Contamination of antibiotics and sul and tet(M) genes in veterinary wastewater, river, and coastal sea in Thailand.

Water systems in Southeast Asia accumulate antibiotics and antibiotic resistance genes (ARGs) from multiple origins, notably including human clinics and animal farms. To ascertain the fate of antibiotics and ARGs in natural water environments, we monitored the concentrations of these items in Thailand. Here, we show high concentrations of tetracyclines (72,156.9 ng/L) and lincomycin (23,968.0 ng/L) in pig farms, followed by nalidixic acid in city canals. The city canals and rivers contained diverse distributions of antibiotics and ARGs. Assessments of targeted ARGs, including sul1, sul2, sul3, and tet(M), showed that freshwater (pig farm wastewater, rivers, and canals) consistently contained these ARGs, but these genes were less abundant in seawater. Although sulfonamides were low concentrations (<170 ng/mL), sul1 and sul2 genes were abundant in freshwater (minimum 4.4 × 10-3-maximum 1.0 × 100 copies/16S), suggesting that sul genes have disseminated over a long period, despite cessation of use of this class of antibiotics. Ubiquitous distribution of sul genes in freshwater appeared to be independent of selection pressure. In contrast, water of the coastal sea in the monitored area was not contaminated by these antibiotics or ARGs. The density of Enterobacteriales was lower in seawater than in freshwater, suggesting that the number of ARG-possessing Enterobacteriales falls after entering seawater. From the pig farms, through rivers/canals, to the coastal sea, the occurrence of tetracyclines and tet(M) exhibited some correlation, although not a strong one. However, no correlations were found between concentrations of total antibiotics and ARGs, nor between sulfonamides and sul genes. This is the first comprehensive study showing Thai features of antibiotics and ARGs contaminations. The pig farm is hot spot of antibiotics and ARGs, and sul genes ubiquitously distribute in freshwater environments, which become less abundant in seawater.

Genetic characterization of coliform bacterial isolates from environmental water in Thailand.

INTRODUCTION: In contrast to the study in other part of the world, information about characteristics of plasmids carrying antimicrobial resistance genes (ARGs) in Enterobacteriaceae derived from environmental water in tropical Asian countries including Thailand is limited. This study, therefore, aimed to gain insight into genetic information of antimicrobial resistance in environmental water in Thailand. METHODS: Coliform bacteria were isolated from environmental water collected at 20 locations in Thailand and identified. Then, susceptibility profiles to ampicillin, cefazoline, cefotaxime, kanamycin, ciprofloxacin, sulfamethoxazole, tetracycline, and nalidixic acid were assessed. In addition, antimicrobial resistant genes integrons, and replicon types were analyzed. And furthermore, plasmids carrying blaTEM and tetM were identified by S1-PFGE analysis and confirmed transmissibility by transconjugation experiments. RESULTS: In 130 coliform bacteria isolated, 89 were resistant to cefazoline while 41 isolates were susceptible. Cefazoline-resistant coliform bacteria were found to be significantly resistant to cefotaxime and tetracycline as compared to susceptible isolates. Hence, blaTEM and tetM correlating with ß-lactam antibiotics and tetracycline, respectively, were analyzed found to co-localize on the IncFrepB plasmids in isolates from pig farms' wastewater by S1-PFGE analysis. And furthermore, transmissibility of the plasmids was confirmed. CONCLUSIONS: Results obtained in this study suggested that ARGs in coliform bacteria may have been spreading on the farm via IncFrepB plasmids. Hence, appropriate use of antimicrobials and good hygiene management on the farm are required to prevent the emergence and spread of resistant bacteria.

Decreased Serum Adiponectin Reflects Low Vitamin D, High Interleukin 6, and Poor Physical Performance in Knee Osteoarthritis.

Obesity is a major contributor to deterioration of physical function toward sarcopenia in knee osteoarthritis (OA) due to its effect mediated through adipokines-derived molecules that have pro-/anti-inflammatory properties. This study aimed to investigate relationships of serum adiponectin, 25-hydroxyvitamin D (25(OH)D), interleukin (IL)-6, and physical performance in knee OA patients. A total of 175 knee OA patients and 52 healthy controls were recruited. Serum adiponectin, 25(OH)D, IL-6, biochemical markers, knee pain and functional scores, muscle strength, physical performance, metabolic parameters, and body composition were evaluated. Serum adiponectin levels were significantly higher in knee OA patients than that in controls, while its serum levels were significantly decreased in obese patients, especially those with sarcopenia. Furthermore, there were independent relationships of serum adiponectin with body composition parameters, knee pain scores, physical function tests, and metabolic parameters in knee OA patients. Besides, serum adiponectin levels were positively associated with 25(OH)D levels, and negatively correlated with C-reactive protein and IL-6 levels in knee OA. Additionally, low serum adiponectin could be used to distinguish knee OA patients with sarcopenic obesity from those without sarcopenic obesity. Circulating adiponectin levels may serve as a possible surrogate biomarker for exacerbated physical function in knee OA patients-particularly sarcopenic obesity.

Association of adiponectin gene polymorphisms with knee osteoarthritis.

AIM: To investigate the possible relationship of adiponectin (ADIPOQ) gene polymorphisms, plasma adiponectin, and the risk of knee osteoarthritis (OA). METHODS: A total of 398 subjects, 202 knee OA patients and 196 healthy individuals, were enrolled in the case-control study. Genotyping at +45T/G (rs2241766) and +276G/T (rs1501299) loci was performed using polymerase chain reaction-restriction fragment length polymorphism. Plasma adiponectin levels were assessed using enzyme-linked immunosorbent assay. OA severity was determined using the Kellgren-Lawrence (KL) grading system. RESULTS: No significant associations were observed in the genotype distributions and allele frequencies at two loci of +45T/G and +276G/T polymorphisms in the ADIPOQ between knee OA patients and control subjects. There was a significant association between genotype distribution of +276G/T polymorphism and KL grade 2, 3 or 4 (P = 0.037, P = 0.046, P = 0.016, respectively). At +45T/G locus, the percentage of GG genotype was notably greater in control subjects (13.40%) compared with OA subjects (1.70%) (P = 0.023). Plasma adiponectin was markedly decreased in OA subjects compared with control subjects (P = 0.03). Likewise, circulating adiponectin in OA subjects was notably lesser than that in control subjects in GG genotype of +45T/G (P = 0.029) and +276G/T polymorphisms (P = 0.012). CONCLUSION: Polymorphisms +45T/G and +276G/T of the ADIPOQ gene might not be responsible for OA susceptibility among Thais.

Vitamin D Supplementation Improves Quality of Life and Physical Performance in Osteoarthritis Patients.

(1) Background: Lower levels of serum 25-hydroxyvitamin D (25(OH)D) are common in osteoarthritis (OA) patients. However, the effect of vitamin D supplementation on muscle strength and physical performance remains unclear. This study will investigate the effects of vitamin D2 supplementation on muscle strength and physical performance in knee OA patients; (2) Methods: One hundred and seventy-five primary knee OA patients with low levels of serum 25(OH)D (<30 ng/mL) received 40,000 IU vitamin D2 (ergocalciferol) per week for six months. Body composition, muscle strength, physical performance, serum 25(OH)D level, leptin, interlukin-6 (IL-6), parathyroid hormone (PTH), protein carbonyl, and metabolic profile were analyzed; (3) Results: Baseline mean serum 25(OH)D levels in knee OA patients was 20.73 ng/mL. Regarding baseline vitamin D status, 58.90% of patients had vitamin D insufficiency, and 41.10% had vitamin D deficiency. After vitamin D2 supplementation for six months, mean serum 25(OH)D level was 32.14 ng/mL. For post-supplementation vitamin D status, 57.10% of patients had vitamin D sufficiency and 42.90% had vitamin D insufficiency. From baseline to six months, there was a significant increase in mean serum 25(OH)D level (p < 0.001), while mean LDL cholesterol (p = 0.001), protein carbonyl (p = 0.04), and PTH (p = 0.005) all significantly decreased. Patient quality of life (SF-12) and pain (visual analog scale, VAS) both improved significantly from baseline to the six-month time point (p = 0.005 and p = 0.002, respectively). Knee OA patients demonstrated significant improvement grip strength and physical performance measurements after vitamin D2 supplementation (p < 0.05); (4) Conclusions: Vitamin D2 supplementation for six months reduced oxidative protein damage, decreased pain (VAS), improved quality of life, and improved grip strength and physical performance in osteoarthritis patients.

Elevated serum leptin levels are associated with low vitamin D, sarcopenic obesity, poor muscle strength, and physical performance in knee osteoarthritis.

CONTEXT: The associations between serum leptin, vitamin D status, sarcopenic obesity, muscle strength and physical performance in osteoarthritis (OA) remain uncertain. OBJECTIVE: To analyse the relationships between serum leptin, vitamin D status, muscle strength and physical performance in OA patients. METHODS: A total of 208 knee OA patients were enrolled. Serum leptin, vitamin D, muscle strength and physical performance were evaluated. RESULTS: OA patients with sarcopenic obesity had significantly higher serum leptin levels than those with non-sarcopenic obesity. In addition, knee OA patients with sarcopenic obesity displayed low grip strength and poor physical performance. Furthermore, high serum leptin was negatively associated with vitamin D and physical performance. CONCLUSIONS: Serum leptin levels were correlated with low vitamin D, reduced muscle strength and functional impairment, suggesting that serum leptin might serve as a biomarker reflecting physical performance in OA patients.

Upregulation of inducible nitric oxide synthase and nitrotyrosine expression in primary knee osteoarthritis.

BACKGROUND: To investigate nitrite and inducible nitric oxide synthase (iNOS) levels in the plasma and synovial fluid of patients with primary knee osteoarthritis (OA) and to determine protein nitrotyrosine in synovial tissue of OA patients. MATERIAL AND METHOD: Thirty patients and 30 healthy controls were recruited into the present study. Plasma and synovial fluid nitrite levels were measured using Griess reaction. Plasma and synovial fluid iNOS concentrations were analyzed by enzyme-linked immunosorbent assay. Nitrotyrosine was detected immunohistochemically in synovial tissue of OA patients. RESULTS: Plasma and synovial fluid nitrite concentration in the OA group were significantly higher than those in the healthy control group were (p = 0.007 and p = 0.012). Furthermore, plasma iNOS levels were significantly higher in the OA group than those in healthy control group were (p = 0.04). Moreover, nitrotyrosine was detected immunohistochemically in macrophages, synovial lining layer and synoviocytes of synovial tissue in the OA group. CONCLUSION: These findings indicate that reactive nitrogen species and nitrotyrosine-containing proteins may be involved in the joint destruction process, and play a potential role in the pathogenesis of knee osteoarthritis.

Serum autotaxin levels correlate with hepatic dysfunction and severity in postoperative biliary atresia.

OBJECTIVE: To investigate correlation of serum autotaxin and disease severity in biliary atresia (BA). METHODS: Eighty postoperative BA patients and 15 controls were recruited. Serum autotaxin levels were determined by enzyme-linked immunosorbent assay. RESULTS: BA patients had greater serum autotaxin and liver stiffness than controls. Serum autotaxin and liver stiffness were markedly elevated in BA patients with jaundice compared to those without jaundice. Furthermore, serum autotaxin was correlated with liver stiffness and biochemical parameters in BA. CONCLUSIONS: Elevated serum autotaxin was correlated with hepatic dysfunction in BA. Accordingly, serum autotaxin is a promising biomarker reflecting the severity in BA.

Association of adiponectin +276G/T polymorphism with knee osteoarthritis.

Osteoarthritis (OA) is a prevalent, degenerative joint disorder leading to the destruction of articular cartilage, osteophyte formation and subchondral bone sclerosis. Genetic and environmental factors are involved in the development of OA. The role of adiponectin gene polymorphisms in OA has not yet been established. The aim of this study was to investigate the association of adiponectin +276G/T (rs1501299) gene polymorphism with knee OA. Genotype distributions and allelic frequencies of adiponectin gene, +276G/T polymorphism were determined in a total of 200 subjects (100 knee OA patients and 100 healthy controls). Single-nucleotide polymorphism (SNP) of the adiponectin +276G/T gene was genotyped by polymerase chain reaction (PCR)-restriction fragment length polymorphism analysis. The genotype distribution of +276G/T SNP was observed in the Hardy-Weinberg equilibrium for OA patients and controls. No statistically significant difference was identified between the two groups with respect to genotype distributions and allelic frequencies (P>0.05). The T- and G-allele frequencies were indicated as 24.5 and 75.5%, respectively, in OA patients, whereas the frequency was 23-70% in the control group. Findings of this study therefore suggest that the +276G/T SNP was not associated with susceptibility to knee OA.

Oxidative stress, vitamin e, and antioxidant capacity in knee osteoarthritis.

BACKGROUND: Osteoarthritis (OA) is a chronic progressive degenerative joint disorder which is characterised by strongly age-related regressive changes in articular cartilage. The objective of this study was to evaluate oxidative stress and antioxidant parameters in plasma and synovial fluid of patients with primary knee osteoarthritis. MATERIAL AND METHODS: Thirty-five OA patients and 35 healthy controls were recruited for this study. Nitrite, malondialdehyde (MDA), vitamin E, Trolox Equivalent Antioxidant Capacity (TEAC), and Ferric Reducing Antioxidant Power (FRAP) levels in plasma and synovial fluid were determined. RESULTS: Plasma nitrite levels in OA patients were significantly higher than those in healthy controls (p = 0.037). Furthermore, plasma MDA levels were significantly higher in OA patients than those in healthy controls (p < 0.001). Moreover, plasma vitamin E levels in OA patients were significantly lower than those in healthy controls (p < 0.001). Synovial fluid vitamin E levels of OA patients were significantly lower than paired plasma samples (p < 0.001). The total antioxidant capacities, as were measured by TEAC and FRAP assays in plasma of OA patients, were significantly lower than those in healthy controls (p < 0.01). MDA concentrations were positively correlated with nitrite concentrations but they were negatively associated with vitamin E and TEAC levels in synovial fluid of OA patients. CONCLUSION: The increased plasma levels of nitrite and MDA and the decreased plasma levels of vitamin E, TEAC, and FRAP indicated that oxidative stress was present in OA patients. These findings suggest that oxidative stress plays a potential role in pathophysiology of knee osteoarthritis.

Correlation of connective tissue growth factor with liver stiffness measured by transient elastography in biliary atresia.

AIM: Biliary atresia (BA) is a neonatal liver disease defined as chronic progressive fibrotic obliteration of extrahepatic bile ducts. The objective of this study was to determine the association of serum connective tissue growth factor (CTGF) with clinical outcome and liver stiffness measurement. METHODS: Eighty-two BA patients post-Kasai operation and 28 healthy controls were recruited. BA patients were categorized into two groups based on their portal hypertension (PH) status. Serum CTGF levels were determined by enzyme-linked immunosorbent assay. Liver stiffness scores were measured by transient elastography. RESULTS: BA patients had greater CTGF levels (905.9 ± 57.7 vs 238.3 ± 23.5 pg/mL, P < 0.001) and higher liver stiffness values than controls (28.2 ± 2.6 vs 5.0 ± 0.5 kPa, P < 0.001). Serum CTGF levels were remarkably elevated in BA patients with PH compared to those without PH (1092.4 ± 73.9 vs 582.6 ± 45.7 pg/mL, P < 0.001). Furthermore, BA patients with PH had significantly higher liver stiffness values compared to those without PH (37.3 ± 3.0 vs 10.6 ± 1.1 kPa, P < 0.001). Additionally, serum CTGF was positively correlated with liver stiffness (r = 0.875, P < 0.001) and total bilirubin (r = 0.462, P < 0.001). There was an inverse correlation between serum CTGF and serum albumin (r = -0.579, P < 0.001). CONCLUSION: High serum CTGF was associated with a poor outcome in BA patients. Accordingly, serum CTGF and transient elastography may serve as non-invasive biomarkers reflecting the disease severity in postoperative BA patients.

+276 G/T single nucleotide polymorphism of the adiponectin gene is associated with the susceptibility to biliary atresia.

BACKGROUND: Biliary atresia (BA) is an intractable neonatal inflammatory and obliterative cholangiopathy, leading to progressive fibrosis and cirrhosis. Adiponectin, an anti-inflammatory adipokine, is known to play a possible role in liver diseases. The objective of our study was to determine the relationship between adiponectin gene polymorphisms and BA susceptibility. METHODS: A total of 106 BA patients and 107 healthy controls were included in this study. Two single nucleotide polymorphisms (SNPs) of the adiponectin gene, +45T/G (rs2241766) and +276G/T (rs1501299), were genotyped by polymerase chain reaction-restriction fragment length polymorphism analysis. RESULTS: Genotype distributions of +45 T/G and +276 G/T SNPs were seen in the Hardy-Weinberg equilibrium for both BA patients and controls. The frequency of the G/G genotype at +276G/T was significantly higher in BA patients than in the controls (P=0.009). Regarding +45T/ G in BA patients, the frequency of the T/T genotype tended to be lower than in the controls, but the difference was not significant. Moreover, the G allele at +276G/T in BA patients was more common than in the controls (P=0.0043). In contrast, the frequency of the T allele at +45T/G was not significantly different between BA patients and the controls. None of the haplotypes studied was found to significantly influence the risk of BA. CONCLUSIONS: +276G/T SNP is strongly associated with BA, particularly with the G allele. We postulate that the +276G/T adiponectin gene polymorphism confers increased susceptibility to BA.

Plasma and synovial fluid connective tissue growth factor levels are correlated with disease severity in patients with knee osteoarthritis.

BACKGROUND: Connective tissue growth factor (CTGF) has been implicated in development of osteoarthritis (OA). OBJECTIVE: To determine the correlation between plasma and synovial fluid CTGF levels and the severity in knee osteoarthritis patients. METHODS: A total of 100 subjects were recruited into this study (75 OA patients and 25 controls). CTGF concentrations in plasma and synovial fluid were analyzed by enzyme-linked immunosorbent assay. RESULTS: Plasma and synovial fluid CTGF concentrations were correlated with radiographic severity. There was a positive correlation between plasma and synovial fluid CTGF levels. CONCLUSION: CTGF could be useful for monitoring the severity and progression of OA.

Correlation between plasma and synovial fluid basic fibroblast growth factor with radiographic severity in primary knee osteoarthritis.

PURPOSE: The aim of this study was to investigate plasma and synovial fluid basic fibroblast growth factor (bFGF) levels in patients with primary knee osteoarthritis (OA) and to evaluate the correlation between bFGF levels and disease severity. METHODS: Thirty-five patients with knee OA and 15 healthy individuals were recruited into this study. Knee OA grading was performed according to the Kellgren-Lawrence classification. bFGF concentrations in both plasma and synovial fluid were determined using enzyme-linked immunosorbent assay. RESULTS: Plasma and synovial fluid bFGF levels in knee OA patients were significantly higher than in controls (P < 0.001). Moreover, plasma and synovial fluid bFGF concentrations were positively correlated with radiographic severity (r = 0.535, P < 0.001 and r = 0.570, P < 0.001, respectively). Further analysis revealed that there was a positive correlation between plasma and synovial fluid bFGF levels (r = 0.674, P < 0.001). CONCLUSIONS: Plasma and synovial fluid bFGF levels were significantly increased in OA patients, and these elevated levels were positively correlated with radiographic severity. These findings indicate that bFGF levels may be a monitor of disease severity and could play an essential part in the pathophysiology of degenerative process in OA.

Ginseng supplementation does not change lactate threshold and physical performances in physically active Thai men.

BACKGROUND: Ginseng has been one of the most popular herbs said to improve human exercise performance. Unclear and anecdotal information is known about the effect of ginseng on lactate threshold and aerobic performance in humans. OBJECTIVE: The purpose of the present study was to investigate the effect of ginseng supplementation on lactate threshold in physically active young men. MATERIAL AND METHOD: Sixty men from the Naval Medical Corps, Royal Thai Navy, aged 17- 22 years old, were randomized into either the ginseng (n = 30) or placebo (n = 30) group. The ginseng group took 3 grams of 100% ginseng orally, while the placebo group took an equal amount of lactose powder each day, for 8 weeks. Blood lactic acid levels for determination of lactate threshold (LT) were measured during an incremental cycle ergometer work. LT exercise performance, and heart rate (HR) responses to exercise were determined at baseline and after 8 weeks of ginseng and placebo consumption. Substrate oxidation rates during steady state exercise were assessed upon study completion. Selected markers for liver and kidney functions, including serum aspartate aminotransferase, alanine aminotransferase, blood urea nitrogen, and creatinine were monitored for possible side effects of ginseng. RESULTS: L T before and after 8 weeks of supplementation in the ginseng group (164.5 +/- 32.8 and 170.9 +/- 26.4 watts), and in the placebo group (163.7 + 25.1 and 163.7 + 17.3 watts) were not different (p = 0.448). Both groups had a similar pattern of exercise heart rate (p = 0.918), total exercise time (p = 0.241), and peak power output (p = 0.411). After 8 weeks, the magnitude of difference between ginseng and placebo groups on oxidation rates of fat (3.82 +/- 10.0 cal. kg(-1). min(-1), p = 0.704) or carbohydrate (4.36 +/- 12.6 cal. kg(-1). min(-1), p = 0.731) was not statistically significant. There were no abnormal changes of markers of liver and renal functions after ginseng administration. CONCLUSION: Daily administration of 3 g of ginseng for an 8-week period did not improve LT nor did it affect physical performances. Therefore, ginseng supplementation did not exert an ergogenic property on aerobic fitness enhancement in well-fit individuals.

Effects of cardiac rehabilitation on fibrinolysis in patients with coronary artery disease.

The aim of the present study was to demonstrate the influence of exercise component of a cardiac rehabilitation program on fibrinolysis in coronary artery disease (CAD) patients. Cardiac rehabilitation program was claimed to have an important role for improving quality of life and reducing the incidence of recurrent disease. The program used in the present study included aerobic exercise for 8 weeks, 4 days per week, 30 minutes per day at light to moderate intensity. Thirty-three male patients with CAD were recruited in the present study. Subjects from Thammasat University Hospital and King Chulalongkorn Memorial Hospital, whose age ranging from 40 to 70 years, were random assigned into 2 groups: control and experimental groups. The results showed that no significant differences in tissue plasminogen activator levels (t-PA) (both antigen and activity), plasminogen activator inhibitor-1 levels (PAl-1) (both antigen and activity) were observed in control and experimental groups after exercise training for 8 weeks as compared to the baseline. However significant improvement of fibrinolysis via a decrease in PAI-1 activity level from 16.3 (3.7) to 14.8 (6.3) AU/ml (p < 0.024) and an increase in t-PA activity from 2.3 (0.8) to 2.7 (0.5) IU/ml and t-PA antigen from 7.5 (2.9) to 9.2 (2.7) ng/ml (p < 0.01) in experimental group were observed when compared between pre and post acute submaximum exercise (65% VO(2 peak)) at the end of the program. In addition the authors found a significant improvement in VO(2 peak) resting heart rate, and serum triglyceride level in experimental group after 8 weeks of exercise training. This study demonstrated that patients with CAD participating in 8 weeks exercise cardiac rehabilitation program at light--moderate intensity could improve physical fitness and physical health although there was no significant change of fibrinolysis. The CAD patients should be advised to enroll in this cardiac rehabilitation program since it did not have any harmful effect due to the fibrinolytic function but it also augmented the patients' physical health.

Low dose creatine supplementation enhances sprint phase of 400 meters swimming performance.

This study demonstrated the effect of low dose creatine supplement (10 g. per day) on the sprinting time in the last 50 meters of 400 meters swimming competition, as well as the effect on exertion. Nineteen swimmers in the experimental group received creatine monohydrate 5 g with orange solution 15 g, twice per day for 7 days and nineteen swimmers in the control group received the same quantity of orange solution. The results showed that the swimmers who received creatine supplement lessened the sprinting time in the last 50 meters of 400 meters swimming competition than the control group. (p<0.05). The results of Wingate test (anaerobic power, anaerobic capacity and fatigue index) compared between pre and post supplementation. There was significant difference at p<0.05 in the control group from training effect whereas there was significant difference at p<0.000 from training effect and creatine supplement in the experiment group. Therefore, the creatine supplement in amateur swimmers in the present study enhanced the physical performance up to the maximum capacity.

Changes of immune system in military recruits after the training program.

The aim of the exercise was to improve body health including the immune system function. However, several studies have observed the suppression of immune response in athletes who were trained with strenuous exercise. Recruits who have been in the recruitment-training program for 8 weeks might also have immune suppression. Twenty males, aged from 21 to 23 years in Chulachomklao Royal Military Academy infantry battalion volunteered to participate in this study. They were asked to wear a Polar Accurex Plus Telemetric heart rate monitor during the training period. Tuberculin skin test which is the screening method for cellular immune responses was significantly decreased at post-training compared to the pre-training (p<0.02). The levels of serum IgG and IgM at pre-training, 4th week, 8th week and one week post-training were significantly decreased at the initiation period, which were compared with pre-training and 4th week of training (p<0.01), and returned to normal at the 8th week of training and one week post-training. The level of circulating immune cells and number of CD markers (CD3, CD4, CD8, and CD56) were not significantly changed throughout the training period. This study demonstrated that subjects who underwent the recruit-training program were able to improve their health after the period of training i.e., in the adaptation period. The decrease in the initiation period could be compensated with proper diet and enough rest.