Biological Effect of Unripe Cheese Produced with Coagulant from Solanum Aethiopicum L
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International Journal of Scientific & Engineering Research Volume 11, Issue 5, May-2020 48 ISSN 2229-5518 Biological effect of unripe cheese produced with coagulant from Solanum aethiopicum L. Shum fruit Valentin Désiré Guiama, Juliette Koubé, Esther Ngah, Jean Marcel Bindzi and Carl Moses Mbofung. Abstract— This study assessed the reaction of rats to the consumption of cheese made with plant coagulant from Solanum aethiopicum L. Shum (SA cheese), compared to calf rennet (CR) cheese and casein/sunflower oil diets. The body weight increased by 73, 80 and 84% in the control, SA cheese and CR cheese diet groups respectively. The consumption of SA cheese had protective effect on blood cells. Iron in serum decreased, with a low risk of iron deficiency anemia. The levels of total cholesterol, triglycerides and LDL-cholesterol in the serum were lower in rats fed SA cheese diet, as well as AST, ALT, suggesting protective effect of cardiovascular system, renal and hepatic functions. The biomarkers of oxidative stress were not changed, as well as bilirubin levels. In addition, SA cheese also improved the lactobacilli colon flora. Therefore, plant coagulant (SA extract) affects biological activity of cheese, and tends to improve its health benefits. Index Terms—Cheese, consumption, health benefits, plant coagulant, Solanum aethiopicum. —————————— —————————— 1 INTRODUCTION merging scientific findings indicate that dairy products contain fat (rumenic acid, butyrate and sphingomyelin), There is evidence that cheese is a probiotic food carrier. Probi- E proteins, peptides, oligosaccharides, vitamins and min- otic bacteria activity is maintained in all the stages of cheese- erals; which provide beneficial effects beyond basic nutritional making, from their manufacture up to their ingestion by the value [1]. Due to their health advantages, dairy products have consumer, and they survive during their transit in the gastro- also served as vehicles for functional food ingredients, such as intestinal tract. Probiotic bacteria are protected in the gastroin- phytosterols, fish fatty acids and various kinds of probiotic testinal tract by buffer created by cheese against the high acid- bacteria. Furthermore, dairy foods have been a rich source for ic environment, its dense matrix and relatively high fat con- the development of a large variety of health promoting ingre- tent. Probiotic unripe cheese (Argentinean fresh cheese) con- dients that are found in the markets as functional foods and taining L. acidophilus A9, B. bifidum A12 and L. paracasei A13 dietary supplements [2]. Among the dairy foods, cheese is demonstrated immunomodulating capacity in mice [6]. known as a concentrated daIJSERiry product and a form of milk Many constituents of cheese are involved both directly and preservation, since milk is highly perishable. Consumption of indirectly in the fight against oxidative stress. Fat soluble vit- cheese has increased consistently in most countries, unrelated amins (A and E) and peptides act directly by preventing the to their socio-economic level of development [3]. formation of radicals, by scavenging radicals or hydrogen Many studies have proven the biological role of cheese in sev- peroxide and other peroxides [7]. Indirectly, amino acids from eral aspects of health [4]. Using rat model, a study on the bi- digestion of casein stimulate the biosynthesis of antioxidant omarkers of metabolic syndrome revealed that consumption enzymes, and their activity depends on some minerals found of cheese was associated with a decrease of the serum low- in cheese. It is also efficient against kidney stones, osteoporosis density lipoprotein, adiponectin, triglyceride and cholesterol and dental caries [1]. levels and weight of the mesenteric adipose tissue [5]. This Cheese has become an important line of defense in the preven- suggests that cheese possess a suppressive effect on ab- tion of chronic diseases. Thus, numerous varieties of cheese dominal adipose accumulation, and prevent the development are being manufactured using coagulants from animal, micro- of metabolic syndromes. bial and plant origin. However, the use of animal and micro- bial coagulants is limited in many parts of the world. The ———————————————— choice of ingredients for cheese making is influenced by many Esther Ngah, PhD, Associate-Professor, Department of Food Science and factors some of which include their availability, diet habits, Nutrition, University of Ngaoundere, Cameroon. E-mail: hadas- ethics, public health, law and religious issues. Therefore, natu- [email protected] ral coagulants from plants are preferred by the people on Valentin Désiré Guiama, PhD, Department of Food Science and Nutrition, whom constrains are imposed by the use of animal and mi- University of Ngaoundere, Cameroon. E-mail: [email protected] Jean Marcel BINDZI is currently pursuing PhD degree program in Food crobial coagulants. Plant coagulants are characterized by non- Science and Nutrition in University of Ngaoundere, Cameroon. E-mail: specific proteolytic action on milk proteins. In addition, their [email protected] toxin content constrains their use as food. The health benefits Juliette Koubé, PhD, Department of Pharmaceutical Science, University of of cheese are hardly known by consumers, but the use of new Douala, Cameroon. Carl Moses Mbofung, PhD, Professor, Department of Food Science and coagulant raises questions about the safety, and biological ef- Nutrition, University of Ngaoundere, Cameroon. fect of new product compared with that obtained using con- ventional coagulants (calf rennet). Different coagulants may IJSER © 2020 http://www.ijser.org International Journal of Scientific & Engineering Research Volume 11, Issue 5, May-2020 49 ISSN 2229-5518 show similar clotting properties and extent of hydrolysis early nity guidelines (Official Journal of European Union L197 vol. in the process, but differ significantly during the continuation 50, July 2007). of cheesemaking [3]. This protease activity of coagulant during cheese-making can affect the biological quality of cheese; due 2.3. Experimental procedure to the variation in milk compounds retained in the curd. Food and water intake, faeces and body weights of each rat Numerous studies in bioactivity of cheese have been essential- were recorded every five days of feeding period. Then, food ly carried out with the cheeses made with animal or microbial efficiency was calculated and expressed as body mass gain (g-1 coagulants. In contrast, very few studies have provided details intake. rat-1). At the end of the feeding period, all the rats were on the bioactivity of cheese produced with plant extract used deprived of food 12 h before sacrifice under anesthesia. Ani- as a coagulant. In this investigation, rat models were used to mals were later dissected and blood samples collected into evaluate the biological effect of cheese made with Solanum two types of tubes: EDTA tubes for haematological analyses aethiopicum L. Shum fruit (mock tomato) extract, which has and dried tubes centrifuged (3,500 rpm, 5°C for 10 min) to ob- been suggested as potential plant coagulant in cheesemaking tain serum (stored at -20°C until biochemical analysis). Colon [8]. Therefore, the present work aimed essentially to study the content was withdrawn for microbiological analysis. The en- effect of feeding rats with cheeses (both made using calf rennet tire organs such as lung, heart, brain, spleen, kidneys and liver and S. aethiopicum extract) on haematological, microbiologi- were excised, weighed and stored at -20°C until analysis. cal and biochemical parameters. In addition, its effect on food efficiency and somatic index was evaluated. 2.4. Analysis 2 Materials and methods 2.4.1. Physicochemical Physicochemical analysis concerned with the diet ingredients, 2.1. Cheesemaking procedure colon content and faeces of rats. Total solids (infra-red drying Solanum aethiopicum L. Shum (SA) coagulant was prepared 105°C to constant weight), total protein (Kjeldahl method), by soaking 12 g of SA fruits (harvested in Ngaoundere area, and ash (ignition at 550°C) contents of cheese and fecal matter Adamawa Region-Cameroon) powder in 100 mL solution 4% of rats were measured [10]. Total lipids were determined us- (w/v) NaCl at 25°C for 20 h. Then, mixture was centrifuged ing organic solvents (dichloromethane/methanol: 2/1) and a and filtered, the supernatant was SA coagulant. Animal rennet gravimetric calculation [11]. Carbohydrate content of cheese was prepared by diluting 1g of calf rennet (80% of chymosin was determined as the difference between total solids and the and 20% of pepsin) powder (BioRen® Naturlabextrakt, Aus- sum of fats, proteins and ash contents [12]. The pH of cheese tria; with declared activity of 15,000 Soxhlet units) in 200 mL and colon content was measured by using a Consort pH-meter of distilled water to obtain the same rennet coagulation time. (Thumout, Belgium) model C831. Raw zebu (Bos indicus) milk (40 kg) from five herds situated around Ngaoundere, Adamawa Region-Cameroon, was ob- 2.4.2. Colon microbiological tained in the morning by manual milking during August 2011. The total viable bacterial count, lactobacilli, moulds and yeasts Zebu milk was mixed andIJSER filtered. Milk was heated with a colonies of the colon content were enumerated. Samples were holding time of 30s at 73°C, cooled at 35°C, then CaCl2, 2H20 serially diluted 109 times, pre-reduced in tryptone water and and starter were added to a final concentration of 0.02% (w/v) inoculated onto a range of agars designed to be selective for and 2% (w/v) respectively. After 15 min, 200 mL of each en- predominant microorganisms: total viable count on Plate zyme solution was added to 20 kg of milk. Forty minutes later, Count Agar (PCA) at 37°C for 48 h, yeast and moulds on the obtained coagula were cut and drained. Resulting unripe Sabouraud Dextrose Agar (SDA) at 37°C for 120 h, Lactobacilli cheeses were analyzed and stored at -20°C until formulation of on Man Rogosa and Sharpe (MRS) agar at 37°C for 48 h. The rat diets. dishes were placed in anaerobic jars for Lactobacilli culture; and the rest were incubated aerobically.