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Kolin Plus™ as a Natural Alternative to Synthetic Choline Chloride : Proof of Concept Studies
Executive Summary
Choline is an essential nutrient for poultry, critical in lipid metabolism, liver health, and overall performance. While synthetic choline chloride (SCC) has been the standard dietary supplement, its limitations—including hygroscopicity, vitamin degradation, and regulatory restrictions in organic farming—necessitate effective natural alternatives. Kolin Plus™, a proprietary polyherbal formulation developed by Natural Remedies Pvt. Ltd., combines Acacia nilotica and Curcuma longa. Kolin plus has been found to be an effective replacement of synthetic choline chloride.This white paper provides an in-depth scientific evaluation of Kolin Plus™, spanning preclinical, zootechnical, biochemical, and genomic evidence to prove its suitability as a natural alternative to synthetic choline chloride.
1. Introduction: Importance of Choline in Poultry Nutrition
- Metabolic Role: Choline acts as a labile methyl donor, supporting methionine synthesis , methylation reactions critical for energy metabolism and most importantly involved in fat mobilization through the formation of VLDL
- Structural Role: Essential for phospholipid formation, maintaining cell membrane integrity, and neuromuscular function via acetylcholine synthesis.
- Deficiency Outcomes: Leads to fatty liver syndrome (FLS), poor feed conversion ratio (FCR), reduced growth, perosis, and higher mortality.
SCC has traditionally bridged dietary gaps but presents practical and regulatory issues. The search for herbal alternatives aligns with industry trends toward sustainability and consumer preference for natural feed additives.
2. Limitations of Synthetic Choline Chloride (SCC)
- Hygroscopicity: Causes feed instability and caking.
- Nutrient Interaction: Accelerates oxidative loss of vitamins and generates trimethylamine (TMA) in the gut, affecting bird health and product quality.
- Organic Production Barrier: Regulatory frameworks often disallow SCC in certified organic poultry.
- Safety Concerns: Handling hazards due to corrosive nature; unstable in premix storage.
These limitations drive the demand for robust, plant-derived lipotropic alternatives.
3. Kolin Plus™: Composition and Rationale
- Botanical Sources:
- Acacia nilotica: Known for hepatoprotective and antioxidant effects
- Curcuma longa: Exhibits anti-inflammatory, antioxidant, and lipid-lowering properties.
- Standardization: Optimized to contain ≥8% polyphenols, ensuring batch-to-batch consistency.
Rationale : The potent anti- lipogenic and lipolysis effect of Kolin plus could replace the functional role of choline, that is fat metabolism . While the structural requirements are met by endogenous or intrinsic choline from diet.
4. Research Evidence
4.1 Preclinical Studies (Chandrasekaran P et al , 2019)
This study evaluates the lipotropic effect of Kolin Plus on hepatic fat accumulation in rats fed a methionine-choline deficient (MCD) diet. Choline deficiency leads to fatty liver and impaired liver function. Kolin Plus was tested in vivo on Wistar rats and in vitro on HepG2 cells. Results showed that Kolin Plus significantly reduced liver lipids, triglycerides, cholesterol, and serum alanine transaminase (ALT) levels, indicating improved liver health and reduced fat accumulation. Histological analysis confirmed restoration of hepatic architecture. In vitro, Kolin plus demonstrated anti-lipogenic activity by reducing oleic acid-induced fat accumulation in HepG2 cells. The findings suggest Kolin Plus as a potential supplement for preventing fatty liver and maintaining liver health, with possible applications in agricultural livestock. Further studies in target species are recommended.
4.2 Zootechnical Trials in Broilers
Choline Deficiency Model (Selvam et al , 2018) : This study aimed to establish a choline deficiency model (CDM) in broiler chickens to evaluate the efficacy of Kolin Plus compared to synthetic choline chloride (SCC). Choline is an essential nutrient for poultry, playing a critical role in growth, liver health, and lipid metabolism. The researchers replaced soybean meal (SBM), a primary choline source, with soy protein isolate (SPI) at varying levels to induce choline deficiency. Results showed that increasing SPI replacement led to linear declines in body weight gain (BWG), feed conversion ratio (FCR), and liver health, confirming the successful induction of CDM. Kolin Plus demonstrated comparable efficacy to SCC in restoring growth performance, liver function, and lipid metabolism. Optimal results were achieved with Kolin plus at 400 g/ton, which effectively mimicked SCC’s biological activities, including hepatoprotective and lipotropic effects. Histopathological analysis revealed reduced liver damage in Kolin Plus -treated birds. The study concluded that Kolin Plus could serve as a natural, greener alternative to SCC in broiler diets, offering similar benefits in growth and liver health while addressing the drawbacks of synthetic choline chloride.
High Energy Diet Model (D’Souza, P et al , 2022) :
This study evaluated the effects of Kolin Plus™, on broilers fed a high-energy diet (HED). The trial involved 500 Cobb 430 male chicks divided into five groups, including a negative control (NC), a positive control with synthetic choline chloride (SCC), and three Kolin Plus -supplemented groups at varying doses (400, 500, and 750 gm/ton). Results showed that Kolin Plus supplementation improved body weight, feed conversion ratio (FCR), livability, and the European Production Index (EPI) in a dose-dependent manner, with the highest dose (Kolin Plus 750g) performing comparably to SCC. Kolin Plus significantly reduced carcass fat accretion and increased serum L-carnitine levels, indicating enhanced fat metabolism and mobilization. The study concluded that Kolin Plus at 750 gm/ton could effectively replace SCC at 1,500 gm/ton in broiler diets, offering a natural alternative to synthetic choline chloride.
4.3 Transcriptomic expression Insights (D’Souza P et al 2019)
Transcriptomic studies done in the chicken with reference to muscle growth.This study investigates the effects of Kolin PlusTM, a polyherbal formulation containing Acacia nilotica and Curcuma longa, on muscle growth in Cobb 430 broiler chickens fed a choline chloride-deficient (CCD) diet. Using microarray analysis, researchers identified 2,900 differentially expressed genes (DEGs) in breast muscle tissue, with 481 genes significantly upregulated and 326 downregulated in the Kolin PlusTM group compared to the CCD group. Key growth-related genes such as CSRP3, BCO1, CTHRC1, LMOD2, PHGDH, and ANKRD2 were upregulated, which are known to enhance muscle growth, protein translation, energy metabolism, and sarcomere assembly. On the other hand, genes like SOX10, CALB1, KLF15, and UTS2R were downregulated, reducing their negative effects on muscle development. Specifically, SOX10 inhibits skeletal muscle differentiation, CALB1 is linked to muscle degeneration, KLF15 promotes muscle atrophy during nutrient deprivation, and UTS2R regulates fat accumulation and muscle atrophy. The modulation of these genes by Kolin PlusTM supplementation highlights its ability to counteract the adverse effects of choline deficiency, leading to improved muscle growth, body weight, and feed conversion ratio in broilers. The study concludes that Kolin PlusTM can effectively replace synthetic choline chloride, offering a natural alternative for enhancing broiler productivity through targeted gene regulation.
Lipid Regulation (Marimuthu, S et al , 2022) :
The study aimed to investigate the effects of Kolin Plus™ (KP), on lipid metabolism in broiler chickens fed a choline chloride-deficient (CCD) diet. Choline is essential for lipid regulation, and its deficiency can lead to fatty liver and growth issues in chickens. KP, containing Acacia nilotica and Curcuma longa, was evaluated as a natural alternative to synthetic choline chloride (CCL). Using transcriptome analysis, the study identified 12,614 differentially expressed genes (DEGs) in the liver, with significant modulation of genes involved in lipid metabolism, fatty acid oxidation, and lipogenesis. KP supplementation regulated key genes such as LIPC, ABCG5, ABCG8, PPARG, ACACA, and ACLY (involved in lipogenesis) and PPARGC1A, CPT1A, PNPLA2, and HADHB (involved in fatty acid oxidation). KP demonstrated a stronger effect than CCL in reversing the negative impacts of the CCD diet, improving performance parameters like body weight and feed conversion ratio, and preventing fatty liver. The study concluded that KP effectively mimics the lipotropic effects of choline by modulating gene expression related to lipid metabolism, making it a promising natural alternative to synthetic choline in poultry diets.
Probable role of Kolin Plus in choline absorption :
Choline absorption is not a very robust effect in monogastrics. Example Choline is estimated to be absorbed in the range of 50 to 70 % from common feed stuffs. The absorption of free choline in the diet is facilitated by choline transporter protein in the gut called SLC44A1. Kolin plus was known to upregulate this transporter protein gene. Further predictive transcriptomics study also revealed that Gallic acid an important active of Kolin plus could augment choline transporter activity.
Conclusion:
Kolin plus is a novel phytogenic product proven to be a non synthetic alternative to choline chloride. This review shows evidence of its efficacy in a choline deficent model which clearly provides the proof of its efficacy. The key mechanism of Kolin plus invoves lipid mobilisation by way of increased lipolysis and reduced lipogenesis. Kolin plus may also improve absorption of free choline by upregulating the choline transporter activity.
References :
Chandrasekaran P. Raja, Murugan S., Richard E. Jothie, Bethapudi B., Purusothaman D., Velusami C. C., D’Souza P., Mundkinajeddu D., & Talkad M. S. (2019). Evaluation of lipotropic effect of herbal formulation on hepatic fat accumulation in rats fed with methionine-choline deficient diet. Pharmacognosy Magazine, 15(66), 476-482.
Selvam, R., Saravanakumar, M., Suresh, S., Chandrasekeran, C. V., & D’Souza, P. (2018). Evaluation of polyherbal formulation and synthetic choline chloride on choline deficiency model in broilers: implications on zootechnical parameters, serum biochemistry and liver histopathology. Asian-Australasian Journal of Animal Sciences, 31(11), 1795-1806.
D’Souza, P., & Selvam, R. (2022). Evaluation of polyherbal formulation in broilers fed high energy diet: Implications on zootechnical parameters, fat accretion, and serum L-carnitine levels. Journal of Advanced Veterinary and Animal Research, 9(1), 166-174.
D’Souza, P., Marimuthu, S., Kaninathan, A., & Selvam, R. (2019). Modulatory effect of Kolin Plus™ , a polyherbal formulation on muscle growth development in choline-deficient diet model of broilers: a microarray approach. International Journal of Scientific Reports, 5(12), 344-350.
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