Dietary Cholesterol Supplementation Research Articles

Enhanced cholesterol utilization and impact on growth through taurine supplementation in high plant-based diets of Pacific white shrimp, Penaeus vannamei in biofloc system

Cholesterol is vital for the optimal growth and physiological function of Pacific white shrimp, Penaeus vannamei. Due to limited availability of fish meal, plant-based ingredients are increasingly added in the shrimp diet which limit the availability and utilization of cholesterol. This study investigated the synergistic effects of dietary taurine and cholesterol supplementation on growth and physio-metabolic responses of Pacific white shrimp, Penaeus vannamei fed with high plant-based protein diets reared in a biofloc system for 60 days. A basal diet (control) comprised 82.55% plant-based ingredients and 36% crude protein. Five isonitrogenous and isolipidic diets were prepared (T1-T5) with cholesterol and taurine supplements at inclusion levels of 0.15% and 0.16% in T1 and T2, respectively. In the rest of the treatments, T3, T4, and T5, the combinations of cholesterol and taurine were 0.15% and 0.16%; 0.05% and 0.24%; 0.25% and 0.08%, respectively. The experimental diets were allocated to triplicate groups of 100 shrimp per tank following a completely randomized design. The results showed that shrimp fed T3 diet have a significantly (P < 0.05) higher percentage of weight gain, specific growth rate (SGR), percentage of survival, and protein efficiency ratio (PER) with efficient feed conversion ratio (FCR) (1.14±0.06) compared to other treatments. The hematological, antioxidant, and digestive enzyme activities, except amylase activity, were significantly (P < 0.05) higher in shrimp fed T1 and T3. Total bile acid content in T3 fed shrimp was higher than in other dietary treatments (P < 0.05). The results indicated that supplementing a combination of 0.15% cholesterol and 0.16% taurine in a high plant-based diet significantly improved growth performance, nutrient utilization, and physio-metabolic responses of P. vannamei.

Cholic acid increases plasma cholesterol in Smith-Lemli-Opitz syndrome: A pilot study

BackgroundSmith-Lemli-Opitz syndrome (SLOS) is an inherited disorder of cholesterol biosynthesis associated with congenital malformations, growth delay, intellectual disability and behavior problems. SLOS is caused by bi-allelic mutations in DHCR7, which lead to reduced activity of 7-dehydrocholesterol reductase that catalyzes the last step in cholesterol biosynthesis. Symptoms of SLOS are thought to be due to cholesterol deficiency and accumulation of its precursor 7-dehydrocholesterol (7-DHC) and 8-dehydrocholesterol (8-DHC), and toxic oxysterols. Therapy for SLOS often includes dietary cholesterol supplementation, but lipids are poorly absorbed from the diet, possibly due to impaired bile acid synthesis. We hypothesized that bile acid supplementation with cholic acid would improve dietary cholesterol absorption and raise plasma cholesterol levels. MethodsTwelve SLOS subjects (10 M, 2F, ages 2–27 years) who had plasma cholesterol ≤125 mg/dL were treated with cholic acid (10 mg/kg/day) divided twice daily for 2 months. Plasma cholesterol, 7-DHC and 8-DHC were measured by GC–MS. Oxysterols were measured by ultra-high-performance LC-MS/MS. Data were analyzed using paired t-tests. ResultsAt baseline, plasma cholesterol was 75 ± 24 mg/dL (mean ± SD; range 43–125, n = 12). After 2 months on cholic acid, mean plasma cholesterol increased to 97 ± 29 mg/dL (p = 0.011). Eleven of 12 subjects showed an increase in plasma cholesterol that varied from 3.8% to 85.7% (mean 38.7 ± 23.3%). 7-Hydroxycholesterol decreased by 20.6% on average (p = 0.013) but no significant changes were seen in 7-DHC or 8-DHC. Mean body weight tended to increase (3.6% p = 0.069). Subjects tolerated cholic acid well and experienced no drug-related adverse events. ConclusionsIn this pilot study, cholic acid supplementation was well tolerated and safe and resulted in an increase in plasma cholesterol in most SLOS subjects. Further controlled longitudinal studies are needed to look for the sustainability of the biochemical effect and possible clinical benefits.

Balancing dietary plant-based lipids and cholesterol to increase fillet omega-3 deposition in rainbow trout (Oncorhynchus mykiss) fed a diet without animal ingredients

We hypothesized that a low phytosterol oil, such as soybean oil, in combination with an α-linolenic acid (ALA) rich oil, such as linseed oil, and supplemental cholesterol could effectively replace fish oil in salmonid feeds. As such, the primary objective was to blend soybean oil with linseed oil and supplemental cholesterol to maintain fish growth and increase de novo production of the omega-3 fatty acids (n-3 FA) eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in trout muscle, since EPA and DHA have been identified as beneficial nutrients in human nutrition, and farmed seafood must meet consumer needs and expectations. To accomplish this, we used soy oil for its low phytosterol properties in combination with linseed oil for its n-3 FA content, with and without cholesterol to improve utilization. Upon conclusion of the study, our results did not support our hypothesis. Trout fed plant-based diets containing any amount of soy oil had reduced growth performance. Only fish fed the plant-based diet with 100% linseed oil plus supplemental cholesterol grew as well as both control groups, fishmeal/fish oil and plant meal/fish oil. Dietary cholesterol supplementation increased plasma cholesterol levels and EPA in fillets (P < 0.05). Two-way ANOVA demonstrated that growth performance and muscle EPA content were significantly increased in fish fed cholesterol-supplemented diets. However, expression of genes involved in fatty acid biosynthesis were not enhanced by cholesterol, suggesting a potential disconnect between gene expression and functional activity. Although our hypothesis failed and soybean oil was found to be less accepted by the fish, resulting in lower growth and feed intake, we were successful in developing a high soy-protein diet (35%) with all-plant ingredients using linseed oil and supplemental cholesterol that resulted in equal fish performance to fish fed a fishmeal/fish oil control diet. More research is needed to identify a cost-effective EPA/DHA source to meet consumer needs and expectations for n-3 FA in trout fillets.

Positive effects of optimal dietary cholesterol levels on the ovary development and health of female Pacific white shrimp, Litopenaeus vannamei broodstock

Cholesterol is an important precursor necessary for the synthesis of steroid hormones and other functional substances. This study conducted a 28-day feeding experiment to assess the effects of different levels of dietary cholesterol (0.2%, 0.8%, 1.4%, 2.5%, or 3.2%) on the ovarian development and health status of female Litopenaeus vannamei broodstock (initial weight 39.5 ± 4.3 g). The results showed that dietary supplementation with 1.4% cholesterol can significantly increase the GSI and HSI of shrimp. Meanwhile, the 1.4% cholesterol group significantly increased the antioxidant capacity of shrimp, and increased mRNA expression of transcription factors (FoxO, GST) was also coordinated to regulate the antioxidant system. The TG and T-CHO contents were significantly accumulated in the hepatopancreas of shrimp fed a 1.4% cholesterol diet compared to other treatments. In addition, in the 1.4% group, the concentration of estrogen (E2, PROG, MF) was significantly increased compared to that in the other experimental feed groups. This was achieved by upregulating the mRNA expression of Vg, Vgr, StAR3, 3β-HSD, 17β-HSD, and ERR associated with the steroid hormone synthesis pathway. Correlation analysis of serum metabolomics and ovarian transcriptomics found that differential metabolites and differentially expressed genes were commonly significantly enriched in the glutathione metabolism pathway. Increased GST activity further enhances shrimp antioxidant capacity, which may also promote steroid hormone synthesis. In conclusion, dietary supplementation with 1.4% cholesterol can significantly promote ovarian development and strengthen the antioxidant system of female L. vannamei broodstock. To improve reproductive performance and health status, it is suggested that the appropriate dietary cholesterol supplementation content for female broodstock of L. vannamei is 1.606%–1.680%.

Dietary Cholesterol Supplementation Inhibits the Steroid Biosynthesis but Does Not Affect the Cholesterol Transport in Two Marine Teleosts: A Hepatic Transcriptome Study.

Cholesterol has been used as additive in fish feeds due to the reduced use of fish meal and fish oil. In order to evaluate the effects of dietary cholesterol supplementation (D-CHO-S) on fish physiology, a liver transcriptome analysis was performed following a feeding experiment on turbot and tiger puffer with different levels of dietary cholesterol. The control diet contained 30% fish meal (0% fish oil) without cholesterol supplementation, while the treatment diet was supplemented with 1.0% cholesterol (CHO-1.0). A total of 722 and 581 differentially expressed genes (DEG) between the dietary groups were observed in turbot and tiger puffer, respectively. These DEG were primarily enriched in signaling pathways related to steroid synthesis and lipid metabolism. In general, D-CHO-S downregulated the steroid synthesis in both turbot and tiger puffer. Msmo1, lss, dhcr24, and nsdhl might play key roles in the steroid synthesis in these two fish species. Gene expressions related to cholesterol transport (npc1l1, abca1, abcg1, abcg2, abcg5, abcg8, abcb11a, and abcb11b) in the liver and intestine were also extensively investigated by qRT-PCR. However, the results suggest that D-CHO-S rarely affected the cholesterol transport in both species. The protein-protein interaction (PPI) network constructed on steroid biosynthesis-related DEG showed that in turbot, Msmo1, Lss, Nsdhl, Ebp, Hsd17b7, Fdft1, and Dhcr7 had high intermediary centrality in the dietary regulation of steroid synthesis. In conclusion, in both turbot and tiger puffer, the supplementation of dietary cholesterol inhibits the steroid metabolism but does not affect the cholesterol transport.

Dietary Cholesterol Differentially Regulates the Muscle Lipidomics of Farmed Turbot and Tiger Puffer.

Exogenous cholesterol has been supplemented into aqua-feeds due to the reduced proportions of fishmeal and fish oil. This study aimed to investigate the effects of dietary cholesterol supplementation on the muscle lipidomics of two marine fish species, turbot and tiger puffer. A 70-day feeding trial was conducted, where two low-fishmeal diets supplemented with 0 or 1% cholesterol were used. The lipidomic analysis with targeted tandem mass spectrometry showed that, in turbot, a total of 49 individual lipids exhibited significant differences in their abundance in response to dietary cholesterol, whereas the number was 30 for tiger puffer. Dietary cholesterol up-regulated the abundance of cholesterol and cholesterol ester in both species. In turbot, the dietary cholesterol also increased the abundance of triacylglycerol and acylcarnitine, whereas in tiger puffer, it primarily regulated the abundance of phospholipids and BMP. This was the first time the responses of marine fish muscle lipidomics to dietary cholesterol supplementation have been investigated.

A High Cholesterol, (n-3) Polyunsaturated Fatty Acid Diet Induces Hypercholesterolemia More than a High Cholesterol, (n-6) Polyunsaturated Fatty Acid Diet in Hamsters

This study was designed to study the effects of (n-3) polyunsaturated fatty acids (PUFA) on plasma and liver lipids, particularly lipoprotein cholesterol concentrations, in hamsters. Diets rich in (n-3) PUFA (21 g/100 g fatty acid) or (n-6) PUFA (37.4 g/100 g fatty acid) with or without 5 g/kg cholesterol (C) supplements were given for 4 wk to male hamsters weighing 70–90 g. The VLDL- and (IDL + LDL)-cholesterol concentrations were 114 and 128% higher in hamsters fed the (n-3) PUFA + C diet than in those fed the (n-6) PUFA + C diet. However, these differences were not observed when cholesterol was not supplemented. Hamsters fed the (n-3) PUFA diet had significantly lower plasma and hepatic triglyceride concentrations were comparable in hamsters fed (n-6) PUFA + C and (n-3) PUFA + C. Hepatic cholesteryl esters were significantly lower, while hepatic microsomal acyl-CoA:cholesterol acyltransferase activity and VLDL cholesteryl esters were significantly higher in hamsters fed the (n-3) PUFA + C diet than in those fed the (n-6) PUFA + C diet. Our results demonstrate that elevation of VLDL- and (IDL + LDL)-cholesterol in hamsters by (n-3) PUFA, compared with (n-6) PUFA, is dependent on dietary cholesterol supplementation and may be due to decreased catabolism of these lipoproteins.

Cholesterol supplementation improved growth performance, cholesterol metabolism, and intestinal health of Pacific white shrimp (Litopenaeus vannamei) fed a low fishmeal diet

An 8-weeks feeding trial was conducted to evaluate the effects of dietary cholesterol supplementation on growth performance, hemolymph biochemical parameters, cholesterol metabolism, intestinal health, and disease resistance of Pacific white shrimp (Litopenaeus vannamei) fed a low fishmeal diet. Four experimental diets including HFM (25% fishmeal), LFM (15% fishmeal+10% soy protein concentrate), LC1 (LFM+0.6 g kg−1 cholesterol), and LC2 (LFM+1.2 g kg−1 cholesterol) were formulated, each diet was fed to quadruplicate tanks with 40 shrimp (0.33 ± 0.03 g) per tank for 56 days. The results shown that cholesterol supplementation in the diet did not affect the growth performance of Litopenaeus vannamei (P > 0.05), but the crude lipid content significantly reduced in the LC2 group than that in HFM group (P < 0.05). In the hemolymph biochemical indices, the content of total cholesterol in LFM and LC1 groups were significantly lower than that in HFM group; the HDL-C content was significantly lower in the shrimp fed low fishmeal based-diets (LFM, LC1, and LC2) than those fed in HFM diet; the TG content of two cholesterol supplemented groups were significantly lower than that of HFM group; the MDA content was significantly higher in LFM group than that in HFM group (P < 0.05). In hepatopancreas lipid metabolism, the shrimp fed LFM diet has higher expression in srebp, fas, and acc than those fed the HFM diet, and two cholesterol groups significant decreased the acc relative expression compare with LFM group; compare with HFM group, the LFM group showed significantly up-regulated hmg-CoA, npc1 relative expression, and LC1 group significantly up-regulated scp relative expression, then LC2 group significantly up-regulated scp, npc2 relative expression of hepatopancreas in cholesterol metabolism; LFM, LC1 group significantly up-regulated beclin1, atg12 relative expression and LC2 group up-regulated beclin1 relative expression than those in HFM group of hepatopancreas in cell autophagy (P < 0.05). LC1 group up-regulated imd, relish, toll, myd88, pelle, tube, and dorsal relative expression and LC2 group up-regulated ikkβ, myd88, tube, and dorsal relative expression in intestinal immunity than those in HFM group (P < 0.05). Cholesterol supplementation increased the height of the intestinal folds of shrimp fed the low fishmeal based-diets (P < 0.05), as well as improved the integrity of the mitochondria and the endoplasmic reticulum. Dietary cholesterol supplementation decreased the abundance of harmful bacteria phylum (Chlamydiae) and increased the abundance of beneficial bacteria genus (Pseudoalteromona, Rhodopirellula, and Ruegeria) compared with LFM group (P < 0.05). Shrimp fed the LC1 diet showed higher resistance to V. parahaemolyticus challenge than those fed the LFM diet (P < 0.05). In conclusion, cholesterol supplementation in a low fishmeal diet significantly enhanced the lipolytic capacity of shrimp, promoted the cellular autophagy, improved the intestinal health, and enhanced disease resistance of Litopenaeus vannamei.

Evaluation of Necessity of Cholesterol Supplementation in Diets of Two Marine Teleosts, Turbot (Scophthalmus maximus) and Tiger Puffer (Takifugu rubripes): Effects on Growth and Lipid Metabolism

Two 70-day feeding trials were conducted to reevaluate the necessity of cholesterol supplementation in diets of two important marine fish species, turbot and tiger puffer. Graded levels (0, 0.5, 1.0, 2.0, and 4.0% of dry matter) of cholesterol were added into the basal diet (with 30% fishmeal but no fish oil) to make five experimental diets, which were designated as control, CHO-0.5, CHO-1.0, CHO-2.0, and CHO-4.0, respectively. The dietary cholesterol concentration was measured to be 0.11%, 0.65%, 1.10%, 2.32%, and 4.59%, respectively. Each group had triplicate tanks, and each tank was stocked with 30 fish. The results showed that compared to the control, dietary cholesterol supplementation had no significant effect on the growth of both turbot and tiger puffer. Nevertheless, excess dietary cholesterol (CHO-2.0 and CHO-4.0) led to significant lower weight gain of tiger puffer compared to CHO-1.0. The feed intake decreased with increasing levels of dietary cholesterol. In general, the cholesterol supplementation decreased the crude lipid content in the liver, whereas 0.5% dietary cholesterol supplementation tended to increase the crude lipid content in the muscle of both species. Dietary cholesterol supplementation significantly regulated the lipid-related biochemical parameters in serum, liver, and muscle, and compared to turbot, tiger puffer lipid compositions had a higher buffering capacity in response to changes in dietary cholesterol level. Dietary cholesterol supplementation increased the 20:4n-6 content in the liver. Dietary cholesterol supplementation significantly downregulated the gene expression of HMG-COAr, upregulated the CYP7A1 expression, and downregulated the expression of lipid absorption and biosynthesis genes, but had no significant effect on gene expression of CPT1 and lipid transport-related genes (ApoA1, ApoA4, ApoB100, and ApoEα) in the liver. In conclusion, under the present experimental condition, dietary cholesterol supplementation had no significant effect on the growth of turbot and tiger puffer. In both species, dietary cholesterol supplementation significantly regulated the lipid accumulation and metabolism.

A New Insight Into the Underlying Adaptive Strategies of Euryhaline Marine Fish to Low Salinity Environment: Through Cholesterol Nutrition to Regulate Physiological Responses.

Salinity is an important environmental factor that can affect the metabolism of aquatic organisms, while cholesterol can influence cellular membrane fluidity which are vital in adaption to salinity changes. Hence, a 4-week feeding trial was conducted to evaluate the effects of water salinity (normal 23 psu and low 5 psu) and three dietary cholesterol levels (CH0.16, 0.16%, CH1.0, 1.0% and CH1.6, 1.6%) on osmoregulation, cholesterol metabolism, fatty acid composition, long-chain polyunsaturated fatty acid (LC-PUFA) biosynthesis, oxidative stress (OS), and endoplasmic reticulum stress (ERS) of the euryhaline fish black seabream (Acanthopagrus schlegelii). The results indicated that in low salinity, fish fed with the CH1.0 diet improved ion reabsorption and osmoregulation by increased Na+ concentration in serum as well as expression levels of osmoregulation-related gene expression levels in gills. Both dietary cholesterol level and water salinity significantly affected most cholesterol metabolic parameters in the serum and tissues, and the results showed that low salinity promoted cholesterol synthesis but inhibited cholesterol catabolism. Besides, in low salinity, hepatic expression levels of LC-PUFA biosynthesis genes were upregulated by fed dietary cholesterol supplementation with contents of LC-PUFAs, including EPA and DHA being increased. Malondialdehyde (MDA) was significantly increased in low-salinity environment, whereas MDA content was decreased in fish fed with dietary CH1.0 by activating related antioxidant enzyme activity and gene expression levels. A similar pattern was recorded for ERS, which stimulated the expression of nuclear factor kappa B (nf-κb), triggering inflammation. Nevertheless, fish reared in low salinity and fed with dietary CH1.0 had markedly alleviated ERS and downregulated gene expression levels of pro-inflammatory cytokines. Overall, these findings demonstrate that cholesterol, as an important nutrient, plays vital roles in the process of adaptation to low salinity of A. schlegelii, and provides a new insight into underlying adaptive strategies of euryhaline marine fish reared in low salinity.

Dietary cholesterol supplementation and inhibitory factor 1 serum levels in two dizygotic Smith-Lemli-Opitz syndrome twins: a case report

BackgroundSmith-Lemli-Opitz syndrome (SLOS) is a rare genetic neurodevelopmental disorder caused by the defect in the 7-dehydrocholesterol reductase. This defect leads to the deficiency of cholesterol biosynthesis with accumulation of 7-dehydrocholesterol. Inhibitory factor 1 (IF1) is a well-known mitochondrial protein. Recently, it has been discovered in the human serum where it is reported to be involved in the HDL-cholesterol intake. Here we report the IF1 presence in the serum of two paediatric SLOS dizygotic twins treated with dietary cholesterol supplementation.Case presentationThe patients showed a typical phenotype. They started dietary supplementation with cholesterol when 2 months old. The cholesterol intake was periodically titrated on the basis of weight increase and the twin 1 required a larger supplementation than the twin 2 during the follow-up. When 6.4-year-old, they underwent IF1 assay that was 7-fold increased in twin 2 compared to twin 1 (93.0 pg/ml vs 13.0 pg/ml, respectively).ConclusionsWe report, for the first time, the presence of circulating IF1 in the serum of SLOS patients, showing different levels among them. Our findings confirm that IF1 could be a novel research target in cholesterol-related disorders and also in SLOS, and could contribute to the general debate on IF1 as a new modulator of cholesterol levels.

A calcified chronic total occlusion preclinical model.

To create an experimental chronic total occlusion (CTO) model with calcification by dietary modification (cholesterol, calcium carbonate, vitamin D) and local injection of pro-calcification factors (dipotassium phosphate, calcium chloride, and bone morphogenetic protein-2 [BMP-2]). Percutaneous revascularization of CTOs frequently fails in heavily calcified occlusions. Development of novel approaches requires a reproducible preclinical model of calcified CTO. CTOs were created in 18 femoral arteries of 9 New Zealand White rabbits using the thrombin injection model. Dietary interventions included a high cholesterol diet (0.5% or 0.25%), calcium carbonate (150 mg × 3-5 days/week), and vitamin D (50,000 U × 3-5 days/week). In selected animals, BMP-2 (1-4 μg), dipotassium phosphate, and calcium chloride were injected locally at the time of CTO creation. Animals were sacrificed at 2 weeks (n = 4 arteries), 6 weeks (n = 4 arteries), and 10-12 weeks (n = 14 arteries). CTOs showed evidence of chronic lipid feeding (foam cells) and chronic inflammation (intimal/medial fibrosis and microvessels, inflammatory cells, internal elastic lamina disruption). In calcium/vitamin D supplemented rabbits, mineralization (calcification and/or ossification) was evident as early as 2 weeks post CTO creation, and in 78% of the overall arteries. Mineralization changes were not present in the absence of calcium/vitamin D dietary supplements. Mineralization occurred in 85% of BMP-treated arteries and 60% of arteries without BMP. Complex mineralization occurs in preclinical CTO models with dietary supplementation of cholesterol with vitamin D and calcium.

Dietary cholesterol level affects growth, molting performance and ecdysteroid signal transduction in Procambarus clarkii

The effects of dietary cholesterol supplementation on growth, molting performance and ecdysteroid signal transduction of juvenile crayfish Procambarus clarkii (initial weight = 3.55 ± 0.03 g) were evaluated over a 6-week feeding trial. The crude protein and lipid level of basal diet was targeted at 27% and 6%, respectively. Five levels of supplemented cholesterol (0, 0.25, 0.50, 0.75, and 1.00%) resulted in diets analyzed to contain 0.06, 0.29, 0.54, 0.81, and 1.05% cholesterol. Final weight, weight gain, feed intake, molting times, molting rate, chitinase mRNA expression and hemolymphatic LDL-C content significantly increased (P < .01) as the cholesterol supplementation level increased from 0.00 to 0.50%, and then significantly decreased (P < .05) as the cholesterol supplementation level increased from 0.50 to 1.00%. P. clarkii fed with 0.25% cholesterol supplementation and without cholesterol supplementation had significantly higher MIH expression level compared to P. clarkii fed with 0.50 and 0.75% cholesterol supplementation. Total cholesterol content in the hemolymph, hepatopancreas, and intestine, as well as hemolymphatic HDL-C content of P. increased significantly as the cholesterol supplementation increased from 0.00 to 1.00%. Taken together, the results presented here indicated that P. clarkii juvenile would grow better when fed with a 27% protein and 6% lipid diet supplemented with 0.50% cholesterol. The reason might be that more cholesterol transported from hepatopancreas resulted in the variation of ecdysteroid signal transduction, ultimately leading to increasing molting times in P. clarkii fed diets with 0.50% cholesterol supplementation.

Study of the requirements of dietary cholesterol at two different growth stages of Pacific white shrimps, Litopenaeus vannamei

Two 8-week feeding experiments were conducted to investigate the cholesterol requirements of Litopenaeus vannamei at two different growth stages—with initial body weights of 0.61 ± 0.00 g (juvenile) and 4.25 ± 0.00 g (subadult). Six diets containing cholesterol (C) 0.03% (FM-fish meal), 0.09% (0.05% C), 0.14% (0.1% C), 0.27% (0.2% C), 0.47% (0.4% C), and 0.90% (0.8% C) were prepared with red fish meal, dehulled soybean meal and peanut meal as the protein source, and fish oil as the fat source. Two levels of phytosterol (P) substitutes—0.05% (0.05% P) and 0.8% (0.8% P)—for dietary cholesterol were evaluated. Results showed dietary cholesterol levels had significant effects on the weight gain rate (WGR), specific growth rate (SGR), and feed conversion ratio (FCR) of juveniles (P < 0.05). Crude fat in the 0.05% C and 0.1% C groups of subadults was significantly higher than that in the FM group (P < 0.05). The contents of CHOL, HDL, LDL, and liver CHOL were significantly associated with the content of cholesterol (P < 0.05). The dietary cholesterol proportion significantly affected serum AST and ALT (P < 0.05). Phytosterol supplementation presented growth-promoting effects for juvenile L. vannamei. A broken-line equation for weight growth showed the optimum dietary cholesterol requirement was 0.16% for L. vannamei juveniles (0.61 ± 0.00 g), and no extra dietary cholesterol supplementation was needed for L. vannamei subadults (4.25 ± 0.00 g).

Survival, molting pattern, and growth of early blue swimmer crab, Portunus pelagicus, juveniles fed diets containing varying levels of cholesterol

AbstractThis study examined the effects of dietary cholesterol on the survival, molting pattern, and growth of early juveniles of the blue swimmer crab, Portunus pelagicus. Seven isocaloric diets were formulated to contain cholesterol at 0 (basal diet), 2.5, 5.0, 7.5, 10, 12.5, and 15 g/kg diet dry weight. Survival ranged from 20 to 47.5%, with the highest survival recorded for crabs fed the diet supplemented with 10 g/kg cholesterol, while the lowest survival was demonstrated by crabs fed the basal diet. The highest occurrence of molt death syndrome was found for crabs fed the diet with cholesterol supplementation of 15 g/kg. A general trend of decreasing development time to the crab 3 (C3) stage was observed with increasing dietary cholesterol supplementation up to a level of 10 g/kg, but this then decreased as dietary cholesterol level was increased to 12.5 and 15 g/kg. The specific growth rates of crabs, calculated for dry weight, carapace width, and carapace length, followed a similar trend. Our results suggest that, under the current feeding conditions, a dietary cholesterol level of 10 g/kg appears to be optimal for the culture of early juveniles of P. pelagicus.

Síndrome de Smith-Lemli-Opitz: Revisión de la literatura a propósito de un caso

Skin reactions induced by drugs in pediatrics are rare entities; However, these are associated Smith-Lemli-Opitz syndrome is an autosomal recessive disorder characterized by multiple congenital malformation, intellectual disability and behavioral problems. It is due to mutations of DHCR7 with a deficiency of 7-dehydrocholesterol reductase. This enzyme catalyzes the reduction of 7-dehydrocholesterol to cholesterol. Therefore, it results inincreased 7-dehydrocholesterol levels and other cholesterol precursors, and decreased cholesterol levels in blood and biological tissues. Its phenotypic spectrum varies broadly with characteristic physical malformations such as microcephaly, characteristic face, genital abnormalities in male patients, intellectual disability and syndactyly of the second and third toes. The clinical diagnosis is confirmed by finding elevated 7-dehydrocholesterol and mutations in DHCR7, and prenatal diagnosis is also possible. The treatment is a symptomatic treatment and dietary cholesterol supplementation is beneficial. On the other hand, other therapies are being studied such as simvastatin treatment and antioxidant supplementation.

Effect of Dietary Cholesterol and Niacin Supplementation on Serum Enzymatic Alterations in Experimentally Induced Renal Dysfunction in Wistar Rats

Effect of Dietary Cholesterol and Niacin Supplementation on Serum Enzymatic Alterations in Experimentally Induced Renal Dysfunction in Wistar Rats

Effect of dietary cholesterol and phospholipids on feed intake, growth performance and cholesterol metabolism in juvenile turbot (Scophthalmus maximus L.)

Dietary phospholipids play important roles in growth and nutrition metabolism in fish. Previous studies showed that moderate dietary cholesterol supplementation improved the growth performance of juvenile turbot fed plant-based or defatted fishmeal diets. Therefore, we examined the effect of dietary cholesterol and phospholipids on feed intake, growth performance and cholesterol metabolism in juvenile turbot (Scophthalmus maximus L.) (initial body weight 5.18 ± 0.01 g) during a 10-week period. Nine isonitrogenous and isolipidic diets were formulated to contain 0.0, 1.0 and 2.0% cholesterol (LC, MC and HC), and each with 0.0, 2.0 and 4.0% phospholipids (LP, MP and HP), respectively. A significant interaction between dietary cholesterol and phospholipids on weight gain rate was shown in the present study (P = 0.049). However, no significant interaction between dietary cholesterol and phospholipids was found in feed intake (P = 0.332) though the expression of the orexigenic factor ghrelin in the intestine was significantly affected by the interaction (P = 0.011). Except for total cholesterol, free cholesterol and cholesterol ester, both high-density lipoprotein cholesterol (HDL-C) (P = 0.012) and low-density lipoprotein cholesterol (LDL-C) (P = 0.002) in serum were significantly affected by the interaction between dietary cholesterol and phospholipids. Regarding the key genes involved in cholesterol and bile acid synthesis, no interaction between dietary cholesterol and phospholipids was found in the expression of 3-hydroxy-3-methylglutaryl-CoA reductase (hmgcr) (P = 0.380) and cholesterol 7α-hydroxylase (cyp7a1) (P = 0.587) in liver. In conclusion, there is a significant interaction between dietary cholesterol and phospholipids on the growth performance and the HDL-C and LDL-C involved in cholesterol transport, while no significant interaction was found on the feed intake in juvenile turbot.

Abstract 441: Zebrafish Larvae as a Model System for High-throughput, Image-based Genetic Screens in Dyslipidemia and Atherosclerosis

Background: Genome-wide association (GWAS) have identified hundreds of loci that are robustly associated with the risk of cardiometabolic risk factors and diseases. With few exceptions, the causal genes through which these loci influence disease risk remain uncharacterized. Since murine model systems are not suitable for systematic characterization of candidate genes in hundreds of loci, novel in vivo model systems are desirable. Methods: My group has developed and validated zebrafish model systems that make optimal use of: 1) the zebrafish’ well-annotated genome, with orthologues of at least 71.4% of human genes; 2) recent developments in multiplex CRISPR-Cas9-based mutagenesis; 3) advances in automated positioning of non-embedded zebrafish larvae; 4) fluorescent transgenes and dyes; and 5) custom-written image-quantification pipelines. Results: Five days of overfeeding, dietary cholesterol supplementation and/or exposure to glucose induce atherogenic and insulin resistant phenotypes (higher/more whole-body LDL cholesterol (LDLc) and triglyceride levels; vascular foam cell formation and inflammation; beta-cell number and volume; subcutaneous and hepatic accumulation of fat) that can largely be prevented by concomitant treatment with lipid-lowering or diabetes medication (N&gt;4000). In line with recent results in humans, treatment with atorvastatin and ezetimibe results in higher whole-body glucose levels (N~835). Each additional mutated allele in zebrafish orthologues of APOE results in higher LDLc levels, while mutations in LDLR affect vascular infiltration by lipids and macrophages (N~330). Ongoing characterization of 37 prioritized genes in GWAS-identified loci for triglyceride levels identified genes that influence triglycerides, LDLc, and/or vascular inflammatory and atherogenic traits, while characterizing prioritized genes in loci associated with heart rate variability yielded genes that influence cardiac rate and rhythm. Conclusions: Systematic characterization of candidate genes for cardiometabolic traits in zebrafish model systems will help increase our understanding of human disease and identify novel targets that can be translated into efficient therapeutics.

Prevention of Retinal Degeneration in a Rat Model of Smith-Lemli-Opitz Syndrome

Smith-Lemli-Opitz Syndrome (SLOS) is a recessive human disease caused by defective cholesterol (CHOL) synthesis at the level of DHCR7 (7-dehydrocholesterol reductase), which normally catalyzes the conversion of 7-dehydrocholesterol (7DHC) to CHOL. Formation and abnormal accumulation of 7DHC and 7DHC-derived oxysterols occur in SLOS patients and in rats treated with the DHCR7 inhibitor AY9944. The rat SLOS model exhibits progressive and irreversible retinal dysfunction and degeneration, which is only partially ameliorated by dietary CHOL supplementation. We hypothesized that 7DHC-derived oxysterols are causally involved in this retinal degeneration, and that blocking or reducing their formation should minimize the phenotype. Here, using the SLOS rat model, we demonstrate that combined dietary supplementation with CHOL plus antioxidants (vitamins E and C, plus sodium selenite) provides better outcomes than dietary CHOL supplementation alone with regard to preservation of retinal structure and function and lowering 7DHC-derived oxysterol formation. These proof-of-principle findings provide a translational, pre-clinical framework for designing clinical trials using CHOL-antioxidant combination therapy as an improved therapeutic intervention over the current standard of care for the treatment of SLOS.