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·सत्यमेव जयते
Indian Council of Medical Research
TECHNICAL REPORT
OF THE
SCIENTIFIC ADVISORY BOARD
FOR THE YEAR
1953
PRICE Re. 11-
NEW DELHI
1954

·INDIAN COUNCIL OF MEDICAL RESEARCH GOVERNING BODY President : {]kumari ANRIT Kaur, Minister for liralth, Goycramcn of India, New Delbi. l'ice-President : Shri V. K. B. PILIA, 3.C..., occretary to the Government of ndir, Ministry of Ilealth, New Delhi. Soonbers : Licut..Colonel M. l.. ILJA, 11.11., 1.11., 11.1)., 15.1'.11., Director, Central Rescarch Institusic, Kauli. Licut. Clonel C, K. LAKSAMANAN, L. & s.. 18.R.C.I., 0.7.3. & ff., D.C.ll., Dircctor-General of Health Scrvices, NCH Delhi, Dr. J. X. Vuxurinji, Dr. S. S. BATXAGANT, 1.c. 1'.8.5., 8.8.1., Director, Council of Scientific and Incurrrial Rescarclı, New Delhii. V.SC., 1.C.S., F.H.A.S.D., P.8.1., Nahar Avcnut, Purancliand Calcutta. Dr. R. X. CILJUDITURI, 99.8., 1l.C.I'., Director, School of .cdicine, Calcutta "Tropical Licut-Colonel T. C. Puri, 1.1.13.5., 1.8.11., D.T.I.A 17., Deputy Director-Gcncral of ficallh Services, New Delhi. Dr. SATYANIX Roy, 3.l'., 3, Gaistaun Sansions, Park Sirect, Calcutta. Maharaja Su kiessa CHANDRA GAjaratın NARAYAN Dio, Vaharaja of Parlikimedi, District G:::າງົວ:ໄມ້. Dr. B. B. Dixsur, $11.11., :1.11.11.6., 1.1.C.1'., Dr. G. L. SHARJA, 31.1., Dean, Mcdical College & Hospital, Nagpur. Surycon General with the Government of Dumbay, Bomby. Dr. S. 1. LULZIR, 11.1., 1/10,388, llimayatnagar, Ilyderabad. Major-Gcncral Sir SAT SINGH SOKIEY, U.SC., M.A., Ch.B., 1.1..1.&., M.1., Clo Haminc Instituit, l'arel, Bombay. Dr. K. V. Krislinar. 15.11.0.9., D.D., M.R.C.1'., :2,5C. F.Haley Director, All-India Instilutc of Hygiene & Public licaltii, Calcutta. Dr, C. Varced, 1.0.0.5., Radha Niwas, Kunnukuzhi, Trivandrum. Secretary Dr. C. G. Paxdir, 16.13.1.S., rh.D., D.P.ll., D.T.J., F.8.1.

·CONTENTS 1. CovrosTION or THE SCIENTIFIC ADVISORY BOARD II. COMPOSITION OF THE ADVISORY COWITTEES III. RESEARCHES CARRIED OUT DURING TIIE YEAR 10 16 CHOLERA. 1. Inquiry on the immunological studies of vibrio-cholere under the Director, Central Research Institute, Kasauli 2. Investigation on cholera endemicity with special reference to Hulsa and other estuarine fishes under Dr. M. N. Laluri at the All-India Institute of Hygiene & Public Health, Calcutta MALARIA. 1. Insecticide and mosquito repellent inquiry under the Director Malaria Institute of India, Delhi 2. Inquiry to study nutritional states and their effects on mammalian malaria under the Director, Malaria Institute of India, Delhi 3. Inquiry on chemotherapy of malaria at the School of Tropical Medicine, Calcutta 4. Inquiry on the effects of surface and surface treatment of the residual toxicity of Sarious formulations of DDT with special reference to conditions in rural India under Shri j. C. Vedamanikkam at the Central Malaria Laboratory; Andras 5. Inquiry for making detailed study of the malaria parasites in the Himalayan flying squirrel (Pelaunsta inomatus) and the Himalayan Langoor (Semnopithecus entellus) under Dr. H. X. Ray at the Indian Veterinary Rescarch Institute, Sukleswar- Kumaun, U.P. 6. Investigation on the possibility of controlling the incidence of lala-azar bs spray-killing of adult sandlics under Dr. B. C. Basu at the School of 'Tropical Medicine, Calcutta.. NUTRITION, 1. Nutrition Rescarch Laboratories, Coonoor 2. Nutrition rescarch unit under Dr. B. C. Guha at the University College of Science & Technology, Calcutta 3. Nutrition rescarch unit under Dr, R. G. Chitre at the Seth G. S. Medical College, Bombay 4: Inquiry on physiological effects of dictary proteins uncler Dr. M. Damodaran at the National Chemical Laboratory of India, Poona Inquiry on the metabolism of vijamin A under Dr. K. Rajagopal at the All-India Institute of Hygiene & Public Health, Calcutta 37 39 42 19 .. 75 85 87 ·00 00 100 103 107 6. Inquiry on the biosynthesis of nicotinic acid under Dr. S. Banerji at the Presidency College, Calcutta 7. Inquiry on the enzymnic role of inositol under Dr. P. S. Sarma at the University Biochemical Laboratory, University of Vadras, Viadras 8. Inquiry on gencralized mechanism of the action of vitamin A in the body inder Dr. D. P. Sadlu at the Presidency College, Calcutta 9. Inquiry on the determination of sodium and potassium of common Indian foodstuffs and preparation of Intra sodium high-potassium dici for differcni clinical conditions under Prof. K. Rajagopal and Dr. Snit.! B. Banerjee at the All-India Instituic of Hygicnc & Public Healii. and Medical College Hospitals, Calcutta 10. Inquiry on human requirements of riboflavin and thiamin under Dr. H. X. De at the VI. G. J. Vicdical College. Indorc 11. Study of the quality of proteins, 00:-pricias and fats or some edible varieties of Bombay fishi under Dr. Smi. Karnala Sohonic at the Departmeni of Biochemistry, Institute of Science, Bombay. 12. Inquiry on nutritional cedema under Siiri X. C., D.atta at the Grant Vicdical College, Bombay. 13. Inquiry on the metabolism offre under Dr. K. P. Basi at he Indian Dairy Research Instique, Bangalore 14. Studies on the infiuence of diciary habits on incidence of goitre and tuberculosis in Darjeeling under Dr. S. R. Son Gupta, .Issistant Director of Health Services, llest Bengal. Calcutta 15. Inquiry on physiological study of human breast milk and its correlation with the sui-nutritional status (specially underweight of the newb0n71 in India under Dr. S. K. Roy and Dr. Imala Ramcharan at the Ramkrishna Mission Sishumangal Pratishthan, Calcutta 16. Studies on energy metabolism under Dr. S. Berji :1 the Presidency College, Calcutta . 113 . 115 118 ... 121 124 LEPROSY. 137 1. Leprosy: inquiry under Di, Dharmendra at the School of Tropical Vedicine, Cileuca 2. Leprosy inquiry under Dr. V. R. Klanolkar at the Tata Víemorial Hospital, Bombay 3. Leprasy inquiry under Dr. Paul 11. Brand at the Christian Sledical College, Vellore ... 1. Leprosy inquiry under Dr. S. D. Desai at the leworth Leprosy Home, Bombay 5. Leprosy inquiry under Dr. B. B. Gokhale at the H. J. Vedical College and Sassen Hospital. Poona ... 139 .

·PLAGUE. 156 1. Plague researches under the Director, Haffkine Institute, Bombay 2. Inquiry into the breeding habits of rat-fleas in relation to the epidemiology of plague under Dr. S. C. Seal at the All-India Institute of Hygiene & Public Health, Calcutta 162 CLINICAL RESEARCH. 168 170 174 ... 176 177 187 189 1. Clinical research unit under Dr. V. R. Khanolkar at the Tata Memorial Hospital, Bombay 2. Neuropathological unit under Dr. V. R. Khanolkar at the Tata Memorial Hospital, Bombay 3. Inquiry on the role of nutritional factors in hepatic cirrhosis under Dr. M. V. Radhakrishna Rao at the Haffkine Institute, Bombay 4. Inquiry into typhus in Bengal under Dr. K. V. Krishnan at the All-India Institute of Hygiene & Public Health, Calcutta 5. Hæmatological unit under Dr C R. Das Gupta at the School of Tropical Medicine, Calcutta 6. Inquiry on the synthesis of anti-folic acid compounds and studies on their chemotherapeutic activities under Dr. S. C. Roy at the University College of Science & Technology, Calcutta 7. Inter-laboratory evaluation of procedures for serological tests for syphilis followed in various laboratories in India under Dr. R. V. Rajam, Director, Upgraded V. D Department, Government Genera. Hospital, Madras 8. Clinical research unit under Dr. R N Chaudhuri at the School of Tropical Medicine, Calcutta 9. Liver diseases research unit under Dr. P. N. Wahi at the S. N. Medical College, Agra 10 Studies of endometrial biopsies in cases of sterility with a view to estimate the incidence of latent tuberculosis of the endometrium under Dr. (Kumari) P, K. Malkani at the Lady Hardinge Medical College, New Delhi 11. Inquiry entitled fungus diseases of skin identification of fungi under Dr. K. N. Saxena at the S, N, Medical College, Agra 12. Inquiry on electro-retinography in vitamin A deficiency under Dr. R. P. Dhanda at the M G. M. Medical College, Indore 13. Inquiry on localization of a “ feeding centre" in hypothalamus under Dr. B. K. Anand at the Lady Hardinge Medical College, New Delhi 14. Inquiry on the possibilities and scope of homologous venous and arterial grafts under Dr. P. K, Sen at the Seth G. S. Medical College, Bombay 192 210 222 228 229 235 !!

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D Research Worker, Government of West Bengal, Calcutta 31. Epidemiological research in tuberculosis under Dr. P V. Benjamin, Adviser in Tuberculosis, Directorate-General of Health Services, New Delhi 32. Inquiry into the incidence of penicillin resistant strains of Staphylococcus pyogenes in Calcutta under Dr B. P. Trbedi at the Medical College, Calcutta 33. Inquiry on the serological classification of Corynebacterium diphtheriæ in Calcutta under Dr. B. P. Tribedi and Dr D Barua at the Medical College, Calcutta 298 299 MATERNITY AND CHILD WELFARE. 1. Inquiry into infant mortality in the Madras State under the Director of Public Health, Miadras 2. 301 T 304 308 311 317 Calcutta. 3. Investigation on the problem of late abortions from four to seven months of pregnancy under Dr H V Tilak at the Topiwala National Medical College, Bombay 4. Inquiry into the incidence and causes of still-births and neonatal deaths in Hyderabad under Dr (Smt.) P. M. Naidu at the Osmania Medical College, Hyderabad-Deccan 5. Inquiry on the structural and functional adaptation of the kidneys in pregnancy under Dr Chunilal Mukherjee at the Medical College, Calcutta 6. Research on radiological and electro-cardiographic changes in normal pregnancy under Dr. (Kumarı) S Padmavati at the Lady Hardinge Medical College, New Delhi 7. Inquiry into the incidence of intra-natal and early rickets and their ætiology with special reference to mother's milk and her diet under Dr B.D Patel, Dr. H. I. Jhala, Dr B.D Patwardhan and Shri N. C. Datta at the Grant Medical College, Bombay 8. Inquiry on the ætiology of toxæmias in the later months of pregnancy under Dr. S C. Bose and Dr.S.C. Roy at the Medical College, Calcutta 317 319 321 324 FILARIASIS. 1. Filariasis control unit (Orissa) under the Director, Malaria Institute of India, Delhi 2. Filariasis inquiry under the Director, King Institute, Guindy, Madras 3. Inquiry on pharmacological and toxicological studies of compounds prepared for chemotherapy of filariasis under Dr. G. Werner at the School of Tropical Medicine, Calcutta 326 327 ·(vi) VIRUS DISEASES. 334 ... 336 .. 1. Rabies research under the Director, Central Research Institute, Kasauli 2. Polio research unit under Dr. P. V. Gharpure at the Grant Medical College, Bombay 3. Inquiry on the antigenic analysis of Indian strains of influenza virus under Dr. I. G. K. Menon at the Pasteur Institute of Southern India, Coonoor 4. Virus Research Centre, Poona 5. Inquiry on the incidence, nature and types of coxsackie virus infections in Bombay City under Dr. D. D. Banker at the Seth G. S. Medical College, Bombay 6. Rabies research under the Director, Pasteur Institute of Southern India, Coonoor 344 351 361 362 INDUSTRIAL HEALTH. 364 377 1. Industrial health research unit at the All-India Institute of Hygiene & Public Health, Calcutta 2. Inquiry into the problem of traffic accidents and accidentprone personnel amongst bus and tram drivers in Calcutta under Shri S. K. Bose at the University College of Science & Technology, Calcutta. 3. Inquiry into the effect of sewage treatment and excreta disposal methods on intestinal parasites under Dr. T. R. Bhaskaran at the All-India Institute of Hygiene & Public Health, Calcutta 4. Inquiry into treatment and hygienic disposal of lac wastes under Dr. T. R. Bhaskaran at the All-India Institute of Hygiene & Public Health, Calcutta 378 381 ... PHARMACOLOGY. 384 ... 386 393 .1. Indigenous drugs inquiry under Dr. G. Werner at the School of Tropical Medicine, Calcutta 2. Indigenous drugs inquiry under Bt.-Col. R. N. Chopra at the Drug Research Laboratory, Jammu 3. Inquiry into the isolation of the active principles of plants of N. 0. Dioscoreacea under Dr. R. N. Chakravarti at the School of Tropical Medicine, Calcutta 4. Inquiry on pharmacological action and biological assay of adrenal cortical hormones under Dr. B. Mukerji at the Central Drug Research Institute, Lucknow 5. Inquiry on the effects of cardio-active drugs, such as digitalis, strophanthine, etc., and related compounds, on various metabolic reactions involved in muscle contraction under Dr. B. N. Ghosh at the University College of Science & Technology, Calcutta 396 397 T.

·(vii) 6. Inquiry on indigenous drugs under Dr. B, C. Bose at the M. G. M. Medical College, Indore 7. Inquiry on the biogenesis of alkaloids in tobacco plants tinder Dr. B. C. Bose at the M. G. M. Medical College, Indore 399 403 ... 4.. IV. ABSTRACTS FROM REPORTS OF I.C.M.R. RESEARCH FELLOWS - 1. Dr. J. M. Senapati 2. Dr. M. R. Rajarama Rao 3. Dr. C. S. Dawn. 4. Dr. K. K. Sikka 5. Dr. Sami Hameed 6. Dr. S. K. Vaish 7. Dr. K. L. Mſukhopadhya 8. Dr. J. K. Biswas 9. Dr. A. P. Chakravarty 10. Shrimati S. S. Rao • 11. Shrimati S. Chandra Sekhar 12. Shri P. A. Kurup 13. Kumari Vedam Jayem 14. Shri K. C. Abraham 15. Shri S Duraiswami 16. Dr. Dhirendra Kumar V. INDIAN JOURNAL OF MEDICAL RESEARCH AND INDIAN MEDICAL RESEARCH MEMOIRS VI. INDIAN JOURNAL or MALARIOLOGY VII. MICROFILM AND PHOTOCOPY SERVICE UNITS VIII. PAPERS PUBLISHED IN THE INDIAN JOURNAL OF MEDICAL RESEARCH AND INDIAN JOURNAL OF MALARIOLOGY DURING THE YEAR 1953 IX. LIST OF PAPERS PUBLISHED BY THE WORKERS OF THE INDIAN COUNCIL OF MEDICAL RESEARCH DURING 1953 408 411 414 415 419 419 422 424 427 429 429 430 431 431 432 432 ... 435 436 437 438 P. 444 ·I. COMPOSITION OF THE SCIENTIFIC ADVISORY BOARD. The Scientific Advisory Board during the year consisted of the following :- Chairman. Lieut.-Colonel C. K. Lakshmanan, 1.M.S., M.R.C.S., D.T.M. & H., D.P.H., Dircctor-General of Health Services, New Delhi. Members. Dr. A. C. Banerjca, M.B.B.S., Dr.P.H., 31, Station Road, Lucknow. Dr. Basudeva Narayana, M.Sc., M.B., Ph.D., F.R.S.F., F.N.I., Principal, P. W. Medical College, Patna. Major-Gencral S. L. Bhatia, M.A., M.D., B.Ch., M.R.C.S., F.R.C.P., . Chairman, Pharmaceutical Inquiry Committee, New Delhi. Dr. R. N. Chaudhuri, M.B., M.R.C.P., T. D.D., F.N.I., Director, School of Tropical Medicine, Calcutta. Dr. R. G. Dhayagude, M.D., Dean, Seth G. S. Medical College, Bombay. The Hon'ble Dr. Jivraj N. Mchta, M.D., M.R.C.P., Minister for Finance, Prohibition and Industries, Government of Bombay, Bombay. Lieut.-Colonel Jaswant Singh, M.B., Ch.B., D.T.M. & H., D.P.H., Director, Malaria Institute of India, Dclhi. Dr. V. R. Khanolkar, B.Sc., M.D., Director, Indian Cancer Research Centre, Tata Memorial Hospital, Bombay. Dr. K. V. Krishnan, M.B.B.S., F.R.C.P., D.B., D.Sc., F.N.I., Director, All-India Institute of Hygiene & Public Health, Calcutta. Dr. P. Kutumbiah, B.A., M.D., M.R.C.P., Principal, Christian Medical College, Vellore. Brigadier S. Narain, M.B., M.R.C.P., D.T.H., D.T.M., A.M.C., Director of Rescarch and Hсalth, Directorate-General of Armed Forces Medical Services, Ministry of Defence, New Delhi. Dr. K. C. K. E. Raja, L.R.C.P. & S., L.R.F.P.S., D.P.H., D.T.M. & H., Oficer on Special Duty, Ministry of Health, New Delhi. Dr. K. C. Sen, Ph.D., D.Sc., F.N.I., Director, Indian Dairy Research Institute, Bangalore. Dr. K. S. Viswanathan, B.A., M.B.B.S., B.Sc., M.P.H., Director of Public Health, Madras. Dr. K. L. Wig, M.B.B.S., M.R.C.P., M.R.C.S., Professor of Medicine, Medical College, Amritsar. Secrclary. Dr. C. G. Pandit, M.B.B.S., Ph.D., D.P.H., D.T.M., F.N.I., Secretary, Indian Council of Medical Research, New Delhi.

·11, COMPOSITION OF THE ADVISORY COMMITTEES. 3. Cholera. Lieut-Colonel M. L. Ahuja, Director, Central Research Institute, Kasauli, (Chairman). Dr. B. C. Das Gupta, Director of Health Services, West Bengal, Calcutta Dr. Gurkripal Singh, Assistant Director, Central Research Institute, Kasauli Dr. K. V. Krishnan, Director, All-India Institute of Hygiene & Public Health, Calcutta. Dr. R. B. Lal, Rasi Kunj, Srinagar Estate, Mussooree. Shri K. K. Mathen, Professor of Statistics, All-India Institute of Hygiene & Public Health, Calcutta Dr. Y. S. Narayana Rao, Director, King Institute, Guindy, Aladras. Dr. P. M. Wagle, Director, Halſkınc Institute, Parel, Bombay Dr. K. V. Venkatraman, Serologist and Chemical Examiner to the Government of India, School of Tropical Medicine, Calcutta (Secretary), 11. Malaria. Lieut.-Colonel Jaswant Singh, Director, Afalaria Institute of India, Delhi. (Chairman). Dr. A. C. Banerjea, 31, Station Road, Lucknow, Major-General S. L. Bhatia, Chairman, Charmaceutical Inquiry Committee, New Delhi. Dr. N. Ganguli, Assistant Director of Malarıology, Directorate of Health Services, West Bengal, Calcutta. Lieut.-Colonel N. D. P. Karani, Professor of Hygiene, Armed Forces Medical College, Poona. Dr. Dev Raj Michte, Afalariologist, Public Health Department, East Punjab, Kasauli. Dr. J. N. Mukherji, D, Puranchand Nahar Avenue, Calcutta. Dr. D. K. Viswanathan, Director of Public Health, Connaught House, Poona Dr. K. S. Viswanathan, Director of Public Health, Madras. Dr. B. Ananthaswamy Rao, Deputy Director, Malaria Institute of India, Delhi. Secretary). 1). Nutrition. Dr. K. Alitra, Assistant Director General of Health Services, New Delhi. (Chairman, Dr. S, T, Achas, Professor of Pediatrics, Aledical College, Madras. Dr. A. C. Banerjea, 31, Station Road, Lucknow. Dr. H. B. Dikshit, Surgeon-General with the Government of Bo Bombay, ay, ·D:. E.C.ka, Esad of the Dance: C.E. CaseyLagu College of Science & Technology: 2 CEPE Cerca Praca Calcuna Dr. VI. V. Fantariksa Pen, siran: Direcur-élags of the Lirca brera de Piator Creedt of Erica, Cis FESTE I Facal, Ertzbar Dr. E. Bajagopal. Profesor sí Eierto II Irctic Hrniere Et Pric Hama Calcio Dr. E C. Sar, Diecar, Ir. Dairy Puzzle Institut Escape Dr. V. Srikrakstarta Diretor, Central Ecce Tece ricerca pe Instítitie Vrore. Dr.V. Fatnard.122, Director. Harition Pearce. Lehcraries, Ccrecer. (Serie- IV. LETOS Dr. Dharmendra, Ir-charge. Lepice Department Scicoi cf Trogir Medicine, Calcutta. Chanat, - Dr. Parl W. Era... Profesor E Orthopadic azd Head of tie Sca Departrcers. Ceritia: Viedeca! Cclesz, Telirre.. Dr. - Fişieredo, Seriaceaente correth Leprosy Hoce Wačala, Erinfiam. Dr. V. P. Fhaholicar, Director. Irin Carcer Research Cerra Tata Verzorial Finspitzl, Erzhar. Dr. P. S. Wardekar, Secretaris Carina Viscrizi Lepram Forcade Saratan, Wardha. Dr. S. SI Charenje, Lepico; Flecach. Departem Sokeri cê Tropica [edicine, Calcutta {SLETE:- V. Piasts. Dr. P. VI. Wagle, Director, Haflare Instituto Farel. Eocav. CE 16T;- D;. P. J. Derran, esítani Director, Hafikine Insturta Pael Brmbar. Dr. A. M. Franck, Sperial Plaque OFCEE, OSCE of the Dream of ErbEC Health, 1, Mount Pesad, Cathedral P. 0. Vadrag. Dr. B. C. Das Gupta, Director of Health Serrices, West Bengal, Caerdz. D:. R. B. Lal, Rar Fun, Srinagar Estate, Vuscorte. Dr. S. Paghavendra Rao, étant Director of Public Health (Epiter... Hydrzabadi. Dr. K.V. Venkatraman, Serologist and Chemical Ezaniner to the Govern ment of India, School of Tropical Medictie, Calcutta. Dr. D). K. Visvanathar, Director of Public Health, Conraight House Poona. Dr. S.C. Scal, Ansistant Professor of Epidemiology. All-India Irastintz CÉ Hygiene & Public Health, Calcutta Secretary ·VI. Clinical Research. + Dr. V. R. Klianolkar, Director, Indian Cancer Research Centre, Tata Memorial Hospital, Parel, Bombay. (Chairman). Dr. R. N. Chaudhuri, Director, School of Tropical Medicine, Calcutta Dr. P. Kutumbiah, Principal, Christian Medical College, Vellore Dr. B. Mukerji, Director, Central Drug Research Institute, Lucknow. Brigadier S. Narain, Director of Research and Health, Directorate-General of Armed Forces Medical Services, Ministry of Defence, New Delhi Dr. R. V. Rajam, Director, Upgraded V. D. Department, Government General Hospital, Madras. Dr. P. K. Sen, Professor of Tuberculosis and Chest Diseases, Medical College, Calcutta. Dr. P. N. Wahi, Professor of Pathology, S. N. Medical College, Agra. Dr. B. B. Yodh, Professor of Medicine, Grant Medical College, 18, Darabshah Road, Bombay. Dr. Arthur E. de Sa, Honorary Surgeon, K. E. M. Hospital, Bombay (Secretary). VII. Maternity and Child Welfare. 1 Dr. B. C. Das Gupta, Director of Health Services, West Bengal, Calcutta. (Chairman). Dr. S. T. Achar, Professor of Pediatrics, Medical College, Madras. Dr. (Smt.) S. Bhatia, Adviser on Maternity and Child Welfare, Directorate- General of Health Services, New Delhi. Dr. C. R. Das Gupta, School of Tropical Medicine, Calcutta. Dr. V. S. Manglik, Dean, King George's Medical College, Lucknow. Shri K. K. Mathen, Professor of Statistics, All-India Institute of Hygiene & Public Health, Calcutta. Dr. (Kumari) H. M. Rajamanikkam, Deputy Director of Public Health, (M. & C. W.), Madras. Dr. R. K. K. Tampan, Superintendent, Government Hospital for Women and Children, Egmore, Madras. Dr. (Smt.) Muktha Sen, Professor of Maternity and Child Welfare, All- India Institute of Hygiene & Public Health, Calcutta. (Secretary). VIII. Filariasis. Dr. B. B. Dikshit, Surgeon-General with the Government of Bombay, Bombay. (Chairman). Dr. N. V. Bhaduri, Officer-in-Charge, Filariasis Research Laboratory, School of Tropical Medicine, Calcutta. Dr. M. O. T. Iyengar, 48, Southern Avenue, Calcutta.' Dr. Y. S. Narayana' Rao, Director, King Institute, Guindy, Madras.

·(xii) Dr. S. S. Pillay, Superintendent, Public Health Laboratory, Trivandrum. Licul.-Colonel Jaswant Singh, Director, Malaria Institute of India, Delhi. Dr. R. N. Sahu, Professor of Clinical Medicine, S. R. C. B. Mcdical Collegc, Cuttack. Dr. N. G. S. Raghavan, Assistant Director, Malaria Institute of India, Delhi. (Secretary). IX. Virus Discases. Licut.-Colonel M. L. Ahuja, Director, Central Research Institutc, Kasauli. (Chairman). Licut.-Colonel S. L. Kalra, Assistant Professor of Pathology, Armed Forces Alcdical College, Poona. Dr. J. A. Kerr, Director, Virus Rcscarch Centre, Poona. Dr. K. V. Krislinan, Director, All-India Institute of Hygiene & Public Hicalth, Calcutta. Dr. R. O. A. Smith, Eintomologist, Typhus Inquiry, All-India Institute of Hygiene & Public Health, Calcutta. Dr. D. Soman, Assistant Director, Haffkinc Institutc, Parci, Bombay. Dr. R. Vccrarahavan, Director, Pastcur Institute of Southern India, Coonoor. Dr. K. V. Venkatraman, Scrologist and Chemical Examiner to the Government of India, School of Tropical Medicine, Calcutta. Dr. R. Sanjiva Rao, Director, Lymph Institute, Belgaum. (Scorelary'). X. Industrial Health. Licut.-Colonel C. K. Lakslimanan, Dircctor-General of Health Services, New Dcihi. (Chairman). Dr. A. C. Banerjca, 31, Station Road, Lucknow. · Dr. H. P. Dastur, Medical Officer, Department of Industrial Medicine, Tata Industrics Ltd., Bombay House, Bombay. Dr. Nils P. V. Lundgren, Professor of Industrial Hygiene, All-India Institute of Hygiene & Public Health, Calcutta. Shri N. S. Mankiker, Chief Advisor on Factories, Government of India, Ministry of Labour, New Delhi. Dr. K. K. Mathur, Medical Officer-in-Charge, Industrial Health Organization, 7/198, Swaroop Vagar, Kanpur. Lieut.-Colonel M. B. Menon, Assistant Director of Medical Services, Ordnance Factorics, Calcutta. Dr. S. Roy, Chief Sanitary Officer, Mines Board of Health, Asansol. The Professor of Sanitary Engincering, All-India Institute of Hygiene & Public Hсalth, Calcutta. Dr. M. N. Rao, Assistant Professor of Physiological and Industrial Hygiene, All-India Institute of Hygiene & Public Health, Calcutta. (Secretary).

·(xiii) XI. Pharmacology, Bt.-Colonel R. N. Chopra, Director, Drug Research Laboratory, Jammu, Tawi. (Chairman) Dr. M. D. Chakravarty, Director, Central Drugs Laboratory, Calcutta. Dr. B. B. Dikshit, Surgeon-General with the Government of Bombay, Bombay. Dr. B. N. Ghosh, Professor of Pharmacology, R. G. Kar Medical College, Calcutta. Dr. G. K. Karandıkar, Professor of Pharmacology, Medical College, Baroda. Dr. B. Mukerji, Director, Central Drug Research Institute, Lucknow. Dr. B. B. Yodh, Professor of Medicine, Grant Medical College, 18, Darabsliah Road, Bombay. Dr. G. Werner, Professor of Pharmacology, School of Tropical Medicine, Calcutta. Shri P. M. Nabar, Drugs Controller (India), Directorate-General of Health Services, New Delhi, (Secrelary).

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·M. TECHNICAL REPORT OF THE RESEARCHES CARRIED OUT DURING THE YEAR 1953. 1 The researches carried out during the year under report were recommended by the Scientific Advisory Board at its meetings held in Jaipur on the 3rd and 4th December, 1952 and were approved by the Governing Body of the Indian Council of Medical Research at its meeting held in New Delhi on the 28th March, 1933. The views expressed by the indırdual workers are not necessarily the riews of the Council. CHOLERA 1. Inquiry on the immunological studies of Vibrio-cholera under the Director, Central Research Institute, Kasauli. AL The purpose of this inquiry is to study questions relating to the immunizing efficiency of cholera vaccines with particular reference to the establishment of international reference preparations for the assay of the potency of cholera vaccine. At the request of the Biological Standardization Committee of the W.H.O., tests were carried out on the comparative protective value of different samples of cholera vaccines of unknoun origin received from the State Serum Institute, Copenhagen. The methods used for the evaluation of the potency of these vaccines were 1. Active immunization of mice followed by test infection with 2. Passive immunization a virulent strain of V. cholere, 3. Assessment of vibriocidal titre after immunization with two doses of vaccine. 4. Serological examination of vaccines using mono-specific Ogawa, Inaba and cholera rough O'sera. The results of tests brought out the following interesting facts (see bles I to IV) :- Two vaccines (Nos. 144 and 145) out of the eight tested can be weeded as unsatisfactory by serological examination alone, provided cholera ch'O' serum is employed for the purpose. Vaccine No 144 has been ared from a strain which is in the smooth-rough phase and vaccine 145 has been prepared from a strain which is almost entirely rough. e vaccines evoke poor vibriocidal response in guinea-pigs and afford protection to mice as evidenced by active and passive immunity tests ther six vaccines, Nos. 141, 142, 143, 147, 738-19 and 73850, appear -e been prepared from smooth strains of V. cholere ; they pass the ical tests, evoke good vibriocidal response and afford considerable Jon to mice. In our opinion, these vaccines are satisfactory products stical purposes. It is interesting to note that vaccines which evoke ibriocidal response, Nos. 144, 145, are also poor in affording sive, and (6) active immunity to mice and further that such can be rejected as unsatisfactory by serological tests involving of cholera rough 'O'serum,

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·3 From the above findings it is obvious that the serological examina of cholera vaccines yields valuable information regarding the quality suitability of the products for immunization purposes. The more elabo active and passive immunity tests do not furnish any additional infor tion which is not obtained from the serological examination alone. do not wish to decry the use of active or passive immunity tests in ass. of relative potencies of cholera vaccines or in the selection of strains use as seed for vaccine manufacture, but, in our opinion, for routine assa of cholera-vaccine brews, prepared from approved smooth strains, t simple serological tests are adequate. We make this statement keeping F view the practical difficulties that many manufacturing concerns an laboratories will experience, if we insist on certain minimum requirement for anti-cholera vaccine in terms of active and passive immunity units The use of animal tests, as a routine, would be advisable if by doing so one could obtain extra information of value which could not be got by serological tests, but frankly we are not convinced that it is so. The results of the planned collaborative assays of the vaccines sent by the Department of Biological Standardization, Copenhagen, have convinced us of the soundness of the practice which we follow at this Institute, viz. the use of serological tests only for 'passing' of finished brews of anti-cholera vaccines and the use of both serological and animal tests for selection of cholera strains to be used as seed for mass manufacture of vaccine. Our minimum requirements at this Institute for (a) passing the vaccines as satisfactory for prophylactic purposes, and (b) for selection of a strain of V. choleræ to be used as seed for vaccine manufacture are as follows: (a) 1. The vaccine must be prepared from smooth strains. It must contain both Inaba and Ogawa sub-types in equal proportion 2. The vaccine must agglutinate with Inaba and Ogawa 'O' type specific sera. 3. The vaccine must not agglutinate with cholera rough 'O' serum. 4. In case of vaccines prepared from agar, the vibrio content must not be less than 8,000 million vibrios per c c. (6) A smooth strain of cholere (G and V'O'group I) isolated Huring the course of an epidemic must be used as the seed. It must aggluinate to full titre with the corresponding type-specific O'serum accine prepared from the strain must evoke a satisfactory vibrocidal esponse in guinea pigs and afford protection to mice against intraperitoneal fection with a heterologous strain of V. cholere. The results of active nd/or passive) immunity tests obtained must compare favourably with ose of the control vaccine maintained at the Institute as reference preration for the purpose. Our findings and comments have been communicated to the Experi mittee on Biological Standardization of WHO and will form the basis iscussion at their next meeting of the difficult problems involved in assay and control of cholera vaccines The 11. MONO-SPECIFIC OGAWA AND INABA AGGLUTINABLE SERA. Type-specific cholera sera prepared at the Central Research Institute, li, some four years ago and stored at the State Serum Institute, Copenfor distribution as international reference preparations were tested ·4 TABLE II. Passive immunity tests of cholera vaccines. Experimental animals, Swiss mice, 17 g. to 22 g. in weight: Immunizing dose given intraperitoneally 4 hours before challenge in a volume of 0.2 C.C. -- . Challenge dose given intraperitoneally in a volume of 0.5 c.c. in 5 per cent mucinized suspension. Period of observation, 72 hours: Ogawa vaccines. DOSE OF IMMUNE SERUM INJECTED (c.c.) 3 Vaccinc number Challenge strain Challenge dose Controls unvaccinated Viable' Count 0.05 0.025 0.0125,0.00625 0.003125.0.0015625 -3 143 10 6/6 6/6 5/6 6/6 5/6 4/6 10: 0/6 10,750,000 10.. 1/6 10,75,000 -4 -3 10 144 1/6 1/6 1/6 1/6 2/6 1/6. 10 3/6 107,500 lio 6/6 | 10,750 V. cholere sub-type Ogawa strain 425/52 -3 10 147 616 -6 10 6/6 1,075 6/6 616 5/6 4/6 3/6 --3 73849 10 5/6 6/6 6/6 6/6 6/6 6/6 Inaba vaccines L. -3 10 1 141 4/6 6/6 6/6 6/6 4/6 5/6 10 0/6 16,800,000 10 1/6 1,680,000 -3 10 142 5/6 6/6 6/6 5/6 6/6 5/6 10 5/6 168,000 V. cholerc sub-type Inaba strain 375/52 10 6/6 16,800 109 145 10 3/6 1/6 1/6 1/6 :-)! 1/6 110 6/6 1,680 to 73850 10 6/6 5/6 6/6 6/6 .5/6 410 Numerator/denominator =Survivals/total.

·5 for type specificities and keeping qualities. No difference in titre between the samples or between the original tests carried out four years ago and the renewed tests were noticed. The sera keep up their agglutinating capacities satisfactorily under cold storage during long periods as well as during postal transmission. These sera have now been accepted as suitable international reference preparations, III, ENZYMES OF V. CHOLFR.E. In our report for 1952 we mentioned that an enzyme capable of dissolving collagen was present in culture filtrates of cholerogenic and noncholerogenic vibrios. Further studies of this enzyme show that it does The presence of elastinase in vibrio-culture filtrates has been confirmed and further work has shown that it is capable of acting both on the native and purified elastin at an optimum pH of 8. The action of this enzyme is inhibited by animal and human sera and by dialysates of ground pancreatic tissue. The mucinolytic, tryptic and elastinolytic properties of vibrio filtrates as well as their lipase, amylase and invertase activities have been found to be due to different enzymic entities. These properties are not peculiar to V. cholere alone but are exhibited by non-cholerogenic vibrios. To assess the possible rôle of these enzymes in cholera pathogenesis, the following experiments were carried out Membrane sacs were made using living pieces of guinea-pig ileum and the effects of vibrio filtrates on permeability of these membranes investigated. It was found that there was a notable increase in permeability when vibrio filtrates were allowed to act on the membranes. Some thermolabile constituent or constituents of the filtrates abolish peristaltic activity. The factors causing the increased intestinal permeability produce their effect independent of the muscular or nervous mechanisms concerned with peristalsis since membranes which have lost their peristaltic capacity are also equally affected by the filtrates. This activity is demonstrable when the filtrate is allowed to act on the intestine from the lumen to the peritoneum or from the peritoneal surface to the lumen. The activity is inhibited by normal serum which as well as a trypsin inhibitor. The ultimate of the filtrate-18 hours-on the intestinal sues. Histamine 1-500 in Ringer-Locke solution does not produce increase permeability of peristalsis changes when tried under identical conditions. 'Filtrates of both V. cholere and NAG vibrios have been observed to give identical results in permeability experiments. 'Membranes consisting of the walls of blood vessels also show, like intestinal membranes, increased permeability when acted on by V cholere filtrates. The effect is observed both when the action takes place on the blood-vessel wall from the periphery to the lumen 'or from the lumen to the periphery, f ·TABLE III. Vibriocidal response lo cholera vaccines. Experimental animals Guinca-pigs 300 g. to 100 6 in wcight. Immunizing dose-Two doses of vaccine (0-5 c.c. and I c.c.) at weckly intervals subcutaneously. Vibriocidal titre tested 10 days after second immunizing dosc. Ogawa vaccines Inaba vaccinics Vilriocidal titre Vibriocidal titre Vaccinc number Vaccinc mmber Inaba Ogawa Inaba Ogawa 143 141 142 1/30 1/405 1/3615 1/3645 1/10,935 1/135 1/32,805 1/32,805 1/10,935 1/10,935 1/15 1/32,805 1/10,935 1/10,935 1/45 1/32,805 147 73,8-19 73,850 *Metliod of icst-As published in Indian Journal of Medical Rescarch, 1948, 36, p. 3. TARLE IV. Agglutination results of cholera raccine suspensions when tested against mono-spoecific cholera 'O' sera and cholera rough 'O' sera. Ogawa vaccines Inaba vaccines Vaccine number Inaba typc 1000 Ogawa Cholera Vaccinc Inaba typc rough number type 1000 1000 1000 Ogawa Cholera type rough 1000 1000 500 143 144 147 73,849 1000 500 500 250 111 142 1.15 73,850 500 500 50 500 50 500 2. Investigation on cholera endemicity with special reference to Hilsa and other estuarine fishes under Dr. M. N. Lahiri, at the All-India Institute of Hygiene & Public Health, Calcutta. Collection of water samples was continued from the surface and bottom of a selected stretch of the river Hooghly, both from the shore and the mid-stream, at three fishing centres, viz. Nawabgunge, Baghbazar and Falta or Diamond Harbour. The last station which is an important endemic centre for cholera was sclected when fishing was closed at Falta. A total of 722 samples of water were examined, and out of these, three samples were positive for cholera vibrios and 306 samples for NAG vibrios. *Ahuja, M.L., and Gurkirpal Singh (1948), Observations on cholera vaccines-Ind. Jour. Aled. Res. 36, I, p. 3.

·For the corresponding period last year, seven positive isolations of cholera vibrios were made. All these isolations were obtained from March to May, but the period of occurrence this year was found to extend up to July from March As during the previous year, all the isolations were made from the upper reaches of the estuary (Nawabgunge) at high tide only. Two isolations were made from the surface and one from the bottom.' Out of the three strains isolated, two were of the Ogawa type and one of the Inaba type. NAG vibrios isolated from water samples were grouped according to Heiberg's classification. Groups I, II and V formed the predominant constituents of the vibrio flora of water The seasonal fluctuations of groups I and II show a definite correlation with the period of cholera epidemics in Calcutta. The pH and salinity of the water samples collected from the three fishing centres were determined with a view to elucidate any probable correlation between these values and the occurrence of vibrios. While the fluctuations in pH are not very marked, the salinity varies to an appreciable extent. Besides the diurnal variations due to tidal influence, seasonal variations are caused by the large quantities of flood water brought down during the S.-W. monsoons and the excessive evaporation during the summer months. As is to be expected, the average salinity is higher at the lower reaches of the estuary at Falta and Diamond Harbour than in the upper reaches at Nawabgunge and Baghbazar. It is interesting to note that isolations of cholera vibrios have been made only from the upper reaches of the estuary. No definite correlation appears to exist between the occurrence of vibrios and the salinity fluctuations of the Hooghly water. The possibility of vibrios being adsorbed by the silt particles in the water and being deposited on the estuarinc bottom, where they thrive, was considered to be a fruitful line of investigation. Pending the development of a technique for the concentration of bacterial flora from a large quantity of silt, the top 1 4.1 sq. ft. was collected by hand, fr which about 10 g. were taken at No Vibrio cholere have so far been isolated, but NAG vibrios have been obtained. Out of a total of 102 samples examined, 44 were positive for vibrios No distinct relationship between the seasonal fluctuations in the occurrence of the two predominant groups, viz. groups 1 and II, and the seasons of cholera epidemics could be traced from the available data. Preliminary experiments were conducted to study the survival of vibrios in silt. "Silt collected from the Hooghly river bed was sterilized and crushed into a fine powder. In test-tubes containing 5 g of this silt, 10 cc. of water were added and live cultures of AG and NAG vibrios were inoculated into them, keeping suitable controls. Significant differences were not found in the survival of vibrios in the water and the silt, but it is necessary to repeat the experiments on a larger scale to arrive at any definite 'conclusions. Different size groups of hilsa ranging from the fry stage to the adult stage were collected during the seasons of their availability in the river Hooghly from the three selected centres every week and their gut-contents and gill-samples bacteriologically examined. During the off-season for. Hilsa, other estuarine fishes available 'were examined. No !''; cholera have so far been isolated, but NAG vibrios were of 204 ·8 samples of Hilsa, consisting of 648 specimens and 55 samples of other, fishes consisting of 268 specimens, the gut-contents of 76 samples of Hilsa and 34 samples of other fishes were positive. The gills of 77 samples of Hilsa and 26 samples of other fishes were also positive for vibrios. NAG vibrios have been found to occur in the fishes, either in the guts or : the gills, throughout the year. Groups I, II and V are the three predominant groups of NĂG isolated from the alimentary tracts of fishes. The seasonal fluctuations in the occurrence of groups 1 and II show a 'definite correlation with the periodicity of cholera in Calcutta. Parallel data for the gill samples also give a picture somewhat similar to the one obtained for the gut-contents. In view of the above relationship it seems that groups I and II vibrios are in some way connected with the maintenance of cholera endemicity. In the absence of better equipment, the pH of the gut-contents of Hilsa was studied by the spot-test method. The stomach-contents were found to be definitely acidic and the intestinal contents showed a pH ranging between 6 and 7. In view of the possibility that the gut-contents may be undergoing some chemical changes during the period that 'elapses between the capture of the fish and their dissection in the Laboratory, thus ' affecting the viability of the vibrios, similar samples of fish' were dissected in the field and in the Laboratory. No significant differences were noticed in the pH or the vibrio flora of these two sets of samples. As a preliminary to artificial infection experiments envisaged in the programme, it was necessary to perfect a method of collection, conditioning and rearing of Hilsa in confined areas. Hilsa is an extremely delicate fish and it was believed that it will die immediately after it strikes the net used for its capture. The settling tanks and filter-beds of the Calcutta Corporation Water Works at Pulta was considered the cleanest and most suitable source from which collections could be attempted. After a good deal of experimentation, it was possible to perfect a method of collecting young Hilsa and rearing them in cement cisterns 10' x 3'3'. Water from the settling tanks was fed into these cisterns and plankton was provided as fish food. In view of the possible difficulties in transporting the fish to Calcutta for the artificial infection experiments, it, was decided to conduct these in Pulta in cement cisterns at the premises of the Central Inland Fisheries Research Station. Before these experiments are undertaken,, it was, however, thought advisable to try some small-scale experiments with hardier fishes in the Laboratory in the form of pilot experiments. Kôi (Anabas testudineus) and Lata (Ophicephalus striatus) were employed for these studies. Specimens of these two species were reared separately, in ,,vibrio-free water in glass, aquaria and fed on vibrio-free food. The water. was, renewed daily and the old water examined bacteriologically, for the presence of vibrios. When the water provęd negative for three consecutive days, they were fed on infected food. Pupäe and larvæ of house-flies bred in the Laboratory and fed on vibrio-free milk were injected with live cultures, of, cholera vibrios and the fish were fed on these. Every day, the fish. were removed to fresh sterilized aquaria, after washing them several times in vibrio-free water. The water from the aquaria was examined daily, but no vibrios were recovered for over 30 days. The fish were then dissected and their intestinal contents bacteriologically examined. Hæmolytic 'non-agglutinable vibrios were recovered. 1 ·9 This result led to an examination of the possibility of the vibrios remaining in the guts without being excreted, as also of the artificially infected specimens having had vibrios in their alimentary tracts before they were infected After a consideration of possible means of checking up the vibrio-free nature of the gut of a'fish before it is artificially infected, it was decided to conduct an experiment to determine this. A sample of thirty Kôi (Anabas testudineus) was reared in a large tub in vibrio-free water which was changed every day. The fish were fed on boiled yolk of eggs regularly. Samples of 1,000 ct of water were examined from the tub daily for the presence of vibrios and after they were negative consecutively for over a week, ten specimens were taken at random and dissected to find out if vibrios were present in their guts One fish showed the presence of inagglutinable vibrios. The experiment is now being repeated with smaller number of fish and lesser quantity of water, so that the whole water in the aquaria can be examined bacteriologically and the possibility of vibrios having remained undetected can be ascertained Particular attention was directed to the study of the cholera cnzyme, mucinase 'present in the filtrates of V. cholere which has been reported to cause sloughing of the mucosa from strips of guinea-pigs ileum (Burnet & Stone, 1947). After a good deal of experimentation a method foi the quantitative titration of the enzyme has been standardized It has been observed that mucinase elaboration and the maintenance of quantitative activity are influenced by several factors, i.e. media, pH, moisture, cic. Twenty-seven 'strains of V. tholere (21 Ogawa sub-type and 6 Inaba sub-type) isolated from cholera patients and from Hooghly river water during the past two years were tested. With each strain the mucinolytic activity was evident at a titre of 1 in 1,600 or above. One strain, however, showed a very high titre of 1 in 25,600, and a second strain tested on four occasions during a period of six months gave a constant titic of 1 in 6,400. The presence of this enzyme, has also been demonstrated in certain intestinal organisms, i.e. B. coli type I, Salm. entertidis, and Sk flexneri, but the titre was of a very low order and never exceeded 1 in 10. In the course of this work a smooth strain of V. cholere became rough, and when tested later its mucinase-forming property was found to be very considerably lowered. It is presumed that this is true for all strains of 1', cholcre, if or when they become rough. Subsequently, investigations were carried out with 18 strains of NĂG vibrios. These were isolated from Hooghly 11... tr.. cruz 1 variation in the elaboration of the enzyme but the titre was low and ranged from 1 in 100 to 1 in 400. Thus, it appears that the intensity of mucinase activity is of greater importance rather than its mere presence. Since high titres have so far 'been obtained with true cholera vibrios, it appears that it may be possible to utilize the test as an adjunct to other known tests 'which characterize such i vibros.

·10 MALARIA. Insecticide and mosquito rcpcllcnt inquiry under the Director, Malaria Institute of India, Delhi. The following investigations were carried out in the Laboratory Il field il. SANOKATORY INVESTIGATIONS : 1. Solubility of DIT crystals in cuticular wax of housc-ſlics and 31/052uitrocs, 2. Resistant strain of Culex Foligans, 3. Susceptibility of DDT-resistant strain of Culex faligans to other rhlorinated hydrocarbxın insecticides. .). Effect of fccding normal and resistant strains of Culex ſaligans on DIT-Icc fowls. 5. Rapid meilios of screening DDT watcr-dispersiblc powder samples. B. FIELD INVESTIGATIONS : 1. Studies on tlic bchaviour of mosquitoes in rclation to insecticidal application. 2. Tivolution of DDT, BHC, DIT and BHC combined and dicldrin residual sprays against mosquitocs in dificrent parts of India. C. CONTIRMATION OF TINDIXGS INSECTICIDE AND MOSQUITO REPLIEST INQUIRY. PunLICATIONS. TIE A. LABORATORY INVESTIGATIONS. 1. Solubility of DDT crystals in cuticular wax of house-flies and mosquitoes-- Pal, (Bull. Ent. Res., 41, 1, pp. 121-139, 1950) sludicd thc wetting of insect cuticle by insecticidal liquids ; as thic cffcctiveness of almost all contact insecticides depends upon it. He found that apart from the chemical and physical naturc of the cuticular lipoids, irrcgularities on the body surface werc important. With the application of residual deposits of insecticides, however, thic position is completely different. Thcinsccticide is not availablc in tlic form of liquid but as crystals. Under such circumstances the rate and degree of dissolution of crystals in the cuticular wax is an important factor afccting thic toxicity of contact insecticides. ·:The object of the present investigation was to study the solubility of DDT in the cuticular wax or insects particularly those of public health importance. Cuticular wax of Musca ncbulo, Anopheles stephensi and Culex faligans were extracted in chloroform by soaking a large number of insccts in sufficicnt quantity of chloroform for 24 hours. The lipoid-chloroform solution was filtered through a cintered glass-funnel and the chloroform was allowed to evaporatc at room tempcrature. One c.c. of I per cent solution of cuticular wax in chloroform was uniformly applicd on the excavated portion of a cavity glass-slide (arca about 2.27 sq. cm.). Technical DDT specially pulverized was dusted on the wax-treated area of the slide after ensuring complete cvaporation of chloroform. .

·11 r.l. . mnen Aficio-photographs of the crystals on wax were taken without disturbing the slide, after 5 and 30 minutes, 1, 3, 6 and 24 hours. It was observed that the dissolution of crystals in the cuticular wax of Alusca nebulo and Anopheles stephensi started almost immediately and they almost completely disappeared in 3 and 6 hours, respectively. In the case of Culex fatigans crystals were observed intact even after 24 hours. This is of considerable interest because these observations are in conformity with the natural biological resistance of Culex fatigans to DDT residual deposits. 2. Resistant strain of Culex fatigans.--Reports of culicine mosquitoes having developed resistance to DDT have been received from several parts of the wold. In those states of India where DDT residual spraying has been in progress for the last six years of so, there is a possibility of certain culicine species having developed resistance to DDT. Investigations were undertaken in Delhi State where DDT has been in use since 1946. Culex fatigans were captured from a sprayed village Khuraiji Khas which had been regularly sprayed with DDT during the past six years, and from an unsprayed village Ram Garhi (U.P.) 10 miles apart. These two strains were reared for one generation simultaneously under identical conditions. The mosquitoes h) coated with er sq. ft. The Cent per cent knock-down of mosquitoes collected from insprayed village was obtained within five hours, whereas only 66 per cent (females) and 79 per cent (males) mosquitoes from sprayed village were knocked-down during that period. Even after 24 hours contact, only 78 per cent (females) and 24 per cent (males) mosquitoes were knocked-down. In subsequent tests the mosquitoes were exposed for half-an-hour to glass-panels sprayed with DDT at the rate of 50 mg./sq. ft. The exposed insects were kept under observation for 48 hours. Twenty experiments were carried out, mortality amongst the mosquitoes from sprayed village was nine per cent (females) and 31 per cent (males) after 48 hours, and the corresponding figures for the mosquitoes from unsprayed village were 90 and 93 per cent, respectively. 3. Susceptibility of DDT-resistant strain of Culex fatigans to other chlortnated hydrocarbon insecticides ---One object of this investigation was to determine whether or not DDT-resistant strain of Culex fatigans was also resistant to other chlorinated hydrocarbon insecticides. Both normal and resistant strains were exposed to glass-panels treated with DDT, chlordane, dieldrin applied at the rate of 50 mg./sq. ft. and BHC gamma isomer at the rate of 10 mg /sq ft. for 5 and 15 minutes, and were then kept under observation in clean cages for 24 hours after which the mortality was recorded. It was found that the mortality rate was comparatively low when the resistant strain was exposed to DDT, whereas in the case of other insecticides the mortality rates were high and almost identical to those of the normal strain. It seems that the DDT-resistant strain of Culex fatigans is initially not cross resistant to other insecticides tested. Terry ning os ·14 + (i) Since 1950, in the field experiments with different insecticides against mosquitoes, total mosquito-catch data from sprayed and unsprayed catching stations of the treated villages was found very useful in evaluating residual sprays (Pal, R. Ind. J. Mal., 5, pp. 195-199, 1951. Entomological observations from comparison village may sometime vary from the experimental villages due to some natural causes and hence are not always helpful in the assessment of results. Other workers (Rao, B. A. 1953, personal communication) have also expressed similar opinion and they also consider that entomological data from a comparison village is unnecessary; though it may provide useful epidemiological data. (iii) Resistance in terms of knock-down or mortality; Pal and Sharma (1951, Proc. IXth Internat. Gong. Entomol.) and Pal et al. (Ind. Four. Malariol., 6, pp. 303-316 1952), in their studies on the development of resistance to insecticides in housflies concluded that resistance of flies; should be specifically mentioned in terms of 'knock-down resistance and' mortality resistance ' because wide variations exist between the two. These findings have been confirmed by Busvine, J. R. (Nature, 171, pp. 118-119, 1953). D. PUBLICATIONS. 2. Inquiry to study nutritional states and their effects on mammalian malaria under the Director, Malaria Institute of India, Delhi. The object of the inquiry is to determine the effect of different nutritional states of the host (rats) on the host-parasite relationships of animals with P. berghei infections. It has been stated in the last year's report that multiplication of this parasite in starved host was not of the order as in normally fed hosts. Three possible reasons were considered, namely an increased resistance in the host to invasion of parasites, starvation of parasites themselves and ketosis in the host. The first possibility was ruled out as the parasites multiplied normally in infected hosts which were fed after a certain period of starvation. The second possibility was perhaps the most probable cause of infection not establishing itself during starvation. Infected animals were fed on PABA during starvation and the parasites became patent though their density was low. This seemed to show that PABA is probably one of the essential nutrilites for P. berghei and that it is not readily available in sufficient quantity to the parasites during starvation of the host. Similar experiments during the current year showed that methionine is another essential nutrilite not available during starvation. Thirdly, a highly ketogenic diet consisting of a high proportion of butter-fat was given to infected animals. The parasitæmia was less than in controls fed on normal diet and showed the probability of ketosis being an additional factor for the restricted parasitic growth in starved hosts. The age of the animal appears to be important in the consideration of its host-parasite relationship during its starvation. In two experiments it was found that when six-week old infected animals were starved, the parasites were patent in considerably higher numbers and for a longer period than in similarly treated adult animals. The high susceptibility, of young animals to infection seems to persist even when they are starved. Feeding infected and starved hosts on chemically purę nutrilites, either singly or in combination, has led to a better understanding of the ·15 physiological needs of parasites. The scope is wide in respect of vitamins, particularly with the availability of specific vitamin an.ogonists. EFFECT OF DIFFTRENT QUANTITIES OF THE SAMC DIET ON THE COURSE OF INFECTION IN AI BINO RAIS Six replicate experiments were carried out in 108 rats. The experimental animals were given half the quantity of standard diet for control animals. Under-nourishment, in so far as malaria is concerned, bears a two-fold effect: one on the malaria parasites and the other on the host's defence mechanism. The parasite in the under-nourished host grows and multiplies to a less extent than in a well-nourished host. An interesting feature is that due to impaired capacity of the host to develop immunity even mild infection tends to end fatally. Under-nourishment seems to greatly affect the capacity of the host to acquire immunity. If an infected under-nourished (or starved) host is fed normally the parasite reaps the benefit more quickly than the development of host-defence mechanism. Under such circumstances, the parasite multiplies rapidly both due to better available nourishment as well as the absence of adequate check by host immunity. More often than not the host dies. Such a sequence of events probably explains the high case mortality during malaria epidemics and in particular the one which occurred during the Bengal famine of 1943 It was noticed that institution of relief measures, at a time when fresh transmission was low or absent, resulted in greater number of deaths due to malaria. + EFFECT OF DIFFERENT QUALITY DIETS ON THE COURSE OF INFECTION IN RATS One hundred and fifty rats were used for the study which consisted of six experiments, some of which were replicates. The animals in each group, experimental as well as control, were mostly of the same age and sex distribution, The different diets used in the experiments were of three types representing vegetarian, lacto-vegetarian and mixed. It was ensured that these diets were not deficient in any of the vitamins or minerals. Vegetarian diets of a high-carbohydrate content, containing rice or wheat gave, rise to a more severe acute infection than normal balanced diet. Very few of the animals on such a diet died of chronic infection and the degree of acquired immunity was sufficient to nullify a challenge inoculation, Lacto-vegetarian diets consisting of milk and rice or wheat were responsible for a milder acute infection as compared to vegetarian diet, though the course of chronic infection was similar in animals on either of the above diets. Animals on a diet with a high proportion of meat suffered from a more severe, acute as well as chronic, infection than those on other diets. It seemed logical that it sould be so in view of the fact that such a diet probably provided, in abundance, all nutrients required by the parasite, far in excess of its capacity to enhance the acquirement of immunity by thc host. Among the different proteins constituting these diets, milk proteju more clearly established a host-parasite relationship in favour of the lost.

·18 alonc. is elic method of choice and in this inquiry attempts have been made to make the sprayinı; of the test pancis conform in a largc mcasure to what is obtained in malaria control operations in the field, without at the sine timc losing sight of the required necessities for laboratory spraying, such as capacity to cleliver cqual doses repcatedly and an assurance that all parts of thc ics panels are fairly uniformly covered. The use of an ordinary Hudson pneumatic sprayer with a fan-shaped nozzic make the equipment very nicar to conditions of spraying.. In the spraying equipment reviscol for spraying thic test pancls, an ordinary Hudson pressure sprayer is used with a 'fai-spray nozzle. The test panels move on canvas bei running over two metal rollers driven by an cicctric motor. The insecticidic is dischargerl at a pressure of 30-1b. to 28-1b, all which pressure the spraying of the panels is done and the pressure is not allowed to fall bclow. 28 lb. The nozzlc is adjusted so that ilu: width of the spray just touches the eyes of the panel and the entire surface is thus assured of a uniform spray. MITH01 01 WORKING. (1) The motor is switched on and the mumber of revolutions donc by thic canvas belt in a second is calculated. (2) The discharge rate of thc pump per sccond is calculated at 30- Ib. to 28-11), pressure. (3) The time taken for the bclt to travel a distancc cqual to the length of the panel is calculated. (4) Thc strength of the spraying fuid is adjusted to see that the required dosage is sprayed over the panel. In this inquiry pancis had to be spayed in two dosages : (i) 50 mg, per square foot. (ii) 200 mg. per square foot. This mcant that the pancls which measured 6" X 5.7" have to be sprayed so as to get 11.8 mg. of DDT for thic lower dosage or 47.2 mg. or DDT for the higher dosage. The time taken for the panel to travel its own length was found to he 0.5 second. In one second the sprayer, at 30-lh. to 28-1b). pressure, discharged 20 c.c. Therefore, in 0.5 sccond, the total discharge will be 10 c.c. Now, this 10 c.c. must be made to contain the 11.8 mg. or 47.27 mg. of DDT as the case may, bc so that the DDT may be uniformly sprayed over the test pancl. In this strength alone, 5,000 c.c. have to be prepared so that the pncumatic sprayer may work satisfactorily. A large zinc tray to contain the pncumatic sprayer and the moving belt was found usclul in collecting the spilt DDT emulsion or suspension at the time of the spray. TEST INSECTS. Test insects were being reared under comparative conditions with the same quantity of food supply and water. The pupã that develop on any one day have been utilized for the test. A. stephensi type has been used for all the tests recorded in this inquiry.

·19 STUDY OF SOIL TYPE. The Madras State is a part of the land surface of Indian peninsula. which has remained stable and undisturbed for untold geological ages The parent rock of the geological formation is responsible for the variable soils present, as for instance the difference between the black and red soils or if they are of alluvial origin on the rock formation in the catchment area of the river bringing them down and depositing them in the deltas The sol _The soil as ordinarily conceived consists of the following (a) Mineral materials. (6) Organic matter, (c) Water (d) Air. These, in the main, are in a fine state of subdivision and are intimately mixed. In fact, the contact is often so close as to render satisfactory scparation rather difficult. The mineral material has its genesis in the regolith or soil material. Some of the soil minerals have persisted more or less unchanged, while others have developed as the regolith weathered either before or during soil formation. Naturally, various sizes of particles occur ranging from those which are coarse, such as gravel and sand, to those, such as silt and clay which are in a fine state of division. The organic matter represent the accumulation of plant and animal residues and is generally in an active state of decay. An examination of any soil will reveal the presence of pore spaces of varying sizes. These occur not only between the large solid particles, but also between 'and within the clumps and aggregates which the fine particles tend to form. The pore spaces variable as to continuity, dimensions and total volume are occupied in large part by water and air, the proportion depending on the character of the soil and'the conditions under which it is functioning. Water, the third component of the soil is held within these pore spaces with varying tenacity due to certain surface forces. Soil water also carries many soluble salts. The principal gases of the soil air, the fourth component, are nitrogen, oxygen and carbon dioxide Volume and composition.—A silt-loam surface soil contains approximately 50 per cent of solids and 50 per cent of pore space. This pore space will be somewhat less for sandy soils and somewhat more for soils of a more clayey nature. The 50 per cent of solid space is occupied by about. 45 per cent of mineral and 5 per cent The proportion of air and water is, of under natural conditions depending on In short, the soil may be visualized as possessing a frame work of mineral matter—in part non-colloidal and inert; in part colloidal and extremely active. In mineral soils clayey materials make up much of this latter friction, but mixed with this viscous, galatinous mineral st ·20 partially adsorberi thercby, is a certain amount of lumus, notably colloidal iind unstable clay and humus, inore or less intimatcly mixed constituto most of the colloidal matter found in soils. This ſincly divided fraction not only promotes inctivity catalytically, but may also participatc intensively in the changes that occur. In fact the latter may react and bc reacıcd upon to the extent of losing its colloidal character in thc production of simple compounds. Adsorption, plasticity, colicsion and ccriain chemical changes are all controlled by substances in the colloidal statc. Whilc humic colliods arc the more active, the presence usually of a larger amount of viscous, gelatinous and more stable claycy matter preserves a balance in their relative importance. In Agriculture: fivc soil types rcferred to below are recognized in the andras Stalowme 1. The red soils. 2. Tlc black soils. 3. Coastal alluvium. 4. Delta soils. 5. Latcritc soils. Types 1, 3 and 5 are predominantly red, reddish or brown in colour, while types 2 and 4 arc dark coloured popularly described as black. Type I occurs over il large part of the Madras State and may be said to be the common type. It occurs in all sorts of situntions ranging from hill slopes to decp vallcys between hills in various parts of the State. Types 3 and 5 are also red or reddish brown, though they form different soil types from the point of vicw of thic soil chemist. Type 2 is a characteristic soil occupying, for the most part, the central platcau of the peninsula. Type , the delta soil, occurs mainly on the deltas of the rivers. The latcritc type 5 occurs in the West Coast and in some parts of thic East Coast. To start with, samples were obtained from sis arcas of the Madras State and the chemical analysis of the soils is furnislıcd in Table I :- TABLE 1. Chemical analysis of samples of earth. Araku, Chaga- Ennore. Dhim Mala- Tonjore Hcads of analysis. per 1111, per per bam, per bar, per per ccnt. cent. cent. ccnt. (a) Chemical analysis : Moisture 0.30 6.02 3.01 4.36 2.82 0.82 Less in ignition 23.95 6.53 1.81 11.61 4:43 3.05 Iron oxide (Fc,2°3) 23.84 3.41 1.96 6:17 4.60 2.18 Alumina (A1203) 46.73 7.60 3.95 12.87 8.71 7.43 Insolubles 5.6 8.5 91.14 68.74 81.98 87.08 (b) Mechanical(analysis : Clay 10.53 47:13 19.45 38.63 21.35 27.33 Silt 8.88 14:10 1:10 11.55 9.22 2.57 Finc sand 8.92 18.06 3.06 14.01 21.06 15.24 Coarse sand 68.92 15.86 75.60 33.48 48.34 56.61 Acid solubles 2.75 4.81 0.79 2.33 0.03 TOTALS 100.00 100.00 100.00 100.00 100.00 101.75 cent. cent, .. ..