Fish Nutrient Supplementation in High-intensity Functional Training

Study Overview
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Brief Summary
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Nowadays, many persons who exercise consume dietary supplements. Previous scientific studies have found that protein supplementation increases muscle mass when combined with an exercise program. In addition, vitamin D probably improves performance and muscle function. Likewise, ω3 fatty acids, apart from improving cardiovascular function, may also increase protein synthesis and performance. Thus, the aim of this study is to investigate the efficacy of a novel aquaculture-based sport supplement of proteins, ω3 fatty acids and vitamin D on physical performance, body composition, and the biochemical profile of persons following high-intensity functional training (HIFT). The efficacy of the experimental supplement (E) will be compared with whey protein (W) and maltodextrin (a carbohydrate) as placebo (P). Thirty healthy trained individuals (15 male and 15 female), aged 18-35, will take E, W, and P during three 6-week periods of HIFT (a different supplement each period) with three workouts a week and a washout (no supplement) period of 2 weeks between supplementation periods. E is a combination of powder, containing protein and vitamin D, and capsules containing ω3 fatty acids. When on E, participants will receive 0.6 g protein/kg body weight, 20 μg vitamin D, and 1.8 g ω3 fatty acids daily. When on W, they will receive 0.6 g protein/kg body weight and, when on P, they will receive 0.6 g maltodextrin/kg body weight daily. The order of E, W, and P will be random and counterbalanced. Throughout the study, participants will be on isoenergetic nutritional plans to avoid differences in energy intake that might compromise the validity of the study. The dietary plans will be individualized and will provided 1.0 g protein/kg body weight/day. Participants will undergo measurements of muscle strength, muscle endurance, aerobic capacity, and body composition prior to the first supplementation period, between periods, and after the third period. Also, blood amino acids, fatty acid acids, vitamin D, hematology, biochemistry, and hormones will be measured. To avoid bias, assessors will not know which supplement each participant is taking during each period. The main hypotheses of the study are: 1) Consumption of E will increase lean body mass compared to W and P; 2) consumption of E will improve muscle strength, muscle endurance, and aerobic capacity compared to W and P; and 3) consumption of E will result in a better blood amino acid, fatty acid, and vitamin D profile.

Official Title
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Effects of a Nutritional Supplement of Proteins, ω3 Fatty Acids and Vitamin D on Physical Performance, Body Composition and Biochemical Profile of Persons Following High-intensity Functional Training

Conditions
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Exercise Training, Dietary Supplements

Intervention / Treatment
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* Dietary Supplement: Fish nutrients
* Dietary Supplement: Whey protein
* Dietary Supplement: Maltodextrin


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Regular training (mixed endurance and resistance training 3-6 times a week, 50 min each session, for the past 4-6 months) as assessed by gym records or questionnaires. Clearance from a pathologist or cardiologist to perform maximal exercise. Mixed isoenergetic diet for the past 4-6 months. Exclusion Criteria: Smoking (even one cigarette or nicotine-containing device over the past 6 months). Any injuries to the musculoskeletal system that could interfere with the execution of training. Chronic disease. Fish or oyster allergy. Milk allergy. Pregnancy, lactation or planning a pregnancy within the duration of the study. Regular use of prescription medicine or supplements that might affect muscle function or recovery over the past month. Intermittent or religious fasting. Any vegetarian, ketogenic and protein diet.

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 35 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Other
Allocation: Randomized
Intervention Model: Crossover Assignment
Masking: Triple


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Other: Fish nutrients - Whey protein - Maltodextrin<br>Participants will receive fish nutrients, whey protein, and maltodextrin in this order. | Dietary Supplement: Fish nutrients<br>* Intake of fish protein, omega-3 fatty acids, and vitamin D<br>Dietary Supplement: Whey protein<br>* Intake of whey protein<br>Dietary Supplement: Maltodextrin<br>* Intake of maltodextrin<br>|
| Other: Fish nutrients - Maltodextrin - Whey protein<br>Participants will receive fish nutrients, maltodextrin, and whey protein in this order. | Dietary Supplement: Fish nutrients<br>* Intake of fish protein, omega-3 fatty acids, and vitamin D<br>Dietary Supplement: Whey protein<br>* Intake of whey protein<br>Dietary Supplement: Maltodextrin<br>* Intake of maltodextrin<br>|
| Other: Whey protein - Fish nutrients - Maltodextrin<br>Participants will receive whey protein, fish nutrients, and maltodextrin in this order. | Dietary Supplement: Fish nutrients<br>* Intake of fish protein, omega-3 fatty acids, and vitamin D<br>Dietary Supplement: Whey protein<br>* Intake of whey protein<br>Dietary Supplement: Maltodextrin<br>* Intake of maltodextrin<br>|
| Other: Whey protein - Maltodextrin - Fish nutrients<br>Participants will receive whey protein, maltodextrin, and fish nutrients in this order. | Dietary Supplement: Fish nutrients<br>* Intake of fish protein, omega-3 fatty acids, and vitamin D<br>Dietary Supplement: Whey protein<br>* Intake of whey protein<br>Dietary Supplement: Maltodextrin<br>* Intake of maltodextrin<br>|
| Other: Maltodextrin - Fish nutrients - Whey protein<br>Participants will receive maltodextrin, fish nutrients, and whey protein in this order. | Dietary Supplement: Fish nutrients<br>* Intake of fish protein, omega-3 fatty acids, and vitamin D<br>Dietary Supplement: Whey protein<br>* Intake of whey protein<br>Dietary Supplement: Maltodextrin<br>* Intake of maltodextrin<br>|
| Other: Maltodextrin - Whey protein - Fish nutrients<br>Participants will receive maltodextrin, whey protein, and fish nutrients in this order. | Dietary Supplement: Fish nutrients<br>* Intake of fish protein, omega-3 fatty acids, and vitamin D<br>Dietary Supplement: Whey protein<br>* Intake of whey protein<br>Dietary Supplement: Maltodextrin<br>* Intake of maltodextrin<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Plasma amino acid profile pre-supplementation | Plasma amino acid profile (that is, the concentration of each individual amino acid) will be determined by liquid chromatography - mass spectrometry. | Within 2 days before the beginning of supplementation. | 
| Plasma amino acid profile post-supplementation | Plasma amino acid profile (that is, the concentration of each individual amino acid) will be determined by liquid chromatography - mass spectrometry. | Within 2 days after the end of supplementation. | 
| Whole-blood fatty acid profile pre-supplementation | Whole-blood fatty acid profile (that is, the concentration of each individual fatty acid) will be determined by gas chromatography. | Within 2 days before the beginning of supplementation. | 
| Whole-blood fatty acid profile post-supplementation | Whole-blood fatty acid profile (that is, the concentration of each individual fatty acid) will be determined by gas chromatography. | Within 2 days after the end of supplementation. | 
| Maximal dynamic strength of shoulder muscles pre-supplementation | Maximal dynamic strength of shoulder muscles will be assessed by measurement of one-repetition maximum. | Within 2 weeks before the beginning of supplementation. | 
| Maximal dynamic strength of shoulder muscles post-supplementation | Maximal dynamic strength of shoulder muscles will be assessed by measurement of one-repetition maximum. | Within 2 weeks after the end of supplementation. | 
| Force-velocity relationship of knee flexors and extensors pre-supplementation | Force-velocity relationship of knee flexors and extensors will be assessed in isokinetic dynamometer. | Within 2 weeks before the beginning of supplementation. | 
| Force-velocity relationship of knee flexors and extensors post-supplementation | Force-velocity relationship of knee flexors and extensors will be assessed in isokinetic dynamometer. | Within 2 weeks after the end of supplementation. | 
| Aerobic fitness pre-supplementation | Aerobic fitness will be assessed by measurement of maximal oxygen uptake through a maximal graded exercise test on treadmill. | Within 2 weeks before the beginning of supplementation. | 
| Aerobic fitness post-supplementation | Aerobic fitness will be assessed by measurement of maximal oxygen uptake through a maximal graded exercise test on treadmill. | Within 2 weeks after the end of supplementation. | 
| Lean and fat mass pre-supplementation | Lean and fat mass of the whole body and its parts (trunk, legs and arms) will be assessed by dual X-ray absorptiometry. | Within 2 weeks before the beginning of supplementation. | 
| Lean and fat mass post-supplementation | Lean and fat mass of the whole body and its parts (trunk, legs and arms) will be assessed by dual X-ray absorptiometry. | Within 2 weeks after the end of supplementation. | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Plasma vitamin D concentration pre-supplementation | Vitamin D will be measured by liquid chromatography - mass spectrometry. | Within 2 days before the beginning of supplementation. | 
| Plasma vitamin D concentration post-supplementation | Vitamin D will be measured by liquid chromatography - mass spectrometry. | Within 2 days after the end of supplementation. | 
| Muscle endurance pre-supplementation | Muscle endurance will be assessed by performing multiple sit-ups, knee flexions and knee extensions. | Within 2 weeks before the beginning of supplementation. | 
| Muscle endurance post-supplementation | Muscle endurance will be assessed by performing multiple sit-ups, knee flexions and knee extensions. | Within 2 weeks after the end of supplementation. | 
| Force-velocity relationship of shoulder muscles pre-supplementation | Force-velocity relationship of shoulder muscles will be assessed by a linear encoder. | Within 2 weeks before the beginning of supplementation. | 
| Force-velocity relationship of shoulder muscles post-supplementation | Force-velocity relationship of shoulder muscles will be assessed by a linear encoder. | Within 2 weeks after the end of supplementation. | 
| Full blood count pre-supplementation | Full blood count will be performed by flow cytometry. | Within 2 days before the beginning of supplementation. | 
| Full blood count post-supplementation | Full blood count will be performed by flow cytometry. | Within 2 days after the end of supplementation. | 
| Biochemical analyses pre-supplementation | Plasma glucose, triacylglycerols, total cholesterol, HDL cholesterol, LDL cholesterol, urea, and creatinine (all in mg/dL) will be measured by chemiluminescence in an automated analyzer. | Within 2 days before the beginning of supplementation. | 
| Biochemical analyses post-supplementation | Plasma glucose, triacylglycerols, total cholesterol, HDL cholesterol, LDL cholesterol, urea, and creatinine (all in mg/dL) will be measured by chemiluminescence in an automated analyzer. | Within 2 days after the end of supplementation. | 
| Plasma enzymes pre-supplementation | Creatine kinase and γ-glutamyltransferase (both in U/L) will be measured by chemiluminescence in an automated analyzer. | Within 2 days before the beginning of supplementation. | 
| Plasma enzymes post-supplementation | Creatine kinase and γ-glutamyltransferase (both in U/L) will be measured by chemiluminescence in an automated analyzer. | Within 2 days after the end of supplementation. | 
| Hormonal analyses pre-supplementation | Plasma cortisol and testosterone (both in μg/dL) will be measured by immunoluminescence in an automated analyzer. | Within 2 days before the beginning of supplementation. | 
| Hormonal analyses post-supplementation | Plasma cortisol and testosterone (both in μg/dL) will be measured by immunoluminescence in an automated analyzer. | Within 2 days after the end of supplementation. | 
| Internal load of exercise pre-supplementation | The internal load of exercise will be assessed by telemetric heart rate sensors and software. | Within the first week of supplementation. | 
| Internal load of exercise post-supplementation | The internal load of exercise will be assessed by telemetric heart rate sensors and software. | Within the last week of supplementation (week 6). |