The somatic and genetic effects of 2.45 GHz Microwave radiation (MWR) on male and female

Sprague Dawley rats were studied. Two hundred rats were used for this study. They were

grouped into control and exposed according to the parameters under consideration. The animals

were exposed to various levels of specific absorption rate (SARs) using the microwave

generator, model ER660E, Serial No MX704CCR from Toshiba UK Ltd. All animals were kept

in healthy, and radiation free environment with water and feed provided ad libitum. The study is divided into three parts; firstly, the effects of MWR on the anxiolytic and exploratory behaviour of the animals. MWR produced dose-dependent significant decrease in the cumulative time spent in the open arms of maze in the first twelve days after exposure and gradually increased towards the control value before the end of two weeks. It also affects the exploratory behavoiur of the animals, for the first two weeks after exposure but by the third week there was no significant difference between the control and the exposed animals. The modifications in the behaviour were not sex dependent as there were no significant differences between male and female rats. Secondly, the effects of MWR on the vital organs of male and female most especially, male reproductive functions were studied. The sperm count, gross motility and sperm morphology were determined using standard methods. The results showed that 2.45 GHz MWR reduced the sperm concentration, gross motility while increasing the percentage abnormal sperm cells. There was a decrease in reproductive organ weights and increased dead sperm cells as large numbers of the sperm cells membranes took up the eosin-nigrosin vital stain in the exposed group. Lastly, the genotoxic effects of MWR were studied using Chromosomal aberrations, single cell gel electrophoresis (comet assay) and direct amplification of length polymorphisms (DALP). There was a significant increase in the frequency of chromosomal aberrations between control and  exposed groups. The exposure of vital mammalian cells to 2.45 GHz radiation altered the band patterns of the exposed animals, especially in the range 40 – 120 bp. Starting with appearance of additional peaks compared with the control which was not seen in their tail DNA before exposure to MW and these were confirmed through the densitometric gel analysis. There was a statistically significant difference in the Olive moment and % DNA in tail of the exposed animals compared with control (p < 0.05). Thus, these findings support the hypothesis that exposure to 2.45 GHz MWR at studied SARs may have potential somatic and genetic effects. 

Title page i

Certification ii

Declaration iii

Dedication iv

Acknowledgements v

Table of contents vi

List of Figures xi

List of Tables xiii

List of Plates xiv

Abstract xvi

11 Preamble 1

12 Statement of the Problem 4

13 Rationale for the study 4

141 Aim and Objectives 4

142 Specific Objectives 5

21 Microwave Origin and Spectrum 6

22 Generation and Uses of Microwaves 7

221 Telecommunication 7

222 Remote Sensing 8

223 Health 9

224 Navigation 9

225 Power 9

23 Interactions of Microwave with Biological Materials 10

 231 Thermal Mechanisms of Interaction 10

232 Athermal (Non-thermal) Mechanisms of Interaction 11 

24 Macroscopic Interactions 13

25 Microscopic Interaction -- Molecular Mechanisms 18

251 Cellular Interactions 22

252 Other Interaction Mechanisms 23

26 Temperature Effects 23

261 Effects: High – Frequency Fields Force 26

27 Radiometry of Non-ionizing Radiation 28

28 Free Radicals 29

281 Hydroxyl Radical 29

282 Nitric Oxide 30

283 Superoxide (O2-) 30

284 Hydrogen Peroxide 30

285 Trichloromethyl (CCI3) 31

286 Free Radicals and Lesion in DNA 31

29 Plausible Biological Mechanism 31

210 Non-human Studies 32

210 1 Thermoregulatory Responses to RF radiation 32

2102 Reproductive Effects 32

2103 Effects on the Blood-forming and Immune Systems 33

2104 Nervous System 35

2105 Behavioural Effects 36

2106 Molecular, Subcellular and Cellular Effects 37

211 Genetic Toxicology 37

212 Epidemiological Investigations 40

31 Calibration of Microwave (MW) Source 42

32 Determination of Specific Absorption Rate (SAR) 42

33 Animal and Sample Preparations 42

34 Behavioural Studies 43

341 Exploratory Behavioural Activity Study 43

342 Anxiolytic Activity Study 43

3421 Elevated Plus Maze Study 43

3422 Y- maze Study 44

35 Fertility Studies 46

351 The Weight of the Specimens 46

352 Semen Collection 46

353 Sperm Motility Study 46

354 Sperm Morphology Study 46

355 Sperm Count 47

356 Histopathological Studies on the Reproductive Organs 47

36 Chromosomal Aberration Study from Rats Bone Marrow 47

37 Deoxyribonucleic Acid (DNA) Assay 48

371 DNA Extraction and Washing of the Blood Cells 48

372 Lysing of WBC and other Organs 48

373 Phenol Extraction and Precipitation of DNA 49

38 Quantification of DNA Sample 49

39 Primers 49

310 Amplification and Electrophoresis 49

311 Single Cell Gel Electrophoresis (Comet Assay) 50

41 Calibration and Determination of Specific Absorption Rate (SAR) 53

42 Effects of Microwave Radiation on the Behavioural Study 53

421 Effects of Microwave on the Exploratory Behaviour 53

422 Effects of Microwave Radiation on the Elevated Plus Maze and Y-Maze Study 54

43 Effects of Microwave Exposure on the Animal Reproductive Organs 66

431 Effects on the Body and Organ Weights of the Wistar Rats after Exposure to MW Radiation

432 Effects of Microwave Radiation on the Spermatozoa Parameters 67

433 Effects MW Radiation on the Histopathology Study 76

44 Effects of Microwave Radiation on the Chromosomal Study 84

45 Genotoxic Effects of Microwave 86

451 Microwave Effects on DNA Direct Amplification of Length Polymorphisms

 (DALP) Analysis 86

452 Microwave radiation Effects on Comet Assay 87

46 Conclusion and Recommendation 105

11 Electromagnetic Spectrum 2

41 Variation of Thermistor Resistance with Temperature (Calibration Curve) 57

42 Variation of Rectal Temperature with Time 58

43 Variation of SAR with Mass 58

44 Microwave Effects on the Exploratory Behaviour in Male Rats 59

45 Microwave Effects on the Exploratory Behaviour in Female Rats 59

46 % Time Spent in Open Arms of EPM by Male Rats 62

47 % Time Spent in Open Arms of EPM by Female Rats 62

48 % Time Spent in Open Arms of Y-Maze by Male Rats 65

49 % Time Spent in Open Arms of Y-Maze by Female Rats 65

410 Variation in Body weights over a period of 4 weeks (Male) 69

411 Variation in Body weights over a period of 4 weeks (Female) 69

412 Relative weight of vital organs four weeks post exposure (Male) 72

413 Relative weight of vital organs four weeks post exposure (Female) 72

414 Variations in the semen cells morphology four weeks post exposure 74

415 Variations in the gross sperm motility four weeks post exposure 74

416 Variations in the % Life/dead Sperm four weeks post exposure 75

417 Variations in the Sperm Counts four weeks post exposure 75

418 Variations of the Chromosomal Aberrations in Male Rats 85

419 Variations of the Chromosomal Aberrations in Female Rats 85

420 Densitometric track analysis of male and female blood DNA Lane a is the

 DNA of the control; b-j are DNA from exposed animals 94

421 Densitometric track analysis of male and female brain DNA Lane a is the

 DNA of the control; b-j are DNA from exposed animals 95

422 Densitometric track analysis of male and female lung DNA Lane a is the

 DNA of the control; b-j are DNA from exposed animals 96

423 Densitometric track analysis of male and female heart DNA Lane a is the

 DNA of the control; b-j are DNA from exposed animals 97 

424 Densitometric track analysis of male and female liver DNA Lane a is the

 DNA of the control; b-j are DNA from exposed animals 98

425A Densitometric track analysis of Ovary DNA Lane a is the DNA of the

 control; b-j are DNA from exposed animals 99

425B Densitometric track analysis of Testis DNA Lane a is the DNA of the

 control; b-j are DNA from exposed animals 99

426 Densitometric track analysis of male and female kidney DNA Lane a is the

 DNA of the control; b-j are DNA from exposed animals 100

427A Densitometric track analysis of male and female Tails DNA before

 exposure having peak 1 only 101

427B Densitometric track analysis of Prostate DNA Lane a is the DNA of the

 control; b-j are DNA from exposed animals 101

428 Densitometric track analysis of male and female Thyroid DNA Lane a is the

 DNA of the control; b-j are DNA from exposed animals 102

429 Densitometric track analysis of male and female Spleen DNA Lane a is the

 DNA of the control; b-j are DNA from exposed animals 103

430 The mean + SD of % DNA in tail after exposure to 245 GHz microwave radiation 104

431 The mean + SD of Olive moment after exposure to 245 GHz microwave radiation 104 

41 Cumulative time spent by the male rats in the Elevated Plus Maze 60

42 Cumulative time spent by the female rats in the Elevated plus Maze 61

43 Cumulative time spent by the male rats in the Elevated Y-Maze 63

44 Cumulative time spent by the female rats in the in Elevated Y-Maze 64

45 Effects of microwave radiation exposure on the average weight of the male rats 68

46 Effects of microwave radiation exposure on the average weight of the female rats 68

47 Effect of exposure on matured male Sprague Dawley to 245 GHz microwave radiation

after four weeks post-exposure period on reproductive organ and other visceral vital

organs weight 70

48 Effect of exposure on matured female Sprague Dawley to 245 GHz microwave

 radiation after four weeks post-exposure period on reproductive organ and other

 vital organs weight 71

49 Effect of exposure of matured male Sprague Dawley to 245 GHz microwave

 radiation after four weeks post-exposure period on semen parameters 73 

41A Micrograph of kidney for control group (female) 78

41B Micrograph of kidney for SAR 048 W/kg (female) 78

41C Micrograph of kidney for SAR 143 W/kg (female) 78

41D Micrograph of kidney for SAR 191 W/kg (female) 78

41E Micrograph of kidney for SAR 239 W/kg (female) 79

41F Micrograph of liver for control group (female) 79

41G Micrograph of liver for SAR 095 W/kg (female) 79

41H Micrograph of liver for SAR 143 W/kg (female) 79

41I Micrograph of liver for SAR 191 W/kg (female) 80

41J Micrograph of liver for SAR 239 W/kg (female) 80

41K Micrograph of ovary for control group (female) 80

41L Micrograph of ovary for SAR 048 W/kg (female) 80

41M Micrograph of ovary for SAR 095 W/kg (female) 81

41N Micrograph of ovary for SAR 143 W/kg (female) 81

41O Micrograph of ovary for SAR 191 W/kg (female) 81

41P Micrograph of ovary for SAR 239 W/kg (female) 81

41Q Micrograph of testis for control group (male) 82

41R Micrograph of testis for SAR 048 W/kg (male) 82

41S Micrograph of testis for SAR 095 W/kg (male) 82

41T Micrograph of testis for SAR 143 W/kg (male) 82

41U Micrograph of testis for SAR 191 W/kg (male) 83

41V Micrograph of liver for control group (male) 83

41W Micrograph of liver for SAR 048, 095 and 143 W/kg (male) 83

41X Micrograph of liver for SAR 191 and 239 W/kg (male) 83

42 Hybridization of the blood DNA in male and female rats Note the

 appearance of other bands in lane b –j (exposed animals) 89

43 Hybridization of the brain DNA in male and female rats Note the

 appearance of other bands in lane b –j (exposed animals) 89

44 Hybridization of the lung DNA in male and female rats Note the

 appearance of other bands in lane b –j (exposed animals) 90

45 Hybridization of the heart DNA in male and female rats Note the

 appearance of other bands in lane b –j (exposed animals) 90

46 Hybridization of the liver DNA in male and female rats Note the

 appearance of other bands in lane b –j (exposed animals) 91

47 Hybridization of the ovary DNA B Hybridization of the testis DNA

 Note the appearance of other bands in lane b –j (exposed animals) 91

48 Hybridization of the kidney DNA in male and female rats Note the

 appearance of other bands in lane b –j (exposed animals) 92

49 Hybridization of the prostate and control tails DNA in male and female rats

 Note the appearance of other bands in lane b –j (exposed animals) 92

410 Hybridization of the Thyroid DNA in male and female rats Note the

 appearance of other bands in lane d –j (exposed animals) 93

411 Hybridization of the spleen DNA in male and female rats Note the

 appearance of other bands in lane b –j (exposed animals) 93