A Brief Talk on the New Star of Functional Evaluation of Health Food——Zebrafish
By: Date: 2021-04-02 Categories: chinesefood Tags: ,
  The vitality of the industry comes from innovation. For the health food industry, what does innovation create? How to create? It has always been the direction that health food people are struggling for. The relevant state supervision and management departments are also working hard to plan for the long-term development of the industry while strictly supervising under the premise of ensuring large food safety. Recently, the State Administration for Market Regulation has publicly solicited opinions on four documents including the”Guiding Principles for Ethical Review of Health Food Crowd Consumption Trials (2020 Edition)” and”Health Food Functional Evaluation Guidelines (2020 Edition) for Solicitation of Comments”. Among them, the”Guiding Principles for Functional Evaluation of Health Foods (2020 Edition)” for comments on the subject content and scope of application of functional evaluation of health foods, the basic requirements for functional evaluation of health foods, basic requirements for animal testing of health foods, and human health food testing Five aspects, including the basic requirements of the test and the factors to be considered when evaluating the functions of health foods, are publicly solicited for comments. Among them, the basic requirements for animal experiments on health food are as follows:
  1 Requirements for experimental animals, feed, and experimental environment
  1.1 According to the specific requirements of each experiment, select experimental animals reasonably. Rats and mice are commonly used, and the strains are not limited. Animal strains suitable for corresponding functional evaluation should be used. Inbred animals are recommended.
  1.2 The sex and age of the animal are selected according to the needs of the experiment. The number of experimental animals is required to be 10-15 mice per group (single sex), and 8-12 rats per group (single sex).
  1.3 Animals and their experimental environment should comply with the relevant national regulations on experimental animals and their experimental environment.
  1.4 Animal feed should provide relevant information such as feed manufacturer. If it is a customized feed, the basic feed formula, preparation method should be provided, and the animal feed test report should be provided.
  2 Requirements for the dose of the test sample
   There should be at least 3 dose groups in various animal experiments, the dose selection should be reasonable, and the lowest effective dose should be found as far as possible. In the three dose groups, one of the doses should be equivalent to 5 times (rats) or 10 times (mouse) the recommended intake of humans (converted to the dose per kilogram of body weight), and the highest dose should not exceed the recommended intake of humans. 30 times the input amount (except for special circumstances), the functional experimental dose of the test sample must be within the safe dose range determined by the toxicological evaluation.
  3 Requirements for the way of giving the test sample
   The test sample must be given orally, and gavage is preferred. When giving the test substance by gavage, a suitable solvent (solvent, suspending agent or emulsifier) ​​should be selected according to the characteristics of the test and the physical and chemical properties of the test substance, and the test substance should be dissolved or suspended in the solvent. Generally, distilled water can be used. , Purified water, edible vegetable oil, edible starch, gelatin, carboxymethyl cellulose, sucrose fatty acid ester, etc. If other solvents are used, the reasons should be explained. The selected solvent itself should not produce toxic effects, and should not have a chemical reaction with the components of the test substance, and maintain its stability, without special pungent taste or odor. If gavage is not possible, it can be given by adding drinking water or mixed with feed, and calculating the dose of the test sample.
   should describe the preparation method, administration method and time of the test substance.
   Here, I will introduce you a new spine model animal star:zebrafish.
  Zebrafish, a slender tropical freshwater fish, is named after its side with zebra-like vertical dark blue and silver stripes. It is 3-4cm long, spindle-shaped, with a small and pointed head and short snout. It is native to South Asian countries such as India, Pakistan, Bangladesh and Nepal in the southern foothills of the Himalayas. It is a sentinel for environmental monitoring, an assistant to a biological developmental scientist, and a rookie in clinical medical research. (Picture 1:Photos of zebrafish)
A Brief Talk on the New Star of Functional Evaluation of Health Food——Zebrafishimage
   Picture 1:Zebrafish photo
   As a new type of model animal, zebrafish has incomparable advantages over other animals. It is listed by the National Institutes of Health as the third largest model organism after mice and rats, and is subject to European and American food and drug supervision Recognized by the management department and passed GLP certification. The OECD recognizes the zebrafish embryo toxicity test as the most sensitive in vivo method for evaluating the toxicity of chemicals. So far, OECD and ISO have issued a total of 10 fish (zebrafish) chemical toxicity testing standards. It is particularly worth mentioning that according to the EU Animal Protection Law, zebrafish embryos and juveniles within 5 days of birth are not animals and can be used to substitute mammals for testing and comply with the 3R (replacement, reduction, optimization) principle.
   Because of the low cost of feeding zebrafish, the large amount of eggs, and the advantages of in vitro fertilization, short development cycle, transparency and easy observation, it has been widely used in many fields such as basic research, drug development, food safety, and environmental protection. In recent years, as an internationally recognized new type of spine model animal suitable for efficacy evaluation, zebrafish can evaluate the safety and other efficacy attributes of various foods.
   The physiology, development and metabolism of zebrafish are highly similar to mammals, and their genes are 87%homologous to human genes, which can reliably simulate and predict human physiology and pathology (Figure 2:Zebrafish genes and human genes) Homology 87%). Moreover, zebrafish is a recognized ecotoxicology model. In the 1970s, foreign countries have begun to use zebrafish to study the acute toxicity of heavy metals and organics. At the same time, in addition to toxicity, zebrafish is also used to evaluate the biological activity of food ingredients. We see that more and more biologically active ingredients are increasingly found in foods, and special foods formed by the proportion of some active ingredients have gradually entered people’s lives. Related to this, there are many problems in the field of food research, such as:how to distinguish active compounds, how to infer whether the active compounds have biological properties, and whether their safety is reliable, etc. Therefore, researching and confirming the effective active ingredients of foods and evaluating their safety is an important content in the current food research field. Including functional tests on foods, food extracts and molecules isolated from foods.
A Brief Talk on the New Star of Functional Evaluation of Health Food——Zebrafishimage(1)
   Figure 2:The homology between zebrafish genes and human genes reaches 87%
   The zebrafish detection technology is a detection technology approved by FDA, EPA, EURL ECVAM and other international organizations, and has been published in”Nature” and other academic journals. Many foreign companies such as Merck, Novartis, Johnson & Johnson have their own zebrafish laboratories for their product development. At present, in China, zebrafish technology has continuously received special funding from the Ministry of Science and Technology of China for”Major New Drug Development”, and 13 standards related to zebrafish experiments have been promulgated, and it has been widely used in food safety evaluation, active ingredient screening, and mechanism research. Application, such as metabolic abnormality, anti-oxidation, immune regulation, neuroprotection and other related nutrition and health food development and detection play an effective role. (Table 1:Health food functions that can be evaluated by zebrafish)
  Table 1:Health food functions that can be evaluated with zebrafish
Health food function
Indices that can be evaluated with the zebrafish model
Enhance immunity
Number of neutrophils, number of macrophages, phagocytic function of macrophages
Improve sleep function
Sedation, sleep improvement effect
Assist in lowering blood sugar
Glucose
Assist in lowering blood lipids
The effect of lowering blood lipids
Slimming function
Inhibition of fat absorption, promotion of lipid metabolism, inhibition of pancreatic lipase activity
Antioxidant function
ROS removal effect
Auxiliary protection for chemical liver injury
Chemical liver injury (alcohol liver) model:degree of liver steatosis, liver size, yolk sac absorption (liver function)
Improve hypoxia tolerance
Improvement of exercise capacity under hypoxic conditions, survival test of sodium nitrite poisoning, and improvement of polycythemia caused by hypoxia
Relieve visual fatigue
Zebrafish eye apoptosis
Increase bone density function
Increase bone calcium content, promote bone development, and promote cartilage development
Intestinal and laxative
Evaluation of bowel motility function
Protect gastric mucosal damage
Improvement of intestinal inflammation, improvement of intestinal mucosa and intestinal cavity expansion
Regulate the intestinal flora
Number of intestinal flora
Improve nutritional anemia function
Number of red blood cells
Clear pharynx function
Anti-inflammatory effect
Promoting lead discharge function
Lead discharge function
Help to improve memory function
Improve motor dysfunction and improve reaction ability
Relieve physical fatigue
Motion function, lactate, ATPase activity
Other new features
Assist in lowering uric acid, anti-haze effect, improve blood viscosity, protect blood vessels, and promote wound healing

With the continuous upgrading of consumer consumption levels, the demand for high-quality and special foods is also increasing. Effective, convenient and economical zebrafish will continue to play an active role in the research on the efficacy of special foods. At the same time, We look forward to more and more uses of zebrafish being mastered by humans, and adding new and effective verification methods to promote human health.