The CD4 means ± standard deviation in test and control group.
Foods supplemented with probiotics enhance athletes’ immune system functions, improve body health and consequently decreases athlete’s health maintenance costs. Probiotics improve immune system function against pathogens via affecting on innate immune system, humeral immunity and cytokines. The effects of consumption of Iranian probiotic Bifidobactrium bifidum (BIB2) on athletes’ immune system functions were evaluated. The results showed studied immune system factors were significantly different between test and control groups, so that IgA, IgM, lymphocyte and monocytes percentage and CD4 measurements of test group were higher than control. The Bifidobacterim bifidum (BIB2) probiotic consumption can affect some immune system factors; therefore its ability to improved general health should be studied more.
- sprint athletes
- Bifidobactrium bifidum (BIB2)
There is a general belief among elite athletes and their coaches that overtraining causes resistance to infection. Epidemiological studies report that symptoms of respiratory tract infection increases in 1–2 weeks after strenuous endurance competitions. The highest percentages of patients were athletes who exceeded their training threshold level that is associated with the training load [1, 2]. The biological balance of body organs improves the health of the host, improving performance and increasing power of the immune system . Probiotics are a group of living microorganisms that improve health by improving biological balance when added to foods or consumed as supplements. These organisms increase immune system function and enhance host defense against harmful microorganisms. The benefits of probiotics such as reducing toxins, increase immunity and resistance to infection, produce vitamins and nutrients, organic acids, reduce allergic reactions, respiratory infections, reduce the symptoms of irritable bowel syndrome, arthritis, rheumatoid and modulating immune responses have been shown in many studies [4, 5]. Overwhelming exercise undertaken by athletes or military personnel diminishes immune system function and increase gastrointestinal complaints, as well as increasing the risk of disease and infections [6, 7, 8]. In many studies, reduced immunity after chronic fatigue has been seen in over trained athletes [9, 10, 11]. Nieman et al. (2000) showed that regular and continuous exercise enhances the strength of athletes’ immune system; while undertaking heavy and alternate physical activities had the reverse impact. Also, some research shows that excessive exercise also damages the immune system is impaired [12, 13].
The humoral immunity in athletes is often studied by mucosal immunoglobulin measurements, especially the changes in the secretion of IgA and IgM from tissue was reported in sporting activities . Due to reduction of immunoglobulins in sport activities (and increased risk of infection in the upper respiratory tract) investigating the IgA and IgM is important . Because the antibodies secretion, the lymphocytes have a very important role in the immune system . Monocytes are the largest cells in the bloodstream and involved in phagocytosis in the early stages of the immune response. Also, monocytes produce the cytokines that activate lymphocytes and consequently stimulate inflammation . Immune system malfunctions might be caused by stress, sleep disorder, exercise, and negative energy balance. Hard exercises raise neutrophil count; however, they decrease lymphocyte count, natural killer cell activity (NK cell) via disturbance with oxidative burst, neutrophil function, immunoglobulin’s level and antimicrobial proteins level in saliva.
The effects of probiotics bacteria in improving immune function and preventing disease have been shown in numerous studies [6, 17, 18, 19]. The
Effects of fruit juices supplemented by
1.1 Blood sample collection
First, prior to the tests, the blood samples were collected from athletes after fasting, then once a month 24 h after 100 meters running, 8 ml of blood were collected from both studied groups in EDTA tubes. The bloods were centrifuged at 5000 rpm for 15 min and serums were collected. The blood cell counting was done using cell counter device (BC-2000 Mindray).
1.2 The IgA and IgM measurements
The IgA and IgM were measured using ELISA methodology using human IgA and IgM ELISA Kit (ab137980 and ab137982) according to protocol suggested by manufacture (abcam Inc., USA) using ELISA reader (Biotech microplate reader ELX800).
1.3 The CD4 cells count measurements
Flow cytometry is a technique for counting microscopic particles. The CD4 cells count was measured by flow cytometry technique using BD FACSCalibur system and anti-CD4 monoclonal antibody and results were reported based on the percent of peripheral blood mononuclear cell (PBMC) in total suspension.
1.4 Statistical analysis
The obtained data were analyzed using Graphpad Prism (version 6.01) software with 0.05 significant levels.
2. Results and discussion
The results of present study showed that consuming probiotic supplemented fruit juice increased serum IgA level and can be boosted immune system activities (Table 1). According to results shown in chart 1, CD4 cells count increased after consuming probiotics (Figure 2, Table 1) and it probably affects immune system ability to defense against antigens.
|Test group||Control group|
|Means ± SD*||Means ± SD|
|Before test||364.954 ± 25.98||356.234 ± 42.36|
|First month||398.065 ± 40.24||350.568 ± 36.78|
|Second month||431.365 ± 37.96||358.653 ± 42.63|
Probiotics or their products could have antimicrobial activities or they can prevent colonization of pathogens . They probably have adjuvant effect so they may be stimulate phagocytosis done by leucocytes or may be increase IgA and defensins secretion. They could be attached to gut immune receptors, thus they inhibit competitively pathogenic virus’s or bacteria’s attachment. In addition, competition occurs for earning foods and nutrition so with suitable colonization of probiotics, we can conquer pathogenic bacteria.
Indigenous bacteria are accepted to add to the immunological protection of the host by making a hindrance against colonization by pathogenic microbes. This hindrance can be upset by sickness and by utilization of antibiotics, in this way, permitting less demanding access of the host gut by pathogens. It is presently trusted that this hindrance can be kept up by giving enhancements containing live ‘alluring’ microbes: such supplements are called probiotics .
n expansion to making a barrier impact, a few of the metabolic products of probiotic bacteria (e.g. lactic acid and a class of anti-microbial proteins named bacteriocins, created by a few bacteria) may hinder growth of pathogenic organisms. Moreover, the alluring bacteria may compete for nutrients with the pathogens. At last, there’s a few prove that probiotic microbes may improve the intestine immune reaction against pathogenic microscopic organisms (Figure 1).
Studies in rats and mice uncover that lactic acid bacteria managed orally increment the numbers of T lymphocytes, CD4þ cells and antibody-secreting cells, counting those within the intestinal mucosa, and improve lymphocyte expansion, normal killer cell activity, IL-1, TNF and IFN-g generation, antibody production (counting secretory IgA), phagocytic activity and the respiratory burst of macrophages and the DTH reaction .
Hard and continues physical activities CAN decrease immune system function and it can lead to infectious diseases . Probiotics have an important role to improve and boost an individual’s health via pathogen growth prevention, amino acids and vitamins production, detoxification, cholesterol reduction and allergic reaction inhibition . In the present study, the effects of
|Time||IgA (μg/dl)||IgM (μg/dl)|
|Test group||Control group||Test group||Control group|
|Means ± SD*||Means ± SD||Means ± SD||Means ± SD|
|Before test||230 ± 47||210 ± 56||143 ± 28||129 ± 73|
|First month||237 ± 48||185 ± 78||147 ± 75||132 ± 84|
|Second month||248 ± 75||193 ± 65||150 ± 62||135 ± 46|
|Third month||253 ± 83||194 ± 42||152 ± 43||133 ± 73|
West et al. reported that probiotic consumption lead to mucosal immune system improvement and also increased CD4 and dendritic cells . In other research conducted by Ohashi et al. in the same field showed that increasing IgA level and cytokine secretion occurred during probiotic consumption . Furthermore, it has been showed that probiotic consumption caused potent increasing in lymphocyte and NK cells count in peripheral blood samples and they improved immune system and general health while NK cells count reduction was determined during hard exercises [31, 32]. These results mentioned above were completely in accordance with our findings. Other studies have been done in the field of relationship between CD4 cells and probiotic consumption [33, 34, 35]. For example, Jensen et al. reported that consumption of
In the present study athletes who consume probiotic showed the monocyte and lymphocyte cells significantly higher than control group (Figures 4 and 5, Table 3). Kekkonen et al. showed that in runners who take probiotics lower respiratory infections and gastrointestinal symptoms were reported, but the study did not find any effect of changes in monocyte cells that are inconsistent with our results .
|Time||Monocyte (%)||Lymphocyte (%)|
|Test group||Control group||Test group||Control group|
|Means ± SD*||Means ± SD||Means ± SD||Means ± SD|
|Before test||3.85± 0.36||3.78 ± 0.42||32.7 ± 5.6||34.3 ± 4.8|
|First month||4.30± 0.65||3.77 ± 0.25||34.7 ± 6.4||33.6 ± 7.2|
|Second month||5.10± 0.42||3.78 ± 0.24||35.5 ± 5.7||33.1 ± 6.4|
|Third month||5.60± 1.2||3.91 ± 0.90||36.2 ± 7.3||33.7 ± 6.8|
West et al. reported the protective effect of probiotics on respiratory infections. In professional athletes who practice a lot in the long and intensive period, body temperature increased, the secretion of IgA confusion is created and therefore interferes in mucosal immunity . Cox investigated the effect of probiotic
Zhao and colleagues showed that
The advantageous impacts of probiotics have been illustrated in numerous diseases. One of the major mechanisms of probiotic activity is through the control of resistant reaction. A few of the prevalently utilized probiotic microorganisms are
As shown below a number of mechanisms are thought to be associated with probiotic beneficial effects:
Production of inhibitory substances such as H2O2, bacteriocins, organic acids, and so on.
Blocking of adhesion sites for pathogenic bacteria.
Compete with and inhibit growth of potential pathogens,
Degradation of toxins as well as the blocking of toxin receptors,
Modulate inflammatory immune responses .
The results of this study show that the probiotic
Margonis K, Fatouros IG, Jamurtas AZ, Nikolaidis MG, Douroudos I, Chatzinikolaou A, et al. Oxidative stress biomarkers responses to physical overtraining: Implications for diagnosis. Free Radical Biology and Medicine. 2007; 43(6):901-910
Fry AC, Schilling BK, Weiss LW, Chiu LZ. β2-Adrenergic receptor downregulation and performance decrements during high-intensity resistance exercise overtraining. Journal of Applied Physiology. 2006; 101(6):1664-1672
Prasanna P, Grandison AS, Charalampopoulos D. Bifidobacteria in milk products: An overview of physiological and biochemical properties, exopolysaccharide production, selection criteria of milk products and health benefits. Food Research International. 2014; 55:247-262
Gareau MG, Sherman PM, Walker WA. Probiotics and the gut microbiota in intestinal health and disease. Nature Reviews Gastroenterology and Hepatology. 2010; 7(9):503-514
Sanders ME, Guarner F, Guerrant R, Holt PR, Quigley EM, Sartor RB, et al. An update on the use and investigation of probiotics in health and disease. Gut. 2013; 62(5):787-796
Kekkonen R, Vasankari T, Vuorimaa T, Ekblom T, Hintsa A, Kautiainen H, et al. The effect of probiotics on respiratory infections and gastrointestinal symptom episodes during the marathon training period in marathon runners. International Journal of Sport Nutrition and Exercise Metabolism. 2007; 17(4):352-363
Cox AJ, Pyne DB, Saunders PU, Fricker PA. Oral administration of the probiotic Lactobacillus fermentum VRI-003 and mucosal immunity in endurance athletes. British Journal of Sports Medicine. 2010; 44(4):222-226
Bishop N, Gleeson M. Acute and chronic effects of exercise on markers of mucosal. Immunity. 2009; 1(14):4444-4456
Svendsen IS, Hem E, Gleeson M. Effect of acute exercise and hypoxia on markers of systemic and mucosal immunity. European Journal of Applied Physiology. 2016:1-11
Reid V, Gleeson M, Williams N, Clancy R. Clinical investigation of athletes with persistent fatigue and/or recurrent infections. British Journal of Sports Medicine. 2004; 38(1):42-45
Cox AJ, Gleeson M, Pyne DB, Callister R, Hopkins WG, Fricker PA. Clinical and laboratory evaluation of upper respiratory symptoms in elite athletes. Clinical Journal of Sport Medicine. 2008; 18(5):438-445
Nieman DC. Is infection risk linked to exercise workload? Medicine and Science in Sports and Exercise. 2000; 32(7 Suppl):S406-S411
Barret S, Storey A, Harrison M. The effect of exercise mode on salivary IgA secretion in high level triathletes. Journal of Science and Cycling. 2014; 3(2):3
Moreira A, Mortatti AL, Arruda AF, Freitas CG, de de Arruda M, Aoki MS. Salivary IgA response and upper respiratory tract infection symptoms during a 21-week competitive season in young soccer players. The Journal of Strength & Conditioning Research. 2014; 28(2):467-473
Barbeau ML. Transactional Distances in an Online Histology Laboratory Course. Ontario, Canada: The University of Western Ontario; 2012
Abbas AK, Lichtman AH, Pillai S. Cellular and molecular immunology. Elsevier Health Sciences. 2014
Spence L, Brown WJ, Pyne DB, Nissen MD, Sloots TP, McCormack JG, et al. Incidence, etiology, and symptomatology of upper respiratory illness in elite athletes. Medicine and Science in Sports and Exercise. 2007; 39(4):577
Lenoir-Wijnkoop I, Sanders ME, Cabana MD, Caglar E, Corthier G, Rayes N, et al. Probiotic and prebiotic influence beyond the intestinal tract. Nutrition Reviews. 2007; 65(11):469-489
Wanke M, Szajewska H. Lack of an effect of Lactobacillus reuteri DSM 17938 in preventing nosocomial diarrhea in children: A randomized, double-blind, placebo-controlled trial. The Journal of Pediatrics. 2012; 161(1):40-43. e1
Servin AL. Antagonistic activities of lactobacilli and bifidobacteria against microbial pathogens. FEMS Microbiology Reviews. 2004; 28(4):405
Gourbeyre P, Denery S, Bodinier M. Probiotics, prebiotics, and synbiotics: Impact on the gut immune system and allergic reactions. Journal of Leukocyte Biology. 2011; 89(5):685
Biswas D, Wideman NE, O’Bryan CA, Muthaiyan A, Lingbeck JM, Crandall PG, et al. Pasteurized blueberry ( Vaccinium corymbosum) juice inhibits growth of bacterial pathogens in milk but allows survival of probiotic bacteria. Journal of Food Safety. 2013; 32(2):204
Arunachalam KD. Role of bifidobacteria in nutrition, medicine and technology. Nutrition Research. 1999; 19(10):1559
Kotani Y, Shinkai S, Okamatsu H, Toba M, Ogawa K, Yoshida H, et al. Oral intake of Lactobacillus pentosusstrain b 240 accelerates salivary immunoglobulin A secretion in the elderly: A randomized, placebo-controlled, double-blind trial. Immunity and Ageing. 2010; 7:11
Glaser R, Kiecolt-Glaser JK. Handbook of Human Stress and Immunity. The University of Michigan: Academic Press; 1994
Gareau MG, Sherman PM, Walker WA. Probiotics and the gut microbiota in intestinal health and disease. Nature Reviews Gastroenterology and Hepatology. 2010; 7(9):503
Sharma R, Kapila R, Kapila S. Probiotics as anti-immunosenescence agents. Food Reviews International. 2013; 29(2):201
Lee J-W, Shin J-G, Kim EH, Kang HE, Yim IB, Kim JY, et al. Immunomodulatory and antitumor effects in vivo by the cytoplasmic fraction of Lactobacillus caseiand Bifidobacterium longum. Journal of Veterinary Science. 2004; 5(1):41
West N, Pyne D, Peake J, Cripps A. Probiotics, immunity and exercise: A review. Exercise Immunology Review. 2009; 15(107):e26
Ohashi Y, Ushida K. Health-beneficial effects of probiotics: Its mode of action. Animal Science Journal. 2009; 80(4):361
Yamada Y, Aoyama A, Tocco G, Boskovic S, Nadazdin O, Alessandrini A, et al. Differential effects of denileukin diftitox IL-2 immunotoxin on NK and regulatory T cells in nonhuman primates. The Journal of Immunology. 2012; 188(12):6063
Golden-Mason L, Cox AL, Randall JA, Cheng L, Rosen HR. Increased natural killer cell cytotoxicity and NKp30 expression protects against hepatitis C virus infection in high-risk individuals and inhibits replication in vitro. Hepatology. 2010; 52(5):1581
Mengheri E. Health, probiotics, and inflammation. Journal of Clinical Gastroenterology. 2008; 42:S177
Schmidt EGW, Claesson MH, Jensen SS, Ravn P, Kristensen NN. Antigen-presenting cells exposed to Lactobacillus acidophilusNCFM, Bifidobacterium bifidumBI-98, and BI-504 reduce regulatory T cell activity. Inflammatory Bowel Diseases. 2010; 16(3):390
Selbovitz M. A review of probiotics studies in HIV research suggests improved immunological presentation and preservation of viral host restrictive factors of TH17 in HIV patients. Retrovirology. 2012; 9(1):1
Herich R, Levkut M. Lactic acid bacteria, probiotics and immune system. Veterinarni Medicina-PRAHA. 2002; 47(6):169-180
Leyer GJ, Li S, Mubasher ME, Reifer C, Ouwehand AC. Probiotic effects on cold and influenza-like symptom incidence and duration in children. Pediatrics. 2009; 124(2):e172
Colbère-Garapin F, Martin-Latil S, Blondel B, Mousson L, Pelletier I, Autret A, et al. Prevention and treatment of enteric viral infections: Possible benefits of probiotic bacteria. Microbes and Infection. 2007; 9(14):1623
Hu X, Zhang H, Lu H, Qian G, Lv L, Zhang C, et al. The effect of probiotic treatment on patients infected with the H7N9 influenza virus. PLoS One. 2016; 11(3):e0151976
Ashraf R, Shah NP. Immune system stimulation by probiotic microorganisms. Critical Reviews in Food Science and Nutrition. 2014; 54(7):938-956
Jespersen L, Tarnow I, Eskesen D, Morberg CM, Michelsen B, Bügel S, et al. Effect of Lactobacillus paracaseisubsp. paracasei, L. casei431 on immune response to influenza vaccination and upper respiratory tract infections in healthy adult volunteers: A randomized, double-blind, placebo-controlled, parallel-group study. The American Journal of Clinical Nutrition. 2015; 101(6):1188-1196
Sakai F, Hosoya T, Ono-Ohmachi A, Ukibe K, Ogawa A, Moriya T, et al. Lactobacillus gasseriSBT2055 induces TGF-β expression in dendritic cells and activates TLR2 signal to produce IgA in the small intestine. PLoS One. 2014; 9(8):e105370
Gao Q , Xiao C, Min M, Zhang C, Peng S, Shi Z. Effects of probiotics dietary supplementation on growth performance, innate immunity and digestive enzymes of silver pomfret, Pampus argenteus. Indian Journal of Animal Research. 2016;(OF)
Gleeson M. Immune function in sport and exercise. Journal of Applied Physiology. 2007; 103(2):693-699
Narayan SS, Jalgaonkar S, Shahani S, Kulkarni VN. Probiotics: Current trends in the treatment of diarrhoea. Hong Kong Medical Journal. 2010; 16(3):213-218