The isolation, characterisation and quantification of the equine plasma lipoproteins

Plasma lipoproteins were isolated from eight Thoroughbred horses and eight Shetland ponies on the basis of particle size by gel filtration chromatography and according to density using rate-zonal ultracentrifugation. Three major classes corresponding to very low density lipoproteins (VLDL), low density lipoproteins (LDL) and high density lipoproteins (HDL) were identified and characterised by their lipid and apolipoprotein compositions. The particle size distributions of each class were determined by electron microscopy and non-denaturing polyacrylamide gradient gel electrophoresis. HDL was found to dominate the equine lipoprotein spectrum, accounting for 61 per cent of the total plasma lipoprotein mass (VLDL 24 per cent, LDL 15 per cent). The VLDL class was isolated as a single population of particles that were triglyceride rich and cholesterol, phospholipid and protein poor. Equine LDL was characteristically cholesterol rich and was found to be polydisperse comprising three subfractions that were discrete with respect to particle size and lipid composition. The HDL class was composed of homogeneous particles that were typically protein rich. Apolipoprotein (apo) B was the major protein of VLDL and LDL, and presented two components on polyacrylamide gel electrophoresis with molecular weights in the region of human apoB-100 and a third in VLDL similar to that of apoB-48. ApoA-I was the predominant protein in equine HDL. Although there were no breed differences in the physical or chemical properties of each lipoprotein class, the Shetland ponies had higher plasma triglyceride and VLDL concentrations than their Thoroughbred counterparts.     

Quantitative analysis of canine plasma lipoproteins

A combined ultracentrifugationl/precipitation method for the measurement of lipoprotein cholesterol concentrations was developed and validated for use with canine plasma. Very low density lipoproteins (VLDL) were isolated by flotation ultracentrifugation and low density lipoproteins (LDL) separated from high density lipoproteins (HDL) by precipitation with heparin-manganese chloride. Effective separation of these classes was confirmed by agarose gel electrophoresis of native lipoproteins and by sodium dodecyl sulphate polyacrylamide gel electrophoresis of their apolipoprotein distributions. There was trace contamination of the LDL precipitate with HDL, but this represented less than 4 and 9 per cent of the total plasma HDL in normo- and hypercholesterolaemic dogs, respectively. The intra-assay and interassay coefficients of variation for LDL- and HDL-cholesterol concentrations were between 3·3 and 6·9 per cent, and 7·2 and 9·0 per cent, respectively, for plasma cholesterol concentrations between 2·67 and 8·14 mmoll/litre. The intra-assay coefficient of variation for VLDL-cholesterol was 53·8 and 18·4 per cent at plasma cholesterol concentrations of 2·67 and 8·14 mmol/litre, respectively. The interassay coefficient of variation for VLDL was 22·5 per cent. Storage of plasma at -20°C for between two and eight weeks did not affect VLDL-cholesterol concentrations, but led to an increase in LDL-cholesterol and a decrease in HDL-cholesterol concentrations of approximately 10 per cent. The method described is appropriate for the measurement of lipoprotein concentrations in plasma from normo- and hypercholesterolaemic dogs, but samples should not be subjected to prolonged storage before analysis.     

The effect of Eimeria acervulina infection on plasma lipids and lipoproteins in young broiler chicks

In young broiler chicks inoculated with 2 x 10(6) sporulated oocysts of Eimeria acervulina per bird, total plasma lipids were significantly depressed compared with controls in the first week after inoculation. The lowest level observed was at 5 days post-inoculation (d.p.i.), at which time the chick host is known to experience malabsorption in the chick host (Ruff and Wilkins, 1980). Analysis of plasma components of infected chicks at 4 and 7 d.p.i. showed that triglycerides, total cholesterol, free fatty acids, pigments and total protein were significantly decreased compared with controls. At 7 d.p.i., reduction of total cholesterol reflected mainly reduction in high density lipoprotein (HDL) cholesterol. However, the ratio of HDL cholesterol/total plasma cholesterol was not significantly different from the control ratio. Density gradient ultracentrifugation of chick plasma separated lipoproteins into three main fractions: portomicrons plus very low density lipoproteins (PM + VLDL), low density lipoproteins (LDL) and HDL. These fractions were analyzed for lipid content. Infection with E. acervulina caused (1) significant reduction in the triglyceride and cholesterol contents of the PM + VLDL fraction at 3 and 5 d.p.i., (2) significant reduction of LDL cholesterol at 9 d.p.i. and LDL phospholipid at 5-9 d.p.i., and (3) significant reduction of HDL cholesterol at 3-9 d.p.i. and HDL phospholipid at 5-9 d.p.i. Starvation of uninfected chicks for 48 h caused significant reduction in plasma triglycerides and phospholipids, but an increase in total cholesterol. Density gradient ultracentrifugation showed that the changes in these components reflected mainly reduction of the lipids in the PM + VLDL fraction. The LDL fractions, however, appeared more intense than those of the controls and contained more cholesterol and phospholipids. These results suggest that changes at 3 and 5 d.p.i. in the plasma lipoprotein pattern of chicks infected with E. acervulina most closely resemble changes seen in chicks starved for 48 h as far as PM + VLDL fraction is concerned. However, changes seen from 7 to 9 d.p.i. involve the LDL and HDL fractions and may reflect alterations in lipid and/or lipoprotein synthesis in the liver and intestine.     

Bovine lipoprotein and apolipoprotein profiles as influenced by sex and growth.

Three (intact) Angus males and females that were half-sibs and born within 21 d of each other were selected for this study. Each animal was bled periodically from birth to slaughter (18 mo) to determine the qualitative composition of plasma lipoproteins and apolipoproteins during its growth and development. Major components observed were: 1) very low-density (VLDL), 2) low-density (LDL), and 3) high-density lipoproteins (HDL). Individual amounts of triglycerides, cholesterol, and proteins for the VLDL were not different (P greater than .05) between sexes at any time during growth and development. At 1 yr of age and 15 mo of age, females had significantly larger (143.4 and 93.5 mg/dl) amounts of protein in the HDL than males (67.0 and 93.5 mg/dl), respectively. Within the male group, the LDL triglyceride concentration of calves was significantly (P less than .05) higher (7.4 mg/dl) than at all other bleeding times. Within the female group, cholesterol values for the VLDL were significantly (P less than .05) larger as calves and weanlings (16.5 and 21.7 mg/dl respectively) than for other bleeding periods. At all stages of growth and development, the HDL apoprotein profiles showed a distinct band with a weight of about 28,000 Da, which represented apolipoprotein-A-I. During the suckling stage, pooling of LDL fractions provided two components on the acrylamide gel (7.5 to 20%), apolipoprotein-B and a low molecular weight band. At 12 and 15 mo, no low molecular weight band was present in the pooled LDL fraction.(ABSTRACT TRUNCATED AT 250 WORDS)     

Corneal lipid deposition in Cuban tree frogs (Osteopilus septentrionalis) and its relationship to serum lipids: an experimental study.

To evaluate the association between corneal lipid infiltration (corneal arcus) and dietary cholesterol in Cuban tree frogs (Osteopilus septentrionalis), 47 wild-caught frogs were fed diets of either regular or high-cholesterol crickets containing 0.7% and 1.7% cholesterol dry matter, respectively. Serum total cholesterol and triglycerides were measured when the frogs were euthanized after 17 mo. In a subsample of frogs, serum lipoproteins were characterized using high-performance liquid chromatography. The first case of corneal lipid deposition occurred in a female frog after 13 mo on the high-cholesterol diet. In the subsequent 4 mo, 5/11 males and 11/35 females developed the disease. Four of these affected frogs were females on the regular diet. Frogs with corneal lipid deposition had elevated serum total cholesterol (27.3 +/- 19.8 mmol/L) and low-density lipoproteins (LDL, 17.8 +/- 18.9 mmol/L) compared with unaffected captive frogs (16.5 +/- 20.4 and 9.0 +/- 7.6 mmol/L, respectively). Corneal lipid deposition was more prevalent in frogs on the high-cholesterol diet, and this group had higher serum total cholesterol (34.1 +/- 15.2 mmol/L in females, 22.8 +/- 14.8 mmol/L in males) than did frogs on the diet of regular crickets (12.3 +/- 8.7 mmol/L in females, 10.4 +/- 3.1 mmol/L in males). Captive frogs on both diets had higher serum total cholesterol than did wild frogs (3.1 +/- 2.1 mmo/L in females, 5.3 +/- 2.6 mmo/L in males). This additional serum cholesterol was primarily carried on very low-density lipoproteins (VLDL) and LDL rather than high-density lipoproteins (HDL), as indicated by the significantly higher ratio of VLDL cholesterol and LDL cholesterol over HDL cholesterol in captive frogs compared with wild frogs. Elevation in this ratio was significantly higher in captive females than in captive males. There was no evidence that increased serum cholesterol in captive females was directly related to the process of vitellogenesis.     

Spectrophotometry and Ultracentrifugation for Measurement of Plasma Lipids in Dogs with Diabetes Mellitus

Background

There are conflicting reports of plasma lipoprotein lipid content in dogs with diabetes mellitus (DM).

Objectives

To determine lipoprotein lipid content of plasma of dogs with DM by spectrophotometry and ultracentrifugation; to compare lipoprotein lipid content in diabetic and healthy dogs; and to quantify apolipoprotein B‐100 (ApoB) in dogs with DM.

Animals

22 dogs with DM and 9 healthy dogs.

Methods

Cross‐sectional study. Triglyceride (TG), total cholesterol (TC), and high‐density lipoprotein cholesterol (HDL‐C) concentrations were measured by spectrophotometry. Very low‐density lipoprotein cholesterol (VLDL‐C) and low‐density lipoprotein cholesterol (LDL‐C) concentrations were calculated after ultracentrifugation. Non‐HDL‐C cholesterol was calculated by subtracting HDL‐C from TC. ApoB was quantified by ELISA. The Mann‐Whitney test was used for comparison of median lipoprotein concentrations, and Spearman's correlation was used to assess associations between ApoB and lipoprotein fractions.

Results

All values are reported in mg/dL. Median TG (122), TC (343.5), HDL‐C, (200), VLDL‐C, (27) LDL‐C (68), non‐HDL‐C (114), and ApoB (320) were significantly higher in dogs with DM, compared to healthy dogs (57, 197, 168, 12, 16, 31, and 258, respectively, P‐values 0.0079, <0.001, 0.029, 0.011, <0.001, <0.001, 0.025, respectively). A significant association was found between ApoB and LDL‐C (Spearman's rho = 0.41, P = 0.022) and between ApoB and non‐HDL‐C (Spearman's rho = 0.40, P = 0.027).

Conclusions and Clinical Importance

Dyslipidemia of dogs with DM is characterized by pronounced increases in LDL‐C and non‐HDL‐C concentrations, although all lipoprotein fractions are significantly increased. Knowledge of specific lipoprotein fraction alterations in dogs with DM can enhance treatment options for diabetic dyslipidemia in dogs.     

Plasma lipids, lipoproteins and post-heparin lipases in ponies with hyperlipaemia.

The metabolic origins of equine hyperlipaemia were investigated by analysing the concentration and composition of plasma lipoproteins in 18 ponies with the condition. The mean concentrations of cholesterol, triglyceride and very low density lipoproteins (VLDL) were increased by 4-, 52- and 19-fold, respectively, compared with a control group of 18 healthy ponies. These increases were due to the appearance of a buoyant VLDL fraction (VLDL1) not present in healthy ponies. The mean diameter of VLDL1 particles was 44% greater than control VLDL, and the particles were enriched in triglyceride and free cholesterol and depleted of cholesteryl esters, phospholipid and protein. The apolipoprotein (apo) B-100 content of VLDL1 was reduced and the ratio of apoB-100 to apoB-48 particles was 1:1, compared with 2:1 in control VLDL. The VLDL1 was also enriched in apoE, but had normal complements of apoC-II and apoC-III. The conventional VLDL (called VLDL2), LDL and HDL fractions were moderately enriched with triglyceride, and HDL contained increased amounts of apoE, apoC-II and apoC-III. The activities of lipoprotein lipase and hepatic lipase, the enzymes responsible for the catabolism of VLDL and their remnants, were increased by 2- and 3-fold, respectively, in response to the increased concentrations of their substrates. The composition of VLDL1 suggested that the liver was maximising the secretion of triglyceride by producing larger number of VLDL particles that accommodated a greater mass of triglyceride by having apoB-48 rather than apoB-100 as their structural protein. Plasma free fatty acid (FFA) concentrations were elevated in 17 of the 18 ponies, suggesting that increased FFA flux might be the stimulus for hepatic triglyceride synthesis and VLDL secretion. We conclude that overproduction, rather than defective catabolism, of VLDL was the cause of the hyperlipidaemia and that lipid lowering agents which reduce VLDL synthesis, by decreasing adipose lipolysis and FFA flux, are candidates for the management of hyperlipaemia.     

Blood lipid concentrations and lipoprotein patterns in captive and wild American black bears (Ursus americanus)

OBJECTIVE: To compare blood lipid concentrations and lipoprotein patterns for captive and wild American black bears (Ursus americanus). ANIMALS: 7 captive and 9 wild adult (> or = 4 years old) black bears. PROCEDURE: Blood was collected from 2 groups of captive black bears (groups A and B) and 1 group of wild black bears (group C). Blood triglyceride (TG) and cholesterol concentrations were compared among groups. Plasma lipoproteins were isolated by use of a self-generating gradient of iodixanol, and lipoprotein patterns were compared between groups A and B. RESULTS: Captive bears (mean +/- SD, 187.8 +/- 44.4 kg) weighed significantly more than wild bears (mean, 104.8 +/- 41.4 kg), but mean body weight did not differ between groups A and B. Mean blood TG concentrations for groups B (216.8 +/- 16.0 mg/dL) and C (190.7 +/- 34.0 mg/dL) were significantly higher than that of group A (103.9 +/- 25.3 mg/dL). Mean blood cholesterol concentration was also significantly higher for group B (227.8 +/- 8.2 mg/dL) than for groups A (171.7 +/- 35.5 mg/dL) or C (190.8 +/- 26.8 mg/dL). Mean very-low-density lipoprotein TG and low-density lipoprotein cholesterol concentrations were 2- and 3-fold higher, respectively, for group B, compared with concentrations for group A. CONCLUSIONS AND CLINICAL RELEVANCE: Blood lipid concentrations vary significantly among populations of black bears. Plasma lipoprotein patterns of captive bears differed significantly between colonies and may have reflected differences in diet or management practices.     

Progesterone production by cultured luteal cells in the presence of bovine low- and high-density lipoproteins purified by heparin affinity chromatography.

The objectives of this study were to separate plasma lipoprotein particles based on the presence (low-density lipoproteins; LDL) or absence of apolipoprotein B (high-density lipoproteins; HDL) and to compare the abilities of bovine LDL and HDL to stimulate progesterone production by bovine luteal cells in culture. Plasma lipoproteins were isolated by ultracentrifugation and separated into LDL and HDL by heparin affinity chromatography. Luteal cultures were treated with LDL or HDL cholesterol for 48 h on d 3 of the culture (d 0 = day of dissociation). Progesterone production by luteal cells increased in a dose-dependent manner with increasing concentrations of either LDL or HDL cholesterol. There were no differences in the ability of LDL or HDL cholesterol to stimulate luteal cells to produce progesterone. Because LDL and HDL were equally potent, and experiment was designed to investigate the ability of modified LDL or reconstituted particles without apolipoproteins to stimulate progesterone production. Stimulation of luteal cell progesterone production by lysine-modified LDL was 70% of unmodified LDL. Progesterone production in the presence of phosphatidylcholine liposomes or BSA containing cholesterol was 50 and 108% of that obtained with HDL or LDL. Evidence indicated that apolipoprotein-free particles that contained free cholesterol but not cholesterol esters stimulated luteal cells to produce progesterone.     

Plasma lipids, lipoprotein composition and profile during induction and treatment of hepatic lipidosis in cats and the metabolic effect of one daily meal in healthy cats

Anorexia in obese cats may result in feline hepatic lipidosis (FHL). This study was designed to determine plasma lipids and lipoprotein profiles in queens at different stages during experimental induction of FHL (lean, obese, FHL), and after 10 weeks of treatment. Results were compared with those obtained from lean queens of same age fed the same diet but at a maintenance level, once a day. Hepatic lipidosis led to an increase in plasma triacylglycerol (TG), very low density lipoprotein (VLDL) and low density lipoprotein (LDL), and an enrichment of LDL with TG and of high density lipoprotein (HDL) with cholesterol, suggesting that VLDL secretion is enhanced, VLDL and LDL catabolism is lowered, and lipoprotein exchanges are impaired in FHL. This study also showed that cholesterolaemia is increased in cats fed at a dietary rhythm of one meal per day compared to ad libitum feeding.     

Serum lipid and lipoprotein parameters of Turkman horses

BACKGROUND: Lipid transport systems in animals have been evaluated both as experimental models for lipid and lipoprotein metabolism in humans and to gain insight into the lipid metabolism of specific breeds. The Turkman horse is a pure-blooded horse that lives on the margins of the Caspian Basin; information about serum lipids and lipoproteins in this breed is lacking. OBJECTIVE: The purpose of this study was to evaluate serum lipid and lipoprotein concentrations in clinically healthy Turkman horses. METHODS: Concentrations of triglyceride (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDLC), low density lipoprotein cholesterol (LDL-C), and very low density lipoprotein cholesterol (VLDL-C), and lipoprotein electrophoretic patterns were determined in serum samples from 48 Turkman horses of both sexes and 0-12 years of age. Results were analyzed by 1-way ANOVA. RESULTS: In both sexes and all age groups, the relative percentages of LDL-C (44.2-64.3%) were higher than those of HDL-C (29.7-47.1%) and VLDL-C (5.9-9.0%). Serum concentrations of TG and TC were significantly higher in male (0.45 +/- 0.12 mmol/L; 2.70 +/- 0.45 mmol/L) than in female (0.34 +/- 0.1 mmol/L; 2.15 +/- 0.3 mmol/L) horses in most age groups. The relative percentages of alpha- and beta-lipoproteins in electrophoretic tracings were 62.77 +/- 5.05% and 36.67 +/- 4.29%, respectively. CONCLUSION: Serum lipid and lipoprotein values in Turkman horses differ from those of other equine breeds, and may be useful for evaluating metabolic diseases in this species.     

Effect of hypothyroidism on the blood lipid response to higher dietary fat intake in mares

Blood lipid and lipoprotein concentrations were measured and compared between euthyroid and thyroidectomized mares on low-fat or high-fat diets to test the hypothesis that hypothyroidism alters the blood lipid response to higher dietary fat intake. Four healthy adult mares and four adult mares that had been thyroidectomized 3 to 6 mo earlier were placed on low-fat or high-fat diets according to a replicated 2 x 2 Latin square design consisting of two 5-wk feeding periods separated by a 2-wk washout interval. Plasma lipid concentrations were measured at 0, 3, 4, and 5 wk, and plasma lipase activities were measured at the end of each 5-wk feeding period. Compared with euthyroid mares (0.46 ng/mL [range 0.34 to 0.68 ng/mL T3], and 21.5 ng/mL [range 18.1 to 25.1 ng/mL T4], respectively), median serum concentrations of T3 and T4 were lower (P = 0.029 and P = 0.021, respectively) in thyroid-ectomized mares (0.26 ng/mL [range 0.23 to 0.26 ng/ mL T3], and undetectable T4). Serum T4 concentrations were below the limits of detection in thyroidectomized horses. Alterations in body weight over 5 wk did not differ between groups. Mean plasma very low density lipoprotein (VLDL) and triglyceride (TG) concentrations were higher (P = 0.045 and 0.034, respectively) in hypothyroid mares (55.42 +/- 35.05 mg/dL and 52.83 +/- 34.46 mg/dL, respectively) compared with euthyroid mares (28.28 +/- 13.76 mg/dL and 23.53 +/- 9.84 mg/dL, respectively). Mean plasma total cholesterol (TC) concentrations increased from 88.73 +/- 25.49 mg/dL at baseline to 103.93 +/- 24.42 mg/dL after 5 wk on the low-fat diet, but increased by a greater magnitude (P = 0.006 diet +/- time interaction) in mares that were on the high-fat diet (81.05 +/- 17.24 mg/dL and 123.84 +/- 32.27 mg/ dL, respectively). Mean plasma TC concentrations were higher (P = 0.099) in hypothyroid mares (116.16 +/- 32.89 mg/dL) than in euthyroid mares (89.56 +/- 14.45 mg/ dL). Higher post-heparin plasma lipoprotein lipase and hepatic lipase activities (P = 0.012 andP = 0.017, respectively) were detected in mares that were on the high-fat diet (2.66 +/- 0.91 micromol FA x mL(-1) x h(-1) and 2.95 +/- 0.49 micromol FA x mL(-1) x h(-1), respectively) vs. a low-fat diet (1.75 +/- 0.55 micromol FA x mL(-1) x h(-1) and 2.27 +/- 0.59 micromol FA x mL(-1) x h(-1), respectively). We conclude that plasma VLDL and TG concentrations are elevated in hypothyroid mares, but the blood lipid response to higher dietary fat intake is not influenced by hypothyroidism.     

Lipoprotein metabolism and hyperlipidaemia in the clog and cat: A review

The term hyperlipidaemia is used to describe raised plasma concentrations of cholesterol and, or, triglycerides. These aqueous insoluble lipids are transported through plasma in special particles called lipoproteins of which there are four main types; chylomicrons, very low density lipoproteins (VLDL), low density lipoproteins (LDL) and high density lipoproteins (HDL). A transient rise in plasma triglycerides occurs following a meal as dietary fat is carried from the small intestine into the circulation by chylomicrons; this is called post prandial hyperlipidaemia. In addition, hyperlipidaemia is caused by defects in the metabolism of one or more of the lipoprotein classes which may be either genetic in origin or, more commonly in the dog and the cat, secondary to diabetes mellitus, hypothyroidism, hyperadrenocorticism, and renal and hepatic disease. Hypertriglyceridaemia is caused by reduced clearance of chylomicrons and VLDL, sometimes with overproduction of VLDL, whereas hypercholesterolaemia results from altered metabolism of LDL and HDL. Raised plasma triglycerides interfere with a number of clinical chemistry tests and may be associated with cutaneous xanthomata, vomiting and diarrhoea, peripheral nerve paralyses, seizures, pancreatitis, hepatosplenomegaly and lipaemia retinalis. The clinical manifestations of hypercholesterolaemia in the dog are few and largely confined to the eye. Diagnostic efforts should concentrate on determining whether the hyperlipidaemia is either genetic in origin or secondary to endocrine and systemic diseases. Plasma lipid concentrations usually return to normal with effective therapy of any underlying disease. Where no such disease can be identified, the hyperlipidaemia should be considered idiopathic in origin and the patient placed on a low fat diet.     

Effect of dietary conjugated linoleic acids on the distribution of fatty acids in serum lipoprotein fractions and different tissues of growing pigs

The effect of conjugated linoleic acid (CLA) on blood lipids [total cholesterol, high-density lipoprotein (HDL), low-density lipoprotein (LDL) and triacylglycerols (TAG)] and the fatty acid distribution of the lipoprotein fractions, backfat, muscle fat, and liver lipids were examined in an experiment with two groups of 40 pigs [Pietrain x (Landrace x Large White)] each. The 20 female and 20 male castrated pigs of each group were fed with isoenergetic and isonitrogenous diets supplemented with either 20 g/kg rapeseed oil (control) or 20 g/kg CLA-TAG. The CLA preparation contained 54.2% pure CLA consisting of approximately two-thirds cis,trans/trans,cis-isomers and one-third trans,trans-isomers. The fatty acids of lipoproteins, backfat, muscle lipids and liver lipids were analysed by gas chromatograph (GC). CLA supplementation did not significantly influence blood lipids and the LDL to HDL ratio. In the CLA-fed pigs the very low-density lipoprotein (VLDL) contained higher saturated fatty acid (SFA) concentrations at the cost of the monounsaturated fatty acids (MUFA). The percentage of polyunsaturated fatty acids (PUFA) remained unchanged. The highest CLA content was analysed in VLDL (4.00%) followed by LDL (2.78%) and HDL (1.45%). The ratio of cis,cis to trans,trans isomers increased from VLDL over LDL to HDL. The content of SFA, probably in backfat and muscle lipids, increased whereas the part of MUFA decreased as a result of reduced Delta9-desaturase activity. The percentage of PUFA (without CLA) was higher in backfat of the control group in accordance with the dietary PUFA supply. This shift in the fatty acid distribution was not observed in the liver lipids. In all the three tissues analysed, the CLA-fed pigs had a significantly increased CLA content: the highest increase was in the backfat (5.65%), followed by liver lipids (2.41%), and muscle lipids (1.47%). An isomer-specific accumulation was observed for cis,cis-CLA isomers in muscle, and for trans,trans-CLA isomers in backfat. We conclude that CLA supplementation results in a higher SFA content in backfat and muscle lipids but not in liver lipids. There is a discrimination of the trans-10, cis-12 isomer and the trans,trans isomers in the formation of the cell membranes.     

Potential use of cholesterol lipoprotein profile to confirm obesity status in dogs

A common sign of obesity, in dogs, is hyperlipidemia, which is characterized by hypercholesterolemia and/or hypertriglycemia. Hyperlipidemia can be caused by a quantitative increase in circulating lipoproteins (LP) or by a higher lipid concentration in the various LP classes. In this study, we sought to determine whether aberrations occur with cholesterol lipoprotein profile, especially with sub HDL-cholesterol fraction % in obese dogs. Using clinically healthy and disease free (no overt signs) body condition score classified obese dogs, of all ages, we attempted to determine the influence of age, gender and obesity status on cholesterol lipoprotein profiling. Overall, no aberration in pattern was observed in obese dogs <8 years of age. However, in older obese animals (≥8 years of age), the general aberration pattern to cholesterol lipoprotein observed was that a significant decrease in HDL2 and 3 fraction % occurs with a concomitant increase in either HDL1-Cho or VLDL and LDL -Cho fraction % depending on gender. Linear regression analysis indicated that obesity status appears to significantly affect total cholesterol, HDL2 and 3-Cho, VLDL and LDL-Cho levels (P = 0.02, 0.046, and 0.045, respectively), whereas it is borderline with HDL1-Cho (P = 0.062). On the other hand, age significantly influenced TG, Total cholesterol, and HDL1-Cho levels (P = 0.009, 0.006, and 0.002, respectively), while gender influenced VLDL and LDL-Cho (P = 0.024) level. Therefore, aberrations in cholesterol lipoprotein profile pattern might be of potential use to assess and diagnose obesity status, in conjunction with BCS, especially of older overweight animals which might be considered borderline obese.     

Effects of rice bran oil on plasma lipid concentrations, lipoprotein composition, and glucose dynamics in mares

Plasma lipid concentrations, lipoprotein composition, and glucose dynamics were measured and compared between mares fed diets containing added water, corn oil (CO), refined rice bran oil (RR), or crude rice bran oil (CR) to test the hypothesis that rice bran oil lowers plasma lipid concentrations, alters lipoprotein composition, and improves insulin sensitivity in mares. Eight healthy adult mares received a basal diet fed at 1.5 times the DE requirement for maintenance and each of the four treatments according to a repeated 4 x 4 Latin square design consisting of four 5-wk feeding periods. Blood samples were collected for lipid analysis after mares were deprived of feed overnight at 0 and 5 wk. Glucose dynamics were assessed at 0 and 4 wk in fed mares by combined intravenous glucose-insulin tolerance tests. Plasma glucose and insulin concentrations were measured, and estimated values of insulin sensitivity (SI), glucose effectiveness, and net insulin response were obtained using the minimal model. Mean BW increased (P = 0.014) by 29 kg (range = 10 to 50 kg) over 5 wk. Mean plasma concentrations of NEFA, triglyceride (TG), and very low-density lipoprotein (VLDL) decreased (P < 0.001) by 55, 30, and 39%, respectively, and plasma high-density lipoprotein and total cholesterol (TC) concentrations increased (P < 0.001) by 15 and 12%, respectively, over 5 wk. Changes in plasma NEFA (r = 0.58; P < 0.001) and TC (r = 0.44; P = 0.013) concentrations were positively correlated with weight gain over 5 wk. Lipid components of VLDL decreased (P < 0.001) in abundance over 5 wk, whereas the relative protein content of VLDL increased by 39% (P < 0.001). Addition of oil to the basal diet instead of water lowered plasma NEFA and TG concentrations further (P = 0.002 and 0.020, respectively) and increased plasma TC concentrations by a greater magnitude (P = 0.072). However, only plasma TG concentrations and VLDL free cholesterol content were affected (P = 0.024 and 0.009, respectively) by the type of oil added to the diet. Mean plasma TG concentration decreased by 14.2 mg/dL over 5 wk in the CR group, which was a larger (P < 0.05) decrease than the one (-5.3 mg/dL) detected in mares that received water. Consumption of experimental diets lowered S(I), but glucose dynamics were not affected by oil supplementation. Addition of oil to the diet altered blood lipid concentrations, and supplementation with CR instead of water specifically affected plasma TG concentrations and VLDL free cholesterol content.     

Effect of hypothyroidism on blood lipid concentrations in horses.

OBJECTIVE: To measure and compare concentrations of selected blood lipids before and after thyroidectomy in horses. ANIMALS: 5 healthy adult mares. PROCEDURE: Mares were confirmed to be euthyroid. Thyroidectomy was performed, and hypothyroidism was confirmed. Selected blood lipid variables were measured before hypothyroidism was induced and weekly for 4 weeks after induction. Plasma concentrations of very low-density lipoprotein (VLDL), low-density lipoprotein (LDL), serum triglyceride (TG), total cholesterol (TC), and nonesterified fatty acid (NEFA) were measured. The composition of VLDL and LDL also was examined. RESULTS: Mean plasma concentrations of VLDL and LDL increased significantly after thyroidectomy. By 4 weeks after thyroidectomy, a ninefold increase in mean plasma concentration of VLDL and a threefold increase in LDL, compared with baseline values, were detected. After thyroidectomy, mean percentage of TG in VLDL increased significantly, whereas free cholesterol and cholesterol ester content decreased. Mean percentage of TG in LDL also increased by 3 to 4 weeks after thyroidectomy. Serum concentrations of TG and TC increased, whereas serum NEFA concentration decreased. CONCLUSIONS: Hypothyroidism significantly alters blood lipid concentrations of horses. After thyroidectomy, markedly high VLDL concentration, appearance of TG-rich VLDL, increased serum concentrations of TG and TC, and decreased blood concentration of NEFA were evident. CLINICAL RELEVANCE: Examination of blood lipid concentrations of horses may be useful for detecting naturally acquired hypothyroidism.     

Effect of hypothyroidism on kinetics of metabolism of very-low-density lipoprotein in mares

OBJECTIVE: To compare kinetics of the metabolism of very-low-density lipoprotein (VLDL) apolipoprotein B (apoB) before and after thyroidectomy in mares. ANIMALS: 5 healthy adult mares. PROCEDURE: Thyroidectomy was performed in euthyroid mares. Kinetics of VLDL apoB metabolism were measured before and after thyroidectomy by use of a bolus IV injection of 5,5,5-2H3 (98%) leucine (5 mg/kg) and subsequent isolation of labeled amino acid from plasma and VLDL. Labeled leucine was quantified by use of gas chromatography-mass spectrometry. Production rate (PR), delay time, and fractional catabolic rate (FCR) were calculated for the 2 forms of equine VLDL, apoB-48 VLDL, and apoB-100 VLDL. Plasma lipid concentrations were measured, and VLDL composition was determined. RESULTS: Physical appearance of horses was not altered by thyroidectomy. Significantly lower mean blood concentrations of thyroid hormones and non-esterified fatty acids were detected following thyroidectomy. Mean percentage of free cholesterol in VLDL was significantly higher after thyroidectomy. Mean plasma VLDL concentration or kinetics of apoB-48 or apoB-100 were not significantly altered by thyroidectomy. Mean +/- SEM PR was significantly lower (8.70 +/- 1.61 mg/kg/d) and mean delay time significantly longer (1.58 +/- 0.12 hours) for apoB-48 VLDL in euthyroid mares, compared with values for thyroidectomized mares (16.15 +/- 2.24 mg/kg/d and 0.93 +/- 0.10 hours, respectively). CONCLUSIONS AND CLINICAL RELEVANCE: Hypothyroidism did not significantly alter plasma VLDL concentrations or kinetics of VLDL apoB metabolism. Metabolism of apoB-48 VLDL differed significantly from that of apoB-100 VLDL in euthyroid mares.     

A review of lipoprotein cholesterol metabolism: importance to ovarian function

Cholesterol utilized for steroid synthesis by ovarian tissue may be derived from de novo synthesis or cellular uptake of lipoprotein cholesterol. The majority of blood cholesterol is transported by either low (LDL) or high (HDL) density lipoproteins, depending on the animal species. Prior to vascularization, only HDL are in follicular fluid and contribute sterol to granulosa cells because other lipoproteins are unable to transverse the basement membrane due to their molecular masses. Following vascularization, both LDL and HDL bathe luteal cells. Most species preferentially use LDL cholesterol as a precursor for ovarian steroid synthesis. The LDL uptake by ovarian tissue occurs by receptor-mediated endocytosis. The receptor recognizes apolipoprotein B of LDL and apolipoprotein E found on some, but not all, HDL. Within a species, a positive relationship may exist between HDL apolipoprotein E content and importance of HDL cholesterol as a precursor for steroidogenesis. A "HDL pathway" exists for uptake of sterol from HDL void of apolipoprotein E. The HDL receptor exhibits broad binding specificity. Unlike LDL, the HDL particle is not internalized, and cholesterol preferentially is taken up relative to other HDL constituents. In most species, lipoproteins, rather than de novo synthesis from acetate, contribute the majority of cholesterol used for steroid production. Trophic hormones increase lipoprotein binding, internalization, degradation and conversion of lipoprotein-derived sterol to steroids, effects that are mediate through cyclic adenosine monophosphate. Knowledge recently acquired regarding lipoprotein sterol utilization by the ovary may be useful in developing nutritional, pharmacological or endocrine manipulations that may positively affect cholesterol clearance by the ovary, steroidogenesis and reproductive performance.     

An investigation of the relationships between body condition and plasma lipid and lipoprotein concentrations in 24 donkeys

Obese donkeys are susceptible to a hyperlipaemic crisis characterised by high plasma triglyceride concentrations. In this study, the relationships between the body condition of 24 donkeys and their basal lipid metabolism were investigated. Plasma cholesterol, triglyceride and lipoprotein cholesterol concentrations were measured in healthy donkeys classified according to their body condition as thin, ideal or obese. There were significant differences between the groups in the concentrations of triglyceride and very low density lipoprotein (VLDL), which increased in concentration with body condition (P less than 0.05). Cholesterol, low density lipoprotein (LDL) and high density lipoprotein (HDL) concentrations were similar in all the groups. Triglyceride and VLDL concentrations were positively correlated with body weight (r = 0.82) and plasma free fatty acid concentration (r = 0.48). There were no significant differences in basal plasma concentrations of insulin or cortisol. These results suggest that obesity in donkeys is associated with changes in lipid and lipoprotein metabolism that might predispose the animals to hyperlipaemia.