THE EFFECTS OF PRETREATMfiNT WKTH EXERCESE 3N MELT MALE ALBERQ RAW EN ACQUERING CROSS-RESISTANCE TQ CQLQ WAfiR iMh‘iEESiON Thesis {or flu: Degree 0‘ M. A. MICHIGAN STATE UNIVERSITY Donald Peter Frank 1963 LIBRARY L Michigan State Univcrsity ' THESE SURE/“U CF CD'W’ATWW'L FF”7ARCH ( :_ c : rim! :_.I-I\ q Ilu. LJ;¢'-t :J.RY EAJF LAHVRNG, A’vHCHi-SAN THE EFFECTS OF PRETREATMENT WITH EXERCISE ON ADULT MALE ALBINO RATS IN ACQUIRING CROSS- RESISTANCE TO COLD WATER IMMERSION By Donald Peter Frank AN ABSTRACT OF A THESIS Submitted to Michigan State University in partial fulfillment of the requirements for the degree of MASTER OF ARTS Department of Health, Physical Education and Recreation 1963 Approved: ABSTRACT THE EFFECTS OF PRETREATMENT WITH EXERCISE ON ADULT MALE ALBINO RATS IN ACQUIRING‘CROSS— RESISTANCE TO COLD WATER IMMERSION by Donald Peter Frank The purpose of the problem was to determine whether adult male albino rats can acquire cross-resistance to cold water immersion by pretreatment with exercise. Thirty-three adult male albino rats of the Carlworth Farm's CFE strain of specific pathogen-free rats were used in this experiment. The animals in group I B were forced to swim daily carrying an additional five per cent of their weight attached to their tails until they reached the crucial point. The animals in group I A were forced to swim carrying a weight that would permit them to swim for a half hour. Group V and the control group were forced to swim only during the all out swims. To determine whether the exercised animals had acquired cross—resistance to cold water the following pro— cedure was used. All animals were loosely taped around the thorax, and a wire inserted into the tape. The wire was used to adjust the animal's depth in the water during the cold water immersion test. Survival time was recorded from the time the animal was placed in the water until drowning occurred. Donald Peter Frank The results indicate that adult male albino rats pre— treated with exercise did not survive significantly longer, during the cold water immersion test, than the control animals. The weight of the control animals was significantly higher than the animals that were exercised to the crucial point daily. A correlation between body weight and survival time was .339, which is statistically significant at the .05 level. This indicates the heavier animals survived longer than the lighter animals. THE EFFECTS‘OF PRETREATMENT WITH EXERCISE ON ADULT MALE ALBINO RATS IN ACQUIRING CROSS— RESISTANCE TO COLD WATER IMMERSION By Donald Peter Frank A THESIS Submitted to Michigan State University in partial fulfillment of the requirements for the degree of MASTER OF ARTS Department of Health, Physical Education and Recreation 1963 Dedication is made to my wife, Marilyn, whose patience and love has given me added purpose and encourage— ment to complete the task. ACKNOWLEDGMENTS The author wishes to acknowledge Wayne Van Huss whose professional assistance has given me confidence in the undertaking and completion of this task. Acknowledgment is also made to Wynn Updyke for his assistance while doing my research. D.P.F. TABLE OF CONTENTS CHAPTER I. THE PROBLEM Introduction. Statement of the Problem. Importance of the Study Limitations of the Study. . . . . Definition of Terms II. REVIEW OF THE LITERATURE, PrewTreatment with Stressors Other Than Exercise PremTreatment with Exercise. III. EXPERIMENTAL METHOD Subjects Treatment of Animals Cold Water Immersion Procedures IV. RESULTS Survival Time Weight of Animals Survival Time and Body Weight Discussion V. SUMMARY, CONCLUSIONS, AND RECOMMENDATIONS Summary Conclusions Recommendations. PAGE JTWRDNRDH \OKO‘\10\. IO 11 12+ 1A 11 16 17 18 18 19 20 CHAPTER PAGE BIBLIOGRAPHY . . . . . . . . . . . . . 21 APPENDIX A--Data Sheets—-Treatment of Animals . . . . 23 B-—Weight of Animals . . . . . . . . . 29 C-—Cold Water Survival Time . . . . . . . 38 D--Water Temperature . . . . . . . . . 40 E—-All Out Swim Time . . . . . . . . . A2 LIST OF TABLES TABLE PAGE I. Analysis of Variance Data for Survival Time During the Cold Water Immersion Test . . . . . . . . . . . . 16 II. Analysis of Variance Data for Weight of Animals . . . . . . . . . . 16 LIST OF FIGURES FIGURE 1. Mean Survival Time Per Group During Cold Water Immersion . 2. Mean Body Weight Per Group Prior to Cold Water Immersion Test 3. Water Temperature PAGE 15 15 Al CHAPTER I THE PROBLEM Introduction The problem of stress, since its formal presentation by Hans Selye (12) in 1936, has been an interesting and thought provoking area of experimentation. This can readily be confirmed by the numerous articles written on the subject of stress. We, in physical education in general, have failed to realize the real implications of the stress theory in the area of physical education, since strenuous exercise is a stressor which produces physiological changes. We, in physical education, state that exercise is good for the normal individual, implying that the physiological changes are beneficial. Many authors (4, 8, 9, IO, 13) have stated the benefits of exercise upon animals or humans. However, few studies have shown whether pretreating an organism with a stressor will increase the organism's resistance to another stressor. The experimental animals in this study were stressed using exercise. When the treatment phases were completed the experimental animals, along with the controls, were exposed to the cold water immersion. The data obtained determined whether pre-treating the adult male albino rats with exercise had elicited cross—resistance to cold water immersion. Statement of the Problem The purpose of the problem was to determine whether adult male albino rats can acquire cross-resistance to cold water immersion by means of pretreatment with exercise. Importance of the Study If it could be proven that exercised animals are able to withstand the stressor cold water significantly better than sedentary animals, then further research, using exer- cise as the means of adaptation to other stressors, might be performed. If animals which have been pre—treated with exercise are able to acquire cross—resistance to the effects of cold water immersion then one should consider the impli- cations of exercise as a means of withstanding other stresses in humans. Limitations of the Study 1. No attempt was made to control the amount of food or water consumed by the animals. 2. No attempt was made to control the humidity in the test rooms. 3. The exercise level might not be effective to bring about cross—resistance to cold water in the experimental animals. 4. The results of animals studied are not directly applicable to humans. Definition of Terms Cross-resistance. Crossmresistance is increased tolerance to a stressor other than that to which the body has adapted. Stress. Stress is the state manifested by the specific syndrome which consists of all the non-specifically induced changes within a biological system. Stressor. A stressor is that which produces stress. Control group. Control group is used to denote that group of animals whose activity was confined to cages, except for the all out swims in phase three (see Appendix for all out swim time). Crucial point. The crucial point is that time during the animal's swim when drowning will take place, according to the observer, if the animal is not rescued. CHAPTER II REVIEW OF THE LITERATURE Selye (12) has stated that bodily changes take place as an organism reacts to stress. "Stress causes certain changes in the structure and chemical composition of the body. Some of these changes are merely signs of damage. ’Others are manifestations of the body‘s adaptive reactions, its mechanism of defense against stress."(l2) The general adaptation syndrome is the total of all the changes that take place as an organism faces stress. This general adaptation syndrome has three stages: (1) the alarm re— action, (2) the stage of resistance, and (3) the stage of exhaustion. When the alarm reaction stage is entered the organism's level of resistance is lowered, due to the effect of the stressor. This stage is manifested by enlarged adrenals, loss of weight, shrinkage of the thymus, stomach ulcers, and changes in the body's chemistry. During this stage one of two alternatives may happen. Either the organism adapts to the stressor and enters the stage of resistance or the stress is so intense that the organism dies. The manifestations of the stage of resistance are quite different from, and in many instances the exact opposite, of those that characterized the alarm reaction stage. The organism's adaptation to the stress during the second stage raises its resistance above normal. Although the organism can live in the stage of resistance, in time, the acquired resistance is lost and the organism enters the stage of ex- haustion. The symptoms of the stage of exhaustion are similar to those of the alarm reaction stage. During the alarm reaction stage the organism is able to adapt to the stress and lives. In contrast in the stage of exhaustion the organ- ism can not adapt to the stress and dies. Such stressors as cold, exercise, irradiation, and drugs can produce the general adaptation syndrome with its three stages. Selye (12) states that any one agent is a stressor in proportion to the degree of its ability to pro- duce stress. In this experiment the stress was produced by the exercise. Numerous articles have been written stating the effects of stress on animals and humans. The review of literature in this study was limited to those articles which dealt with the effects of cross-resistance. According to the definition of cross-resistance it is feasible for an organism to increase its tolerance to stress by pre-treatment with any stressor. The review of the literature dealt with the following: (1) pre—treatment with stressors other than exercise, and (2) pre-treatment with exercise. Pre—treatment with Stressors Other than Exercise Heroux and Hart (5) have demonstrated that adaptation to the stressor cold provided a degree of resistance to restraint hypothermia. Bajusz and Selye (2) found that cardiac necrosis pro- duced under experimental conditions can be prevented by pre-treatment with cold bath, noradrenaline, and restraint. Egolinskii and Bogorad (15) established that training animals by regular cooling of the body increases the resistance of rats to hypoxia and irradiation. Korobkov, Golavacheva, and Shkurdoda (7) state that increased resistance to irradiation can be acquired by injections of ”dibazol.” Most studies found that cross-resistance can be acquired by pre—treating the animals with stressors other than exercise, however, some authors found that pre-treating animals with certain stressors would not produce CPOSS- resistance. Egolinskii and Bogorad (15) stated that cooling did not increase rats' cross-resistance to heat. Selye (ll) found that pre-treatment of animals with corn oil prevented the cardiotoxic action of subsequent treatment with the same substance, but pre—treatment with corn oil did not prevent other stressors from eliciting cardiac necrosis. Premtreatment with Exercise Bartlett,(3) using a group of twenty control animals and a group of twenty forced exercised animals, found there was a highly significant difference in the response of the two groups disclosing that the daily exercise produced a degree of inhibition to restraint hypothermia. Bajusz and Selye (2) found that animals which were gradually pre-treated with exercise had developed cross- resistance to cardiac necrosis when exposed to the full pathogenic dose of other stressors. Korobkov, Golovacheya, and Shkurdoda (7) state that rats trained to climb a pole 1.5 cm. in diameter, in which small circular steps were out every 5 cm., exhibited a greater tolerance to hypoxaemia at a height of 13,000 meters than control animals. Zimnitskay (15) found that rats trained by swimming are more capable of withstanding the effects of cooling and heating than control animals. Zimkin and Korobkov (15) found that ratsE resistance to the action of trichlorethylamine was increased by muscular training. Eighty—six per cent of fifteen trained rats survived the poison, whereas only fifty—eight per cent of the twelve untrained rats survived. They further state that of the animals which died the trained animals survived longer. Shernyakov,(l5) studying whether the body can increase its resistance to certain infective illness through exercise, 8 found that medical assistance was sought by fifty-eight per cent of individuals not doing any exercise, thirty-eight per cent by individuals who did exercise irregularly, and twenty— one per cent by individuals doing regular exercise. Trifonov (15), experimenting with rats, indicates that moderate physical impositions (swimming or maintaining position on a pole for 50-60 minutes daily over a period of six to eight weeks), increased crossmresistance to subsequent irradiation much more than an increase of three hours daily swimming. Kimeldorf and Jones (6) found that exercised animals did not survive any longer than controls animals when faced with the stressor, irradiation. Kudryashov and Nikolayeva (15) found no difference be- tween trained and untrained animals when faced with the stressors strychnine and carbon monoxide. The same authors found that trained rats were less resistant to cyanide poisoning than untrained animals. CHAPTER III EXPERIMENTAL METHOD The purpose of this experiment was to determine whether forced exercise could produce cross-resistance to cold water immersion in adult male albino rats. The thirty~three animals in this experiment were part of a group of 120 male albino rats which were used in an- other study. The Carlworth Farm’s CFE strain of specific pathogenuhwfiarats were used in this experiment. All animals were born on October 24, 1961. These animals were 59 days old when they were received in the laboratory, 61 days old at the beginning of phase one, 67 days old at the beginning of phase two, 151 days old at the beginning of phase three, and 227 days old when thirty~three of the animals were used in the cold water immersion test. The thirty~three animals used in the cold water imm mersion test were previously treated according to the treat- ment of animal data sheets (see Appendix). After the treat- ment phase the animals used in the cold water immersion test were 227 days old. The animals used in the cold water im- mersion test were loosely taped around the thorax and a wire was inserted in the tape. The wire was used to adjust the 10 animals’ depth in the cold water and to prevent the animal from submerging. Survival time was recorded from the time the animals were placed into the water until drowning occurred. Treatment of Animals The diet of the animals consisted of Dietrich and Gambrill pathogenmfree rat and mouse biscuits which were manufactured by Dietrich and Gambrill, Inc., Frederick, Mary- land. Food was placed on the bottom of the animal's cage. Water was supplied to the animals by means of an inverted bottle which was fastened to the outside of the cage. A metal tube extended from the bottle into the cage from which the animals received water by placing their mouths at the end of the tube. Food and water were permitted ad libitum. The animals were housed in individual rectangular cages which measured ten inches long, eight inches wide, and seven inches high. These cages were made of four by four hardware cloth and were fitted into a large mobile rack which provided housing for sixty animals. Because the animals were used in another study the experimenter feels that an understanding of the animals' treatment prior to their cold water immersion test is imper- ative. The treatment the animals received was divided into three phases. Each phase is summarized on the animal data sheets (see Appendix). Along with the summary there is a statement of the objectives of each phase. 11 The rats which were forced to exercise were swum according to the exercise schedule (see Appendix). When the animals were forced to exercise a weight was attached to their tails by means of waterproof tape. The weight con— sisted of lead sinkers attached to wires. The dates when the animals were forced to swim are listed in the treatment of animal data sheets (see Appendix). The rats swam in a galvanized tank that measured three feet wide, eight feet long, and thirty inches deep. Inside the tank were twenty~four plastic swimming sections each twelve inches wide, twelve inches long, and thirty inches deep. These sections prevented the animals from inter— fering with one another while swimming. The water tempera— ture was kept between 35 and 37 degrees centigrade. After completing the forced exercise the animals were placed in various cages to dry. The twenty-four drying cages were placed thirty inches beneath four 250 watt Ken— Rad infra~red heat lamps. The animals, after thirty minutes in the drying cages, were placed into their regular cages. Cold Water Immersion Procedures On June fifth, when the animals were 227 days old, the cold water immersion experiment was conducted. Group IA, IB, V, and a random selection of ten control animals were used. At this time there were six animals in group IA, nine animals in group 1B, and eight animals in group V. 12 The experimental procedures were conducted in the following way: A wooden frame was hung from the ceiling, thirty inches above the eight by four foot immersion tank. After weighing an animal a ten by two inch strip of water- proof adhesive tape was loosely placed under the animal and brought up on both sides of the thorax to the nap of the animal‘s neck (see Appendix for weight of animals). The two inches of excess tape were taped together above the animal's neck. ‘ The ends of a ten inch piece of wire were bent into the shape of hooks. One hook was inserted into the excess tape of the animal. The other hook was attached to the hardware cloth. The hardware cloth was connected to a wire which was suspended from the wooden frame. The animal‘s depth in cold water was adjusted by placing the wire at dif— ferent positions on the hardware cloth. The animals were attached to wires and then placed in water which was kept between fourteen and sixteen degrees centigrade. (See Appendix, Figure l, for the cold water temperature chart.) In order to keep the animal's body below the surface of the cold water a twenty~eight gram weight was attached to the animal‘s tail. After placing an animal in the cold water the wire was adjusted periodically so that the animal‘s head was kept above the cold water. No animal had to swim to keep its head above the cold water: however, every animal swam quite rapidly for about twenty minutes after being placed in the water. 13 While the animals had control of their physiological functions they could keep their heads out of the water. When the animals began to lose control of these functions, because of the anesthetic effect of the cold water, their noses would drop into the water. The animal's nose dipped into the water five to seven times before the rat could no longer lift its nose out of the water for the purpose of breathing. When an animal‘s nose would dip into the water, the time was recorded. If the animal's nose remained below the surface of the water for five minutes the animal was considered unable to withstand the stressor cold water. The survival time was measured from the time the rat was placed into the cold water until the animal could no longer raise its head to inhale. The survival time is recorded in minutes (see Appendix). It should be noted that the length of the animal’s survival time was related to its ability to withstand the cold water stressor; how- ever, actual death occurred because of drowning. CHAPTER IV RESULTS The following groups were used in this experiment to determine whether pre~treatment with exercise will elicit CPOSS~P€SiStanCG to the stressor cold water immersion: Group IA, six animals; Group IB, nine animals; Group V, eight animals; and ten control animals. Survival Time Survival time was measured according to the procedure discussed earlier. The null hypothesis was tested on the basis that no difference existed between the groups. The variance, due to the treatment, was not significant at the five per cent level (F=2.20), therefore, the null hypothe- sis was accepted. Data are presented in Figure 1, and the analysis of variance in Table I. Weight of Animals All animals were weighed, on June sixth, prior to the cold water immersion test (see Appendix). The null hypothe— sis was tested on the basis that no difference exists between groups. The variance due to the treatment was highly significant at the .001 level (F-7.6A), therefore, the null hypothesis was rejected. The data is presented in Figure 2, and the analysis of variance in Table II. These results Survival Time in Minutes Weight in Grams 15 121.5 190 1” 107.7 110 -_ lOO a-h— 90 __ 90-1 89.6 f 80 4.- IA Controls V IB F GrOUps E Fig. 1. Mean survival time per group during cold water immersion. 550 .... 549.0 525 —~ 500 -— 498.3 47 __ 5 465.1 450 -- AA2 .2 A25 4— Controls V IA IB Controls Fig. 2. Mean body weight per group prior to cold water immersion test. l6 confirm previous work indicating the greater the exercise stress the lower the weight of the animal. TABLE I ANALYSIS OF VARIANCE DATA FOR SURVIVAL TIME DURING THE COLD WATER IMMERSION TEST Source of Sum of Degree of Variance Variance Squares Freedom Estimate F F.05 Total 27382, 32 Between Means 5072.A9 3 1690.83 Within Groups 22311. 29 769.3 2.20 2.93 TABLE II ANALYSIS OF VARIANCE DATA FOR WEIGHT OF ANIMALS Source of Sum of Degree of Variance Variance Squares Freedom Estimate F F.001 Total 134,598 32 Between Means 59,419 3 19,806.33 7.6a 7.12 Within Groups 75,179 29 2,592.37 Survival Time and Body Weight In order to determine if a correlation between survival time and body weight exists the Pearson ”r" statistical method was used. The correlation between body weight and l7 survival time was .339, Which is statistically significant at the .05 level. The heavier animals survived longer than \ the lighter animals. Discussion The results indicate that no difference exists between pre-treated exercised rats and control adult male albino rats in their ability to withstand the effects of cold water immersion. At the time of the cold water immersion test the con- trol rats were significantly heavier than the experimental rats which were trained to the crucial point daily. Their weight was not significantly different from the other groups. When the weights and immersion test survival times for the animals in all groups were pooled, a significant relation— ship was found between weight and survival time. Considering the insulating properties of fat this was an expected finding. CHAPTER V SUMMARY, CONCLUSIONS, AND RECOMMENDATIONS Summary The purpose of this experiment was to determine whether exercised adult male albino rats can acquire cross—resistance to cold water immersion. The animals used in this experiment were theCkuflmmnufliFarm's CFE strain of specific pathogen-free rats. The animals were treated in another study prior to the cold water immersion test. The rats in Group IA were forced A to swim for thirty minutes, carrying an overload which was gradually increased daily. The rats in Group IB were forced to swim, carrying an additional five per cent of their body weight, until they reached the crucial point. The rats in Group V were exercised during phase two, but during phase three their activity was confined to cages except for the three all out swim tests. The rats in the control group were confined to cages during phase two and three except for the three all out swim tests. The cold water immersion test was conducted on June sixth using thirty-three adult male albino rats. The groups used in the cold water immersion test were: group IA, six animals; group IB, nine animals; group V, eight animals; and ten control animals. 19 In conducting the experiment a wooden frame was hung from the ceiling, thirty inches above the immersion tank. Wires were suspended from the wooden frame. At the end of each wire was a strip of hardware cloth enabling the experi- menter to adjust the animals' depth in the water. Each rat was weighed prior to the cold water immersion test. After weighing the animals, a ten inch strip of water— proof tape was placed around the rats' bodies to the nape of their necks. A wire was hooked through the excess tape, the Opposite end of the wire being hooked to the hardware cloth. The wire was adjusted periodically so that the animals' bodies were submerged except for their heads. The animals had a twenty-eight gram lead sinker attached to their tails to keep their bodies from floating. The water temper- ature was kept between fourteen and sixteen degrees centigrade. The survival time was measured from the time the rat was placed into the cold water, until the rat could no longer raise its head to inhale. The results indicate that no significant difference exists between exercised rats and sedentary control rats in their ability to survive during cold water immersion. Conclusion Adult male albino rats preetreated with the stressor exercise did not survive significantly longer during the cold water immersion test than control animals. 20 Recommendations Research should be done to find out if there is an optimum exercise level which will produce cross-resistance in adult male albino rats. Research should be done to determine whether other forms of exercise will bring about cross-resistance in adult male albino rats. Research should be done using larger numbers of animals in each group when using swimming as a means of exercise. Animal studies should be conducted in sanitary areas where.heat and humidity can be controlled. Visitors must be prohibited in animals quarters while an eXperiment is being conducted. V! V ' I" '1. ' IO. II. BIBLIOGRAPHY Adolph, E. F. ”Lethal Limits of Cold Immersion in Adult Rats,” American Journal of Physiology, 155:378-387, December, 19A8. Bajusz, Eors and Hans Selye. "Adaptation to the Cardiac necrosis—eliciting effects of Stress," American Journal of Physiology, 199:453-456, Sept- ember, 1960. Barlett, Roscoe G. "Stress Adaptation and Inhibition of Restraintmlnduced (Emotional) Hypothermia,” Journal of Applied Physiology, 8 661—663, May, 1956. Eckstein, R. W. ”Effect of Exercise and Coronary Artery Narrowing on Coronary Collateral Circulation,’ Circulation Research, 5:230~235, 1957. Heroux, 0. and J. S. Hart. ”Restraint Hypothermia and its Inhibition to Cold Acclimation,” American Journal of Physiology, 177:219-222, May, 1955. Kimeldorf, D. J. and D. C. Jones. "The Relationship of Radiation Dose to Lethality Among Exercised Animals Exposed to Roentgen Rays," American Journal of Physiology, 1672626—632, December, 1951. Korobkov, A. V., D. A. Golovacheva, and V. A. Shkurdoda. ”Effects of Muscular Training and Toning Substances on Non—Specific Endurance and Work Capacity in Rats,” Sechenov Physiological Journal of the U.S.S.R, Vol. 47:29-38 (No. l), 1961. Maison, G. L. and A. G. Broeker. ”Training in Human Muscles Working With and Without Blood Supply,” American Journal of Physiology, 132 390, 1941. Mayer, Jean. ”The Role of Physical Activity in Body Weight,” FIEP Bulletin, 226—8, 1958. Morris, J. N. and Margaret D. Crawford. "Coronary Heart Disease and Physical Activity of Work," British Medical Journal, July-December 20, 1958, 1E85—1496. Selye, H., E. Bajusz, and R. Strebel. ”Cross—Resistance to Cardio Toxic Agents," Canadian Journal of Bio— chemistry and Physiology, 39:519—529, March, 1961. 12. 13. 14. 22 Selye, Hans. The Stress of Life. New York: McGraw— Hill Book Company, 1956, pp. 1—326. Steinhaus, Arthur H. "Chronic Effects of Exercise," Physiological Reviews, 13, No. 1, January, 1933. Walker, Helen M. and Joseph Lev, New Statistical Inference. New York: Henry Holt and Company, 1953. pp. 1-510. Zimkin, N. V. ”Stress During Muscular Exercise and the State of Non-Specifically Increased Resistance," Sechenov Physiological Journal of the U.S.S.R., V01. A7281A~825 (No. 6), 1961. APPENDIX A DATA SHEETS-~TREATMENT OF ANIMALS 2A DATA SHEET-~TREATMENT OF ANIMALSl Summary of Phase I Date All Animals N—12O 12/23/61 Handled 12/24/61 Handled 12/25/61 No Treatment 12/26/61 Five minute swim 12/27/61 Ten minute swim, two per cent of body weight was attached (a lead sinker). 12/28/61 Two fifteen minute swims (A.M.), (P.M.), with two per cent of body weight attached. Objective of Phase I To accustom the animals with the experimental procedures. 1The treatment in phases I, II, and III were part of another study. The actual treatment the animals received is summarized. 25 DATA SHEET—~TREATMENT OF ANIMALS Summary of Phase II Dates Experimental Animals Controls N-lOO N-2O 12/29/61 Experimental animals were forced No to to exercise by attaching a lead Treatment l/l7/62 sinker which was four per cent of their body weight to their tails. Time was to be increased until animals could carry four per cent for thirty minutes. 1/18/62 Animals are losing weight, coughing No to and have bloody noses, therefore, Treatment 2/25/62 a rest period was necessary. 2/26/62 Resumed exercising animals. No to Animals were forced to swin with Treatment 3/20/62 one per cent of body weight attached. Weight was increased if animals could swim for thirty minutes. The purpose of this phase was to have the animals reach the objective of Phase II. Because the objective of this phase was not reached the main— tenance study was cancelled. Objective of Phase II To train the animals to swim for thirty minutes carrying four per cent of their body weight attached to their tail. 26 .pCHog Hafiosgo 05p zomog moSp Hands soreness oemaoz hpon mo 9:00 pom 0>Hm on 9509 Spas magpozom omflogoxm 0p mcflpgoo .xmp omflogoxo msoa>mgm Lo>o psmfloz CH ommogocfl Hddpmgw a spa: noessas sesaeh low omfiohoxm mgsogwnlm pom pomflogoxo mmsogwnx< Amazv maogpcoo Amzzv mflogpcoo mm pcmEpmogp oEwm > mm pcoEpmogp 08mm > 0-2 asap oH Asosm >H Am-zvasoe oH ssosm >H . m-z asap m ssosm HHH goa-zW asap m ssosm HHH floa-z ngmo.m spasm HH Aoa-z asap m stash HH auzv xmp mho>m H AOHIZV amp hgo>m H *@ gsogw :< Qsogu ofizponom omflogwmm .oapflmmoa mm hagwoc mm pounded mm: masogw ogp p50 Ham pmgfim ogp mo wo oEHp Eflzm cmoE one .Eazm .mEHBm p30 Ham pom mammp The :0 mgeopm ocflc oucH popa>flp who; mameflcm m©\mm\m pqooxo .pcoEpmogp oz prcoeflgogxo one .%©5pm wcflcflmgp m ma cmflmop 362 ow ma-z mm-z mh\mm\m .Eflzm p50 HHm 05p wcHLSG pfimfloz mpon go pcoo Log m.m pmflggmo mHmEHcm Hoppzoo pcw Hmuzoeflgodxo one "EHZm p50 HH< pmgflm ma-z mm-z m©\am\m .mHogpcOo mHmEHC< Hopcoeflgogxm opmm HHH ommcm mA0LQEH go mpogpoe 039 Spawn OB .%pHHHQm wcfleeflzm CH ocflfioop Log mcommop Lo>oomflp OB HHH eased do osaeoomno .maflwp Aflozp Op pozowppm pgwfloz mpon mo pcoo Log m.m poflsgwo maogpcoo pew mm\m\m mfimpzoeflgomxm .m©\m\© so Eflzm p50 Ham phage .m©\m\m :0 pounds mHmEflc< 0p wfllZ iNIZ N©\N\© .mHSGogom omflogoxo op mgflpgooom msflcflwga mm\H \w Op agiz omsz mm\mm\s .maflmp Aflocp 0p pozomwpm pgmaoz hpop mo psoo hog m.m poflggmo mfiogpcoo pzw m©\wm\: mampgofiflgoaxm .mm\wm\: co EHZm p50 Ham pcooom .m©\wm\: :0 poemog mHmEHC< 0p mazz omnz mp\sm\e mm\pm\e pCoEpwogp oz .oHSUonom omeogoxo on mcflpgooom wgficflmge 0p SHIZ mm-z mm\sm\m maogpcOo mHmEHC< amazoeflgodxm opwa A Aposcflpcoov HHH ommgm 28 DATA SHEET-~TREATMENT OF ANIMALS Phase IV Date Experimental Animals 6/5/62 Group IA N—6 The animals in Phase IV are Group IB N-9 the same animals that received Group V N—8 exercise under the exercise Controls N—IO)* schedule which is listed under Phase III. All animals were placed in water which was 150 centigrade on 6/5/62. Survival time in cold water was recorded. *The ten controls were a random selection from the control animals in Phase III. Objective of Phase IV To determine whether adult male albino rats can acquire cross—resistance to cold water through exercise. APPENDIX B WEIGHT OF ANIMALS 1 1n“_ 3O WEIGHT OF ANIMALS BY DATES Dates Group 12/30/61 1/3/62 1/13/62 1/17/62 1/20/62 IA 2 261 314.5 363.5 360 378.5 3 348.5 354 370.5 358 363 4 288 296.5 293.5 369.5 385 7 253 288.5 287.5 306.5 320.5 8 316 334.5 343 344 356.5 9 264 279 280 291.5 299 IB 7 1 253.5 284 268.5 300 317.5 2 273 259.5 275 287.5 297.5 3 320.5 334 335 278 287 A 329 363.5 378.5 399 414 5 276 301.5 307 302.5 312 6 320.5 352 5 374 397 415 7 290 291.5 292 313 324.5 8 310.5 317.5 343.5 344 347 9 294.5 317.5 332 5 292 305 v 1 307.5 346 378 371.5 382 2 272.5 291 5 326 324 327 3 334.5 311 333 340 355 4 268 294 310.5 321 321 6 273.5 289 287.5 263.5 260 7 304 273.5 304.5 316.5 324.5 8 279 306.5 322 334 340 9 247.5 272 283 281 285 Controls / l 257 286 330.5 357 3A5 2 313 342.5 258 342.5 355.5 3 284 309.5 350.5 372.5 372.5 4 2A9.5 293.5 328.5 347 332.5 5 302.5 327 373.5 400 408.5 6 288.5 320 350 378.5 379.5 7 339 379 422 432.5 441 8 316.5 344 382 403.5 412 9 332 375.5 All 434.5 437 10 270 296.5 335 3A6 330 31 WEIGHT OF ANIMALS BY DATES (continued) Dates Group 1/24/62 1/27/62 1/31/62 2/3/62 2/10/62 IA 2 405 412 427.5 437 465.5 3 349.5 336 5 328 332 355 4 410 416.5 430.5 455 481 7 290 310 322 340 375 8 373.5 386.5 414.5 429.5 461 9 309.5 323.5 336 347.5 373 IB 1 338.5 355.5 363 375 393.5 2 319 334.5 357.5 371 377.5 3 304.5 315 5 311.5 333 355 4 435 444 454 469 496 5 321 327.5 341 354 382 6 441 446 445 484.5 489.5 7 337.5 343.5 355 382 403.5 8 367 366.5 381.5 389 415 9 341 351 365 387.5 400 V 1 394.5 402.5 406.5 417 430 2 338 327 332 346.5 368 3 380 380.5 394 396 420 4 310.5 316 316 5 333.5 361 6 266 278 284.5 340.5 332 7 345 351 374 396.5 414 8 363 371 390 405 430 9 300 307.5 314.5 329.5 351 Controls 1 36:.5 370.5 387.5 390.5 405.5 2 382 389 413.5 416 432 3 384.5 382 399.5 405.5 421.5 4 356.5 385.5 363 360 361 5 404 417 409.5 404 377 5 6 388 366 333 318 353 7 452.5 420 388.5 380 350 8 415 406 403.5 403.5 399 9 456.5 458 47 .5 487.5 505 10 348.5 328.5 315.5 --- -—— 32 WEIGHT 0F ANIMALS BY DATES (continued) Dates Group 2/17/62 2/24/62 2/28/62 3/3/62 3/7/62 IA 2 483.5 513 496 477 481 3 395 410.5 422 414.5 432 4 408 510.5 525 502.5 507 7 400.5 433 437.5 433 444 8 468 485 483 482 477.5 9 396 405.5 410 403 410 IB 1 420 437 437 430.5 438 2 415 433 441 437.5 436 3 370 384.5 392.5 385.5 391.5 4 506 517 526.5 508 510.5 5 412 424 431 420 455 6 543 574.5 575.5 554.5 557.5 7 428 451 452 442.5 455.5 8 438.5 461 465 446.5 460 9 405 425 435 423.5 445 v 1 465 469 466 460 479 2 392 406 418.5 408 420 3 435 460.5 463 451 448 4 35 378.5 378 382 393 6 351 377 385 375 391 7 426 440 450 433.5 451.5 8 446.5 456.5 462.5 458 440 9 374 389.5 393 5 384.5 397.5 Controls 1 411.5 421 432 444 2 449 460 477.5 484 4 386 401 408 417 5 355 381.5 422 444 6 368 395 416 437 7 400 441 463 497.5 8 370 406 450 471 9 529 537.5 542.5 565.5 10 --- --- --— 455.5 33 WEIGHT OF ANIMALS BY DATES (continued) Dates Group 3/10/62 3/12/62 3/17/62 3/21/62 3/27/62 IA 2 480 491.5 491 492 499.5 3 426 428.5 464 411 428.5 4 499.5 518 507 502 501 7 437 447.5 448.5 448.5 454.5 8 460 466.5 460 462 462.5 9 401 416 402 5 398 395.5 IB 1 434.5 442 443.5 444.5 446.5 2 419.5 431 432 433.5 441 3 389.5 398 387.5 398.5 400 4 507.5 503.5 503 507 520 5 452 456 461 464.5 464.5 6 557.5 554.5 662 553.5 550 7 436.5 443 436.5 442 449 8 447.5 453.5 454.5 454.5 465 9 445 457 456 461 455 V 1 471 480 484 421.5 415.5 2 410 421.5 420.5 421 436 3 442 457 457.5 464 475 4 394 410 421 410 428 6 391 397 394 397 414 7 437.5 449 447 445 463 8 435 438 442.5 443.5 449 9 400 409.5 420 419 446 Controls 1 449 452.5 460 467 466 2 494 497 5 508 500 475 3 482 491.5 499 493.5 503.5 4 426 428.5 441.5 438.5 447 5 455 465 473 475.5 492.5 6 449 458.5 464 460.5 514 7 505 519 528 520 434.5 8 479 499.5 507.5 506.5 527 9 567 572 577.5 584 585 10 468 470 482.5 465 5 480 34 WEIGHT OF ANIMALS BY DATES (continued) Dates Group 3/31/62 4/4/62 4/7/62 4/11/62 4/14/62 A 2 480 466 457.5 447.5 448 3 423.5 436.5 425.5 425.5 420 4 487 487.5 469 467 461.5 7 446.5 453.5 456 449 454 8 458 463.5 459 460 468.5 9 394 401.5 400 398 397.5 IB 1 437.5 447 442 444.5 446 2 436 438 432 427 431 3 388 389.5 385 376 382 4 513 522 520.5 515 517 5 454 464.5 461 462 462 6 529 518.5 507.5 509 507 7 442 448 440 432.5 436 8 453 456 449.5 447.5 449 9 459.5 456 456 460 466 v l 409 412.5 414 413.5 417 2 448 458 465.5 472 481.5 3 470 473.5 463.5 467.5 468.5 4 435 441.5 441 449 454.5 6 417 432 433 5 441 450 7 467.5 480 485 493.5 504.5 8 457 470 471 476 483 9 448 461 466 470 474 Controls 1 479 481 490 491.5 491.5 2 475 482 5 486.5 494.5 495 3 506 520 517 521.5 524 4 457.5 473 475.5 485.5 491 5 500 506.5 510.5 510.5 513 6 518.5 524.5 532 532 537 7 430 449 448 5 454 462 8 530.5 550 556.5 562.5 577 9 584.5 599.5 600 608.5 611 10 484 494.5 497.5 511 515.5 35 WEIGHTS OF ANIMALS BY DATES (continued) Dates Group 4/18/62 4/21/62 4/25/62 4/28/62 5/3/62 IA 2 460 458.5 413.5 457.5 460 3 413.5 404.5 400 387 378 4 473 466.5 464.5 474 474.5 7 452.5 451.5 451 461 458 8 475 476 482.5 487 483 9 405.5 400 396.5 403.5 409 1B 1 453 447 446 445.5 451.5 2 436 426 424 430 423.5 3 387 385.5 387.5 388.5 394 4 518 515.5 504 504.5 494.5 5 466 460 465 458 456.5 6 502.5 490 486 492 493 7 434 433.5 432 439 430 8 450 445 446.5 453.5 452 9 469 466 5 468 474.5 475 V 1 413.5 419 418.5 422.5 418 2 493 5 493 5 498.5 505 510 3 480 480 467 452 437 4 465 465 475.5 477 475.5 6 457.5 457.5 465 466.5 473.5 7 513.5 513.5 520 533 528 8 495.5 495.5 496 499.5 498 9 489 489 499 507 503 Controls 1 598.5 501 501.5 506.5 507 2 506.5 512.5 520 525 522 3 533.5 540 551 550 559.5 4 49 8.5 502 5 505 510 506 5 520 524 527 522 509.5 6 548 545.5 549.5 559 555.5 7 475 476 481 479 481 8 588 588 598 600 598 9 622 623.5 619 610.5 618 10 523 523.5 534.5 538.5 537.5 36 WEIGHTS OF ANIMALS BY DATES (continued) Dates Group 5/5/52 5/9/62 5/12/62 5/16/52 5/19/62 I A 2 461 458 461.5 469 475 3 380 388 386.5 388 393 4 477 490 478.5 474 495 - 7 455.5 460 454.5 462.5 460 8 486 484 486 494 497 9 403 413 413.5 410 417 TB 1 449 450 448 5 453 458 2 425.5 428.5 425 5 426 428.5 3 389.5 389.5 386 390 391 4 488.5 478 470 469 466 5 457 460 457 468.5 467 6 500 509 506.5 511.5 513 7 428.5 427 426 428 431 8 446 446 449 452 455 9 476 481 475.5 480 478.5 V 1 414.5 430 424.5 431.5 428 2 511.5 516 514 521 520 3 441.5 444 434 430 424 4 483.5 497 494 5 484 507 6 472 482 474 481 480 7 534 538 539 552 561.5 8 505.5 509.5 504 509 513 9 511 517 514.5 523 528 Controls 1 509.5 514 508 516.5 525.5 2 524.5 535 534.5 534 541 3 561.5 573.5 572 576 581 4 507 500 510 508 511.5 5 508 507 494.5 490 470 6 561 474.5 566 574 575.5 7 480 492 492 496 501 8 601.5 624 613 616 627 9 613.5 626.5 604.5 610 593 10 537.5 1 534.5 551 554.5 560 37 WEIGHTS OF ANIMALS BY DATES (continued) Dates Group 5/23/62 5/26/62 5/30/62 6/2/62 6/5/62 IA 2 467 470 472 473.5 481 3 401 401 404.5 407 408.5 4 430 460 495 501.5 501.5 7 457 459 463 458 473 8 494 494.5 493 492.5 501 9 415 421.5 420 419 427.5 IB 1 459 454.5 457 451.5 455.5 2 428 429.5 433 430 439.5 3 393 394.5 396.5 392.5 401 4 486.5 427.5 410 397.5 392 5 460 469 469 473.5 380 6 511 510 510 505 508 7 433.5 435.5 439 439.5 443 8 449.5 454.5 449 450 460 9 485.5 487.5 492 490 502 V l 429 434 421 416.5 414 2 524.5 529 531.5 533 531 3 411 5 414 404 403 396.5 4 508 520 517.5 521 510 5 6 472.5 482 457 484 478.5 7 560 565.5 566.5 568.5 596 8 518 523.5 525.5 523.5 519.5 9 529.5 540 547 550 542 Controls 1 521 529 532 538.5 538.5 2 539.5 542.5 544.5 549 545 3 580 585 590.5 587 589 4 512.5 517.5 514 520 519 5 471 473.5 460 458 455 6 575 578 584.5 585.5 582.5 7 504 510 510 516 506 8 627.5 627 629 632 612 9 595 584 594 589 589 10 559 563 569 567 555 APPENDIX C COLD WATER SURVIVAL TIME DATA SHEET—eCOLD WATER SURVIVAL TIME 39 Group Animal Minutes IA IB Controls H OKOCDNChUl-P‘UUIUH «100404:me \OCIJN'JmUI-C-‘UUIUH \OCIDN-P‘WR) 125 73 126 134 117 154 88 86 65 55 97 59 147 I25 85 66 88 55 84 117 100 95 116 133 I05 113 133 71 143 157 137 84 I01 APPENDIX D WATER TEMPERATURE Time 12 I2: 12: I2 11 11: 11: 11: IO: 10: 10: 10: (IDKOKOKO :00, :45- :30- :15- :00- :45- :OO- :45- :45.- :30- :15- :00—7 :45- :30- :15:- 30-4 15.. 30- 15 - 00 - 45- 30 .. 15 .. OOY- _L I 14 Temperature I I 5 16 41 APPENDIX E ALL OUT SWIM TIME 43 DATA SHEET--ALL OUT SWIM TIMEl Time Group Animal 3/22/62 4/28/62 6/3/62 IA 2 8:00 4:30 3:30 3 6:15 9:00 31:55 4 4:40 5:15 5:00 7 4:05 5:30 5:30 8 4:00 4:00 21:15 9 3:20 3:45 3:45 IB 1 6:00 4:45 8:45 2 9:50 4:00 5:15 3 4:45 8:00 6:45 4 4:50 7:45 67:45 5 4:00 3:45 3:50 6 3:30 9:00 63:30 7 4:10 7:00 5:30 8 4:30 6:00 5:30 9 2:30 4:50 4:00 V 1 69:45 4:45 51:00 2 9:25 6:00 13:30 3 5:15 5:15 32:40 4 3:45 6:00 17:00 6 4:05 4:45 5:10 7 2:45 3:45 3:25 8 3:50 1:45 2:35 9 3:10 4:00 4:30 Controls 1 Pilot 4:00 7:20 2 205:30 62:30 72:30 3 Pilot 2:30 2:25 4 3:45 11:00 63:40 5 114:45 63:00 41:10 6 163:20 7:00 5:25 7 Pilot 6:30 56:35 8 94:25 3:15 5:40 9 3:05 2:30 4:15 10 5:50 3:15 62:30 I all out swims. All animals carried 3.5% of their body weight during Ml HIGAN STATE UNIVERSITY LIBRARIES 0 3 1293 3056 6446 C