At present, the use of DNA extracted from non-invasive samples, especially feces, is common practice in the discipline of conservation genetics. Even though DNA analysis is often difficult mainly due to the composition and preservation of feces has been shown in many species with the development of suitable extraction methods is possible to maximize the reliability and effectiveness of such procedures. The objective of this study has been optimization of a protocol for the extraction of DNA from vicuña feces (Vicugna vicugna mensalis), species classified as Near Threatened in Peru, in order to obtain DNA of sufficient quality for molecular analysis. A total of 51 samples were taken from vicuña dung piles in two communities located 4600 meters above sea level in the dry Puna ecosystems of the regions of Junin and Moquegua between August and September 2008. After observing defecation, sa...

Actualmente es posible determinar el estatus de especies silvestres mediante el análisis de ADN extraído de muestras no invasivas como las heces. Aunque dichos análisis suelen ser dificultosos debido principalmente a la composición y conservación de las heces, se ha demostrado en muchas especies que con el desarrollo de métodos adecuados de extracción es posible maximizar la fiabilidad y eficacia de este procedimiento. Por tanto, el objetivo del presente estudio fue optimizar un método de extracción de ADN de heces de vicuña Vicugna vicugna mensalis, especie clasificada como casi amenazada en el Perú, con la finalidad de obtener ADN de suficiente calidad para análisis moleculares. Se recolectaron 51 muestras de heces durante los meses de agosto y septiembre de 2008 en dos comunidades vicuñeras ubicadas a más de 4600 msnm en los departamentos de Junín y Moquegua. Luego de o...

The spectacled bear (Tremarctos ornatus) is a vulnerable species because of hunting and destruction of its environment. Also, it is difficult to monitor in the wild hampering the management and research efforts and therefore, the use of non-invasive samples such as faeces becomes a useful tool. The aim of this study was to develop a technique for sexing by the Polymerase Chain Reaction (PCR) using DNA extracted from exfoliated colon cells present in faecal samples. Seventeen samples (from 10 females and 7 males) were collected from bears of known sex reared in captivity in four institutions in Lima Peru. DNA extraction was done by using a specific kit for faecal samples. The concentration of extracted DNA varied between 20 to 30 ng. Using PCR, two gene sequences were evaluated: amelogenin with primers SE47-SE48 and sex-determining region in the Y chromosome (SRY) with primers SRYB5-SRYB3...

El oso de anteojos (Tremarctos ornatus) se encuentra vulnerable debido a la caza y destrucción de su medio ambiente y es de difícil observación en vida libre, lo que obstaculiza su manejo e investigación, por lo que el uso de muestras no invasivas como heces se convierte en una gran herramienta. El objetivo del presente estudio fue adaptar una técnica de sexado mediante la reacción en cadena de la polimerasa (PCR) de ADN extraído de células exfoliadas del colon presentes en muestras de heces. Se utilizaron 17 muestras (10 de hembras y 7 de machos) de osos de sexo conocido y criados en cautiverio en cuatro instituciones de la zona de Lima, Perú. Para la extracción de ADN se utilizó un kit comercial específico para heces. Se logró extraer de 20 a 30 ng de ADN de cada muestra. Mediante PCR se evaluaron dos secuencias de genes: amelogenina con los cebadores SE47-SE48 y la regió...

Determination of sex in wild birds is crucial in developing conservation plans for threatened species. The absence of sexual dimorphism in birds makes impossible to determine sex based on physical characteristics and sexing has traditionally depended upon surgical methods which are highly stressful for the animals. The present study had the purpose of developing of a noninvasive DNA test for sex determination in macaws (guacamayos). Blood samples from five species (Ara ararauna, Ara chloroptera, Ara macao, Ara militaris y Propyrrhura couloni) were studied. DNA was extracted and specific primers were used to amplify both exons and introns of the chd (chromo-helicase- DNA-binding) gene located on the sex chromosomes of all birds. Females are identified by chd-ZW on the W chromosome and males by chd-ZZ on the Z chromosome. The amplification was carried out by the optimization of the conditi...

A case of ovine pulmonary adenocarcinoma is reported in a five month-old indigenous lamb of the department of Puno, Peru. At clinical inspection, the animal presented dyspnea, body weight loss and abundant lung secretion. The necropsy revealed in both diaphragmatic lobes the presence of two grayish white nodular formations with plenty of white foamy fluid emanating from the trachea. At the cut surface of the tumor was observed irregular micronodules with a tendency to coalesce. Histologically, the tumors were composed of columnar or cuboidal cells isolated by a fibrotic stroma and surrounded by alveoli flooded with macrophages of vacuolar cytoplasm. Intraluminal polyps were observed in the terminal bronchioles compromising the bronchiolar epithelium. Molecular analysis of lung tissue and fluid contained the exogenous genetic sequences of type D retrovirus. Retroviral infection with multi...

The aim of this study was to determine the allelic and genotypic frequency of kappacasein gene (κ-CN) in creole cattle. For this purpose, 48 blood samples were collected from animals in nine villages of the Bambamarca district, Cajamarca, Peru. DNA was extracted and genotyping was performed by PCR-RFLP analysis of κ-CN gene. Genotype frequencies of 0.56, 0.27 and 0.17 for the AB, AA and BB genotypes were found, while the allele frequencies were 0.552 and 0.448 for alleles A and B respectively. It was concluded that the tested group had a low proportion of individuals with the desired BB genotype that favors the production of cheese.

The aim of this study was to identify immunoprotective antigens in the genomic sequence of Mannheimia haemolytica serotype A2 isolated from sheep and available in databases that can be used as potential vaccine candidates against pneumonia in alpacas. To do this, the complete genome sequence of this organism available in databases was used and by using bioinformatics tools was proceeded to search for and identification of candidate genes, functional annotation, predicting the subcellular localization and identification of sequence motifs including transmembrane domains, secretion signal peptides and lipoproteins. Virulence factors were identified in the database of virulence factors (VFDB) and the microbial database of virulence factors (MvirDB). Selection criteria included conserved proteins, secreted and associated surface with transmembrane helices <1. Homology search of genes was ...

This paper reviews the existing information on the group of birds known as vultures, birds specialized in ingesting carrion, distributed in almost all continents and with similar physical characteristics and behaviors. The objective of the review is to present the global problems and threats to which this group of birds is subjected, mainly due to the anthropic effect, loss of their natural habitat, lead poisoning, and provoked or accidental poisoning, among other factors, that have caused their disappearance in various countries and have placed them on the brink of extinction. The current state of conservation and main threats of the two large groups, vultures of the Old and New World, are reviewed.

La determinación del sexo en aves de especies silvestres es de vital importancia para tomar medidas de conservación. En especies donde no existe dimorfismo sexual, es necesario contar con una prueba de sexaje alternativo a los métodos quirúrgicos. En este sentido, con la finalidad de estandarizar una prueba para la determinación del sexo en guacamayos, se desarrolló y evaluó una prueba molecular que consistió en el análisis de ADN mediante la Reacción en Cadena de la Polimerasa (PCR). Para ello, se procedió a la extracción de ADN a partir de muestras de sangre cinco especies de guacamayos (Ara ararauna, Ara chloroptera, Ara macao, Ara militaris y Propyrrhura couloni). A través de la técnica de PCR y el uso de primers específicos, se amplificó regiones conservadas (exones) y secuencias de regiones no conservadas (intrones) del gen chd (Chromodomainhelicase- DNA-binding pro...

A case of ovine pulmonary adenocarcinoma is reported in a five month-old indigenous lamb of the department of Puno, Peru. At clinical inspection, the animal presented dyspnea, body weight loss and abundant lung secretion. The necropsy revealed in both diaphragmatic lobes the presence of two grayish white nodular formations with plenty of white foamy fluid emanating from the trachea. At the cut surface of the tumor was observed irregular micronodules with a tendency to coalesce. Histologically, the tumors were composed of columnar or cuboidal cells isolated by a fibrotic stroma and surrounded by alveoli flooded with macrophages of vacuolar cytoplasm. Intraluminal polyps were observed in the terminal bronchioles compromising the bronchiolar epithelium. Molecular analysis of lung tissue and fluid contained the exogenous genetic sequences of type D retrovirus. Retroviral infection with multi...

El presente estudio tuvo como objetivo determinar la frecuencia alélica y genotípica del gen kappa-caseína (κ-CN) en ganado bovino criollo. Se recolectaron 48 muestras de sangre de bovinos de nueve centros poblados del distrito de Bambamarca (Cajamarca, Perú). Se extrajo el ADN y la genotipificación se realizó mediante análisis de RFLP-PCR del gen κ-CN. Las frecuencias genotípicas encontradas fueron de 0.56, 0.27 y 0.17 para los genotipos AB, AA y BB, respectivamente, en tanto que las frecuencias alélicas resultaron ser de 0.552 y 0.448 para los alelos A y B, respectivamente. Se concluye que los animales evaluados presentan una baja proporción de individuos con el genotipo deseado BB, genotipo que favorece la producción de queso.

El objetivo del presente trabajo fue identificar antígenos inmunoprotectivos en la secuencia genómica de Mannheimia haemolytica serotipo A2 Ovino, aislado de ovino y disponible en las bases de datos, que puedan ser utilizados como potenciales candidatos vacunales contra la neumonía en alpacas. Para ello, se empleó la secuencia completa del genoma de este microorganismo disponible en las bases de datos y mediante el uso de herramientas bioinformáticas se procedió a la búsqueda e identificación de genes candidatos, la anotación funcional, la predicción de la localización subcelular y la identificación de motivos de secuencias, incluyendo dominios transmembrana, peptidos señal de secreción y lipoproteínas. Los factores de virulencia fueron identificados en la base de datos de factores de virulencia (VFDB) y en la base de datos microbiana de virulencia (MvirDB). Los criterios ...

Gross lesions, histopathology and microbiological analysis of samples from 27weaned alpaca tuis from southern Peru that died from acute pneumonia are described. In situ viral antigens [Parainfluenza 3 (PI3), bovine respiratory syncytial (BRSB) and bovine adenovirus type 3 (BAdV-3)] were searched in 15 of the 27 samples by direct immunofluorescent test and bacteriology cultures were performed on all 27 samples. The gross alterations corresponded mostly to extensive multilobar pneumonia (11/27) associated with fibrinous pleuritis (8/27), and congestion and pulmonary edema (8/27). With the exception of peribronquial lymphocitic aggregates observed in one case, the histopathological changes were similar to those described in the literature for acute pneumonia in neonatal cases. In 9 out of 15 pneumonic tissues, 14 viral antigens were detected (5 as single, 3 dual and 1 triple infections). BR...

Gross lesions, histopathology and microbiological analysis of samples from 27weaned alpaca tuis from southern Peru that died from acute pneumonia are described. In situ viral antigens [Parainfluenza 3 (PI3), bovine respiratory syncytial (BRSB) and bovine adenovirus type 3 (BAdV-3)] were searched in 15 of the 27 samples by direct immunofluorescent test and bacteriology cultures were performed on all 27 samples. The gross alterations corresponded mostly to extensive multilobar pneumonia (11/27) associated with fibrinous pleuritis (8/27), and congestion and pulmonary edema (8/27). With the exception of peribronquial lymphocitic aggregates observed in one case, the histopathological changes were similar to those described in the literature for acute pneumonia in neonatal cases. In 9 out of 15 pneumonic tissues, 14 viral antigens were detected (5 as single, 3 dual and 1 triple infections). BR...

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 objective of this study was to develop a PCR technique to determine the sex of South American camelids (CSA) using Zinc Finger Protein (ZF) sequences from blood and fecal samples, as well as cells from alpaca embryos. A total of 28 alpaca, llama and vicuña blood samples, 20 vicuña and guanaco fecal samples, and 22 alpaca embryos collected between 72 and 96 hours postcopula were used. The fecal and embryo samples were preserved in 96% and 70% ethanol respectively. DNA was extracted from blood and feces using commercial kits. Three methods (boiling, proteinase K and phenol-cloroform) were used to extract DNA from alpaca embryos. Two PCR techniques were developed to analyze DNA: multiplex (for fecal and blood sample DNA) and heminested PCR (for embryo cell DNA). The multiplex PCR accurately determined the sex in 100% of the DNA samples extracted from blood, in 87.5% of the samples ext...

The aim of this study was to establish the frequency of resistance and susceptible genotypes to scrapie in the Junin sheep breed. For this, 56 blood samples were collected in a sheep farm in the central highlands of Peru. The DNA was extracted and the PCR products were analyzed by enzymatic digestion using restriction endonucleases that recognize the PrP gene polymorphisms. Four of the 5 haplotypes reported for the PrP gene were identified: ARR (48.2%), ARQ (35.7%), AHQ (10.7%) and VRQ (5.4%). In addition, 7 of the 15 reported genotypes for the Prion gene were found: ARR/ARR (17.9%), ARR/AHQ (10.7%), ARR/ARQ (42.9%), ARQ/AHQ (10.7%), ARQ/ARQ (7.1%), ARR/VRQ (7.1%) and ARQ/VRQ (3.6%). Finally, 82.1% of the animals had genotypes resistant to the disease and 17.9% showed genotypes of susceptibility.

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...

Se reporta el desarrollo y optimizaciones de técnicas moleculares (PCR simple, múltiple y semi-anidada) para determinar el sexo de camélidos sudamericanos (CSA) amplificando la secuencia del gen Zinc Finger Protein (ZF). La técnica utilizó ADN obtenido de 28 muestras de sangre de alpacas, llamas y vicuñas, 20 muestras de heces de vicuñas y guanacos conservadas en etanol al 96%, y 22 embriones de alpaca colectados entre 72 y 96 horas postmonta y preservados en etanol. Las muestras de ADN de sangre y heces fueron extraídas usando kits comerciales, y las de embriones aplicando tres métodos (ebullición, proteinasa K y fenol-cloroformo). Una vez optimizada la PCR simple para la detección de los genes ZFY y ZFX, se implementó la PCR múltiple para ADN de sangre y heces y la PCR semi-anidada para ADN de embriones. La técnica de PCR múltiple determinó el sexo correctamente en el 1...