COMPOSICIÓN NUTRICIONAL DE LA CARNE DE GUANACO DE LA PATAGONIA ARGENTINA / NUTRITIONAL COMPOSITION OF GUANACO MEAT FROM PATAGONIA, ARGENTINA

Introducción: Se considera a la carne de guanaco y sus derivados un producto cárnico consumible (Art. 261, CAA). El objetivo fue evaluar los aspectos nutricionales en cortes de uso comercial (lomo y peceto) de la carne de guanaco (Lama guanicoe guanicoe), del departamento Escalante (Chubut, Argentina) durante los años 2022 y 2023. Materiales y métodos: Se obtuvieron las muestras de 10 ejemplares machos adultos. Se determinó humedad (AOAC 950.46), proteínas (AOAC 928.08), grasas totales (AOAC 960.39), cenizas (AOAC 920.153), carbohidratos (cálculo por diferencia) y colesterol (Wiener lab®). Resultados: La composición centesimal de lomo y peceto, expresada en g/100 g en base húmeda, y el contenido de colesterol, expresado en mg/100 g en base húmeda, para el año 2022, en animales faenados en la estancia La Paulina, fue: humedad 75,1±0,13 y 5,2±0,06; proteínas 22,2±0,23 y 22,3±0,18; grasas totales 0,69±0,01 y 0,68±0,02; cenizas 0,85±0,03 y 0,88±0,04; carbohidratos 1,14 y 0,95; y colesterol 58,9±4,52 y 64,4±4,20, respectivamente. No se observaron diferencias estadísticas entre la composición de ambos cortes (p>0,05). La composición centesimal de lomo y peceto, expresada en g/100 g en base húmeda, y el contenido de colesterol, expresado en mg/100 g en base húmeda, para el año 2023 fue: humedad 76,6±0,14 y 76,7±0,17; proteínas 20,6±0,21 y 20,5±0,25; grasas totales 0,58±0,03 y 0,53±0,03; cenizas 1,07±0,04 y 1,02±0,02; carbohidratos 1,21 y 1,19; y colesterol 43,5±3,17 y 37,8±2,10, respectivamente. Se observó un mayor contenido de grasa en el corte lomo (p<0,05), mientras que no se observaron diferencias estadísticas (p>0,05) para el resto de los nutrientes. Conclusiones: El guanaco es una especie abundante y silvestre, muy atractiva para el consumo humano por sus cualidades nutritivas. Este alimento autóctono representa una opción alimentaria regional, con un potencial posicionamiento en el mercado nacional e internacional como una carne exótica. La carne de guanaco presenta un contenido de proteínas ligeramente mayor, un porcentaje de grasas totales significativamente menor y un nivel de colesterol ligeramente inferior, respecto a otras carnes rojas de consumo habitual

Introduction: Guanaco meat and its derivatives are considered a consumable meat product (Art. 261, CAA). The objective was to evaluate the nutritional aspects of commercially used cuts (loin and round) of guanaco meat (Lama guanicoe guanicoe) from the Escalante department (Chubut, Argentina) during the years 2022 and 2023. Materials and Methods: Samples were obtained from 10 adult male specimens. Moisture (AOAC 950.46), protein (AOAC 928.08), total fat (AOAC 960.39), ash (AOAC 920.153), carbohydrates (calculated by difference), and cholesterol (Wiener lab®) were determined. Results: The proximate composition of loin and round, expressed as g/100 g on a wet weight basis, and the cholesterol content, expressed as mg/100 g on a wet weight basis, for the year 2022, in animals slaughtered at La Paulina ranch, was as follows: moisture 75.1±0.13 and 75.2±0.06; protein 22.2±0.23 and 22.3±0.18; total fat 0.69±0.01 and 0.68±0.02; ash 0.85±0.03 and 0.88±0.04; carbohydrates 1.14 and 0.95; and cholesterol 58.9±4.52 and 64.4±4.20, respectively. No statistical differences were observed between the composition of both cuts (p>0.05). The proximate composition of loin and round, expressed as g/100 g on a wet weight basis, and the cholesterol content, expressed as mg/100 g on a wet weight basis, for the year 2023 was as follows: moisture 76.6±0.14 and 76.7±0.17; protein 20.6±0.21 and 20.5±0.25; total fat 0.58±0.03 and 0.53±0.03; ash 1.07±0.04 and 1.02±0.02; carbohydrates 1.21 and 1.19; and cholesterol 43.5±3.17 and 37.8±2.10, respectively. A higher fat content was observed in the loin cut (p<0.05), while no statistically significant differences were observed (p>0.05) for the rest of the nutrients. Conclusions: The guanaco is an abundant and wild species, highly attractive for human consumption due to its nutritional qualities. This native food represents a regional dietary option, with potential positioning in the national and international markets as an exotic meat. Guanaco meat has a slightly higher protein content, significantly lower total fat percentage, and a slightly lower cholesterol level compared to other commonly consumed red meats

Preliminary Study on Screening and Identification of Lewis a Antigen Mimic Epitope in Alpaca Phage Display Nanobody Library / 中国实验血液学杂志

OBJECTIVE@#To establish a new method for synthesizing Lewis blood group antigens, that is, the mimotopes of Lewis blood group antigens were screened by using an alpaca phage display nanobody library.@*METHODS@#We selected mimotopes of the Lewis a (lea) antigen by affinity panning of an alpaca phage display nanobody library using a monoclonal anti-lea antibody. Enzyme-linked immunosorbent assay (ELISA) was used to test the affinity of the positive clones for the monoclonal anti-lea antibody, and the high-affinity positive clones were selected for sequencing and synthesis. Finally, the sensitivity, specificity and reactivity of the synthesized lea mimotope in clinical samples were verified by ELISA.@*RESULTS@#A total of 96 phage clones were randomly selected, and 24 were positive. Fourteen positive clones with the highest affinity were selected for sequencing. The result showed that there were 5 different sequences, among which 3 sequences with the highest frequency, largest difference and highest affinity were selected for expression and synthesis. The sensitivity and specificity of lea mimic antigen by ELISA showed that, the minimum detection limit of gel microcolumn assay (GMA) and ELISA method were 25 times different, and the lea mimic antigen had no cross reacted with the other five unrelated monoclonal antibodies(P<0.001). Finally, 30 clinical plasma samples were analyzed. The mean absorbance of the 15 positive plasma samples was significantly higher than that of the 15 negative plasma samples (P=0.02). However, the positive signal values of the clinical samples were much lower than those of the monoclonal antibodies.@*CONCLUSION@#A new method of screening lea mimic antigen by using alpaca phage nanoantibody library has been established, which is expected to realize the screening of lea mimotopes, thus realizing the application of high-sensitivity detection methods such as ELISA and chemiluminescence in blood group antibody identification.

Estudio morfométrico y ecuaciones de predicción del peso corporal en llamas (Lama glama) Ch'aku y Q'ara / Morphometric study and body weight prediction equations in llamas (Lama glama) Ch'aku and Q'ara

RESUMEN: En el estudio se utilizó una población de 180 llamas de ambos sexos (machos y hembras) clasificados por edades (dientes de leche; dos dientes y cuatro dientes). Se registraron datos del peso corporal y las siguientes medidas morfométricas: En la cabeza: Longitud de la cabeza (LC); ancho de cabeza (ACa) y longitud de oreja (LO). En el cuello: Longitud del cuello (LCU); Perímetro de cuello anterior (PCuA) y perímetro de cuello posterior (PCuP). En el tronco y extremidades: Altura a la cruz (AC); altura de la grupa (AGRU), diámetro dorso esternal (DDE), perímetro torácico (PT), longitud del cuerpo (LOCU), longitud de la cruz a la grupa (LCG), longitud de grupa (LGRU), perímetro de caña anterior (PCA); perímetro de caña posterior (PCP). Los datos fueron procesados utilizando el software estadístico R. Concluyendo que los rasgos de conformación en llamas (Lama glama) Ch'aku y Q'ara están influenciados por la edad, sexo y raza. Los modelos de regresión simple y múltiple para la predicción del peso corporal fueron significativos.

SUMMARY: The study used a population of 180 llamas of both sexes (males and females) classified by age (milk teeth; two teeth and four teeth). Body weight data and the following morphometric measurements were recorded: Head: Head length (CL); head width (ACa) and ear length (LO). On the neck: Neck length (LCU); anterior neck circumference (PCuA) and posterior neck circumference (PCuP). On the trunk and limbs: Height at withers (AC); height at rump (AGRU), sternal dorsal diameter (DDE), thoracic perimeter (PT), body length (LOCU), length from withers to rump (LCG), rump length (LGRU), anterior canine perimeter (PCA); posterior canine perimeter (PCP). The data were processed using R statistical software. It was concluded that conformation traits in Ch'aku and Q'ara llamas (Lama glama) are influenced by age, sex and breed. Simple and multiple regression models for body weight prediction were significant.

Exportación y sanitación internacional en empresas productoras de prendas de alpaca de Huancayo y Lima / Exportation and international sanitation in companies producing alpaca garments in Huancayo and Lima

El Perú se considera un país potencia en cuanto a la exportación textil de la alpaca, cuya fibra natural es muy fina, ligera y con alto valor a nivel mundial. Para el año 2019, la exportación total de fibra de alpaca en Perú llegó a un total de 23 países, incluyendo mercados en Europa, Asia y Norteamérica. La tendencia en el comercio exterior incluyendo las empresas textiles de alpaca, se ha dirigido a garantizar el cuidado del medio ambiente y la inocuidad de los productos respecto a la salud humana. Se realizó un estudio desde el paradigma post-positivista, con enfoque cualitativo y el método hermenéutico, para interpretar la situación de exportación y sanitación internacional en empresas peruanas productoras de prendas de alpaca en Huancayo y Lima. Se obtuvo información a partir de cuatro informantes voluntarios de cada empresa acerca de aspectos relacionados con exportación de las prendas de alpaca: competitividad, promoción, proveedores, producción, calidad de producto, valor agregado, calidad de servicio, mercados destino, demanda y herramientas de inteligencia comercial, entre otros. Asimismo, se indagó sobre la concepción y cumplimiento del Reglamento Sanitario Internacional. Las empresas evaluadas, emplean excelente materia prima, garantizan un control de calidad de sus productos ofreciendo textiles competitivos que se distribuyen en diversos mercados internacionales, emplean herramientas de comercialización que favorecen la identificación de regulaciones arancelarias y la capacitación de su personal. No obstante, muestran carencias en el cumplimiento del RSI, requiriendo adoptar medidas que garanticen la salud pública al comercializar sus productos(AU)

Peru is considered a powerful country in terms of alpaca textile exports, whose natural fiber is very fine, light and highly valued worldwide. For the year 2019, the total export of alpaca fiber in Peru reached a total of 23 countries, including markets in Europe, Asia and North America. The trend in foreign trade, including alpaca textile companies, has been aimed at guaranteeing care for the environment and the safety of products with respect to human health. A study was carried out from the post-positivist paradigm, with a qualitative approach and the hermeneutical method, to interpret the export situation and international sanitation in Peruvian companies that produce alpaca garments in Huancayo and Lima. Information was obtained from four volunteer informants from each company about aspects related to the export of alpaca garments: competitiveness, promotion, suppliers, production, product quality, added value, service quality, destination markets, demand and marketing tools. commercial intelligence, among others. Likewise, the conception and compliance with the International Health Regulations were inquired about. The evaluated companies use excellent raw material, guarantee quality control of their products by offering competitive textiles that are distributed in various international markets, use marketing tools that favor the identification of tariff regulations and the training of their personnel. However, they show deficiencies in compliance with the RSI, requiring the adoption of measures that guarantee public health when marketing their products(AU)

Learning from the llama: on the broad contours of cultural contributions and geographic expansion / Aprendendo com a lhama: sobre os amplos contornos de contribuições culturais e expansão geográfica

Abstract The llama (Lama glama) is the largest domesticated animal species from South America and is today found worldwide. Andean peoples have used the llama for millennia for meat, wool, packing, spiritual etc. In order to know the history of the llama, we must learn about the peoples that have known the animal and the ways those relationships have changed over time. While also considering closely related species, including alpaca, guanaco, and vicuña, this article posits three eras of llama/human entanglements: the era of domestication in pre-Columbian Andean sites; the era of dispersal and co-mingling, from 1530s to the 1890s; and finally popular fads and global appeal.

Resumo A lhama (Lama glama) é a maior espécie animal domesticada da América do Sul e atualmente é encontrada em todo o mundo. Os povos andinos a utilizam há milênios para transporte de carga, obtenção de lã e carne, uso espiritual etc. Para conhecer sua história, precisamos aprender sobre os povos que conhecem e se relacionam com esse animal, e como esses relacionamentos se transformaram ao longo do tempo. O artigo considera ainda outras espécies relacionadas, incluindo alpaca, guanaco e vicunha, e apresenta três eras da interação lhamas/humanos: a da domesticação em sítios andinos pré-colombianos; a da dispersão e mistura, dos anos 1530 à década de 1890; e, finalmente, modismos e interesse global.

Neutralización de la actividad letal del veneno de serpiente Bothrops atrox por suero hiperinmune de llama (Lama glama) / Neutralization of the lethal activity from Bothrops atrox venom by hyperimune llama serum (Lama glama)

RESUMEN Objetivos: Evaluar la capacidad del suero hiperinmune de llama (Lama glama) para neutralizar la letalidad del veneno de la serpiente Bothrops atrox en ratones de laboratorio. Materiales y métodos: Se calculó la dosis letal media (DL50) de un pool de venenos de serpientes de Bothrops atrox de Perú, y se midieron los títulos de anticuerpos por ensayo ELISA; así como la potencia de neutralización del suero inmune por el cálculo de la dosis efectiva media (DE50) durante el periodo de inmunización. Resultados: La DL50 del veneno fue de 3,96 µg/g, similar a otros trabajos realizados en Bothrops atrox en Perú. Los títulos de anticuerpos contra el veneno se incrementan rápidamente en la llama mostrando una rápida respuesta inmune; sin embargo, la capacidad de neutralización se incrementa más lentamente y requiere de varias dosis y refuerzos de las inmunizaciones alcanzado una DE50 de 3,30 µL/g ratón y una potencia de neutralización 3,6 mg/mL después de 15 inmunizaciones. Conclusiones: El suero hiperinmune de llama es capaz de neutralizar la letalidad del veneno de la serpiente Bothrops atrox de Perú en ratones de laboratorio.

ABSTRACT Objectives: To evaluate the capacity of the hyperimmune llama serum (Lama glama) to neutralize the lethal activity of Bothrops atrox venom in laboratory mice. Materials and methods: Mean lethal dose (LD50) was calculated from a Bothrops atrox venom sample pool from Peru. The antibody titers were measured by ELISA assay; and the immune serum neutralization potency was measured by calculating the mean effective dose (ED50) during the immunization period. Results: The venom's LD50 was 3.96 μg/g; similar to what was found in other studies about Bothrops atrox carried out in Peru. The titers of antibodies against the venom increased rapidly in the llama, demonstrating a fast immune response; however, the neutralization capacity increased slowly and required several doses and immunization reinforcements, obtaining a ED50 of 3.30 μL/g mouse and a neutralization potency of 3.6 mg/mL after 15 immunizations. Conclusions: The hyperimmune llama serum is able to neutralize the lethality of the Bothrops atrox venom from Peru in laboratory mice.

Características fisicoquímicas, mecánicas y sensoriales de salchichas secas tipo cabanossi elaboradas con carne de llama (Lama glama) y cerdo (Sus scrofa domestica) / Physicochemical, mechanical and sensory characteristics of cabanossi-type dry sausages made with llama (Lama glama) and pork (Sus scrofa domestica) meat

RESUMEN Se estableció la caracterización de salchichas secas tipo cabanossi con carne de llama, cerdo y muestras comerciales mediante un análisis proximal (contenidos de grasa: 18,6 a 29,6% y proteína: 19,4 a 36,2%), color (C*: 15,8 a 33,7), actividad de agua (0,770 a 0,960), pH (5,2 a 6,5) y propiedades mecánicas como dureza (47,8 a 124 N) y masticabilidad (9,4 a 33,2 N). Se obtuvieron 19 descriptores a partir de la opinión de 83 consumidores; luego, se empleó el método CATA (Check-all-that-apply) para describir las características sensoriales del cabanossi empleando 55 consumidores. Se evidenció que el cabanossi con carne de llama presentó el menor contenido de grasa total y ácidos grasos saturados, y mayor contenido de ácidos grasos poliinsaturados en comparación a las otras muestras. Los descriptores más utilizados fueron: olor característico/embutido, sabor a especias/condimentos, picante y sensación residual picante. Los consumidores indicaron como atributos relevantes para la aceptabilidad al color rojo claro, blando/suave, masticable y picante. La utilización de carne de llama constituye una alternativa en el desarrollo de nuevos productos cárnicos basados sobre las características presentadas.

ABSTRACT The chemical and physical characterization of cabanossi-type dry sausages made with lama meat, pork and commercial samples were performed. Remarkable differences were found in terms of fat (18.6 to 29.6%), color (15.8 to 33.7), water activity (0.770 to 0.960), pH (5.6 to 6.5), hardness (47.8 to 124 N) and chewiness (9.4 to 33.2 N). Nineteen descriptive terms based on the opinions of 83 consumers were used. Then, 55 consumers used the CATA (Check-all-that-apply) method to describe the sensory traits of cabanossi . Sausage from lama meat had the lowest total fat and saturated fatty acid content, and the highest polyunsaturated fatty acid content in comparison to the other samples. The most utilized sensory terms were: characteristic odor/smell of sausage, spicy flavor and spicy after taste. For sensory acceptability, consumers selected light red color, soft/tender, chewable and spicy as the most relevant traits. The application of lama meat represents an alternative to new meat products developed based on the characteristics studied.

A key for hypoxia genetic adaptation in alpaca could be a HIF1A truncated bHLH protein domain / Una clave para la adaptacion genética de la hypoxia en las alpacas podria ser un dominio de proteina bHLH truncado con HIF1A

Animals exposed to hypobaric hypoxia triggers a physiological hypoxia response via Hypoxia Inducible Factor (HIF) proteins that functions as transcriptional complexes. As the South American camelids inhabit at high Andean altitudes we have asked if they have developed genetic adaptations to live at high altitudes. In the present study we investigate genetic structures of the HIF1A proteins carried by members of the superorder Cetartiodactyla. During our investigation we discovered the existence of a genetic event that caused the loss of most of the bHLH domain in the proteins borne by the Alpaca and other members of the Cetartiodactyla superorder; we designate them as bHLH short sequences. Further analysis at the nucleotide level revealed in the 12 short sequences included in the study the presence at the 5´end of the bHLH domains stop codons. Seven out of the 12 short HIF1A proteins, have an identical or almost identical nucleotide sequence at their 5´end with a same TAA stop codon and at the same position. As the mutations affects to both the Artiodactyls and Cetaceans, we postulate that the mutation(s) occurred before their divergence about 55 million years ago. The relevance of these findings for genetic adaptation of Alpacas to hypobaric hypoxia of high altitude conditions is discussed.

Los animales expuestos a hypoxia hipobárica generan una respuesta hipóxica fisiológica debido a unas proteinas de Factor-Hipoxia Inducible (HIF) que funcionan como complejos transcripcionales. Debido a que los camelidos Americanos habitan en las grandes alturas andinas, nos hemos preguntado si han desarrollado una adaptación genética para vivir a grandes alturas. Eneste estudio hemos investigado la estructura genética de las proteinas HIF1A que llevan consigo los miembros de la superorden de los cetartiodáctilos. Durante nuestra investigación, descubrimos la existencia de un evento genético que causó la perdida de la mayoría del dominio bHLH en las proteinas transmitidas por la alpaca y otros miembros de la superorden de los cetartiodáctilos; las hemos designado como secuencias cortas de bHLH. Análisis posteriores a nivel nucleótido revelaron que en la doceava secuencia corta incluida en el studio, hubo presencia de codones de terminación en el extreme 5' del dominio de bHLH. Siete de las doce proteinas cortas HIF1A, tiene una secuencia idéntica o casi idéntica de nucleotidos en su extremo 5', con el mismo codón de terminación TAA y en la misma posición. Debido a que la mutación afecta tanto a Artiodáctilos como Cetáceos, proponemos que la mutación(es) ocurrió antes de su divergencia hace unos 55 millones de años. Analizamos la relevancia de estos descubrimientos sobre la adaptación genética de las alpacas a la hipoxia hipobárica en condiciones de grandes alturas.

First characterization of preantral follicles in the ovaries of adult alpaca / Primera caracterización de folículos preantrales de ovario de alpaca adulta

It is a fact that while even basic reproductive information on alpacas is unavailable, the normal ovarian reserve of this species in comparison to other species is also unidentified. In this study, the ovarian preantral follicles in healthy adult alpacas were characterized in order to establish a general model to in vitro studies. Ten ovaries were collected from five adult alpacas. The ovarian cortex samples were fixed with paraformaldehyde and histological analysis was done. Normal and degenerated follicles percentages were determined. The normal follicles were measured and classified in primordial, transitional, primary and secondary stages. Most of the preantral follicles present in the ovarian cortex of alpacas were primordial and transitional stages; primary (6.10%) and secondary (0.37%) follicles were rarely found. The primary and secondary follicles were larger in diameter when compared with the primordial and transitional follicles. The largest oocyte diameter was recorded in the secondary follicles (P < 0.05). This study serves to establish a biological model for future reproduction studies in Alpacas or as possible biological model for studies of folliculogenesis in humans(AU)

Es un hecho que, si bien no se dispone de información reproductiva básica sobre las alpacas, la reserva ovárica normal de esta especie en comparación con otras especies tampoco está identificada. En este estudio, se caracterizaron los folículos preantrales ováricos en alpacas adultas sanas. Se recogieron diez ovarios de cinco alpacas adultas. Las muestras de la corteza ovárica se fijaron con paraformaldehído y se realizó un análisis histológico. Se determinaron los porcentajes de folículos normales y degenerados. Los folículos normales se midieron y clasificaron en estadios: primordiales, de transición, primarios y secundarios. La mayoría de los folículos preantrales presentes en la corteza ovárica de las alpacas eran estadios primordiales y de transición; Raras veces se encontraron folículos primarios (6.10%) y secundarios (0.37%). Los folículos primarios y secundarios tenían un diámetro mayor en comparación con los folículos primordiales y de transición. El mayor diámetro de ovocitos se registró en los folículos secundarios (P <0.05). Este estudio sirve para establecer un modelo biológico para futuros estudios de reproducción en alpacas o como posible modelo biológico para estudios de foliculogénesis en humanos(AU)

Myocardial Infarction Associated with Anomalous Origin of the Left Coronary Artery from the Pulmonary Artery and Double Right Coronary Artery / 대한내과학회지

Anomalous origin of the left coronary artery from the pulmonary artery (ALCAPA) is a rare coronary artery anomaly and double right coronary artery (RCA) is a very rare coronary anomaly. Because patients with ALCAPA usually die within 1 year of being born due to myocardial infarction (MI) and heart failure, ALPACA is very rarely seen in adults. Here, we report an extremely rare asymptomatic case of MI, presumably caused by ALCAPA and double RCA, and provide a review of the literature. This is the first reported case of coronary artery anomaly that had both ALCAPA and double RCA.

Anticuerpos de cadena única de alpaca para la detección de antígenos de Fasciola hepática / Single-chain antibodies from alpaca for the detection of Fasciola hepatica antigens

RESUMEN Objetivo. Producir anticuerpos recombinantes de cadena única de alpaca que se unan con alta afinidad y especificidad al antígeno excretado-secretado (ES) de Fasciola hepatica para el desarrollo de tecnologías nuevas de diagnóstico de fascioliasis humana y animal. Materiales y métodos. Se ha construido una genoteca de cADNde los dominios variables de anticuerpos de cadena única pesada, conocidos como VHH, a partir de células mononucleares de sangre periférica de una alpaca inmunizada con el antígeno ES de F. hepatica. La genoteca fue tamizada con el antígeno ES por despliegue diferencial de fagos (phage display), seleccionando diez VHH que se unen específicamente a ES. El VHH anti ES fue clonado en un vector de expresión, la proteína recombinante (VHH-ES1) de 15,3 kDa fue producida por fermentación en E. coli y purificada a homogeneidad por cromatografía de afinidad. La unión del VHH-ES1 al antígeno ES fue evaluada por ELISA usando VHH-ES1 como anticuerpo de captura, antisuero policlonal anti-ES de conejo y conjugado anti IgG de conejo con peróxidasa de rábano. Resultados. Se ha identificado y producido un VHH-ES1 recombinante que se une al antígeno ES (VHH-ES1) que correspondía a un anticuerpo de la subclase IgG2 de bisagra larga. La unión del anticuerpo VHH-ES1 al antígeno muestra linealidad respecto a la concentración de ES en el rango de 50-5000 ng/mL y el valor límite de detección del antígeno está en el rango de 30-170 ng/mL de ES (R2=0,99). Conclusión . El VHH-ES1 se une con afinidad y especificidad al antígeno ES de F. hepatica y es un anticuerpo promisorio a evaluar para el desarrollo de nuevas tecnologías de diagnóstico de fascioliasis.

ABSTRACT Objectives. To produce recombinant single-chain antibodies from alpaca that will bind to the excreted-secreted (ES) Fasciola hepatica antigen with high affinity and specificity, so as to develop new diagnostic technologies of human and animal fascioliasis. Materials and Methods. A gene bank of DNA of the variable dominions of heavy single-chain antibodies (VHH) has been created, based on mononuclear cells of peripheral blood of an alpaca immunized with the ES antigen of F. hepatica. The gene bank was screened with the ES antigen by differential phage display, selecting ten VHH that bind specifically to ES. The anti-ES VHH was cloned in an expression vector, the recombinant protein (VHH-ES1) of 15.3 kDa was produced by fermentation in E. coli and purified to homogeneity by affinity chromatography. The binding of VHH-ES1 to the ES antigen was evaluated by ELISA using VHH-ES1 as capture antibody, policlonal anti-ES serum of rabbit and conjugated rabbit anti IgG with radish peroxidase. Results. A VHH that binds to the ES antigen (VHH-ES1) has been identified through differential phage display and produced by fermentation in E. coli; this corresponds to an antibody of the long-hinge IgG2 subclass. The binding of the VHH-ES1 antibody to the antigen shows linearity with respect to the concentration of ES in the 50-5,000 ng/mL range and the limit of detection value of the antigen is in the 30-170 ng/mL range of ES (R2=0.99). Conclusions. The VHH-ES1 binds with affinity and specificity to the ES antigen of F. hepatica and is a promissory antibody to be assessed for the development of new fascioliasis diagnostic technologies.

Aspectos patológicos da intoxicação por aceturato de diminazeno em camelídeos sul-americanos / Pathological aspects of diminazene aceturate toxicosis in american camelids

Aceturato de diminazeno é um fármaco quimioterápico sintético comumente usado na medicina veterinária para o tratamento de doenças causadas por parasitos hematozoários. Entretanto, seu uso pode levar a efeitos colaterais, como alterações neurológicas graves e morte. A criação de camelídeos é uma atividade recente no Brasil, fazendo-se necessário conhecer mais sobre as doenças que acometem essas espécies. De dez camelídeos (seis lhamas e quatro alpacas) da propriedade, seis tiveram sinais clínicos e, destes, apenas uma lhama com manifestações leves recuperou-se. Os sinais clínicos incluíam apatia, andar cambaleante, fraqueza, sialorreia, cabeça baixa e pendida lateralmente, dificuldade em levantar e dispneia, observados a partir de 18 horas após o uso do medicamento. À necropsia e ao exame histopatológico foram observadas alterações de encefalopatia hemorrágica bilateral e simétrica, mais graves em tronco encefálico e tálamo. Este trabalho descreve as principais lesões observadas em um surto de intoxicação por diminazeno em alpacas (Lama pacos) e lhamas (Lama glama) e alerta criadores e veterinários sobre o risco de intoxicação por aceturato de diminazeno em camelídeos sul americanos.(AU)

Diminazene aceturate is a synthetic chemotherapeutic drug commonly used in veterinary medicine for the treatment of diseases caused by hematozoan parasites. However, side effects as severe neurological disorders and death can occur. The raising of american camelids is a recent activity in Brazil, requiring knowledge about diseases that affect these species, in order to avoid misguided conducts. In a herd of ten camelids (six llamas and four alpacas) six showed clinical signs and five died; only a llama with mild signs recovered. The clinical signs included apathy, difficulty to stand up, staggering gait, weakness, down head and drooping the head laterally, dyspnea and drooling of saliva, observed from 18 hours after use of the drug. At necropsy and histopathological examination was found bilateral and symmetrical hemorrhagic encephalopathy, more severe in brainstem and thalamus. This paper describes the main lesions observed in an outbreak of diminazene aceturate poisoning in alpacas (Lama pacos) and llamas (Lama glama) and alert breeders and veterinarians about the risk of poisoning by this drug in american camelids.(AU)

Segment­ specific expression of MMP/TIMP in the oviduct of llama (Lama glama) and gelatinolytic activity in the oviductal fluid / Expresión segmento específica de MMP/TIMP en el oviducto de llama (Lama glama) y actividad gelatinolítica en el fluido oviductal

Oviductal molecules have the potential to improve the reproductive biotechnologies. In camelids, knowledge and assessment of the oviductal environment are necessary to successfully develop species-specific reproductive technologies, especially because of the camelids reproductive particularities. Among the oviductal factors, the matrix metalloproteinases/tissue inhibitor of matrix metalloproteinases system (MMPs/TIMPs) should be investigated more thoroughly due to their participation in reproductive processes. Consequently, the current study assayed gene and protein expression of MMPs throughout the llama oviduct. MMPs zymogen and active forms in the oviductal fluid were also characterized. MMP2 and MMP9 transcripts were detected in ampulla, isthmus, utero-tubal junction and papilla, being MMP2 and MMP9 2.15 and 1.10 folds higher in papilla than in ampulla, respectively. In addition, differences in immunolocalization of MMP2 and MMP9 between the epithelial mucosa layers of the oviductal segments were observed. The presence of MMPs in the epithelium suggests their secretion into the oviductal lumen. Coincidently, bands of 62 and 94 kDa, corresponding to MMP2 and MMP9 were detected by zymography in the oviductal fluid. Treatment with an exogenous activator (APMA) suggests that they are present as proMMPs. TIMP2 and TIMP1, the specific inhibitors of MMP2 and MMP9, respectively, were expressed in each oviductal segment, indicating a well-regulated control of MMP proteolytic activity in the oviduct. These findings prove that the llama oviduct produces and secretes MMPs into the oviductal lumen, suggesting that these enzymes may have an unknown role in the preparation of the oviductal environment for gametes, fertilization and early embryo development in camelids.

Las moléculas oviductales tienen el potencial para mejorar las biotecnologías reproductivas. En los camélidos, debido a sus peculiares características reproductivas, el conocimiento del ambiente oviductal constituye una herramienta útil para el desarrollo de tecnologías reproductivas específicas para estas especies. Entre los factores oviductales de interés se encuentran las metaloproteasas de matriz (MMPs) y sus inhibidores específicos (TIMPs), los cuales han sido involucrados en diferentes procesos reproductivos. Por estas razones, en este trabajo se caracterizó la expresión génica y proteica de MMP2 y MMP9 en el oviducto de llama. Además, se analizó la presencia de las formas activas e inactivas (zimógenos) de estas enzimas en el fluido oviductal. Se observó que todos los segmentos oviductales, ámpula, istmo, unión útero-tubal y papila, expresan MMP2 y MMP9, siendo los niveles de expresión de MMP2 y MMP9 más elevados en papila respecto a ámpula; 2,15 y 1,10 veces respectivamente. Asimismo, se observaron diferencias en la distribución de las MMPs a nivel de la mucosa entre los segmentos oviductales. Consecuentemente, bandas con actividad gelatinolítica de 62 y 94 kDa, se detectaron en el fluido oviductal, las cuales corresponderían a las formas inactivas de la MMP2 y la MMP9, respectivamente. Los inhibidores específicos de MMP2 y MMP9; TIMP2 y TIMP1, también se detectaron en los segmentos oviductales, indicando su probable participación en la regulación de la actividad proteolítica de las MMPs en el oviducto de llama. En conjunto, los datos de este trabajo demuestran que el oviducto de la llama produce y secreta MMPs al lumen oviductal; sugiriendo que estas enzimas pueden participar en la preparación del ambiente oviductal para la recepción de los gametos, la fecundación y el desarrollo embrionario temprano en camélidos.

Infecção por Mycobacterium sp. em herbívoros selvagens de cativeiro no Rio Grande do Sul: estudo retrospectivo e detecção imuno-histoquímica (2003-2015) / Mycobacterium sp. in captive-reared wild herbivores in Rio Grande do Sul: a retrospective study and immunohistochemical detection (2003-2015)

A tuberculose é uma enfermidade infectocontagiosa, debilitante, causada por bacilos álcool-ácido resistentes (BAAR), pertencentes ao complexo Mycobacterium tuberculosis (CMT). As micobacterioses têm importância em Medicina Veterinária devido ao seu potencial zoonótico e sua distribuição mundial, afetando todas as classes de vertebrados. Em animais selvagens as micobacterioses têm sido um problema relatado principalmente em cativeiro. Contudo, há relatos de sua ocorrência também em animais de vida livre, colocando em risco e dificultando os programas de erradicação da tuberculose em animais de produção. O diagnóstico nas espécies selvagens em geral é post mortem, uma vez que o teste de tuberculina não está padronizado para essas espécies, assim como não é confiável para triagem. São consideradas para o diagnóstico lesões de necropsia, observação microscópica de BAAR na coloração de Ziehl-Neelsen (ZN) e, principalmente, isolamento e identificação do agente. No entanto, apenas os achados morfológicos macro e microscópicos não permitem distinguir a espécie de Mycobacterium envolvida. A técnica de imuno-histoquímica (IHQ) com anticorpo policlonal anti-M. tuberculosis confirma a infecção pelo CMT, mas não é específica, pois pode ocorrer marcação de outras micobactérias. As características histológicas, os achados na coloração de ZN e na IHQ de 13 casos de herbívoros selvagens diagnosticados com tuberculose no Setor de Patologia Veterinária da Universidade Federal do Rio Grande do Sul (SPV-UFRGS) no período de 2003 a 2015 são descritos. A partir das amostras em blocos de parafina foram confeccionadas novas lâminas histológicas, coradas com hematoxilina e eosina (HE) e ZN. Cortes foram submetidos à técnica de IHQ para detecção do CMT. Todos os animais eram adultos, provenientes de cativeiro e incluíram lhama (5/13), cervo sambar (4/13), camelo (1/13), cervo vermelho (1/13), anta brasileira (1/13) e antílope Nilgai (1/13). Na IHQ observou-se imunomarcação acentuada (4/13), moderada (4/13) ou discreta (4/13), exceto em um caso, em que não havia quantidade suficiente de material. As características histológicas, bem como os achados na coloração de ZN e na técnica de IHQ confirmaram o diagnóstico de infecção por Mycobacterium sp. e foram considerados métodos rápidos e eficientes, de forma que podem ajudar na prevenção da disseminação da doença em animais.(AU)

Tuberculosis is a debilitating infecto-contagious disease, caused by an acid-fast bacillus (AFB) that belong to different species of the Mycobacterium tuberculosis complex (MTC). Mycobacteriosis are important diseases in veterinary medicine because of their zoonotic potential and worldwide distribution, affecting all classes of vertebrates. In wild animals the mycobacteriosis have been reported mainly as a problem in captivity. There are also reports in free-ranging wildlife, endangering and hampering tuberculosis erradication programs in animal production. The diagnosis of the disease in wildlife is usually postmortem, because the tuberculin test is not standardized for wildlife species, and also it is not reliable for screening. The postmortem diagnosis is based on macroscopic findings, microscopic detection of AFB at Ziehl-Neelsen staining (ZN), and mainly isolation and identification of the agent. However, only gross and microscopic exams do not allow to distinguish the species of Mycobacterium involved. The immunostaining with polyclonal anti-Mycobacterium tuberculosis confirms tuberculosis infection, but is not specific; there may be marking of other mycobacteria. The aim of this study was to describe the histologic findings, of ZN staining and immunohistochemistry technique (IHC) of 13 cases of wildlife herbivores diagnosed with tuberculosis in the Setor de Patologia Veterinária of the Universidade Federal do Rio Grande do Sul (SPV-UFRGS) during the 2003- 2015 period. Formalin fixed paraffin embedded tissues were recut, stained with hematoxylin and eosin and ZN, and samples were submitted to the IHC (anti-M. tuberculosis marking, streptavidin-biotin peroxidase method). All animals included were adults living in captivity and belonged to the following species: llama (5/13), sambar deer (4/13), camel (1/13), red deer (1/13), Brazilian tapir (1/13) and Nilgai antelope (1/13). The IHC revealed immunostaining of accentuated (4/13), moderate (4/13) and discrete (4/13) intensity, except in a case with insufficient material. The histological features, findings in ZN staining and IHC in the wild herbivores with tuberculosis lesions allowed the diagnosis of infection with Mycobacterium sp., turning into fast and efficient methods of diagnosis, which can help to prevent the spread of this disease in animals from the same herd.(AU)

Detección molecular de Sarcocystis aucheniae en sangre de llamas de Argentina / Molecular detection of Sarcocystis aucheniae in the blood of llamas from Argentina

Sarcocystis aucheniae are apicomplexan protozoa that infect South American camelids (SACs), giving rise to macroscopic cysts similar to rice grains in skeletal muscles. Visual detection of macrocysts in slaughtered animals hampers commercialization of SAC meat, a highly relevant economic exploitation for Andean rural families. Importantly, the consumption of undercooked S. aucheniae-infested meat causes gastroenteritis. A carnivore definitive host, possibly the dog, acquires the parasite when feeding on infected SAC meat, and later eliminates infective oocysts in its feces. The parasite cycle is completed when SACs ingest contaminated water or pastures. We hypothesized that parasite DNA can be detected in SAC blood using molecular methods. In order to test this hypothesis, a seminested PCR format was specifically designed to target the hypervariable 18S rRNA gene region of S. aucheniae. PCR conditions were optimized using genomic DNA extracted from macrocyst bradyzoites. A detection limit of up to 1 parasite in 10 μl of llama blood was established based on DNA samples extracted from aliquots of S. aucheniae bradyzoite-spiked non-infected llama blood. The seminested PCR allowed to detect natural infections of S. aucheniae in llama blood samples originating in the Andean flatlands of Argentina. Specific amplification of S. aucheniae DNA was corroborated by amplicon sequencing. This is the first report of S. aucheniae detection in llama blood, which provides a valuable diagnostic tool for epidemiological studies and for the evaluation of the efficacy of control measures for this parasitosis.

Sarcocystis aucheniae es un protozoo apicomplexa que infecta a camélidos sudamericanos (CS), dando lugar a la formación de quistes macroscópicos similares a granos de arroz en los músculos esqueléticos. La detección visual de macroquistes en animales faenados dificulta la comercialización de la carne de CS, una explotación de gran relevancia para la economía de las familias rurales andinas. Es importante destacar que el consumo de carne infectada con S. aucheniae no suficientemente cocida causa gastroenteritis. Un hospedador definitivo carnívoro, posiblemente el perro, adquiere el parásito cuando se alimenta de carne de CS infectada y luego elimina ooquistes infectivos en las heces. El ciclo del parásito se completa cuando un CS ingiere agua o pasturas contaminadas. Hemos hipotetizado que es posible detectar ADN del parásito en la sangre de CS usando métodos moleculares. Para poner a prueba esta hipótesis, se diseñó una PCR semianidada que utiliza como blanco una región del gen 18S ARNr específica para S. aucheniae. Se optimizaron las condiciones de la PCR usando ADN genómico extraído de bradizoítos presentes en macroquistes. Se estableció un límite de detección de un parásito en 10 μl de sangre de llama, basado en muestras de ADN extraído de alícuotas de sangre de llama no infectada a las que se agregaron cantidades conocidas de bradizoítos de S. aucheniae. Más aún, la PCR semianidada permitió la detección de infecciones naturales por este parásito en muestras de sangre de llama de la Puna argentina. La amplificación específica de ADN de S. aucheniae fue corroborada por secuenciación de los productos de amplificación. Este es el primer reporte de la detección de S. aucheniae en sangre de llama. Además, este estudio contribuye una herramienta diagnóstica valiosa para estudios epidemiológicos y para la evaluación de la efectividad de medidas de control para esta parasitosis.

Variabilidad genética y detección de error en filiación utilizando microsatélites en dos rebaños de alpacas Huacaya (Vicugna pacos) / Genetic variability of Peruvian Alpacas (Vicugna pacos) by using microsatellite markers

Determinar la variabilidad genética y evaluar la utilidad de microsatélites (STR) en la determinación de paternidad en alpacas blancas huacaya, pertenecientes al Centro Piloto de Mejoramiento Genético Munay Paqocha y el Fundo Itita, de la Sociedad Peruana de Criadores de Alpacas y Llamas (SPAR) Puno. Realizar la genotipificación y selección de marcadores STR útiles para la asignación de paternidad y parentesco. Metodología: Se evaluaron 10 marcadores STR a partir de ADN aislado de sangre y de folículos pilosos de 183 individuos colectados al azar procedentes de dos rebaños. Resultados y Conclusiones: Se observó un alto nivel de variabilidad alélica en el total de individuos analizados, y la presencia de alelos exclusivos entre poblaciones, con frecuencias menores al 1,5% en los loci LCA37, LCA90, LCA5, VOLP92, YWLL36, YWLL44 y YWLL08. Se propone la incorporación de tres marcadores adicionales, VOLP92, LCA94 y LCA90 para los análisis de variabilidad genética en alpacas. Los valores de FIS (0,016), y FST (0,003) reflejaron bajo niveles de endogamia. El rebaño del Fundo Itita presentó una mayor Ho (0,858) respecto a la He (0,848), mientras que por el contrario el rebaño del Centro Munay Paqocha presentó un menor valor de la Ho (0,815) respecto a la He (0,848), con una tendencia al déficit de heterocigotos. Los 10 marcadores presentaron una probabilidad de exclusión de parentesco adecuada, con un valor superior al 99,9%, cuando se conoce el genotipo de ambos padres, y un poder de discriminación mayor a 0,90...

To determine the genetic variability and the selection of STR markers useful for the evaluation of inbreeding, assignment of paternity and kinship, and the genotyping of two breeding herds of white huacayas alpacas Vicugna pacos, from the Pilot Center for Genetic Improvement Munay Paqocha and Fundo Itita, in Puno Perú. Methodology: 10 STR markers were assessed in 183 individuals, randomly selected. Results and Conclusions: We observed a high level of allelic variability in the total individuals, and unique alleles among populations with frequencies lower than 1.5% in loci LCA37, LCA90, LCA5, VOLP92, YWLL36, YWLL44 and YWLL08. We propose the addition of three markers, VOLP92, LCA94 and LCA90 for the genetic variability analysis in alpacas. FIS (0.016) and FST (0.003) values reflected low levels of inbreeding. Fundo Itita herd showed higher Ho (0.858) than He (0.848), while the herd of Munay Paqocha showed lower Ho (0.815) respect to He (0.848), with trending heterozygote deficit. The 10 markers showed an appropriate exclusion relationship probability, with a value greater than 99.9% when the genotype of both parents was known, and a power of discrimination greater than 0.9...

Relevamiento serológico de anticuerpos contra enfermedades virales de interés sanitario en llamas (Lama glama) de la provincia de Jujuy, Argentina / Serological survey of antibodies aainst vital diseases of public health interest in llamas (Lama glama) from Jujuy rpovince, Argentina

Las poblaciones de llamas de Argentina se concentran principalmente en la provincia de Jujuy; su explotación representa un importante recurso económico de las comunidades altoandinas. El objetivo de este trabajo fue evaluar la seroprevalencia de anticuerpos contra algunos agentes virales asociados a enfermedades de impacto productivo en rodeos de llamas de Jujuy. Se analizaron 349 sueros de llamas adultas de 6 departamentos de la puna jujeña ubicados por encima de los 3300 msnm. Se obtuvo una prevalencia del 100 % para rotavirus grupo A y del 70 % para el virus parainfluenza-3 bovino, mientras que no se detectaron reactores para herpesvirus bovino 1, virus de la diarrea viral bovina, influenza A humana (H1N1) e influenza equina (H3N8). Los resultados obtenidos confirman la amplia distribución de rotavirus y virus parainfluenza y la baja susceptibilidad a herpesvirus y pestivirus en las tropas de llamas de la puna jujeña

Llama population from Argentina is mainly concentrated in the Andean Puna, Jujuy. Llamas represent an important economic resource for the Andean communities. The aim of this study was to investigate the prevalence of antibodies against viral antigens associated to viral diseases of economic impact (neonatal diarrhea, reproductive and respiratory syndromes). A total of 349 serum samples from adult llamas were analyzed. The obtained antibody prevalence was 100 % for Rotavirus A and 70 % for Bovine parainfluenza virus 3. In contrast, no reactors were detected to Bovine herpesvirus 1, Bovine viral diarrhea virus 1, Human influenza A virus (H1N1) and Equine influenza virus (H3N8). These results confirm the wide circulation of rotavirus and parainfluenza virus in Argentinean llamas and suggest that susceptibility to infection with bovine herpesvirus, pestivirus and influenza A viruses is low. This serologic survey provides novel information regarding the epidemiology of viral diseases affecting llamas from the Argentinean Andean Puna

Isolation and characterization of recombinant variable domain of heavy chain anti-idiotypic antibodies specific to aflatoxin B1 / 生物医学与环境科学（英文）

Some unique subclasses of Camelidae antibodies are devoid of the light chain, and the antigen binding site is comprised exclusively of the variable domain of the heavy chain (VHH). The recombinant VHHs have a high potential as alternative reagents for the next generation of immunoassay. In particular, they might be very useful for molecular mimicry. The present study demonstrated an alpaca immunized with the F(ab')2 fragment of anti-aflatoxin B1 mAb and developed an important anti-idiotypic (anti-Id) responses. Antigen-specific elution method was used for panning private anti-Id VHHs from the constructed alpaca VHH library. The selected VHHs were expressed, renatured, purified, and then identified by a competitive enzyme-linked immunosorbent assay (ELISA). Our findings indicated that the VHH would be an alternative tool for haptens mimicry studies.

A case of nasal myiasis due to Oestrus ovis (Diptera: Oestridae) in a llama (Lama glama) / Um caso de miíase nasal devido a Oestrus ovis (Diptera: Oestridae) em lhama (Lama glama)

Infection by the larval form of Oestrus ovis (sheep bot fly) is common in many areas of Peru. This is an obligate parasite of sheep and goats, and it is the cause of oestrosis, or nasal myiasis, which can lead to severe clinical manifestations in livestock. A case of myiasis caused by O. ovis in a llama (Lama glama) in Cuzco, Peru, is reported here. This llama presented with respiratory distress and died due to bilateral hemorrhagic pneumonia. During the necropsy, six intact dipterous larvae were recovered from the nasal fossae and cranial sinuses being identified as O. ovis. This is the first report of nasal myiasis in llamas due to O. ovis in Peru.

Infecção pela forma larval de Oestrus ovis (bicho da cabeça) é comum em muitas regiões do Peru. Este é um parasito obrigatório de ovinos e caprinos, e é a causa de oestrose ou miíase nasal, que pode conduzir à manifestações clínicas graves nos animais. Relatou-se um caso de miíase causada por O. ovis numa lhama (Lama glama) em Cuzco, Peru. A lhama apresentou insuficiência respiratória e morreu de pneumonia bilateral hemorrágica. Durante a necropsia, seis larvas intactas do díptero foram recuperadas das fossas nasais e dos seios cranianos e identificadas como O. ovis. Este é o primeiro relato de miíase nasal em lhamas por O. ovis no Peru.

Inervación de las regiones escapular, braquial y antebraquial de la llama (Lama glama) / Innervation of the scapular, brachial and antebrachial of the llama (Lama glama)

La llama (Lama glama) es un mamífero cuya producción ha crecido exponencialmente en las últimas décadas, sobre todo en la región noroeste y patagónica de Argentina, Chile, Bolivia y Perú. Asociada a este crecimiento, la clínica médica veterinaria ha tenido que enfrentarse con el desafío de nuevos diagnósticos neuroanatómicos, así como también con maniobras semiológicas y quirúrgicas específicas para esta especie. El objetivo del presente trabajo es realizar una detallada descripción de la inervación de las regiones de la espalda, el brazo y el antebrazo, aportando conocimientos que sean de utilidad para que el veterinario clínico pueda interpretar alteraciones funcionales o para la realización de tratamientos quirúrgicos, terapéuticos y/o diagnósticos. Se incluye además la descripción de las relaciones musculares y vasculares de los nervios.

The llama (Lama Glama) is a mammal whose production has grown exponentially in the last decades, especially in the Northwest and Patagonian region of Argentina, Chile, Bolivia and Peru. Associated with this growth, veterinary medical clinic has had to deal with the challenge of new neuroanatomical diagnostic, as well as surgical and semiological maneuvers specific to this species. This work aims to make a detailed description of the innervation of the regions back, arm, and forearm, providing knowledge that is useful to the clinical veterinarian to interpret functional alterations or for the performance of surgical, therapeutic, treatment and/or diagnosis. It also includes the description of the muscular and vascular relationships of nerves.