomycin and unchallenged and another untreated and unchallenged). The doses of the inoculum were 107, 108, 109/ml of S. Thiphimurium per group, respectively, and the animals were monitored for 30 days. Collective stool samples per group were collected to determine the excretion time of S. Typhimurium...

The purpose of this study was to characterize strains of Pasteurella multocida isolated from lungs of 1-2 months old alpacas with signs of pneumonia, collected in 2014. The BOX-PCR technique was used to demonstrate genetic diversity among strains. Twenty-four strains of P. multocida were isolated from 46 animals through biochemical identification tests. The BOX-PCR analysis showed two amplified band patterns, where 22 strains were grouped in one cluster and two strains in another cluster. The present study showed genetic homogeneity in most of the strains, evidencing a common source of infection that affected the animals.

El objetivo del estudio fue la detección de parvovirus canino tipo 2 (CPV-2) en perros jóvenes con/sin sintomatología clínica compatible con parvovirosis mediante la técnica de PCR, usando cebadores que pueden permitir la amplificación de un fragmento del gen codificante de la proteína VP2. Se colectaron hisopados rectales de 78 perros menores a un año y sin historia de vacunaciones previas, de los cuales 39 individuos tuvieron un diagnóstico clínico de parvovirosis canina y los otros 39 fueron animales clínicamente sanos. Para la extracción de ADN viral se usó el método fast boiling, donde las muestras fueron sometidas a un hervido a 100 °C por 10 minutos con posterior centrifugación para extraer el sobrenadante, el cual fue usado como molde para la reacción de PCR. Se usaron cebadores específicos que amplifican un fragmento de 1316 pares de bases del gen VP2 del virus...

A total of 478 fecal samples taken from healthy alpaca crias aged 1-90 days at Centro de Investigación y Producción (CIP - La Raya), Universidad Nacional del Altiplano, Puno, were processed by the sedimentation and flotation technique in saturated sugar solution to detect oocysts and by the McMaster method to determine oocysts per gram (OPG) of fecal sample. Eimeria spp were present in 418 (87.5%) of the samples with a mean of 24,017 OPG (CI of 7,534 and range 50-1,202,400). The most frequent species were E. lamae (60.4%), E. macusaniensis (50.4%), E. alpacae (45.6%), E. punoensis (30%), and E. ivitaensis (6.24%). Parasite load increased with age, 50% of 24 samples at 1-30 days were positive with an average load of 17,216 OPG, at 31-45 days 93% of 82 (28,501 OPG), at 46-60 days 85% of 144 (34,731 OPG), at 61-75 days 94% of 183 (6,564 OPG) and at 76-90 days 80% of 45 with 17,376 OPG. E....

The aim of the study was to evaluate the in vitro immunogenic activity of a recombinant P6-like protein from a culture of Pasteurella multocida isolated from pneumonia cases in young alpacas. Blood peripheral mononuclear cells (PBMCs) were challenged with a recombinant P6-like protein at a concentration of 10 ng per sample, from 3 to 72 hours. Total RNA was extracted to perform the real-time RT-PCR test to observe cytokine expression levels of the Th1 immune response (TNF-α, IFN-γ and IL-2) and Th2 (IL-10 and IL-4) in the established times. Both Th1 and Th2 profile cytokines expressed a greater number of times than PBMC not exposed to the recombinant protein, where at 24 and 48 hours showed the greater expressions. Likewise, an apparent trend toward the Th2 profile was found, but at levels that did not influence the expression of cytokines in the other profile.

The aim of this study was to perform a fungal count and detect mycotoxins in feed supplies and balanced feed from pig farms in four districts of Coronel Portillo, Ucayali. Fifty samples were collected (35 of balances feeds [three rearing stages] from 20 farms and 15 of feed supplies from 10 of these farms). The fungal count was performed using the plate count method and the detection of mycotoxins (aflatoxin B1, ochratoxin A and zearalenone) using commercial ELISA kits. Results showed that 50% of farms had at least one sample of unacceptable quality. Likewise, 6/15 (40%) and 5/35 (14.3%) of feed supplies and balanced feed samples, respectively, had unacceptable quality. The starter feed had the highest average fungal count (9.6 x 104 CFU/g). There was an association between quality and type of food, but none between quality and origin. Half of the farms had at least one feed sample with ...

Se analizaron 478 muestras fecales de crías (1-90 días de edad), aparentemente saludables, en el Centro de Investigación y Producción (CIP - La Raya) de la Universidad Nacional del Altiplano, Puno. Las muestras fueron procesadas mediante la técnica de sedimentación y flotación (solución saturada de azúcar) buscando ooquistes y por el método de McMaster para determinar la carga parasitaria (ooquistes por gramo de heces- OPGH). En 418/478 (87.5%) de las muestras se detectaron ooquistes de Eimeria spp (24,017 OPGH, IC 7,534; rango: 50-1´202,400), preferentemente E. lamae (60.4%), E. macusaniensis (50.4%), E. alpacae (45.6%), E. punoensis (30%) y E. ivitaensis (6.24%). La infección parasitaria se incrementó con la edad. Inicialmente se encontró en el 50% de 24 muestras de 1-30 días de edad (17,216 OPGH), y luego de 93% de 82 crías de 31-45 días (28,501 OPGH), 85% de 144 crí...

The aim of the study was to evaluate the in vitro immunogenic activity of a recombinant P6-like protein from a culture of Pasteurella multocida isolated from pneumonia cases in young alpacas. Blood peripheral mononuclear cells (PBMCs) were challenged with a recombinant P6-like protein at a concentration of 10 ng per sample, from 3 to 72 hours. Total RNA was extracted to perform the real-time RT-PCR test to observe cytokine expression levels of the Th1 immune response (TNF-α, IFN-γ and IL-2) and Th2 (IL-10 and IL-4) in the established times. Both Th1 and Th2 profile cytokines expressed a greater number of times than PBMC not exposed to the recombinant protein, where at 24 and 48 hours showed the greater expressions. Likewise, an apparent trend toward the Th2 profile was found, but at levels that did not influence the expression of cytokines in the other profile.

The results of our recent research work on enterotoxemia in Peruvian alpacas are presented. Microbiological and molecular analyses found that the majority of the isolates corresponded to Clostridium perfringens and contained the cpa coding gene for α toxin (A genotype) while 0.4% contained both the cpa and cpb genes of the α and β toxins (C genotype). A parallel study revealed that 8.5% of the genotype A isolates also had cpb2, but the cpe (enterotoxin) gene was absent in all cases. These results highly suggest that the exotoxins secreted by C. perfringens are the virulent factors in enterotoxemia, rather than the endogenous enterotoxin. Additionally, an histopathological study of intestinal samples from fatal cases showed that 30.6% had abundant immature structures of Eimeria macusaniensis affecting deep mucosa and cryptic gland epithelia, primarily in the jejune and ileum, suggestin...

The objective of the present study was to determine the levels of serum immunoglobulin G (IgG) in neonatal alpacas (5-23 days of age) killed by enterotoxemia and in animals of similar ages, but clinically healthy. In a first phase, a standard curve of physiological degradation of serum IgG was established from blood sera of six apparently healthy neonates from day 2 to 21 days of age (n=9). In a second phase, serum IgG concentrations were determined in 17 dead neonatal alpacas with lesions compatible with enterotoxemia and in 26 animals of similar ages, apparently healthy. The concentrations of IgG, determined by the Radial Immunodiffusion test, showed that all the animals at 48 hours of birth had adequate concentrations of IgG, while only three of the animals killed by enterotoxemia had IgG levels below the obtained standard curve, although only one of them with levels below 900 mg/dl a...