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Journal of Food Science and Technology logoLink to Journal of Food Science and Technology
. 2023 Feb 6;60(3):1175–1184. doi: 10.1007/s13197-023-05671-w

Combined antidiabetic potential of camel milk yogurt with Cinnamomum verum and Stevia rebaudiana by using rodent modelling

Aneela Hameed 1,✉, Farheen Ishtiaq 1, Muhammad Zeeshan 1, Saeed Akhtar 1, Tariq Ismail 1, Rabia Shabir Ahmad 2, Mamoona Amir 1, Muhammad Junaid Anwar 1
PMCID: PMC9998783  PMID: 36908342

Abstract

This study aimed to evaluate anti-diabetic potential of functional camel milk yogurt enriched with Cinnamomum verum and Stevia rebaudiana that not only mask its peculiar flavour rather have an antidiabetic effect as well. Sixty-three pathogen free STZ-induced albino Wistar rats were categorized into 7 groups on the basis of treatments. From each treatment group three animals were dissected periodically at 0, 7 and 21 days of study to determine the effect of all treatments on physicochemical parameters, serum glucose, serum chemistry and haematology. The study revealed that the mean blood glucose level in the untreated control group was within normal range (100–108 mg/dl) while there was noticeable decrease in mean blood glucose level of all the treated groups during three weeks’ trial. Decline in blood glucose level (46%) was higher in animal group containing functional camel milk yogurt (T4) at 3rd week of trial as compared to other treatments.

Keywords: Albino Wistar rats, Camel milk, Diabetes, Functional yogurt, Streptozotocin

Introduction

Diabetes mellitus—type 2 is a metabolic syndrome characterized by chronic hyperglycaemia and disturbances in carbohydrates, lipids and proteins metabolism. Globally there are 536.6 million diagnosed and undiagnosed cases of diabetes at present while the burden of type 2 diabetes is increasing significantly worldwide (Sun et al. 2022). Treatment regimens and therapies currently available for the management of diabetes are expensive for majority of populations inflicted with diabetes particularly in the developing countries. Ineffectiveness and side-effects of existing oral hypoglycemic agents and insulin have compelled researchers of this era to develop nascent functional foods and herbal remedies as an alternative to chemical medications to prevent diabetes mellitus (Nkambule et al. 2021). Present era demands the exploitation of some innovative, naturally occurring, nutritive, cost effective and viable foods to mitigate the impact of diabetes mellitus for improving livelihood standard of diabetic patients, especially among the malnourished and economically oppressed populations of the world.

Utilisation of dairy based food products such as yogurt known to contain valuable microbiota i.e. probiotics are considered helpful to cope the diabetes mellitus. A number of previous studies validate the beneficial impacts of yogurt consumption in reducing the risk of diabetes mellitus as well. Camel milk has long been used as a natural remedy for effectively managing various diseases such as diabetes type 2 (Kaskous 2016). Camel milk is considered a nutritious food as it is known to hold low levels of sugars and cholesterol while on the other hand camel milk is known to contain high levels of minerals such as sodium, potassium, iron, copper, zinc and magnesium along with certain vitamins such as vitamin C. Camel milk is well recognized as an effective food in reducing the glycated haemoglobin levels in the blood (Agrawal et al. 2003). Presence of insulin like proteins in the camel milk has been found to elicit beneficial effect for the treatment of diabetes mellitus. A clinical trial on diabetic patients has even reported 30–35% reduction in insulin doses in patients of type I diabetes on the raw camel milk consumption. Yet, despite its high nutritional significance and sufficient production in some parts of world, no attention has ever been given to dromedary milk and milk based products (Ahmad et al. 2010).

Exploitation of natural sweeteners in place of the sugar is increasing day by day owing to high rates of diabetes mellitus. These low caloric natural sweeteners have a tendency to provide greater sweetness levels such as that derived from stevia leaves is 300 times sweeter than the natural sugar, has no after-taste and is safe and non-toxic (Adesh et al. 2012). A plethora of research suggests that stevia holds an ability to increase the glucose tolerance and decrease postprandial blood glucose levels. Moreover, stevia compounds such as steviol and stevioside have been shown to exert direct effect on β-cells to produce insulin from pancreas (Goyal et al. 2010).

Therefore, stevia containing stevioside has hypotensive and hypoglycemic characteristics and might substantially substitute simple sugars (Lemus-Mondaca et al. 2012). Stevia as a natural sugar has been exploited by various researchers in camel milk Yogurt as an ingredient to mask the natural salty taste of camel milk and provide a substitute for sucrose (Prakash and Dubois 2013).Cinnamon (Cinnamomum verum) herbal spice has a wide acceptance as an important ingredient in the foods and traditional medicine since ancient times. Cinnamon contains various bioactive compounds including proanthocyandins such as cinnamaldehyde, that act as a natural antidiabetic agent (Hafizur et al. 2015).Cinnamon in its various forms has been found to bear anti-mutagenic and antioxidant activities as well as improve the insulin sensitivity and help in lowering down the absorption of carbohydrates in the small intestine thus ultimately manage diabetes (Abd et al. 2010). Moreover, Cinnamon is also known for its typical aroma and flavour that masks the subtle undesirable flavour and odour of camel milk (Anderson et al. 2004).

Researchers from the past have investigated the use of various natural herbs in distinguished foods to reduce diabetes mellitus’ impact on human health. Hence, the utilization of the natural herbs such as cinnamon and stevia in the camel milk Yogurt may be a cumulative, viable and adjunctive therapy to reduce diabetes mellitus in comparison with the artificial anti-diabetic agents. To test the hypothesis, whether functional camel milk yogurt has antidiabetic potential or not, the primary objective of this study was to develop cinnamon and stevia based functional camel milk yogurt to manage diabetes mellitus in animal modelling i.e., albino rat model.

Materials and methods

Procurement of sample and chemicals

Camel milk was procured from local market. Collected milk samples were thoroughly mixed and divided into three lots each representing a sample. Samples were immediately transferred to the laboratory of institution in ice box to maintain the quality of milk. Milk samples thus collected were stored in refrigerator (4–6 °C) for further analysis. Analytical grade chemicals of Sigma-Aldrich, USA were used in the whole research work. Diagnostic kits for serum analysis were purchased from Diasys®, Lahore. Stevia i.e., Stevia rebaudiana (Stevia liquid concentrate, planetary herbals, was imported from United State of America (USA). Cinnamon herb i.e., Cinnamon verum and commercial yogurt culture of Streptococcus thermophilus and Lactobacillus bulgaricus (1:1) were purchased from local market.

Physicochemical analysis of milk

Camel milk was analysed for its chemical composition including pH, acidity, fat, protein, ash content, solids not fat content (SNF) and total solids according to the methods described by AOAC method No 981.12 and 925.23 (1990) respectively (AOAC 1990).

Preparation of standardized camel milk yogurt

Standardized camel milk yogurt was prepared by following the procedure as described by Ahmad (2011). Milk samples were cooled up to 40 °C after pasteurization (68 °C for 20 min), added cinnamon water extract (1:10) (cinnamon: distilled water) (w/v) and 1% gelatin (w/v) as a stabilizer to improve the texture of yogurt and inoculated with commercial yogurt culture Lactobacillius bulgaricus and Streptococcus thermophiles (5%). The samples were then transferred to plastic cups and incubated at 43 °C for 18 h. Both yogurt samples (Standardized and Functional) were stored at 4 °C until analysed for further physicochemical evaluation at different time intervals (0, 10 days).

Physicochemical analysis of yogurt

pH (HANNA-pH 210, Germany), Total protein content (Micro Kjeldahl method by multiplying N with the factor 6.38), total titratable acidity, total proteins and total soluble solids (gravimetric method) of the samples were estimated by using AOAC (1990) and AOAC (2012)respectively. Ash contents were determined by dry ashing of the samples for 24 h at 550 °C in muffle furnace (Ney® VULCAN—550) and moisture was calculated by drying samples overnight at 105 °C in hot air oven (Memmert 91126, Germany) (Longvah and Deosthale 1998). Fat content of the samples were estimated using the standard Gerber method using (Funke Gerber 3670 Nova Safety Centrifuge) (Nielsen 1998). All of the analyses were performed in triplicate.

Efficacy study

Diet preparation

Diet of the rats was prepared according to Lavrat–Vernny (1999) model. The standard diet recipe was followed as corn starch (65.5%), casein (10%), cellulose (10.5%), corn oil (10.00%), mineral mixture (3.00%) and vitamin mixture (1%).

Acclimatization of rats

Sixty-three (63) pathogen free albino Wistar rats weighing 160–200 g were purchased from the rat centre of pharmacy department. Rats were kept in the efficacy room of institute and maintained at 20 ± 2 °C temperature and at 60% R.H. (relative humidity) with day and 12 h night cycle. Rats were acclimatized to the experimental room conditions for one week and standard diet was fed to the rats for a period of seven days followed by feeding treatment diets for the next 21 days (Table 1). Safe and clean water for drinking was facilitated to the experimental rats. Water and feed of each rat was monitored and quantified daily during the whole research period. Body weights with the glucose levels were also estimated daily of each rat and was noted weekly.

Table 1.

Diet plan for different groups of Albino Wistar rats

Sr no. Ingredient T0− T0+ T1 T2 T3 T4 T5
1 Corn starch (g) 57 57 57 57 57 57 57
2 Corn oil (g) 10 10 10 10 10 10 10
3 Casein (g) 20 20 20 20 20 20 20
4 Bran (g) 5.25 5.25 5.25 5.25 5.25 5.25 5.25
5 Mineral Texture (g) 1.6 1.6 1.6 1.6 1.6 1.6 1.6
6 Cellulose (g) 5.3 5.3 5.3 5.3 5.3 5.3 5.3
7 Vitamin Mixture (g) 1 1 1 1 1 1 1
8 Insulin (units/Kg by weight/day) – 6 – – – – –
9 Standardized camel milk yogurt (g) – – 20 – – – –
10 Pure cinnamon extract (mg/kg) – – – 23 – – –
11 Fresh camel milk (mL) – – – – 100 – –
12 Functional camel milk yogurt (stevia + cinnamon) (g) – – – – – 20 –
13 Pure stevia extract (mg/kg) – – – – – – 25
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Level of Significance: P < 0.05, T0−: Control group, T0+: Insulin treated group, T1: Fresh camel milk, T2: Pure cinnamon extract treated group, T3: Standardized camel milk yogurt treated group, T4: Functional camel milk yogurt treated group (stevia + cinnamon), T5: Pure stevia extract treated group

Rat categorization and diabetes induction

Rats were categorized into seven different groups namely untreated controls (T0), insulin treated diabetic (T0+), fresh camel milk (T1), pure cinnamon extract (T2), standardized camel milk yogurt (T3), functional camel milk yogurt (T4) and pure stevia extract fed rats group (T5) each treatment group was comprised of nine rats. The albino rats were first subjected to fasting and then diabetes was induced using freshly made streptozotocin (55 mg/kg body weight) via peritoneal injection in 0.1 M citrate buffer of 4.5pH (Khan et al. 2013).

Blood chemistry and haematological test

Three animals from each group were randomly selected and sacrificed periodically at 0, 7th and 21st day of study to ascertain the effect of all the treatments on serum glucose, serum triglycerides, total cholesterol, creatinine and enzymes including ALT and AST by using serum chemistry analyser. Blood was collected in Ethylene Diamine Tetra Acetic acid (EDTA) containing vials to avoid the coagulation of the blood samples. Serum was separated from the blood collected via micro centrifuge machine in Eppendorf tubes. Blood serum and separated tissues thus collected were stored at − 80 freezers (Ultra-Low Temperature Freezer MDF-U33V) in labelled Ziploc bags for final analysis. Haematological analysis was performed on the blood samples of albino wistar rats. Samples were run in the haematology analyser following the procedure of Dacie and Lewis (1984). In total blood count following tests were performed; WBC, RBC, haemoglobin concentration, HCT/PCV, MCV, MCHC, MCH, platelets, monocytes and lymphocytes.

Statistical analysis

Biological activities were statistically analysed using two-way analysis of variance technique (ANOVA). Pearson correlation was used to establish correlation among the variables. Least significant difference (LSD) test was performed to compare the means and p < 0.05 was considered level of significance. Each analysis was replicated and results were expressed as mean ± S.D (Wu and Hamada 2011).

Results

Physicochemical analysis

Present research revealed a significant reduction in pH of T1 and T2 treatments with storage days whereas the acidity of T1 and T2 treatments increased as the storage duration enhanced. Statistical results for fat in the current study (Table 2) exhibited that standardized camel milk yogurt (T1) contains 3.79% and functional camel milk yogurt (T2) contains 3.86% fat at 0th day storage while on the other hand 3.54% and 3.51% were present in T1 and T2 respectively on 10th day of storage. Mean values for the fat content of T1 and T2 treatments were significantly (p < 0.05) declined along with the storage days up to the final day of study period. Inclusion of cinnamon verum and stevia rebaudiana in the camel milk yogurt had no effect on the solid not fat (SNF) contents of yogurt. An inclined trend in protein contents were observed in both treatments T1 and T2 along with storage days.

Table 2.

Effect of storage on physicochemical properties of camel milk yogurt

Parameters (%) Yogurt type Storage days Overall mean
0 10
pH T1 4.62 ± 0.10a 4.05 ± 0.04b 4.33 ± 0.32a
T2 4.54 ± 0.09a 4.09 ± 0.10b 4.32 ± 0.26a
Acidity T1 0.50 ± 0.01d 0.84 ± 0.01b 0.58 ± 0.09b
T2 0.67 ± 0.01c 0.97 ± 0.01a 0.90 ± 0.07a
Fats T1 3.79 ± 0.01b 3.54 ± 0.01c 3.66 ± 0.13b
T2 3.86 ± 0.01a 3.51 ± 0.01d 3.68 ± 0.19a
Proteins T1 4.81 ± 0.02b 5.03 ± 0.05a 4.92 ± 0.12a
T2 4.96 ± 0.15ab 5.11 ± 0.09a 5.03 ± 0.13a
Total Solids T1 14.82 ± 0.11a 14.91 ± 0.09a 14.86 ± 0.10a
T2 14.74 ± 0.57a 14.63 ± 0.56a 14.69 ± 0.51a
SNF T1 11.04 ± 0.05a 11.35 ± 0.05a 11.20 ± 0.17a
T2 11.32 ± 0.50a 11.42 ± 0.04a 11.37 ± 0.32a
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Statistically non-significant means in rows and columns share same letters (p < 0.05), Superscript letters (a,b,c,d etc.) are used to indicate pairwise comparisons, whether such comparisons are statistically different or not

T1: Standardized camel milk yogurt, T2: Functional camel milk yogurt

Effect of various treatments on Serum glucose

Highly significant response (p < 0.05) of the study period i.e., 14 and 21 days was recorded on the serum glucose level of rats treated with various extracts and standardized formulas as mentioned in Table 3. A consistent decline was observed in the glucose level of diabetic rats treated with fresh camel milk (T1), pure cinnamon extract (T2), standardized camel milk yogurt (T3), functional camel milk yogurt (T4), stevia extract (T5) and insulin treated group (T0+). Rate of serum glucose reduction was highest in T0+ and T4 treated groups followed by T3, T1, T5 and T2. T0− (control group) showed non-significant changes in the serum glucose concentration i.e., 108, 100 and 105 mg/dL at 0, 14 and 21 days of study, respectively. While diabetic control group (T0+) showed 60% decline in serum glucose level as compared to negative control diabetic rats in 21 days’ trial. 46%reduction was noticed in serum glucose of functional camel milk yogurt (T4) fed rats group as compared to that of insulin treated group (T0+). Thirty-seven percent, 35%, 29% and 20% reduction in serum glucose were observed in standardised camel milk yogurt (T3), fresh camel milk (T1), stevia extract (T5) and cinnamon extract (T2) fed rats groups, respectively.

Table 3.

Effect of treatments on Serum glucose (mg/dL) levels of Albino Wistar rats

Groups Treatments Mean ± SD (Day 0) Mean ± SD (Day 14) Mean ± SD (Day 21) % Decrease
Control T0− 108 ± 6.8k 100 ± 4.5j 105 ± 5.0j
T0+ 380 ± 8.5ab 209 ± 5.5h 153 ± 10.3i 60
Treated T1 375 ± 7.6ab 301 ± 10.1d 242 ± 8.5fg 35
T2 374 ± 7.6a 338 ± 17.6c 301 ± 10.5d 20
T3 380 ± 9.5ab 296 ± 16.0d 240 ± 11.5g 37
T4 390 ± 8.7a 263 ± 10.6e 209 ± 16.0h 46
T5 364 ± 10.1b 291 ± 6.5d 258 ± 6.8ef 29
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Level of Significance: P < 0.05, Superscript letters (a,b,c,d etc.) are used to indicate pairwise comparisons, whether such comparisons are statistically different or not, T0−: Control group, T0+: Insulin treated group, T1: Fresh camel milk, T2: Pure cinnamon extract treated group, T3: Standardized camel milk yogurt treated group, T4: Functional camel milk yogurt treated group (stevia + cinnamon), T5: Pure stevia extract treated group

Effect of various yogurt treatments on serum chemistry and haematology

The diabetic rats showed significant changes in serum blood chemistry and haematological parameters (Tables 4, 5) as the storage duration enhanced. Treated groups of STZ-induced diabetic rats showed significant effect on serum levels of proteins, albumin, triglycerides, cholesterol, urea, creatinine, ALT and AST to normalize these changes (Table 4). Treatments T4 and T5 showed a positive effect on STZ-induced diabetic rats to reduce the Triglycerides to normal level. The functional camel milk yogurt treated (T4) groups of STZ-induced diabetic rats exhibited a reduction in cholesterol to controlled levels. Moreover, the rats fed on functional camel milk yogurt (T4) showed a significant reduction in urea and creatinine levels. It was observed that activities of ALT and AST enzymes in STZ-induced diabetic rats that increased due to diabetes were dramatically shifted towards normal values by the help of treatment T4. Results in Table 5 showed that administration of camel milk functional yogurt to STZ-induced diabetic rats helped to manage the normal levels of WBCs, lymphocytes, RBCs, haemoglobin, platelets, MCV, MCH, MCHC, PCV and monocytes as well.

Table 4.

Effect of treatments on Serum Chemistry of Albino Wistar rats

Parameters (%) Yogurt type Storage days Overall mean
0 7 21
Protein T0− 6.90 ± 0.01a 6.87 ± 0.02a 6.86 ± 0.03a 6.88a
T0+ 5.05 ± 0.04g 5.85 ± 0.01e 6.71 ± 0.01b 5.87e
T1 5.33 ± 0.21f 6.04 ± 0.05cd 6.82 ± 0.02ab 6.06c
T2 5.43 ± 0.20f 6.11 ± 0.02c 6.90 ± 0.01a 6.14b
T3 5.13 ± 0.02g 5.96 ± 0.02de 6.81 ± 0.02ab 5.97d
T4 5.09 ± 0.02g 5.92 ± 0.02de 6.79 ± 0.01ab 5.93de
T5 3.28 ± 0.38de 5.39 ± 0.47c 6.32 ± 0.26ab 5.00c
Albumin T0− 3.81 ± 0.01a 3.81 ± 0.01a 3.82 ± 0.02a 3.81a
T0+ 2.56 ± 0.02k 3.04 ± 0.03h 3.54 ± 0.02d 3.05f
T1 2.64 ± 0.02j 3.29 ± 0.02f 3.72 ± 0.01b 3.22c
T2 2.73 ± 0.02i 3.43 ± 0.03e 3.80 ± 0.01a 3.32b
T3 2.53 ± 0.05k 3.28 ± 0.01f 3.70 ± 0.01b 3.17d
T4 2.57 ± 0.03k 3.13 ± 0.02g 3.64 ± 0.01c 3.11e
T5 1.57 ± 0.35f 3.26 ± 0.61bcd 3.57 ± 0.56b 2.80b
Triglyceride T0− 72.33 ± 1.52l 73.33 ± 1.52l 71.00 ± 1.00l 72.22f
T0+ 111.67 ± 3.05gh 100.00 ± 1.00i 89.33 ± 2.51k 100.33e
T1 112.00 ± 4.35fgh 133.00 ± 2.00b 112.67 ± 1.52efg 119.22b
T2 116.33 ± 2.51e 150.67 ± 2.08a 120.33 ± 1.52d 129.11a
T3 113.67 ± 2.51efg 127.67 ± 2.51c 108.67 ± 1.52h 116.67c
T4 115.67 ± 3.05ef 110.00 ± 1.00gh 95.67 ± 1.52j 107.11d
T5 174.33 ± 6.65c 127.33 ± 11.84e 101.33 ± 2.08f 134.33c
Cholesterol T0− 71.33 ± 2.08k 71.67 ± 2.51k 71.67 ± 2.08k 71.56f
T0+ 95.67 ± 1.52hi 109.33 ± 2.51f 101.00 ± 2.00g 102.00e
T1 93.00 ± 2.00ij 130.67 ± 2.08b 120.00 ± 1.00d 114.56b
T2 97.00 ± 2.00h 143.67 ± 2.08a 130.00 ± 1.00bc 123.56a
T3 94.33 ± 1.52hi 127.33 ± 1.52c 112.67 ± 2.08e 111.44c
T4 91.00 ± 3.60j 118.67 ± 1.52d 104.00 ± 1.00g 104.56d
T5 166.67 ± 5.03bc 112.33 ± 6.65d 95.00 ± 3.46d 124.67c
Urea T0− 20.78 ± 0.72j 21.49 ± 0.54j 21.05 ± 0.78j 21.10f
T0+ 26.77 ± 0.39h 30.91 ± 0.17f 24.71 ± 0.41i 27.46e
T1 31.95 ± 0.12e 35.52 ± 0.51c 29.03 ± 0.30g 32.17c
T2 30.90 ± 0.80f 39.34 ± 0.39a 34.27 ± 0.35d 34.83a
T3 29.33 ± 0.56g 37.05 ± 0.15b 31.67 ± 0.49ef 32.68b
T4 29.34 ± 0.60g 34.47 ± 0.45d 27.19 ± 0.26h 30.33d
T5 35.39 ± 3.55bc 25.79 ± 1.79de 24.01 ± 1.53ef 28.39c
Creatinine T0− 0.27 ± 0.05l 0.28 ± 0. 7l 0.28 ± 1.00l 0.27e
T0+ 0.96 ± 0.02j 1.31 ± 0.02e 0.87 ± 0.02k 1.05d
T1 1.09 ± 0.01gh 1.46 ± 0.02c 1.05 ± 0.04hi 1.20b
T2 1.07 ± 0.04hi 1.60 ± 0.02a 1.21 ± 0.02f 1.29a
T3 1.03 ± 0.04hi 1.53 ± 0.08b 1.14 ± 0.03g 1.23b
T4 1.03 ± 0.04i 1.38 ± 0.03d 0.96 ± 0.02j 1.12c
T5 1.73 ± 0.49bc 1.24 ± 0.14ef 0.96 ± 0.23f 1.31b
ALT T0− 36.00 ± 1.00j 37.00 ± 2.00j 36.00 ± 2.00j 36.33d
T0+ 54.00 ± 2.00i 102.33 ± 3.05c 74.00 ± 2.00g 76.77c
T1 61.33 ± 1.52h 110.33 ± 1.52a 83.00 ± 2.00de 84.88a
T2 58.67 ± 1.52h 105.67 ± 2.08b 85.67 ± 1.52d 83.33a
T3 59.00 ± 2.00h 107.00 ± 2.00b 77.00 ± 2.00fg 81.00b
T4 51.00 ± 1.00i 104.00 ± 1.00bc 80.00 ± 2.00ef 78.33c
T5 124.33 ± 4.16b 91.67 ± 6.02ef 70.00 ± 1.00h 95.33b
AST T0− 75.33 ± 1.52k 77.00 ± 1.00k 76.67 ± 2.08k 76.33e
T0+ 93.00 ± 2.00hi 118.00 ± 2.00d 89.00 ± 2.00j 100.00d
T1 95.00 ± 2.00gh 134.00 ± 2.00b 110.33 ± 1.52e 113.11a
T2 90.00 ± 3.00ij 137.67 ± 1.52a 113.33 ± 1.52e 113.67a
T3 94.00 ± 1.00gh 130.00 ± 2.00c 105.00 ± 2.00f 109.67b
T4 90.00 ± 1.00ij 128.67 ± 1.52c 97.00 ± 2.00g 105.22c
T5 116.67 ± 2.08bc 103.00 ± 1.73ef 93.67 ± 3.05g 104.44b
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Level of Significance: P < 0.05, T0−: Control group, Superscript letters (a,b,c,d etc.) are used to indicate pairwise comparisons, whether such comparisons are statistically different or not, T0+: Insulin treated group, T1: Fresh camel milk, T2: Pure cinnamon extract treated group, T3: Standardized camel milk yogurt treated group, T4: Functional camel milk yogurt treated group (stevia + cinnamon), T5: Pure stevia extract treated group

Table 5.

Haematological Analysis of Albino Wistar rats

Parameters (%) Yogurt type Storage days Overall mean
0 14 21
White blood cells (WBC’s) T0− 7.06 ± 0.25cde 6.47 ± 0.51ef 6.76 ± 0.45def 6.76d
T0+ 10.86 ± 0.15a 7.76 ± 0.15bcd 6.63 ± 1.50ef 8.42bc
T1 10.47 ± 0.82a 8.00 ± 0.10bc 8.30 ± 0.98b 8.92ab
T2 11.20 ± 0.43a 8.10 ± 0.10bc 8.53 ± 0.79b 9.27a
T3 11.23 ± 0.55a 8.16 ± 0.61b 6.87 ± 0.55def 8.75ab
T4 10.60 ± 0.62a 7.50 ± 0.62bcde 5.98 ± 0.20f 8.02c
T5 9.53 ± 0.70a 9.50 ± 0.65bcd 7.23 ± 0.80f 8.75ab
Lymphocytes T0− 68.00 ± 1.00ab 66.75 ± 0.25abc 68.50 ± 0.50ab 67.75a
T0+ 35.10 ± 3.72d 59.66 ± 3.78bc 59.33 ± 1.52bc 51.36d
T1 38.83 ± 2.70d 60.66 ± 3.05bc 63.00 ± 14.00ab 57.50bc
T2 44.28 ± 3.44d 62.66 ± 5.68a 66.33 ± 16.01a 57.76b
T3 42.90 ± 3.50d 58.66 ± 8.96bc 61.66 ± 8.96a 54.41bc
T4 40.17 ± 0.97d 57.10 ± 3.05c 59.66 ± 3.21bc 52.31cd
T5 30.85 ± 0.58f 43.83 ± 0.94cde 46.16 ± 4.41cd 40.28b
RBC’s T0− 7.20 ± 0.10abc 7.40 ± 0.10a 7.23 ± 0.49a 7.27a
T0+ 4.63 ± 0.30k 5.81 ± 0.46hij 6.50 ± 0.67defg 5.64d
T1 5.25 ± 0.27ijk 6.80 ± 0.45abcde 7.13 ± 0.15abcd 6.39b
T2 5.87 ± 0.21ghi 6.57 ± 0.56bcdef 6.53 ± 0.50def 6.32b
T3 5.17 ± 0.17jk 6.02 ± 0.24fgh 6.42 ± 0.04efgh 5.87cd
T4 4.83 ± 0.30k 6.56 ± 0.66cdef 7.21 ± 0.37ab 6.20bc
T5 4.98 ± 0.44ef 5.66 ± 0.25cd 6.13 ± 0.25bc 5.59bc
Hemoglobin (HB) T0− 14.80 ± 0.10ab 14.90 ± 0.10a 15.40 ± 1.30a 15.03a
T0+ 11.11 ± 0.83fgh 11.20 ± 0.81hij 11.81 ± 0.47ghij 11.37d
T1 11.80 ± 0.65fghi 12.60 ± 0.36def 12.96 ± 0.32cde 12.45bc
T2 12.06 ± 0.35efg 13.20 ± 0.43cd 13.83 ± 0.30bc 13.03b
T3 10.83 ± 0.70ij 12.64 ± 0.97def 13.80 ± 0.95bc 12.42c
T4 10.50 ± 0.10j 11.40 ± 0.45ghij 12.60 ± 0.45def 11.50d
T5 10.34 ± 0.62j 12.45 ± 0.67d 13.40 ± 0.89bc 12.23d
Platelets T0− 865.33 ± 45.00a 852.00 ± 60.10ab 783.33 ± 104.08bcd 826.78a
T0+ 782.00 ± 13.89bcd 688.33 ± 76.54efg 603.00 ± 60.00h 691.11c
T1 819.33 ± 11.01abc 763.33 ± 30.55cde 696.67 ± 48.95efg 759.78b
T2 865.33 ± 21.57a 732.00 ± 38.69def 641.33 ± 5.50gh 746.22b
T3 823.00 ± 31.57abc 678.33 ± 57.95fgh 666.00 ± 34.17fgh 722.44bc
T4 780.00 ± 10.00bcd 657.33 ± 30.02fgh 663.67 ± 28.14fgh 700.33c
T5 870.67 ± 6.35a 682.67 ± 54.37efg 666.67 ± 32.71fg 740.00bc
Mean corpuscular volume (MCV) T0− 56.00 ± 0.500abc 53.83 ± 0.76cd 53.33 ± 4.72cde 54.38ab
T0+ 52.10 ± 0.26def 49.96 ± 0.75fg 49.73 ± 0.37fg 50.60c
T1 57.46 ± 1.40a 54.53 ± 0.50bcd 49.83 ± 2.02fg 53.94ab
T2 58.00 ± 0.50a 54.13 ± 0.32cd 54.16 ± 2.56cd 55.43a
T3 57.00 ± 2.85ab 53.13 ± 0.70de 51.06 ± 0.50efg 53.73b
T4 53.00 ± 1.00de 48.93 ± 0.40g 50.26 ± 1.10fg 50.73c
T5 49.96 ± 0.83bc 49.93 ± 1.10bc 49.36 ± 1.15bc 49.75b
Mean corpuscular hemoglobin (MCH) T0− 21.00 ± 0.10b 20.90 ± 0.10b 22.33 ± 0.57a 21.41a
T0+ 20.00 ± 0.100cd 19.00 ± 0.10fg 18.43 ± 0.51g 19.14d
T1 20.00 ± 0.50cd 19.53 ± 0.55def 19.10 ± 1.01efg 19.54cd
T2 22.0 ± 0.10a 20.00 ± 0.10cd 18.33 ± 0.57g 20.11b
T3 20.73 ± 0.92bc 19.96 ± 0.40cde 18.66 ± 0.57fg 19.78bc
T4 19.50 ± 0.57def 19.43 ± 0.73def 19.33 ± 0.50def 19.42cd
T5 21.50 ± 0.10b 19.43 ± 0.45def 19.10 ± 0.57efg 20.01b
Mean corpuscular hemoglobin concentration (MCHC) T0− 36.50 ± 0.50a 34.43 ± 0.51bc 31.66 ± 0.57ef 34.20ab
T0+ 32.00 ± 0.10def 30.23 ± 0.25f 30.66 ± 1.15f 30.96c
T1 31.83 ± 2.02ef 34.50 ± 1.30bc 34.10 ± 1.21bc 33.47ab
T2 35.10 ± 0.65ab 32.96 ± 0.15cde 35.43 ± 1.06ab 34.50a
T3 34.03 ± 0.90bc 33.90 ± 1.21bc 33.73 ± 2.87bcd 33.88ab
T4 33.83 ± 0.76bcd 31.43 ± 0.51f 34.30 ± 0.60bc 33.18b
T5 34.10 ± 0.51bc 33.75 ± 1.23bcd 33.45 ± 1.06ab 33.76ab
Packed cell volume (PCV) T0− 49.00 ± 1.00a 47.00 ± 1.00abcd 48.66 ± 3.05ab 48.22a
T0+ 39.36 ± 1.30hi 41.33 ± 2.51ghi 45.16 ± 2.84def 41.95c
T1 40.00 ± 1.00hi 41.66 ± 1.15gh 46.66 ± 1.52abcd 42.77bc
T2 41.00 ± 1.00ghi 45.33 ± 2.08cde 46.00 ± 1.00bcde 44.11b
T3 38.66 ± 2.08i 43.66 ± 1.52efg 48.30 ± 0.60abc 43.54bc
T4 39.66 ± 3.05hi 42.33 ± 1.52fgh 44.66 ± 1.15def 42.22c
T5 40.10 ± 1.15hi 45.30 ± 2.10cde 46.70 ± 1.64abcd 44.03b
Monocytes T0− 2.30 ± 0.60bcde 2.66 ± 0.57abc 2.80 ± 0.26ab 2.58a
T0+ 1.06 ± 0.12g 2.00 ± 0.40cde 2.33 ± 0.57bcd 1.79b
T1 1.83 ± 0.25def 2.63 ± 0.40abc 2.66 ± 0.57abc 2.37a
T2 1.60 ± 0.45efg 2.61 ± 0.51abc 3.23 ± 0.32a 2.48a
T3 1.03 ± 0.35g 2.03 ± 0.06cde 2.66 ± 0.57abc 1.91b
T4 1.20 ± 0.20fg 2.13 ± 0.68bcde 2.10 ± 0.17bcde 1.81b
T5 1.50 ± 0.26d 1.90 ± 0.10cd 2.33 ± 0.15bc 1.91b
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Level of Significance: P < 0.05, Superscript letters (a,b,c,d etc.) are used to indicate pairwise comparisons, whether such comparisons are statistically different or not, T0−: Control group, T0+: Insulin treated group, T1: Fresh camel milk, T2: Pure cinnamon extract treated group, T3: Standardized camel milk yogurt treated group, T4: Functional camel milk yogurt treated group (stevia + cinnamon), T5: Pure stevia extract treated group

Discussion

This study evaluated the antidiabetic potential of functional camel milk yogurt enriched with Cinnamomum verum and Stevia rebaudiana on Sixty-three pathogen free albino Wistar rats categorized into 7 groups of 9 rats on the basis of treatments. For which the effect of all the treatments on physicochemical parameters, serum glucose, serum chemistry and haematology was examined.

The study showed a significant decrease in pH of T1 and T2 treatments with storage days. The results for pH of the present study are similar to the study of Shori and Baba (2013a, b, c) where pH of cinnamon enriched functional camel milk yogurt and standardized camel milk yogurt decreased with storage time. An increase in the acidity of yogurt was observed with the storage duration, which may be associated with conversion of lactose into lactic acid (Ibrahim and Khalifa 2015). Whereas non-significant differences (p < 0.05) have been noticed in another study (Shori and Baba 2013a, b, c). The statistical results for fat in the current study exhibited a significant decline as the storage duration enhanced and are in line with Shori and Baba (2013a, b, c). Moreover, the addition of cinnamon had no effect on the fat percentage of yogurt (Qureshi et al. 2011; Shori and Baba 2013a, b, c). The inclusion of cinnamon verum and stevia rebaudiana in the camel milk yogurt also had no effect on the solid not fat (SNF) contents of yogurt which is also reported in the study of Shori and Baba (2013a, b, c). Protein percentage in both treatments T1 and T2 was increased with storage days in camel milk yogurt. This finding is also supported by the study of Ahmad (2011) who reported that the protein contents were increased from 2.90 ± 0.01 to 3.99 ± 0.02% during the storage period of 15 days.

This study revealed a highly significant response (p < 0.05) of the study period i.e., 14 and 21 days on the serum glucose level of rats treated with various extracts and standardized formulas. Highest reduction was noticed in serum glucose of functional camel milk yogurt (T4) fed rats group as compared to that of insulin treated group (T0+). The results of treatments T1, T3 and T4 are similar to the study of Khan et al. (2013) who reported that camel milk significantly decreases the glucose level of STZ induced diabetic rats. The findings of Al-Numair (2010) and Meena et al. (2016) are also similar to the results of current research wherein treatment of diabetic rats with camel milk decreased the level of serum glucose due to the high concentration of insulin like proteins i.e., 45–128 units/litre. Results of treatments T4 are similar to some extent with the findings of Shivanna et al. (2013) according to which stevia improves the glucose level of diabetic rats and Kim et al. (2006) who reported that cinnamon extract imparts significant reduction in serum glucose concentration in diabetic models. Glucose mediating responses of pure cinnamon extracts and cinnamon and stevia functional Yogurt were also similar to some extent with the findings of Kim et al. (2006) who observed that cinnamon lowered the blood glucose concentration of diabetic rats.

Due to oxidative stress induced by diabetes, significant changes in the serum blood chemistry and haematological parameters were observed in diabetic rats. Treated groups of diabetic rats exhibited a significant effect on serum levels of proteins, albumin, triglycerides, cholesterol, urea, creatinine, ALT and AST to normalize these changes. Treatments T4 and T5 helped reduce the Triglycerides towards normal level and the treatment T4 also aided in the reduction of cholesterol as well as the activities of ALT and AST enzymes to controlled levels in diabetic rats. Moreover, improved kidney health was also observed in the rats fed on functional camel milk yogurt (T4) because of significant reduction in urea and creatinine levels. The study showed that camel milk functional yogurt also helped manage the normal levels of WBCs, lymphocytes, RBCs, haemoglobin, platelets, MCV, MCH, MCHC, PCV, monocytes in diabetic rats. These results are also supported by the findings of Abdel-Reheim and Hosni (2013) and Mansour et al. (2017) according to which camel milk supplementation helps ameliorate serum biochemical and haematological measurements that are altered after diabetes induction.

Conclusion

Present study concludes that functional camel milk yogurt contains hypoglycaemic properties. Individual and combined effects of stevia, cinnamon and camel milk exhibited remarkable reduction in the blood glucose levels and diabetes related complications among the streptozotocin induced Albino Wistar rats. This research work reveals that the functional camel milk yogurt may serve as a helpful functional food to reduce the risk of diabetes. Functional camel milk yogurt may be a good addition to pharma-nutrition and dairy sector for the promotion of the health of the populations at commercial levels.

Acknowledgements

Authors are thankful to institution for providing facilities of research.

Abbreviations

RH

Relative humidity

T0−

Control group

T0+

Insulin treated group

T1

Fresh camel milk

T2

Pure cinnamon extract treated group

T3

Standardized camel milk yogurt treated group

T4

Functional camel milk yogurt treated group (stevia + cinnamon)

T5

Pure stevia extract treated group

USA

United State of America

SNF

Solids not fat

°C

Degree Celsius

g

Gram

%

Percentage

mg

Milligram

Kg

Kilogram

ALT

Alanine Aminotransferease

AST

Aspartate Aminotransferase

EDTA

Ethylene diamine tetra acetic acid

WBC

White blood cells

RBC

Red blood cells

HCT

Hematocrit test

PCV

Packed-cell volume

MCV

Mean corpuscular volume

MCHC

Mean corpuscular hemoglobin concentration

MCH

Mean corpuscular hemoglobin

ANOVA

Analysis of variance

LSD

Least significant difference

SD

Standard deviation

STZ

Streptozotocin

Author contributions

AH convinced and supervised the work, FI and MZ conducted the research work, SA and TI reviewed the manuscript, RSA applied state, MA and MJA write original manuscript.

Funding

Not applicable.

Data availability

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

Code availability

Statistix 8.1 software is used for the result evaluations.

Declarations

Conflict of interest

The authors report no conflict of interest.

Ethical approval

All applicable international, national and/or institutional guidelines for the care and use of animals were followed.

Consent to participate

All authors were voluntarily agreed to take part in this study.

Footnotes

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Contributor Information

Aneela Hameed, Email: draneela@bzu.edu.pk.

Muhammad Zeeshan, Email: Mr.zee7777@gmail.com.

Saeed Akhtar, Email: saeedakhtar@yahoo.com.

Tariq Ismail, Email: tariqismail@bzu.edu.pk.

Rabia Shabir Ahmad, Email: rabiyaseen@yahoo.com.

Mamoona Amir, Email: mamoonaameer@bzu.edu.pk.

Muhammad Junaid Anwar, Email: Junaidfst786@gmail.com.

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Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Data Availability Statement

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

Statistix 8.1 software is used for the result evaluations.

Articles from Journal of Food Science and Technology are provided here courtesy of Springer

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