Circadian rhythm of salivary and serum urea concentration in alpacas and sheep receiving diets with different levels of protein.

All vertebrates possess a daily rhythm, encompassing a comprehensive set of physiological, cognitive, and behavioral patterns that manifest throughout a 24-hour period. The aim of this study was to compare the effect of crude protein (CP) levels in the diet on the daily rhythm of urea in serum (US) and saliva in alpacas and sheep. Ten alpacas and ten sheep, adult, clinically healthy males, were used; they were maintained in natural light conditions and fed ad libitum with two levels of CP (16 and 7%). Blood and saliva samples were taken every 4 h for 48 h. A two-way ANOVA was conducted to analyze the parameters, including adjusted mean rhythm (MESOR), amplitude, and acrophase. It was found that both US and saliva urea exhibited circadian rhythms with a peak during the midday (10:59 - 12:16 h). The MESOR with low CP diets was higher (P < 0.05) in alpacas. The MESOR of urea levels in saliva was greater (P < 0.05) at the highest level of CP in the diet, with no differences between alpacas and sheep (P > 0.05). The amplitude was greater (P < 0.05) in alpacas and at the high level of CP compared to the low level of CP in the diet. Our findings reveal that both serum and saliva urea levels in alpacas and sheep follow a daily rhythm and the MESOR of US was higher in alpacas when they consume food with low CP content, and this difference disappears when CP levels are increased in the diet.

Parasitism-Induced Changes in Microbial Eukaryotes of Peruvian Alpaca Gastrointestinal Tract.

Alpacas, important genetic resources in the Andean region of Peru, are vulnerable to diarrhea caused by pathogenic parasites such as Eimeria lamae and Giardia sp., which can be fatal, especially in neonates, due to their physiological immaturity and limited adaptability. The study investigated the diversity and abundance of intestinal fungi and protists in alpacas infected with Eimeria lamae and Giardia sp. compared to healthy alpacas. A total of 19 alpacas, aged between one and two months, were included. They were divided into two groups, one with pathological conditions (nine) and the other healthy (ten). Parasitological analyses for the detection of parasites and subsequent molecular analysis were performed on the collected fecal samples. The results revealed a greater diversity and abundance of protists in infected alpacas in comparison with healthy alpacas, while the fungal composition did not show significant changes. Therefore, parasitic infections affect the protist component of the alpaca gut microbiota. Also, it was observed that Blastocystis was identified in all healthy alpacas, serving as a possible marker of the health of the intestinal microbiota; in addition, Prussia and Pichia are beneficial fungi that help control diseases. This groundbreaking study in neonatal alpacas is the first to explore potential changes in the intestinal microbiota during an infectious state, underscoring the importance of further research to comprehend its effects on alpaca health and immune responses.

Diet Digestibility and Partitioning of Nutrients in Adult Male Alpacas Fed a Blend of Oat Hay and Alfalfa Pellets at Two Levels of Intake.

Alpacas are well adapted to consume the poor-quality forages present in the arid conditions of the Andean Altiplano. However, studies focusing on understanding the relationship between diet digestibility and intake are scarce. The aim of this study was to evaluate the effects of two levels of dry matter intake (DMI) on a metabolic body weight (BW0.75) basis. The effects of a maintenance level of intake and an ad libitum level of intake (40 and 50 g of dry matter (DM) per kg of BW0.75, respectively) on the apparent diet digestibility and partitioning of specific nutrients (energy, carbon (C), and nitrogen (N)) of alpacas that were fed a blend of oat hay and alfalfa pellets (70:30 ratio as a percentage on a fed basis) were evaluated. Five adult intact male alpacas (BW = 62.9 ± 8.09 kg at the beginning of the study) were fed with the experimental diet and trained to be allocated in metabolism crates for 30 days. After the completion of this phase, two separate experiments for each level of intake were carried out, each lasting for twenty-six days (with the final five days taken for samples and raw data collection). In both experiments, the animals responded differently in terms of nutrient supply and changes in BW (-140 and 100 g/d for the maintenance and ad libitum levels of intake, respectively). Oat hay consumption was rather similar in both experiments, which may be explained by a reduced ingredient selectivity at the ad libitum level of intake. Diet digestibility was similar in both experiments, despite the greater fecal output of nutrients with the increased level of diet intake. In line with this, diet metabolizability, calculated as the ratio between metabolizable energy (ME) and gross energy (GE) contents, indicated a similar energy utilization of the diet. The apparent digestibility of the organic matter (OMD) ranged from 655 to 669 g/kg DM. Water consumption at the ad libitum level of intake was 21% higher than the observed mean at the maintenance level of intake. Fecal outputs of dietary energy, C, and N accounted for the largest source of excreted nutrients, regardless of the level of intake. The N retention increased from 0.439 at the maintenance level of intake, to 0.473 g of DM/kg BW0.75 when the alpacas were fed ad libitum.

Divergent Genotype of Hepatitis A Virus in Alpacas, Bolivia, 2019.

Hepatitis A virus (HAV) is a common human pathogen found exclusively in primates. In a molecular and serologic study of 64 alpacas in Bolivia, we detected RNA of distinct HAV in ≈9% of animals and HAV antibodies in ≈64%. Complete-genome analysis suggests a long association of HAV with alpacas.

Age-Related Differences in Testosterone Concentration and Its Relation to Testicular Biometrics, Hemodynamics, and Fertility in Alpacas (Vicugna pacos).

The goal of this study was to investigate the age-related differences in testosterone concentration and its relation to testicular biometrics, testicular blood flow, and fertility in alpacas (Vicugna pacos). Fifteen alpaca males with different ages (young (YM; ~12-14 mo.), n = 5; intermediate (IM; ~24 mo.), n = 5; and old (OM; ≥36 mo.), n = 5) were enrolled in the study. Blood samples were taken from each alpaca male and the circulating plasmatic testosterone concentration (TC; ng/mL) was determined using ELISA analysis. The testicular traits related to bio-morphometric parameters (the length (L), width (W), area (A), and volume (TV)) were assessed using B-mode ultrasonography. Pulse-wave/power Doppler ultrasonography was used to obtain the circulatory dynamic values (testicular hemodynamics) before the beginning of natural service mating. Significant differences were observed in TC among the age groups, increasing as the age of the males increased (2.47 ± 0.31, 8.45 ± 1.53, and 22.66 ± 2.15 for YM, IM, and OM, respectively; p < 0.05); however, no differences were observed regarding the testicular B-mode ultrasonographic parameters (L, W, and A) (p > 0.05). Positive correlations were observed between TV and testicular L, W, and A (r = 0.96, r = 0.95, and r = 0.96, respectively; p ≤ 0.001). Pulse-wave-Doppler-derived parameters such as the pulsatility index (PI) and the resistive index (RI), as well as the total vascularity area (TVA) assessed by power Doppler, were similar in all of the age groups studied (p > 0.05). General linear model (GLM) analysis showed a relationship between TC and TV (OR = 0.95; p = 0.04), as well as between TC and TVA (OR = 0.99; p= 0.02). Finally, no differences were observed regarding the pregnancy rate among the different age groups (p > 0.05). In conclusion, TC increased as the age of the alpaca males increased. Although TC was related to TV and TVA, the pregnancy rates obtained from individuals belonging to the different age groups were similar, indicating that TC, TV, and TVA were not determining factors in assessing the potential age-related fertility differences in alpaca males.

Influencia de la castración en la composición química de la carne de alpacas (Vicugna pacos)

RESUMEN Objetivo. Determinar la influencia de la castración de alpacas jóvenes sobre la composición centesimal y contenido de colesterol de su carne. Materiales y métodos. Con 20 animales machos, de 1.5 años de edad, de raza Huacaya se formaron grupos de 10 animales: experimental y control. Los del experimental, fueron sometidos a castración bilateral y los del control permanecieron "enteros". La alimentación de los animales fue en base a pastos naturales de la zona, bajo un sistema de crianza extensiva. Luego de seis meses, los animales fueron beneficiados y se tomaron muestras de tejido muscular de cinco regiones anatómicas: cuello, brazo, costillar, lomo y pierna. La humedad se determinó por desecación en estufa hasta peso constante, la grasa total por el método de Soxhlet, la proteína bruta por el método Kjeldahl, las cenizas por calcinación en mufla y el colesterol por el método colorimétrico, previa extracción de grasas con cloroformo/metanol (2:1). Resultados. El contenido de grasa es superior en los castrados (2.28%) que en los enteros (1.82%) (p≤0.01) y varía entre las distintas regiones anatómicas (p≤0.05); la humedad es mayor en enteros (76.14%) que en castrados (75.45%) (p≤0.01), pero similar entre las distintas las regiones anatómicas (p>0.05); no hay diferencia estadística en el contenido de proteínas, cenizas y colesterol (p>0.05), siendo el promedio general de 20.39%, 1.13% y 55.56 mg/100 g, respectivamente. Conclusiones. Se evidenció efecto de la castración sobre la composición centesimal de la carne de alpaca de machos jóvenes al incrementar el contenido graso y disminuir la humedad.

ABSTRACT Objective. Determine the influence of bilateral castration in young alpacas over the centesimal chemical composition and content in their meet. Materials and methods. A total of 20 male Huacaya breed animals aged about 1.5 years old, were studied in two groups: experimental and control with 10 animals each. The experimental one was subjected to bilateral castration and the control one remained "intact". The animals' feeding was based on natural pastures from the surrounding area under an extensive rearing system. After six months, the animals were slaughtered and muscle tissue samples were taken from five anatomical regions: neck, arm, ribs, back, and leg. Moisture was determined by oven drying to constant weight, total fat by Soxhlet method, crude protein by Kjeldahl method, ashes by muffle calcination, and cholesterol by the colorimetric method after fat extraction with chloroform/ methanol (2:1). Results. Fat content was higher in castrated animals (2.28%) than in the intact ones (1.82%) (p≤0.01) and varies among different anatomical regions (p≤0.05); humidity is higher in intact animals (76.14%) than in castrated ones (75.45%) (p≤0.01), but it was similar among different anatomical regions (p>0.05); there is no statistical difference in protein, ash, and cholesterol content (p>0.05) with a general average of 20.39%, 1.13%, and 55.56 mg/100 g, respectively. Conclusion Castration effect was evidenced on the centesimal composition of alpaca meat from young males by its fat content increase and humidity decrease.

Development of a Mobile Open-Circuit Respiration Head Hood System for Measuring Gas Exchange in Camelids in the Andean Plateau.

Peru has the largest inventory of alpacas worldwide. Despite their importance as a source of net income for rural communities living at the Andean Plateau, data on energy requirements and methane (CH4) emissions for alpacas are particularly lacking. In 2019, the International Panel on Climate Change (IPCC; 2006, and Refinement 2019) outlined methods for estimating CH4 emissions from enteric fermentation and no methane (CH4) conversion factors were reported for camelids. IPCC has since updated its guidelines for estimating CH4 emissions from the enteric fermentation of livestock at a national scale. For greenhouse gas (GHG) inventory purposes, conversion factors were developed for ruminants but not for domestic South American camelids (SAC), with this category including alpacas. A mobile open-circuit respirometry system (head hood) for the rapid determination of CH4 and CO2 production, O2 consumption, and thereafter, heat production (HP) for camelids was built and validated. In addition, an experimental test with eight alpacas was conducted for validation purposes. The average HP measured by indirect calorimetry (respiratory quotient (RQ) method) was close to the average HP determined from the carbon-nitrogen balance (CN method); 402 kJ/kg BW0.75 and 398 kJ/kg BW0.75, respectively. Fasting HP was determined by the RQ method and 250 kJ/kg BW0.75 was obtained. The metabolizable energy requirement for maintenance (MEm) was calculated to be 323 kJ/kg BW0.75 with an efficiency of energy utilization of 77%. When intake was adjusted to zero energy retention by linear regression, the MEm requirement increased to 369 kJ/kg BW0.75 and the efficiency decreased up to 68%. The CH4 conversion factor (Ym) was 5.5% on average. Further research is required to gain a better understanding of the energy requirements and CH4 emissions of alpacas in conditions of the Andean Plateau and to quantify them with greater accuracy.

Gastrointestinal parasite diversity of South American camelids (Artiodactyla: Camelidae): First review throughout the native range of distribution.

In South America inhabit an endemic group of ungulates adapted to extreme environments: the South American camelids (SAC), a key component of the Andean biocultural heritage. Until today, SAC are the most important factor of Andean economies and social and ritual life. SAC include two wild species, the guanaco (Lama guanicoe) and the vicuña (Vicugna vicugna), and two domestic species, the llama (Lama glama) and the alpaca (Vicugna pacos). Endoparasitosis are one of the most common diseases in SAC, and have great economic and health relevance. Despite this, there is a lack of knowledge on this concern. The main objective of this work was to conduct the first systematic review of the diversity of gastrointestinal parasites of SAC throughout the entire native range of distribution and to identify several gaps in knowledge. The PRISMA protocol was performed and a total of 101 documents were summarized. At least 36 parasitic helminths and five Eimeria spp. were registered. This work highlights the need for a greater number of works to know with more certainty the parasitic fauna of camelids in the past and present, in order to achieve predictions that allow proper management of camelids for their future conservation. Furthermore, concerted research efforts are needed to understand the biology, epidemiology, diagnosis and distribution of the parasitosis of SAC along the entire distribution range to guide conservation decisions.

Enteric Viral Infections among Domesticated South American Camelids: First Detection of Mammalian Orthoreovirus in Camelids.

Enteric infections are a major cause of neonatal death in South American camelids (SACs). The aim of this study was to determine the prevalence of enteric viral pathogens among alpacas and llamas in Canchis, Cuzco, located in the southern Peruvian highland. Fecal samples were obtained from 80 neonatal alpacas and llamas and tested for coronavirus (CoV), mammalian orthoreovirus (MRV), and rotavirus A (RVA) by RT-PCR. Of the 80 fecal samples analyzed, 76 (95%) were positive for at least one of the viruses tested. Overall, the frequencies of positive samples were 94.1% and 100% among alpacas and llamas, respectively. Of the positive samples, 33 (43.4%) were monoinfected, while 43 (56.6%) had coinfections with two (83.7%) or three (16.3%) viruses. CoV was the most commonly detected virus (87.5%) followed by MRV (50%). RVA was detected only in coinfections. To our knowledge, this is the first description of MRV circulation in SACs or camelids anywhere. These data show that multiple viruses circulate widely among young alpaca and llama crias within the studied areas. These infections can potentially reduce livestock productivity, which translates into serious economic losses for rural communities, directly impacting their livelihoods.

Diarrheagenic Escherichia coli isolates from neonatal alpacas mainly display F17 fimbriae adhesion gene.

Neonatal domestic South American Camelid llamas and alpacas suffer from an enteric disease complex characterized by abdominal distention, lethargy, dehydration, and eventual fatal septicemia. Analysis of rectal swabs from neonatal alpacas suffering clinical diarrheas has constantly isolated Escherichia coli, mainly the EPEC and EHEC pathotypes. The present communication reports the results of molecular analysis of 226 E. coli strains from neonatal alpaca rectal swabs. The isolates were initially tested by multiple PCR, to identify E. coli virulence genes eae, bfp, Lt, Stx1, Stx2, sta, stab, and lt genes and a similar test to detect F4, F5, F6, F17, and F41fimbriae adhesin genes. Forty-two of the 226 (18.5%) isolates tested positive for at least one pathogenic gene, 25 of the 42 were classified as EPEC (3 positives for only eae and 22 for both eae and bfp) and the remaining 17 were classified as EHEC. Twenty-four (57%) of the 42 isolates tested positive to F17 adhesin while one was positive for both F6 and F17.

Modifications of extracellular matrix features in the left and right uterine horns during the embryo pre-implantation period in Vicugna pacos.

More than 98% of the pregnancies in South American camelids is carried out in the left uterine horn (LUH). Hence, embryos originated from right-ovary ovulations have to migrate to the contralateral or left uterine horn (LUH) to implant and survive. A reason for this unique pattern of embryo implantation has not been elucidated yet. In general, embryo implantation involves an extensive extracellular matrix (ECM) remodeling within the endometrium, in which collagen and matrix metalloproteinases (MMPs) play an essential role. Deregulation of collagen and MMPs has been related to embryo implantation failure, miscarriage, and infertility. Therefore, we hypothesized that ECM components in camelids could be involved in differential embryo implantation and consequently the high incidence of left horn gestations. The aim of this study was to describe and compare changes in ECM components in the left and right uterine horn of non-pregnant and 15 days pregnant alpacas. To test this hypothesis, the collagen content was evaluated by specific staining with Picrosirius Red and using ImageJ 1.42q software. Subsequently, gene expression of the following components of the MMP pathway was determined: MMP-2, -3, -7, -9, and -14, MMP substrates (COL1A2 and COL3A1), MMP inhibitors (TIMP1 and TIMP2), LGMN, an MMP activator, and EMMPRIN, an extracellular matrix metalloproteinase inducer. Uterine horns of pregnant alpacas exhibited a marked decrease in collagen content. In contrast, transcript expression of COL1A2 and COL3A1 was higher in the LUH of pregnant alpacas. Gene expression of MMP-3, -7, -9, -14, LGMN, and EMMPRIN were also higher in the LUH of pregnant animals, whereas MMP-2 gene expression was higher in the LUH of both pregnant and non-pregnant alpacas. Expression of TIMP1 and TIMP2 increased during pregnancy, with higher values in the LUH. In conclusion, expression of ECM components displayed a specific pattern depending on the uterine side and the physiological status (pregnant vs non-pregnant) of the animal. The increased expression of ECM transcripts in the left uterine horn during early pregnancy in alpacas suggests the involvement of these molecules in a highly regulated process leading to the implantation process.

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.

Ultrastructural Description of Sarcocystis Sp. in Cardiac Muscle of Naturally Infected Alpacas (Vicugna pacos).

BACKGROUND: Recently, it was proposed the name of Sarcocystis masoni n. sp. for the Sarcocystis that causes microcyst in skeletal muscle of South American camelids. However, there are no ultrastructural reports of microcysts of Sarcocystis in cardiac muscle of alpacas. This study reports ultrastructural features of microcysts of Sarcocystis sp. from cardiac muscle of naturally infected alpacas. METHODS: Thirty alpacas (age range: three to five years) from the province of Junin, Peruvian Central Andes, were included in this study in January 2015. Cardiac muscle samples were evaluated by histology and transmission electron microscopy. RESULTS: Bradyzoites in cysts had typical characteristics of Apicomplexa including organelles, a large nucleus, micronemes, dense bodies, and polysaccharide granules. Moreover, cysts had a thin wall with numerous, short, finger-like shapes with rounded tip protrusions (0.51 × 0.17 µm). CONCLUSION: Sarcocystis sp. from the heart and S. masoni n. sp. from the skeletal muscle have similar ultrastructural characteristics.

Changes in mucins and matrix metalloproteases in the endometrium of early pregnant alpacas (Vicugna pacos).

South American Camelids (SAC) have unique reproductive features, one of which is that 98% of the pregnancies develop in the left uterine horn. Furthermore, early pregnancy is an uncharacterized process in these species, especially in regard to the ultrastructural, biochemical and genetic changes that the uterine epithelial surface undergoes to allow embryo implantation. The present study describes the uterine horn luminal surface and the characteristics of the mucinous glycocalyx in non-pregnant and early pregnant (15 days) female alpacas. In addition, the relative abundance of Mucin 1 and 16 genes (MUC1 and MUC16) was determined, as well as the relative mRNA abundance of matrix metalloproteinases (MMPs) that could be involved in MUC shedding during early pregnancy. Noticeable changes were detected in the uterine luminal epithelium and glycocalyx of pregnant alpacas in comparison to non-pregnant ones, as well as presence of MUCs and MMPs in the endometrial environment. The decrease in glycocalyx staining and in the relative abundance of MUC 1 and MUC 16 transcripts in pregnant females would allow embryo attachment to the luminal epithelium and its subsequent implantation, as has been described in other mammals. These results suggest a crucial role of MUC1 and MUC16 and a possible role of MMPs in successful embryo implantation and survival in alpacas.

Detection and genetic characterization of Pasteurella multocida from alpaca (Vicugna pacos) pneumonia cases.

Pasteurella multocida is a common constituent of upper respiratory tract microbiota but is frequently isolated of alpaca lung tissues from pulmonary infections. Despite its importance, very little is known about this bacteria at molecular level. In order to characterize P. multocida isolates, 24 isolates recovered from 46 mortal acute cases in young alpacas with suspected pneumonia were analyzed, using biochemical and molecular tests for capsule and LPS typing, virulence factors detection, and ERIC-PCR genetic diversity analysis. All the P. multocida isolates belonged to the capsular type A, LPS genotype L6 (related to serotypes 10, 11, 12, and 15), and possessed virulence factors gene toxA and tbpA. ERIC-PCR analysis revealed two electrophoretic profiles, and the majority of isolates (23/24) shared the same fingerprint, indicating strong evidence that there was a common source of infection for all the affect animals. This study revealed the detection of P. multocida type A, LPS genotype L6, and toxA+ and tbpA+ from dead young alpacas with pneumonia in Peru.

Vitrification of in vitro mature alpaca oocyte: effect of ethylene glycol concentration and time of exposure in the equilibration and vitrification solutions.

The effect of different ethylene glycol concentrations, times of exposure and vitrification procedure on viability, cleavage and blastocyst rate of in vitro matured alpaca oocytes chemically activated after vitrification was analyzed. In Experiment 1, oocytes were incubated for 12-15 min with different concentrations of ethylene glycol (EG) in the equilibration solution (ES) followed by chemical activation and in vitro cultured for 8 days to determine oocyte viability, cleavage and blastocyst rates. In Experiment 2, oocytes were incubated in the equilibration solution containing 4% of EG for 12-15 min and then randomly assigned to vitrification solutions containing 25, 35 or 45% of EG for 30s, vitrified and stored at -196°C. In Experiment 3, oocytes were incubated in the equilibration solution containing 4% of EG for 12-15 min and then randomly assigned to the vitrification solution containing 35% of EG for 15, 30 or 45s, vitrified and stored at -196°C. For Experiments 2 and 3, non-vitrified and vitrified oocytes were activated and cultured in vitro. In Experiment 1, oocyte viability was lowest at concentrations of 6 or 8%, intermediate at 2 or 4% and highest at 0% of EG. Oocyte viability and cleavage rate were affected by EG concentration, time of exposure in the vitrification solution or vitrification procedure in Experiment 2 and 3. Alpaca oocytes were viable after vitrification, given that oocyte viability, cleavage and blastocyst rate were affected by the vitrification procedure, EG concentration and time of exposure in the equilibration and vitrification solutions.