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.     

Serum lipid profile in Iranian fat-tailed sheep in late pregnancy, at parturition and during the post-parturition period

Blood samples were obtained from 12 Iranian fat-tailed sheep during 7 weeks pre-partum, at parturition and 7 weeks post-partum. The lipids measured were cholesterol, triglyceride, total lipid, high-density lipoprotein (HDL)-cholesterol, low-density lipoprotein (LDL)-cholesterol, and very low-density lipoprotein (VLDL)-cholesterol. The concentrations of cholesterol, triglyceride, HDL-cholesterol and VLDL-cholesterol during the 7 weeks pre-partum, at parturition and the 7 weeks post-partum were significantly different (P < 0.05). One week before parturition, the concentrations of cholesterol, triglyceride, HDL-cholesterol and VLDL-cholesterol were higher (P < 0.05) than at other periods. The lowest concentrations of these parameters were observed 2-3 weeks after parturition. In this study, significant positive correlations were observed between the time of sampling (pre-partum, parturition and post-partum) and serum cholesterol (r = 0.22; P < 0.01) and HDL-cholesterol (r = 0.25; P < 0.01).     

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.     

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.     

Composition and electrophoretic mobility of plasma lipoproteins of dromedary camels (Camelus dromedarius)

OBJECTIVE: To determine the lipid composition and electrophoretic pattern of plasma lipoproteins in samples obtained from healthy 1-humped camels (Camelus dromedarius). ANIMALS: 34 healthy camels raised under similar farming and dietary conditions. PROCEDURES: Plasma samples were subjected to density-gradient ultracentrifugation for separation of plasma lipoproteins, including very-low-density lipoproteins (VLDL), low-density lipoproteins (LDL), and high-density lipoproteins (HDL). Purity of the separation was assessed by use of polyacrylamide gel disk electrophoresis. Concentrations of triglycerides, cholesterol, and phospholipids were measured in each lipoprotein fraction, and lipoprotein electrophoretic patterns were determined in plasma samples. RESULTS: Phospholipid was the major constituent of VLDL (mean +/- SD concentration, 10.62 +/- 1.2 mg/dL), LDL (24.66 +/- 3.12 mg/dL), and HDL (38.08 +/- 0.76 mg/dL). Low-density lipoprotein, VLDL, and HDL were important plasma lipoprotein carriers for cholesterol (67.94 +/- 9.51%), triglyceride (55.83 +/- 7.81%), and phospholipid (51.91 +/- 1.55%), respectively. On the basis of electrophoresis results, relative percentages of alpha- and beta-lipoproteins were 31.72 +/- 4.88% and 68.3 +/- 4.68%, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: The lipoprotein profile in 1-humped camels differed substantially from that of other ruminants. Results may be useful in the evaluation of metabolic disorders in camels.     

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, 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.     

Changes in the plasma concentrations of lipids and lipoprotein fractions in dogs infected with Leishmania infantum.

A study was made of serum concentrations of cholesterol, phospholipids, triglycerides, cholesterol bound to high-density lipoproteins (HDL), HDL1-cholesterol, HDL2-cholesterol and cholesterol bound to low-density lipoproteins (LDL) in 16 dogs naturally infected with Leishmania infantum (ZMON-1) taken from an endemic focus. Results were compared with those of a control group of ten healthy dogs. Statistically significant increases in cholesterol, triglyceride and LDL-cholesterol levels were observed. There was, however, a statistically significant decrease in HDL-cholesterol level, mainly at the expense of the HDL2-cholesterol subfraction. Cholesterol transport is therefore shown to undergo changes which may be attributed to the consumptive evolution of the disease, immunocomplex deposits in cells, hepatic disorders and interactions between the parasite and the normal cholesterol metabolism of the host.     

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.     

Dietary pea protein stimulates bile acid excretion and lowers hepatic cholesterol concentration in rats

It has been shown that some dietary plant proteins beneficially influence lipid metabolism in animals. The effect of pea protein in this respect however has not yet been investigated. Therefore, we studied the effect of purified pea protein on the lipid metabolism in rats. Twenty-four rats received diets with either 200 g/kg of casein or purified pea protein for 16 days. Concentrations of triacylglycerols in liver, plasma and lipoproteins did not differ between both groups of rats. However, rats fed the pea protein diet had a lower concentration of total cholesterol in the liver and the very low density lipoproteins (VLDL) fraction than rats fed the casein diet (p < 0.05); cholesterol concentration in plasma, low density lipoproteins (LDL) and high density lipoproteins (HDL) did not differ between both groups. Rats fed pea protein moreover had an increased mRNA concentration of cholesterol-7α-hydroxylase in the liver and an increased amount of bile acids excreted via faeces compared with rats fed casein (p < 0.05). Concomitantly, mRNA concentrations of sterol regulatory element-binding protein (SREBP)-2 and its target genes 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase and LDL receptor in the liver were increased in rats fed pea protein (p < 0.05). The data of this study suggests that pea protein stimulates formation and excretion of bile acids, which leads to a reduced hepatic cholesterol concentration and a reduced secretion of cholesterol via VLDL. An increased gene expression of SREBP-2 and its target genes HMG-CoA reductase and LDL receptor may be a means to compensate for the increased loss of cholesterol for bile acid synthesis.     

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)     

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.     

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.     

Changes in lipoprotein composition in horses fed a fat-supplemented diet

This study was designed to compare the effects of a fat-supplemented diet versus a traditional diet on the lipoprotein (LP) content in horses. In the first of two trials, eight two-year old horses were fed a basal diet, in which 80% of the digestible energy was supplied as chopped alfalfa hay. One group of four horses (fat-supplemented group, FS) was fed the remaining 20% of the digestible energy as corn oil, while the other group of four horses (control) was fed rolled corn to complete their diet. Blood samples were collected at the start of the experiment and every 2.5 weeks thereafter for 10 weeks. Total serum lipids were measured in both groups of horses and the lipoproteins were fractionated into very low density LP (VLDL), low density LP (LDL), and high density LP (HDL) using ultracentrifugation and agarose-column chromatography. Each LP fraction was measured for protein, cholesterol (CH) and triglyceride TG) content. Total serum lipids were increased in the FS group above the control (9.16 vs. 4.65 mg/ml, week 5). Serum cholesterol and triglyceride concentrations were elevated in the FS group, but were highly variable. Variations in lipid concentrations may have been due to variation in time of sampling during the day. In the second trial, four of the eight horses were used and divided into the same two groups; FS vs. control. After an initial sample, postprandial serial blood samples (1 hour intervals for 8 hours) were drawn at 2, 4 and 6 weeks. Upon examination of the data, Hour 3 post-feeding was chosen to represent postprandial LP values. In both trials, the horses were able to adapt to the added dietary fat. Maintenance of body weight and increased speed of lipid clearance from the blood by the end of each trial in the FS group support this statement. In the FS group, there was an increase in VLDL TG concentration, but not in LDL. This indicates an increase in VLDL TG clearance from the circulation, presumably by increased lipoprotein lipase (LPL) activity. Cholesterol concentrations were also increased in the FS horses in the LDL and HDL. The rise in cholesterol may be attributed to endogenous recycling of liver products such as bile salts, which aid in digestion and absorption and are cholesterol based. In addition, there were greater quantities of LDL and HOL produced in the FS horses as supported by the increased protein concentrations as well as larger peaks for the eluate from the gel filtration column.     

Effects of supplemental copper and chromium on the serum and egg traits of laying hens

1. This study investigated the effects of supplemental copper and chromium on the serum and egg traits of laying hens using 144 forty-five-week-old White Leghorn layers. The hens were randomly assigned into 9 groups to conduct this 3 copper (0, 125 and 250 mg/kg as copper sulphate) x 3 chromium (0, 800 and 1600 microg/kg as chromium picolinate) factorial trial. 2. Egg yolk cholesterol was significantly reduced by supplementary copper (Cu) and chromium (Cr) and there was an interaction between Cu and Cr supplementation. Egg production, egg weight, eggshell strength and eggshell thickness were not influenced by Cu or Cr supplementation. 3. Serum cholesterol concentration was decreased by Cu supplementation and very low-density lipoprotein (VLDL) was reduced markedly by both Cu and Cr. On the other hand, high-density lipoprotein (HDL) was significantly increased by Cu and Cr. Interaction occurred between Cu and Cr in the VLDL and HDL parameters. Supplmentary Cu also reduced VLDL-cholesterol (VLDL-C) and enhanced HDL-cholesterol (HDL-C), while supplemental Cr had a contrary effect on these two parameters. 4. If minimum egg cholesterol content is the criterion, this study suggests that 125 mg/kg of Cu together with 800 to 1600 microg/kg of Cr provide adequate supplementary concentrations for laying hens.     

Evaluation of a commercially available apparatus for measuring the acute phase protein serum amyloid A in horses

The aim of this study was to investigate the reliability of an immunoturbidometric assay for measuring the acute phase protein serum amyloid A (SAA) in horses in clinical practice. The assay was compared to a previously validated assay, and overlap performance was assessed by measuring the concentration of SAA in clinically healthy horses and horses with inflammatory and non-inflammatory diseases. In pools of serum with low and high SAA concentrations the assay's intra-assay coefficients of variation were 11.7 per cent and 4.6 per cent, and its interassay coefficients of variation were 9.1 per cent and 5.6 per cent, respectively. Slight inaccuracies were observed, but they were negligible in comparison with the range of the SAA response. The assay systematically underestimated the concentrations of SAA in comparison with the results of the validated assay. The assay detected the expected difference in SAA concentrations between the healthy and diseased horses.     

Serum Lipids and Lipoproteins in Clinically Healthy Male Camels (Camelus dromedarius)

Blood samples were collected from the jugular vein of 87 Iranian male dromedary camels of different age groups (<3, 3–5, 5–6 and 6–8 years). Variations in the serum concentrations of cholesterol, triglyceride, total lipid, very low-density lipoprotein (VLDL-cholesterol), low-density lipoprotein (LDL-cholesterol) and high-density lipoprotein (HDL-cholesterol) were investigated. The concentrations of cholesterol, triglyceride, total lipid, HDL-cholesterol, LDL-cholesterol and VLDL-cholesterol in the different age groups were significantly different (p<0.05). With an increase in the age of the animals, there were highly significant increases in the cholesterol, triglyceride, HDL-cholesterol and VLDL-cholesterol concentrations, whereas the concentration of LDL-cholesterol decreased (p<0.001).     

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.     

Paraoxonase (PON) 1, 2 and 3 Expression in Granulosa Cells and PON1 Activity in Follicular Fluid of Dairy Cows

Normal metabolic activity in ovarian follicles may result in oxidative stress and damage to oocytes. The aim of this study was to evaluate expression of the natural anti‐oxidants paraoxonase (PON) 1, 2 and 3 in granulosa cells and PON1 activity in follicular fluid (FF) and plasma of dairy cows. For the first experiment, ovaries were collected from cows at slaughter, after which follicles were dissected and classified as oestrogen active (EAF) or atretic (ATF). Expression of PON1, PON2 and PON3 mRNA was evaluated in granulosa cells, and activity of PON1 was measured in FF. PON1 mRNA was undetectable in granulosa cells, PON2 mRNA expression was not different between follicle types, and PON3 mRNA tended to be higher in EAF (p = 0.11). The activity of PON1 in FF was higher (p = 0.01) for EAF (82.6 ± 8.0 kU/L) than ATF (53.9 ± 6.8 kU/L), as were high‐density lipoproteins (HDL), low‐density lipoproteins (LDL) and total cholesterol concentrations. In the second experiment, we aimed to compare plasma and FF PON1 activity in early lactation Holstein cows (n = 15) with pre‐ovulatory EAF. Activity of PON1 was twofold higher (p < 0.0001) in plasma (122.5 ± 11.1 kU/L) than in FF (61.4 ± 5.2 kU/L). Plasma concentrations were also higher (p < 0.0001) for HDL, LDL and total cholesterol when compared to FF. In conclusion, FF concentrations of PON1, HDL, LDL and total cholesterol were higher in healthy oestrogen active bovine follicles than in atretic follicles. PON1 was not expressed by granulosa cells indicating that high PON1 activity in bovine FF is apparently derived by transfer from blood in association with HDL.     

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.