Effects of Branched-Chain Fatty Acids Derived from Yak Ghee on Lipid Metabolism and the Gut Microbiota in Normal-Fat Diet-Fed Mice.

Branched-chain fatty acids (BCFAs) are natural components with a variety of biological activities. However, the regulation of lipid metabolism by BCFAs is unknown. It was dedicated to examining the impacts of BCFAs inferred from yak ghee on the expression of qualities related to lipid metabolism, natural pathways, and intestinal microbiota in mice. The treatment group (purified BCFAs from yak ghee) exhibited a decrease in cholesterol levels; a decrease in HMGCR levels; downregulation of FADS1, FADS2, ACC-α, FAS, GAPT4, GPAM, ACSL1, THRSP, A-FABP, and PPARα gene expression; and upregulation of SCD1, ACSS1, FABP1, CPT1, and DGAT-1 gene expression. Gut microbiota 16S rDNA sequencing analysis showed that the treatment group improved the gut microbiota by increasing the relative abundances and increasing the short-chain fatty acid levels produced by the genera Akkermansia, Clostridium, Lachnospiraceae, Lactobacillus, Anaerotaenia, and Prevotella. After adding BCFAs to cultured breast cancer cells, pathways that were downregulated were found to be related to fatty acid degradation and fatty acid metabolism, while 20 other pathways were upregulated. Our results suggest that BCFAs reduce body fat in mice by modulating intestinal flora and lipid metabolism and modulating fatty acid metabolism in breast cancer cells.

Effects of diets rich in ghee or olive oil on cardiometabolic risk factors in healthy adults: a two-period, crossover, randomised trial.

This study aimed to evaluate the cardiovascular health-related effects of consuming ghee in the usual diet. Thirty healthy men and women were studied in a free-living outpatient regimen. The participants were instructed for the isoenergetic inclusion of ghee or olive oil in their diets for 4 weeks using a randomised crossover design. At the end of run-in (baseline), 2-week wash-out and interventions, fasting blood samples were drawn. In addition, 2-h postprandial blood samples were collected after ingestion of a meal containing olive oil or ghee at week 4 of each dietary intervention. Body weight was not different between the two interventions. Compared with the olive oil, the diet with ghee increased fasting plasma apo-B (apo B) (0·09, 95 % CI 0·02, 0·17 g/l, P = 0·018), non-HDL-cholesterol (non-HDL-cholesterol) (0·53, 95 % CI 0·01, 1·05 mmol/l, P = 0·046) and LDL-cholesterol did not differ significantly between diet groups (0·29, 95 % CI -0·05, 0·63 mmol/l, P = 0·092), but had no significant effect on total cholesterol:HDL-cholesterol ratio (0·75, 95 % CI - 0·24 to 1·74 mmol/l, P = 0·118). No significant difference was observed in fasting as well as 2-h postprandial plasma TAG, glucose, insulin and plasminogen activator inhibitor-1 concentrations. This study showed that ghee that is predominantly saturated fats had an increasing effect on plasma apo B and non-HDL-cholesterol compared with olive oil, adding further evidence to the existing recommendations to replace dietary fats high in SFA with dietary fats high in unsaturated fats to reduce CVD risk.

Characterization of Aerosol Properties from the Burning Emissions of Typical Residential Fuels on the Tibetan Plateau.

The Tibet Autonomous Region in China is a unique place with high altitude and special Tibetan culture. The residents have different living habits and domestic fuels from those in other parts of China, however, knowledge on the emission characteristics of local residential fuels remain poorly understood until now. In this study, nine popular residential fuels in the Tibet are burned in situ to study the aerosol chemical compositions, mass spectral signatures, and emission characteristics from their burning emissions. Overall, emissions of particulate and gaseous pollutants depend strongly on the burning conditions, in addition to the fuel constituents themselves. Burning the biofuels of yak dung, WeiSang mixture fuels, and two powdery Tibetan incenses with relatively low combustion efficiencies can emit large amounts of CO and aerosols, especially organic aerosols (>90%) with large diameters. In contrast, burning of wood, coal, ghee lamp, stick-like Tibetan incense, and diesel can release abundant CO2 but fewer aerosols from their flaming combustion. A comprehensive database consisting of the high-resolution mass spectra of organics and emission factors of multiple chemical components are established. Distinctly different mass spectral signatures are found among the different fuels, in particularly those unique Tibetan biofuels. All these findings have significant implications for the identification of aerosol sources, compilation of pollutant emission inventories, and assessment of potential environment effects in this remote region.

Detection of soybean oil adulteration in cow ghee (clarified milk fat): an ultrafast study using flash gas chromatography electronic nose coupled with multivariate chemometrics.

BACKGROUND: Cow ghee is one of the expensive edible fats in the dairy sector. Ghee is often adulterated with low-priced edible oils, like soybean oil, owing to its high market demand. The existing adulteration detection methods are time-consuming, requiring sample preparation and expertise in these fields. The possibility of detecting soybean oil adulteration (from 10% to 100%) in pure cow ghee was investigated in this study. The fingerprint information of volatile compounds was collected using a flash gas chromatography electronic nose (FGCEN) instrument. The classification results were studied using the pattern recognition chemometric models principal component analysis (PCA), soft independent modelling of class analogy (SIMCA), and discriminant function analysis (DFA). RESULTS: The most powerful fingerprint odor of all the samples identified from FGCEN analysis was acetaldehyde (Z)-4-heptenal, 2-propanol, ethyl propanoate, and pentan-2-one. The odor analysis investigation was accomplished with an average analysis time of 90 s. A clear differentiation of all the samples with an excellent classification accuracy of more than 99% was achieved with the PCA and DFA chemometric methods. However, the results of the SIMCA model showed that SIMCA could only be used to detect ghee adulteration at higher concentration levels (30% to 100%). The validation study shows good agreement between FGCEN and gas chromatography-mass spectrometry methods. CONCLUSION: The methodology demonstrated coupled with PCA and DFA methods for adulteration detection in ghee using FGCEN apparatus has been an efficient and convenient technique. This study explored the capability of the FGCEN instrument to tackle the adulteration problems in ghee. © 2022 Society of Chemical Industry.

Regenerative repair of full thickness skin wound assisted by dual crosslinking percolative gel casting maneuvered alginate hydrogel embedded with honey ghee blend resembles standard cutaneous properties.

We report synergism in scarless cutaneous wound repair by alginate hydrogel (HGSAG) embedded with an optimized blend of characterized Jamun honey and characterized indigenously prepared ghee. Thorough screening and characterization of honey and ghee are carried out followed by obtaining a novel dual crosslinking percolative gel casting fabrication method to come up with HGSAG showing superior chemical stability, and mechanical strength (Nanoindentation study; lowest stiffness: 0.71 ± 0.19 µN/nm), and surface morphology (SEM; highest roughness: 0.13 ± 0.04 µm) to other variants. In vitro swelling study and degradation behavior study show intermediate swelling (swelling index: 0.59 ± 0.008 in 98 h) and required restricted degradation (PBS: 73.38 ± 0.55%, DMEM: 83.48 ± 0.69% in 10 days) for HGSAG which is necessary for providing nutrients to cells and in vivo therapeutic efficacy. We observe the remarkable antibacterial efficacy of HGSAG against Staphylococcus mutans and Escherichia coli. This particular substrate also shows decent 3T3 fibroblasts viability, cell-cell communication followed by cell-matrix interaction, and proliferation compared to other variants. Molecular gene expression studies by quantitative RT-PCR technique reveal strong upregulation of collagen I, CD26, and TGF-ß3 while downregulation in the case of TGF-ß1 which eventually substantiates scarless wound healing potential of HGSAG. Wound closure kinetics is most rapidly and successfully underpinned by HGSAG while compared to other alternatives including marketed healing patches. Regular close monitoring using histopathological studies and real-time imaging by Swept-Source Optical Coherence Tomography of in vivo wound model treated with HGSAG come up with the fascinating result of scarless healing (HGSAG treated epithelial thickness: 62.96 ± 0.67 µm, unwounded akin epithelial thickness: 62.56 ± 0.34 µm) within 12 days of wounding. Thus, the work highlights modified and stabilized alginate hydrogel embedded with honey and ghee blend as a potential scarless full-thickness cutaneous wound healing bio-scaffold.

Fluorescence Spectroscopy Based Detection of Adulteration in Desi Ghee.

Desi ghee, obtained by buffalo and cow milk, is highly expensive because it contains valuable vitamins and conjugated linoleic acid (CLA). Its high demand and cost result in to its adulteration with inferior banaspati ghee. In this study, Fluorescence spectroscopy along with multivariate analysis has been utilised for the detection and quantification of adulteration. Spectroscopic analysis showed that buffalo ghee contains more vitamins and CLA than cow, whereas cow ghee is enriched with beta-carotene. For multivariate analysis, principle component analysis (PCA) and partial least square regression (PLSR) have been applied on the spectral data for the determination of adulteration. PLSR model was authenticated by predicting 23 unknown samples including 3 commercial brands of desi ghee. The root mean square error in prediction (RMSEP) of unknown samples was found to be 1.7 which is a reasonable value for quantitative prediction. Due to non-destructive and requiring no sample pre-treatment, this method can effectively be employed as on line characterization tool for the food safety assurance.

Role of Omega-3 Fatty Acids and Butter Oil in Targeting Delivery of Donepezil Hydrochloride Microemulsion to Brain via the Intranasal Route: a Comparative Study.

In order to investigate the possible role of butter oil (BO) and omega-3 fatty acids-rich fish oil (O3FO) in the delivery of donepezil hydrochloride microemulsion (DH-ME) to the brain via intranasal route, the present study was conducted. DH:BO and DH:O3FO binary mixtures (9:1 to 1:9) were prepared by simple physical mixing and subjected to in vitro diffusion study. Ratios of DH:BO and DH:O3FO, which showed the highest diffusion, were selected for further development of microemulsion (ME). Globule sizes of DH-BO-ME and DH-O3FO-ME were found to be 87.66 ± 5.23 nm and 88.59 ± 8.23 nm, respectively. Nasal histopathological study and in vitro cytotoxicity study revealed the safety of the formulation. Higher percentage of nasal diffusion was found with DH-BO-ME (71.22 ± 1.21%) and DH-O3FO-ME (62.16 ± 1.23%) in comparison to DH-ME (59.69 ± 1.74%) and DH solution (55.01 ± 1.19%), which was further supported by in vitro cell permeability study. After intranasal administration, %bioavailability of drug in the rat brain (Sprague-Dawley rats)(on the basis of DH-ME IV) was higher with DH-BO-ME (313.59 ± 12.98%) and DH-O3FO-ME (361.73 ± 15.15%) in comparison to DH-ME (168.62 ± 6.60%) and DH solution (8.960 ± 0.23%). The results of ex vivo diffusion study and in vivo pharmacokinetic study suggested that BO and O3FO helped in enhancing the nasal permeability and the brain uptake of drug when administered intranasally.

Characterization of Desi Ghee Extracted by Different Methods Using Fluorescence Spectroscopy.

In the current study, the effect of ghee extraction methods (direct cream DC, milk butter MB and milk skin MS) on its molecular composition has been investigated using Fluorescence spectroscopy. The excitation wavelength of 300 nm was found the best to produce pronounced spectral signatures of beta-carotene, vitamins and conjugated linoleic acid (CLA) in both cow and buffalo ghee types. Principal component analysis (PCA) has been applied on the spectral data to visualize the classification among ghee samples extracted by three methods. Both cow and buffalo ghee contain spectral signatures of vitamin A, E, K, D and CLA which has been verified through plotting loading vectors. The analysis of loading plots has been suggested that for cow ghee, MS extraction method conserve relatively higher concentration of beta carotene while DC and MB methods are a good choice for preserving relatively more concentrations of vitamins D, E and K. Similarly, for buffalo ghee, MS extraction method appear with higher concentration of CLA, whereas DC extraction method looks to preserve relatively higher concentration of vitamin A while MB method retains relatively low concentration of CLA and vitamins as compared to other two methods.

Cow ghee fortified ocular topical microemulsion; in vitro, ex vivo, and in vivo evaluation.

Aim: Utility of cow ghee (CG) as permeation enhancer in development of topical ocular microemulsion (ME) for delivery of fluocinolone acetonide (FA) to posterior eye. Methods: For ME preparation, oil, surfactant and cosurfactant were screened based on solubility of FA. Pseudoternary phase diagrams were constructed to determine their ratios. The developed MEs were characterised for their physicochemical properties like size, polydispersity index, zeta potential, and stability etc. They were evaluated for ex vivo permeation and irritation. In vivo pharmacokinetic studies were performed on Sprague dawley rats. Results: Lauroglycol as oil, labrasol as surfactant and Transcutol as cosurfactant were selected. The optimised ratio of oil:surfactant:cosurfactant:water was 4:23:23:50. The developed FA loaded ME fortified with CG was characterised. Ex vivo study revealed higher permeation and non-irritancy. In vivo pharmacokinetic study showed retention of CG fortified ME in posterior rat eye. Conclusion: Present investigation established CG as permeation enhancer for ocular topical formulation.

Microemulsion-based delivery of triamcinolone acetonide to posterior segment of eye using chitosan and butter oil as permeation enhancer: an in vitro and in vivo investigation.

The aim of the study was to formulate a microemulsion (ME) using chitosan (CH) and the butter oil (BO) as a permeation enhancer for targeting drug to the posterior segment of the eye, via topical route. Triamcinolone acetonide (TA) was selected as the model drug since it undergoes extensive first-pass metabolism, leading to poor oral bioavailability of 23%. For optimisation of BO concentration, different ratios of TA:BO were prepared by simple physical mixing in the ratio of 1:9 to 9:1 and diffusion study was performed. MEs containing TA, TA:BO and TA CH ME were formulated by water titration method. Globule sizes of TA ME, TA:BO ME and TA CH ME were found to be 66.06 ± 0.32 nm, 78.52 ± 1.50 nm and 97.30 ± 2.50 nm, respectively. In ex vivo diffusion studies using goats eye, TA:BO ME (31.33 ± 0.46 and 33.98 ± 0.23) and TA CH ME (24.10 ± 0.41 and 27.00 ± 0.18) showed higher percentage of drug diffusion in comparison to TA ME (13.29 ± 0.41and 15.56 ± 0.34) and TA solution (8.20 ± 1.04 and 10.39 ± 0.22) in presence and in absence of vitreous humour. Fluorescence intensity of coumarin-6 (as a marker) loaded ME with BO and CH was found to be higher, confirming their role in altering membrane permeability and facilitating coumarin-6 diffusion to the posterior chamber. Overall, it was concluded that BO enhances the bioavailability of TA across the retina, thereby proving its potential as permeation enhancer in facilitating drug delivery to the posterior segment of the eye.

Ghee Butter as a Therapeutic Delivery System.

Solid lipid nanoparticles carrying a chemotherapeutic payload (i.e., temozolomide, TMZ) were synthesized using ghee, a clarified butter commonly used in traditional medicine and food products. Ghee solid lipid nanoparticles (GSLN) were characterized through dynamic light scattering, scanning electron microscopy, and UV-visible spectrometry. Formulations were generated with varying ratios of surfactant to lipid, resulting in a maximum TMZ entrapment efficiency of ˜70%. Optimal formulations were found to have an average size and polydispersity of ˜220 nm and 0.340, respectively. Release kinetics revealed TMZ-loaded GSLN (TMZ@GSLN) retained 10% of its pay-load at 2 h with ˜53% released in 5 h. Metabolic activity on human umbilical vein endothelial cells (HUVEC) revealed GSLN treatment resulted in an increase in viability following 3 d while treatment of glioblastoma LN-229 cells with TMZ@GSLN resulted in a significant decrease. Evaluation of diffusion of TMZ across a reconstructed HUVEC monolayer demonstrated TMZ@GSLN resulted in a significantly higher diffusion of drug when compared to free TMZ. This data suggests GSLN pose a promising delivery vehicle for TMZ-based therapeutics. Collectively, this data demonstrates GSLN exhibit favorable drug carrier properties with anti-proliferative properties in glioblastoma cancer cells.

Pragmatic selection of cooking oils.

Fats and oils are one of the very important components of diet. However excess of either overall fat or certain kind of fats in the diet may result in negative health impacts including risk of obesity, dyslipidaemia, cardiovascular diseases and certain malignancies. It is thus important to have an optimum amount of fat in the diet, and also important to choose appropriate sources of fat in the diet. In this mini review we suggest pragmatic selection of cooking oils for optimum health benefits.

Comparative studies on rabbit plasma lipid profile fed with Silybum marianum oil, sunflower oil and vegetable ghee.

Present work is aimed to compare the physicochemical characterization and biochemical effects of oil extracted from Silybum Marianum and Sunflower oil, collected from Peshawar (Pakistan). To investigate the comparative effects on the body weight, organ weight and lipid profile, the crude oil of Silybum marianum, edible sunflower oil and vegetable ghee were given to three groups of rabbits under study. Percent proximate composition and food consumption of all rabbits were determined which showed no significant statistical variation. There is no data available about Silybum marianum oil on animal model in literature. This study clearly revealed that oil from Silybum marianum significantly reduces plasma cholesterol level in rabbits. A threefold higher Triglyceride levels was observed in vegetable ghee feeding groups compared with the sunflower and Silybum marianum oil feeding groups. The crude oil of Silybum marianum was found to be safe in rabbits compared with sunflower oil and vegetable ghee. The results of these studies revealed most valuable information and also support the refining and purification to convert this non-edible oil to edible oil.

Modulation of brown adipocyte activity by milk by-products: Stimulation of brown adipogenesis by buttermilk.

Brown adipocytes dissipate chemical energy in the form of heat through the expression of mitochondrial uncoupling protein 1 (Ucp1); Ucp1 expression is further upregulated by the stimulation of ß-adrenergic receptors in brown adipocytes. An increase in energy expenditure by activated brown adipocytes potentially contributes to the prevention of or therapeutics for obesity. The present study examined the effects of milk by-products, buttermilk and butter oil, on brown adipogenesis and the function of brown adipocytes. The treatment with buttermilk modulated brown adipogenesis, depending on the product tested; during brown adipogenesis, buttermilk 1 inhibited the differentiation of HB2 brown preadipocytes. In contrast, buttermilk 3 and 5 increased the expression of Ucp1 in the absence of isoproterenol (Iso), a ß-adrenergic receptor agonist, suggesting the stimulation of brown adipogenesis. In addition, the Iso-induced expression of Ucp1 was enhanced by buttermilk 2 and 3. The treatment with buttermilk did not affect the basal or induced expression of Ucp1 by Iso in HB2 brown adipocytes, except for buttermilk 5, which increased the basal expression of Ucp1. Conversely, butter oil did not significantly affect the expression of Ucp1, irrespective of the cell phase of HB2 cells, ie, treatment during brown adipogenesis or of brown adipocytes. The results of the present study indicate that buttermilk is a regulator of brown adipogenesis and suggest its usefulness as a potential food material for antiobesity.

Lipoid Pneumonia After Prolonged Inhalation of Clarified Butter Made from the Milk of a Buffalo or Cow (Ghee).

Lipoid pneumonia is a rare form of pneumonia caused by inhalation or aspiration of fat containing substances. It can present acutely or more commonly presents as an insidious onset chronic respiratory illness. It requires a high degree of suspicion with great emphasis on history. It can mimic tuberculosis, malignancy or interstitial lung disease. We report the case of a 31-year-old male with a history of sniffing hydrogenated oil, presenting with a non-resolving pneumonia.

Occurrence and spatial distribution of pesticide residues in butter and ghee (clarified butter fat) in Punjab (India).

The present study was undertaken to monitor organochlorine, organophosphate, and synthetic pyrethroid pesticide residues in butter (n = 55) and ghee (n = 56) samples collected from three different regions of Punjab. The estimation of pesticide residues was done by multiple residue analytical technique using gas chromatography equipped with GC-ECD and GC-FTD. The confirmation of residues was done on gas chromatography mass spectrometry in both selective ion monitoring (SIM) and scan mode. Results indicated the presence of hexacholorocyclohexane (HCH) and p,p' DDE as predominant contaminant in both butter and ghee. Residues of HCH were detected in 25 and 23% samples of butter and ghee, respectively, while residues of p,p' DDE were recorded in 29 and 25% of butter and ghee samples, respectively. None of the butter and ghee sample violated the MRL values of 200 ng g(-1) for HCH and 1250 ng g(-1) for dichorodiphenyl tricholorethane (DDT). The presence of endosulfan, cypermethrin, fenvalerate, deltamethrin, and chlorpyrifos were observed in a few butter and ghee samples at traces. The spatial variation for comparative occurrence of pesticide residues indicated higher levels in the south-western region of Punjab. Additionally, the temporal variation indicated the significant reduction of HCH and DDT levels in butter and ghee in Punjab.

Effects of High-Butterfat Diet on Embryo Implantation in Female Rats Exposed to Bisphenol A.

Bisphenol A (BPA) is an endocrine disruptor associated with poor pregnancy outcomes in human and rodents. The effects of butterfat diets on embryo implantation and whether it modifies BPA's actions are currently unknown. We aimed to determine the effects of butterfat diet on embryo implantation success in female rats exposed to an environmentally relevant dose of BPA. Female Sprague-Dawley rats were exposed to dietary butterfat (10% or 39% kcal/kg body weight [BW]) in the presence or absence of BPA (250 µg/kg BW) or ethinylestradiol (0.1 µg/kg BW) shortly before and during pregnancy to assess embryo implantation potentials by preimplantation development and transport, in vitro blastulation, outgrowth, and implantation. On gestational day (GD) 4.5, rats treated with BPA alone had higher serum total BPA level (2.3-3.7 ng/ml). They had more late-stage preimplantation embryos, whereas those receiving high butterfat (HBF) diet had the most advanced-stage embryos; dams cotreated with HBF and BPA had the most number of advanced embryos. BPA markedly delayed embryo transport to the uterus, but neither amount of butterfat had modifying effects. An in vitro implantation assay showed HBF doubled the outgrowth area, with BPA having no effect. In vivo, BPA reduced the number of implanted embryos on GD8, and cotreatment with HBF eliminated this adverse effect. HBF diet overall resulted in more and larger GD8 embryos. This study reveals the implantation disruptive effects of maternal exposure to an environmentally relevant dose of BPA and identifies HBF diet as a modifier of BPA in promoting early embryonic health.

Effect of Maillard reaction on the quality of clarified butter, ghee.

In Ayurveda, a traditional Indian medicine system, clarified butter is called ghee and is used for food and medicinal purposes. Since butter is subjected to heat to prepare ghee, the heating process affects the ghee quality, such as oxidation, flavor, nutritional value, and biological activity. Therefore, this study focused on the Maillard reaction progress and free-radical scavenging activity with temperature and time during ghee preparation. First, ghee was prepared at low to high temperatures, and its quality (milk fat content, retinol, α-tocopherol, peroxide value, Maillard reaction progress, and free radical scavenging activity) was evaluated. Maillard reaction progress was enhanced at medium and high temperatures (120-160 â„ƒ), and the free radical-scavenging activity of ghee corresponded to the Maillard reaction progress. Since ghee is often reheated during use, we further evaluated the effect of the reheating process. The reheating process did not alter the Maillard reaction progress or the free radical scavenging activity. Our findings serve as good quality control measures for ghee preparation.

Histological and Microscopic Analysis of Fats in Heart, Liver Tissue, and Blood Parameters in Experimental Mice.

The intake of various types and amounts of dietary fats influences metabolic and cardiovascular health. Hence, this study evaluated the impact of routinely consumed Pakistani dietary fats on their cardiometabolic impact. For this, we made four groups of mice, each comprising 5 animals: (1) C-ND: Control mice on a normal diet, (2) HFD-DG: High-fat diet mice on a normal diet plus 10% (w/w) desi ghee, (3) HFD-O: Mice on normal diet plus 10% (w/w) plant oil (4) HFD-BG: Mice on normal diet plus 10% (w/w) banaspati ghee. Mice were fed for 16 weeks, and blood, liver, and heart samples were collected for biochemical, histological, and electron microscopic analysis. The physical factors indicated that mice fed on HFD gained more body weight than the C-ND group. Blood parameters do not show significant differences, but overall, the glucose and cholesterol concentrations were raised in the mice fed with a fat-rich diet, with the highest concentrations in the HFD-BG group. The mice fed with HFD-BG and HFD-O had more lipid droplets in the liver, compared to HFD-DG and C-ND.

Evaluating the in-vivo effects of olive oil, soya bean oil, and vitamins against oxidized ghee toxicity.

The aim of this study was to examine the protective role of various lipids (olive and soya oil) and vitamin (E and C) against the toxicity of thermally oxidized ghee in rabbits. Vanaspati ghee was thermally oxidized on a hot plate at 100°C for ten consecutive hours, and the oxidized ghee was stored in a refrigerator at -20°C until administration. Thirty male rabbits were purchased as experimental animals at a local market and were divided into ten corresponding groups of three based on their body weight. The blood samples of 5 ml were collected on day 0, 7, and 14 of the experiment for the analysis of hematological and biochemical serum parameters. We observed that oxidized ghee significantly elevated ALT level by affecting liver hepatocytes. Furthermore, vitamin E rapidly decreased the ALT levels compared to vitamin C and other oils. The oxidized ghee caused a significant increase in cholesterol compared to the other groups. Vitamin E and C showed the best antioxidant activity and decreased cholesterol levels to normal. Histopathological examinations of the normal rabbits' liver sections revealed no significant histological abnormality. The liver of the rabbits fed with oxidized ghee had an intact lobular architecture but the portal tracts showed inflammation and mild fibrosis, the bile ducts showed proliferation, and the hepatocytes showed feathery degeneration. In the liver sections from the groups fed with oxidized ghee and different doses of olive oil inflammation in portal tracts and large vacuoles in the hepatocytes were observed. The group fed with oxidized ghee and vitamin E had intact lobular architecture with no significant histological abnormality in portal tracts but fatty changes were present in the hepatocytes. These findings support the antioxidant activity of vitamins C and E as they reduced liver infection caused by oxidized ghee. It was concluded that oxidized ghee was highly toxic and not safe for consumption. The present study indicated that soya bean oil and vitamin E were more effective in protecting against the toxicity of thermally oxidized ghee than olive oil and vitamin C.