Llama Research Articles

Serum and urine analyte comparison between llamas and alpacas fed three forages.

Serum and urine analytes were measured in 4 healthy adult llama and alpaca geldings housed in metabolism crates and fed three diets consisting of alfalfa (AH), barley (BH) or grass (GH) hay and water ad libitum. This comparative study was conducted to determine if differences exist in serum metabolites and urinary indices in llamas and alpacas consuming the same forages of differing protein and carbohydrate quality. Daily feed intake was determined and concurrent serum and urine samples were obtained at 4-hr intervals on d13 and d14 for analysis of electrolytes, urea N, and creatinine. AH was consumed more than BH or GH by both species, but llamas consumed more forage on both a body weight (BW) and metabolic BW (MBW, kg .75 ) basis. Serum electrolytes were similarly affected by diet between species, indicating that renal homeostatic mechanisms were functional and analogous in llamas and alpacas. Serum and urine urea N were affected by forage (P<0.05). Urine volume was highest for both camelid species when consuming AH (P<0.05). Mean urine electrolyte excretion only differed by diet. Dissimilarities between these species was evidenced by differences in renal excretion of urea N and differences in urine volume on a MBW (kg .75 ) basis.

Aspectos histológicos del aparato digestivo de la alpaca

Se ha realizado el estudio de la anatomía macroscópica y microscópica normal, del aparato digestivo de la Alpaca en 11 animales, clínicamente normales. El aspecto microscópico ha revelado variación estructural en los proventrículos de estos animales, con respecto a los demás rumiantes por nosotros conocidos.

Novel camelid antibody fragments targeting recombinant nucleoprotein of Araucaria hantavirus: a prototype for an early diagnosis of Hantavirus Pulmonary Syndrome.

In addition to conventional antibodies, camelids produce immunoglobulins G composed exclusively of heavy chains in which the antigen binding site is formed only by single domains called VHH. Their particular characteristics make VHHs interesting tools for drug-delivery, passive immunotherapy and high-throughput diagnosis. Hantaviruses are rodent-borne viruses of the Bunyaviridae family. Two clinical forms of the infection are known. Hemorrhagic Fever with Renal Syndrome (HFRS) is present in the Old World, while Hantavirus Pulmonary Syndrome (HPS) is found on the American continent. There is no specific treatment for HPS and its diagnosis is carried out by molecular or serological techniques, using mainly monoclonal antibodies or hantavirus nucleoprotein (N) to detect IgM and IgG in patient serum. This study proposes the use of camelid VHHs to develop alternative methods for diagnosing and confirming HPS. Phage display technology was employed to obtain VHHs. After immunizing one Lama glama against the recombinant N protein (prNΔ85) of a Brazilian hantavirus strain, VHH regions were isolated to construct an immune library. VHHs were displayed fused to the M13KO7 phage coat protein III and the selection steps were performed on immobilized prNΔ85. After selection, eighty clones recognized specifically the N protein. These were sequenced, grouped based mainly on the CDRs, and five clones were analyzed by western blot (WB), surface plasmon resonance (SPR) device, and ELISA. Besides the ability to recognize prNΔ85 by WB, all selected clones showed affinity constants in the nanomolar range. Additionaly, the clone KC329705 is able to detect prNΔ85 in solution, as well as the native viral antigen. Findings support the hypothesis that selected VHHs could be a powerful tool in the development of rapid and accurate HPS diagnostic assays, which are essential to provide supportive care to patients and reduce the high mortality rate associated with hantavirus infections.

Energy metabolism and methane production in llamas, sheep and goats fed high- and low-quality grass-based diets

This study aimed to test whether the digestive and metabolic characteristics of pseudo ruminants provide superior ability to utilise low-quality diets compared to true ruminants. A total of 18 mature, non-pregnant, non-lactating female animals, including six llamas (Lama glama), six Danish Landrace goats and six Shropshire sheep, were used in a crossover design study. The experiment lasted for two periods of three weeks. Half of the animals were fed either high-quality grass hay (HP) or low-quality grass seed straw (LP) during each period. Animals were placed in metabolic cages during the last 5 d, and gaseous exchange was measured by open-circuit indirect calorimetry for 22 h. Metabolisable energy for maintenance (MEm) and fasting energy expenditure (FEExp) were estimated by regression approach. Dry matter (DM) intake per kg0.75 was substantially reduced in llamas and sheep, but not in goats, on the LP compared to HP diet. Llamas had lower daily energy expenditure (324 kJ · kg−0.75) than sheep (416 kJ · kg−0.75) and goats (404 kJ · kg−0.75) on the LP diet. Llamas in comparison with sheep and goats had lower methane emission (0.83 vs 1.34 and 1.24 l · d−1 · kg−0.75, p < 0.05), lower MEm (328 vs 438 and 394 kJ · d−1 · kg−0.75, p < 0.05) and lower FEExp (246 vs 333 and 414 kJ · d−1 · kg−0.75, p < 0.05), respectively. In conclusion, llamas had lower basal metabolic rate and hence maintenance requirements for energy.

Evaluation of the specificity of intradermal tuberculin and serological tests for diagnosis of tuberculosis in alpaca (Vicugna pacos) and llama (Lama glama) herds under field conditions in Peru

Diagnosis of tuberculosis (TB) in camelids is still a challenge mainly due to the lack of standardised diagnostic assays and the limitations regarding sensitivity (Se) and specificity (Sp) of the...

First llama (Lama glama) pregnancy obtained after in vitro fertilization and in vitro culture of gametes from live animals

The aim of this study was to evaluate the developmental competence and pregnancy rate of llama hatched blastocysts produced in vitro using gametes from live animals and two different culture conditions. Fifteen adult females were superstimulated with 1500IU of eCG, eleven (73%) responded to the treatment and were used as oocyte donors. Follicular aspiration was conducted by flank laparotomy. Semen collections were performed under general anesthesia by electroejaculation of the male. Sixty-six COCs were recovered from 77 aspirated follicles (86% recovery) and were randomly placed in Fertil-TALP microdroplets with the sperm suspension (20×106live spermatozoa/ml). After 24h, they were placed in SOFaa medium supplemented with FCS and randomly assigned to one of two culture conditions. Culture condition 1 (CC1) consisted of 6 days of culture (n=28) and culture condition 2 (CC2) consisted of renewing the culture medium every 48h (n=35). In CC1, the blastocyst rate was 36% (10/28) and the hatched blastocyst rate was 28% (8/28) whereas in CC2, the blastocyst rate was 34% (12/35) and the hatched blastocyst rate was 20% (7/35) (p>0.05). No pregnancies were obtained after embryo transfer (ET) of CC1 blastocysts (0/8) while one pregnancy was obtained (1/7) after transferring a hatched blastocyst from CC2. Forty-two days after the ET, the pregnancy was lost.This study represents the first report of a pregnancy in the llama after intrauterine transfer of embryos produced by in vitro fertilization using gametes from live animals.

Detection of the matrix metalloproteinases MMP-2 and MMP-9 and tissue inhibitors of metalloproteinases TIMP-1 and TIMP-2 in llama (Lama glama) oviduct.

Matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) are involved in several reproductive events like oocyte-spermatozoa interaction and semen liquefaction. In order to study their role in the llama oviductal reproductive process, MMP activity in oviductal fluid (OF) was assayed. Considering that llama genome sequences are partially known, a strategy to procure cDNA sequences of MMP-2, MMP-9, TIMP-1 and TIMP-2 was designed. Afterwards, their expression patterns in the different llama oviductal segments were assayed. Gelatine zymograms detected 62 and 94 kDa protease activities that matched MMP-2 and pro-MMP-9, respectively. Expression pattern analysis showed that MMP and TIMP mRNAs were present in ampulla, isthmus, utero-tubal junction (UTJ) and papilla. Altogether, these findings support the argument that MMPs/TIMPs are produced in the oviduct and secreted into the oviductal lumen. Our results encourage further studies to elucidate the role of these proteins in reproductive oviductal events.

Genetic diversity of GH1 and LEP genes in Argentine llama (Lama glama) populations

Compared to other domestic species, little is known about variability of genes related to energetic metabolism and growth in camelids. Here, we have analyzed leptin (LEP) and growth hormone (GH1) genes and characterized their variability in three local llama (Lama glama) populations from the Argentine Northwest. Eleven novel SNPs and one indel were identified in the Lep gene. In total, eight haplotypes were found for LEP and seven for GH1. Although geographical origin clustering was not observed, SNP and haplotype frequencies varied significantly among populations. Based on variation of both loci, we detected significant genetic differentiation measured through Fst values.Network analysis of LEP gene supported the well-documented history of hybridization in camelids previously identified by mtDNA analysis.

Methane emission by camelids.

Methane emissions from ruminant livestock have been intensively studied in order to reduce contribution to the greenhouse effect. Ruminants were found to produce more enteric methane than other mammalian herbivores. As camelids share some features of their digestive anatomy and physiology with ruminants, it has been proposed that they produce similar amounts of methane per unit of body mass. This is of special relevance for countrywide greenhouse gas budgets of countries that harbor large populations of camelids like Australia. However, hardly any quantitative methane emission measurements have been performed in camelids. In order to fill this gap, we carried out respiration chamber measurements with three camelid species (Vicugna pacos, Lama glama, Camelus bactrianus; n = 16 in total), all kept on a diet consisting of food produced from alfalfa only. The camelids produced less methane expressed on the basis of body mass (0.32±0.11 L kg−1 d−1) when compared to literature data on domestic ruminants fed on roughage diets (0.58±0.16 L kg−1 d−1). However, there was no significant difference between the two suborders when methane emission was expressed on the basis of digestible neutral detergent fiber intake (92.7±33.9 L kg−1 in camelids vs. 86.2±12.1 L kg−1 in ruminants). This implies that the pathways of methanogenesis forming part of the microbial digestion of fiber in the foregut are similar between the groups, and that the lower methane emission of camelids can be explained by their generally lower relative food intake. Our results suggest that the methane emission of Australia's feral camels corresponds only to 1 to 2% of the methane amount produced by the countries' domestic ruminants and that calculations of greenhouse gas budgets of countries with large camelid populations based on equations developed for ruminants are generally overestimating the actual levels.

The arteries on the base of the brain in the camelids (Camelidae)

The aim of this study was to analyse and compare the cerebral arterial circle, its anastomoses, its branches and the manner of blood supply in 20 individuals of the Camelidae family. The Camelidae family was represented by Camelus bactrianus (n = 8), Camelus dromedarius (n = 2), Lama guanicoe (n = 5) and Lama glama (n = 5). We performed the analyses on post-mortem material obtained from Polish zoological gardens. Arteries of the heads were injected with pigmented acetone solution of vinyl superchloride. Arteries of the brain base in Camelus and Lama species are similar. In contrast, the pattern of brain base arteries in Camelidae differs from the patterns described in other species of the Artiodactyla order. Specific traits of the base brain arterial pattern in Camelidae may prove useful as a taxonomic exponent of the animals within the Artiodactyla order.

Combining proteomic tools to characterize the protein fraction of llama (Lama glama) milk

Llamas belong to the Camelidae family along with camels. While dromedary camel milk has been broadly characterized, data on llama milk proteins are scarce. The objective of this study was thus to investigate the protein composition of llama milk. Skimmed llama milk proteins were first characterized by a 2D separation technique coupling RP-HPLC in the first dimension with SDS-PAGE in the second dimension (RP-HPLC/SDS-PAGE). Llama milk proteins, namely caseins (αs1 -, αs2 -, β-, and κ-caseins), α-lactalbumin, lactoferrin, and serum albumin, were identified using PMF. Llama milk proteins were also characterized by online LC-ESI-MS analysis. This approach allowed attributing precise molecular masses for most of the previously MS-identified llama milk proteins. Interestingly, α-lactalbumin exhibits distinct chromatographic behaviors between llama and dromedary camel milk. De novo sequencing of the llama α-lactalbumin protein by LC coupled with MS/MS (LC-MS/MS) showed the occurrence of two amino acid substitutions (R62L/I and K89L/I) that partly explained the higher hydrophobicity of llama α-lactalbumin compared with its dromedary counterpart. Taken together, these results provide for the first time a thorough description of the protein fraction of Lama glama milk.

Pituitary Null Cell Adenoma in a Domestic Llama (Lama glama)

Pituitary gland neoplasia has been reported rarely in camelids. A 12-year-old neutered male llama (Lama glama) presented with lethargy, inappetence and neurological signs. On physical examination, the llama was mentally dull and exhibited compulsive pacing and circling to the left. Complete blood count and serum biochemistry revealed haemoconcentration, mild hypophosphataemia, hyperglycaemia, hypercreatininaemia and hyperalbuminaemia. Humane destruction was elected due to rapid clinical deterioration and poor prognosis. Post-mortem examination revealed a pituitary macroadenoma and bilateral internal hydrocephalus. Microscopically, the pituitary tumour was composed of neoplastic chromophobic pituitary cells. Ultrastructural studies revealed similar neoplastic cells to those previously described in human null cell adenomas. Immunohistochemically, the neoplastic cells were strongly immunoreactive for neuroendocrine markers (synaptophysin and chromogranin A), but did not exhibit immunoreactivity for epithelial, mesenchymal, neuronal and all major pituitary hormone markers (adrenocorticotropic hormone, follicle stimulating hormone, growth hormone, luteinizing hormone, melanocyte-stimulating hormone, prolactin and thyroid stimulating hormone), consistent with the diagnosis of a pituitary null cell adenoma. This is the first report of pituitary neoplasia in a llama.

Pastoralism in Northern Peru during Pre-Hispanic Times: Insights from the Mochica Period (100–800 AD) Based on Stable Isotopic Analysis of Domestic Camelids

Llama (Lama glama) and alpaca (Vicugna pacos) are the only large domesticated animals indigenous to the Americas. Pastoralism occupies a fundamental economic, social and religious role in Andean life. Today, camelid livestock are confined to the ecozone of the puna (above 3,500 masl), while their presence on the Pacific coast during pre-Hispanic times is attested by archaeological skeletal remains. This study aims to document herding practices on the northern Peruvian coast during the Early Intermediate Period (200 BC-600 AD) by gaining insights into diet, location of breeding and mobility of archaeological camelids from the funerary and ritual contexts of two Mochica sites, Uhle Platform in Huacas de Moche and El Brujo. The three first early years and the long-term life histories of the animals were documented by the combined bulk analysis of bone collagen (δ13Ccol and δ15Ncol) and bone structural carbonate (δ13Cbone and δ18Obone) and the serial analysis of structural carbonate of molar tooth enamel (δ13Cenamel and δ18Oenamel). Mochica camelids were bred in the low and/or middle valleys, unlike their modern counterparts, who are restricted to highland puna C3 pastures. Archaeological camelids had diverse and complex life histories, usually with substantial maize foddering. An ontogenetic switch in diet and possible residential mobility during the course of life were identified for some specimens. Although the inference of geographic origin from δ18Obone and δ18Oenamel values was limited because of the lack of understanding of the influence of environmental and biological factors, tooth enamel analysis has great potential for exploring camelid herding practices and Andean pastoralism. Our study suggested that Mochica herders adapted their practices to the difficult lowland environment and that herding practices were varied and not restricted to breeding at higher altitudes. The role of maize in different aspects of the economic life of the Mochicas is also underlined.

A note on the effects of pre-slaughter operations of llamas (Lama glama) on the concentrations of some blood constituents related to stress and carcass quality

El objetivo de este trabajo fue evaluar el efecto de las operaciones presacrificio sobre indicadores fisiologicos de estres (constituyentes sanguineos) y de calidad de la canal (pH y contusiones) en llamas bajo condiciones comerciales. Treinta llamas criadas a pastoreo, de 18-24 meses de edad y peso vivo 54,2±6,4 kg fueron transportadas durante 3 h en un vehiculo a matadero. Se tomaron muestras de sangre a las llamas en predio una hora antes de cargar, despues de descargar, despues del reposo en matadero (18 h) y durante la exsanguinacion. Se encontro un aumento significativo (P < 0,05) de cortisol despues del transporte (16,9 ng/mL), como tambien de actividad de CK despues del reposo (528,4 UI/L); la concentracion de β -hidroxibutirato se mantuvo en los muestreos. Se registraron 19 contusiones en 9 de las 30 canales de llamas. Las lesiones superficiales, pequenas y de color rojo brillante, fueron las mas frecuentes. La region del lomo presento mas contusiones (57,89%), seguida del torax (21,05%). Las lesiones de forma irregular fueron mas frecuentes (81,25%), seguidas de circulares (18,75%). Se concluyo que las operaciones presacrificio de llamas bajo las condiciones comerciales del estudio produjeron cambios fisiologicos similares a los observados en otras especies, que caen dentro de limites aceptables para su bienestar; sin embargo, el estres y los efectos adversos en las canales, como las contusiones, podrian minimizarse mediante diseno de estructuras apropiadas para el manejo, aplicacion de las recomendaciones de OIE para el bienestar durante el sacrificio y capacitacion del personal que maneja las llamas a lo largo de la cadena carnica en Bolivia

Identification of Lamanema chavezi Becklund 1963 infection in a llama (Lama glama) in the United States

Infection with Lamanema chavezi, a parasitic nematode of New World camelids, was diagnosed by examination of feces and formalin-fixed liver from a 14-month-old female llama (Lama glama) that died after a 6-week illness. Infection with L. chavezi was initially suspected when a granuloma containing an unidentified nematode was detected microscopically in the hepatic parenchyma from a necropsy specimen. The subsequent diagnosis of L. chavezi infection was based on the morphologic features of 2 immature nematodes dissected from individual hepatic granulomas, characteristics of eggs detected in feces of the llama by centrifugal flotation in sugar solution (specific gravity: 1.30), development of third-stage larvae within the eggs after incubation of the llama feces at room temperature for ≥30 days, and the morphology of third-stage larvae released from the embryonated eggs. Collectively, these findings indicate that the llama, born and raised in Oregon, harbored an autochthonous L. chavezi infection. Eggs identified as L. chavezi were also detected by centrifugal flotation of pelleted feces from 3 of 7 herd mates of the llama indicating this parasite is endemic in the Oregon herd. The findings reported herein serve to alert diagnosticians and veterinary practitioners to the occurrence of L. chavezi in New World camelids in the United States and describe diagnostic features of this potential pathogen.

Sarcocystis spp. in llamas (Lama glama) in Southern Bolivia: A cross sectional study of the prevalence, risk factors and loss in income caused by carcass downgrades

Llamas (Lama glama) are intermediate hosts of the protozoan parasite Sarcocystis spp. This parasite is described as causing economic losses in the production of llama meat in South America. The aim of this study was to estimate prevalence, identify risk factors and explore spatial patterns of Sarcocystis in llamas in an area of the Bolivian High Plateau including estimating financial losses due to carcass downgrades as a result of the presence of Sarcocystis cysts.Information was collected from a local abattoir between 2006 and 2011 on 1196 llamas. Sarcocystis status was determined at meat inspection where any carcasses with one or more visible cysts were deemed Sarcocystis positive. A high prevalence was found, estimated to vary between 23.4% (95% CI 16.6–30.1) in 2007 and 50.3% (95% CI 44.4–56.3) in 2011. Period prevalence between 2006 and 2011 was estimated at 34.1% (95% CI 31.4–36.8). Age, sex and type (analogous to breed) were identified as risk factors for Sarcocystis using a mixed-effects logistic regression model adjusting for clustering by community and owner. Llamas over 4.5 years of age had an increased odds of being Sarcocystis positive (OR 19.31, 95% CI 9.10–40.98) as well as females (OR 1.75, 95% CI 1.13–2.68) and long haired type llamas (OR 1.90, 95% CI 1.26–2.87). An interaction between age and sex was detected indicating that the increased odds of disease from the youngest age group to the 2.5–4.5 years group was much more pronounced in females than in males. Spatial patterns of Sarcocystis were explored at district level by means of Standardised Morbidity Ratios and some spatial heterogeneity was revealed. Estimates of financial loss due to the disease were calculated using the difference in price paid for Sarcocystis positive and negative meat. Loss due to Sarcocystis varied per year but could be up to 20% of the annual income generated through the abattoir by sale of meat.Overall this study shows a high prevalence of Sarcocystis in the study area with some heterogeneity between districts. It also identifies some previously unknown risk factors for Sarcocystis and gives financial estimates of the cost of the disease as a result of carcass downgrades. We hoped these findings will add to the understanding of Sarcocystis in llamas in Southern Bolivia and will be useful when considering if controls are necessary, worthwhile and practical.

A case of nasal myiasis due to Oestrus ovis (Diptera: Oestridae) in a llama (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.

Molecular identification of Sarcocystis aucheniae as the macrocyst-forming parasite of llamas

The domestic South American camelids (SACs), llama (Lama glama) and alpaca (Lama paco), are frequently found to be infected with Sarcocystis parasites. Infections give rise in skeletal muscle to macroscopic cysts (1–5mm long) that resemble rice seeds, each containing several million living bradyzoites. The finding of cysts prevents commercialization of SAC meat, an important source of income for rural families in the Andean flatlands. Thus, development of diagnostic methods to facilitate the control of these infections is highly desirable, and the first step to this end is the unequivocal species identification of the causative agent. Based on the cyst form and size, the infecting parasite has been described as Sarcocystis aucheniae; however, this traditional approach is not reliable as similar cysts may contain different species. To date, molecular identification has been done for a single isolate of S. aucheniae from an alpaca in Australia. In order to verify the identity of the species present in SACs of South America, the complete 18S rRNA gene was PCR-amplified and sequenced from macrocyst DNA obtained from three llamas of the Andean flatlands. A phylogenetic Bayesian analysis was carried out using the analyzed and available 18S rRNA sequences of Sarcocystis spp. In the constructed tree, all of the new 18S rRNA gene sequences segregated in a single clade together with the 18S rRNA gene sequence reported from an alpaca in Australia, demonstrating that the isolated parasite is S. aucheniae, and that this parasite indiscriminately infects both domestic SACs. This work represents the first molecular identification of the causative agent of SAC sarcocystiosis in South America, and can contribute to the development of control methods for this neglected parasitosis.

Hunting, Herding, and Caravanning: Osteometric Identifications of Camelid Morphotypes at Khonkho Wankane, Bolivia

AbstractKhonkho Wankane is a ceremonial center located in the southern Lake Titicaca Basin, Bolivia. During the Late Formative period (AD 1–500), its residents practiced agropastoral lifeways and participated in the rise of the state at Tiwanaku. Like at many Andean sites, bones from the family Camelidae are the most abundant large mammal in domestic contexts. Identifying camelid morphotypes represented by these bones carries far‐reaching implications for understanding past hunting, herding, and caravanning practices, and their roles in larger social and economic webs. Identifications were based on a locally focused reference collection, including llamas (Lama glama) from the immediate vicinity of the site, as well as Andean guanacos (Lama guanicoe), a much smaller morphotype than the Patagonian guanacos used in many osteometric studies. Multivariate statistical analyses and incisor morphology identified all four camelid. Different analyses suggest that the crux of osteometry lies in the reference collection, not the statistical test. An additional, very large morphotype likely corresponds to a castrated llama, the preferred cargo animal among modern drovers. The presence of these animals is interpreted as evidence that groups hunted vicuña (Vicugna vicugna) and guanaco, which are not currently present around the site, herded llamas and alpacas (Vicugna pacos), and perhaps organized caravans with castrated llamas. Copyright © 2013 John Wiley &amp; Sons, Ltd.

Meat quality, proximate composition and muscle fatty acid profile of young llamas (Lama glama) supplemented with hay or concentrate during the dry season

Thirty llamas were used to study the effect of a 90day feed supplementation on meat quality, chemical composition and muscle fatty acid profile. Treatments were: GR=llama on native pasture until slaughter; GR+SH=like GR, but with overnight free access to barley/alfalfa hay; and GR+SC=like GR, but with overnight free access to a wheat bran/sorghum grain concentrate. The supplementation had no effect on postmortem pH and temperature decline in the Longissimus lumborum muscle (LLM), cooking losses nor Warner–Bratzler shear force values (P>0.05). Meat from GR+SC llama had higher fat content in LLM (P<0.05) compared to GR and GR+SH llama. Intramuscular fat from GR+SH llama showed higher (P<0.01) proportions of polyunsaturated fatty acids, higher (P<0.05) polyunsaturated fatty acids/saturated fatty acids and desirable fatty acids ratio, lower (P<0.05) omega-6/omega-3 (n−6/n−3) ratio, and higher (P<0.01) conjugated linoleic acid.