Skin sensitivity of schoolchildren in Yugoslavia to different mycobacterial sensitins*

Bull. Org. inond. Sante 1971, 45, 657-666 Blill. Wid Hlth Org. Skin Sensitivity of Schoolchildren in Yugoslavia to Different Mycobacterial Sensitins* L. A. SIMEONOV I & Z. JAREC 2 Schoolchildren were tested on one arm with human sensitin RT 23 and simultaneously on the other arm with one of the sensitins prepared from atypical mycobacteria: non- chromogen, RS 10; photochromogen, RS 30; or scotochromogen, RS 64. Testing was made with low and with high doses of sensitins. Because of the extensive BCG vaccination programme in the country the number of persons not previously vaccinated with BCG in the study was very small. However, no indication of the prevalence of higher skin sensitiv- ity to atypical sensitins than to human sensitin wasfoun . Almost all the persons previously vaccinated with BCG who reacted to human sensitin showed a cross-reaction to the atypical sensitins. The cross-sensitivity was more pronounced in persons who were tested with scotochromogen sensitin and when they were tested with higher doses. This study represents one of a number of investi- gations of the prevalence of skin sensitivity to dif- ferent tuberculins (sensitins) amongst the population of the region of Bosnia, Yugoslavia. The results of previous investigations have been published else- where (Simeonov, 1964; Simeonov, 1967; Bleiker, 1969). Bosnia is a mountainous region with an average altitude of 500-1 000 m and has a population ofabout 4 million. The prevalence of bacillary pulmonary tuberculosis has been estimated, on the basis of a sample survey, as 2.7 per thousand, a higher figure than the average rate for the whole country of 2.4 per thousand (Teofanovic & Simeonov, 1967). The prevalence of tuberculosis infection is not easy to establish because of the extensive BCG vaccina- tion programme that started with the 1948-51 mass campaign assisted by the International Tuberculosis Campaign, in which about 2 million of the suscep- tible population of Yugoslavia (10% of the present population) were vaccinated (WHO Tuberculosis Research Office, 1952). Since the completion of the campaign a maintenance programme of BCG vac- * This study was partly supported by a grant from the Fund for Scientific Research of Bosnia and Herzegovina, Yugoslavia. I Epidemiologist, Tuberculosis Institute in Sarajevo, Yugoslavia. Present address: WHO Regional Office for South-East Asia, Delhi, India. 2 Senior Public Health Nurse, Epidemiological Depart- ment, Tuberculosis Institute in Sarajevo, Yugoslavia. cination and revaccination has been carried out and the coverage of the eligible population (tuberculin- negative, less than 25 years of age) at any point of time has been estimated as 62 % in the whole country and 60% in Bosnia (Vlajinic, 1967). Information on the spread of atypical mycobac- terial infection in Yugoslavia is very limited. Todo- rovic (1963) found that more than 50% of 328 flocks of chickens had been infected with Mycobacterium avium and that the percentage of reactors in poultry was 7% in the plains of Yugoslavia. He found stronger skin sensitivity to avian tuberculin in a considerable proportion of cattle in the same area. In some parts of Bosnia a large number of infected and sick fowls were found and Myco. avium was isolated from them (H. Huremovic, personal com- munication, 1968). Two strains of mycobacterium that showed characteristics of Myco. avium were isolated in a hospital from patients living in a region geographically resembling Bosnia. The patients were suffering from a lung disease with all the charac- teristics of pulmonary tuberculosis (Kozamernik & Gasi, 1965). Whereas previous studies had tried to establish the prevalence of infection with Myco. avium, 2 addi- tional sensitins were used in this study. Thus all the 4 main groups of mycobacteria were represented (Wayne, Runyon & Kubica, 1969). A preliminary report on the same study has already been presented elsewhere (Simeonov et al., 1969). 2758 - 657- 658 L. A. SIMEONOV & Z. JAREC MATERIAL AND METHODS Sensitins and doses The following sensitins were used in this study: (1) Human sensitin RT 23 with 0.005% Tween 80, in two doses: 0.02 ,g/ml (1 TU) (low dose) and 0.2 ,ug/ml (10 TU) (high dose). (2) Avian (non-chromogen) sensitin RS 10 with 0.005% Tween 80, in two doses: 0.1 ,ug/ml (low dose) and 1.0 [kg/ml (high dose). (3) Photochromogen sensitin RS 30 with 0.005 % Tween 80, in two doses: 0.1 ,ug/ml (low dose) and 1.0 ,ug/ml (high dose). (4) Scotochromogen sensitin RS 64 with 0.005% Tween 80, in two doses: 0.1 ,ug/ml (low dose) and 1.0 jug/ml (high dose). All the low doses were considered equipotent, as were all the high doses (Magnusson, 1961). All the sensitins were supplied by the Statens Seruminstitut, Copenhagen, in solution ready for application. They were refrigerated until they were taken into the field and they were used before their expiry date. The properties of the sensitins have been described by Guld et al. (1957) and by Magnusson (1961). Techniques All the tests were given intradermally using Omega Pasaic syringes with leakproof rubber piston rings. For each sensitin and for each solution a separate set of syringes and a number of platinum needles were used. All the tests were done by the same skilled nurse who read the tests 72 hours after the injections. Pairs of sensitins were tested simultane- ously, one on each forearm; one of each pair was RT 23. Low doses were used at first and persons who showed indurations less than 12 mm wide to both sensitins were retested with the high doses of the same sensitins. Retests were given in another area of the forearm. The diameters of the indurations were measured and registered as the size of reaction. For calculations of the mean size of reaction in persons not vaccinated with BCG, only;" reactors" (with indurations > 5 mm) were taken into account, whereas all the reactions of BCG-vaccinated persons were included, regardless of reaction size. ' 1 Technical guide for assessment of BCG vaccination pro- grammes, Geneva (unpublished document WHO/TB/Techn. Guide/2 Rev. 4.65). Population Schoolchildren aged 7-14 years from two villages close to each other were tested. The number of children tested with the different sensitins is shown in Table 1. Altogether 1 735 children were tested Table 1. Number of schoolchildren tested simultaneously with human sensitin RT 23 and one of the non-mammalian sensitins RS 10, RS 30, or RS 64 Test sensitin Persons Persons BCG status adeditin to tested with Itested withaddition to low doses high doses Unvaccinated RS 10 53 31 RS 30 69 41 RS 64 53 27 Sub-total 175 99 Vaccinated RS 10 554 393 RS 30 519 336 RS 64 487 290 Sub-total 1 560 1 019 Total 1 735 1 118 with low doses of sensitins and 1 118 with high doses. Only 175 children had not been previously BCG-vac- cinated; all the others had typical scars on the left shoulder following previous BCG vaccination. The children were randomly allocated to different sensitins, by school classes. RESULTS Persons not vaccinated with BCG The reactions to low doses of human sensitin RT 23 showed a bimodal distribution with a clear dis- tinction between non-reactors and reactors (Fig. 1A). Persons tested with high doses and who had indura- tions of 0-11 mm did not show a clear delimitation between reactors and non-reactors. The reactions to each atypical sensitin and correlations between these reactions and those to RT 23 are not shown because of the small numbers of unvaccinated per- sons. The distribution of reactions to RS 10 and RS 30 is shown in Fig. 1B. SKIN SENSITIVITY TO DIFFERENT MYCOBACTERIAL SENSITINS Fig. 1. Frequency distribution (%) of reaction sizes in persons not vaccinated with BCG, after testing with (A) human sensitin RT 23, (B) atypical sensitins RS 1 0 or RS 30. BCG-vaccinated persons whereas the mean reaction to ] The correlation between reactions to RT 23 and below that to RT 23 (Table 2). to atypical sensitins given in low doses was rather Reactions to high doses of poor (Appendix tables 1-3). Mean reactions to RS 10 fairly well with the reactions to and RS 30 were much lower than reactions to RT 23, sitins, in particular with that to RS 64 was not far RT 23 correlated all the atypical sen- RS 64, and the dif- Table 2. Mean diameters of reactions to different sensitins Low dose Neronso Mean persons (imm)e High dose No. of MeanS.D. esrs diameterpe n (mm) Persons not vaccinated with BCG 175 18.10 5.00 122 13.88 3.66 99 13.24 4.48 72 10.90 3.00 BCG-vaccinated persons 1 560 8.80 554 2.72 519 6.70 3.40 3.62 4.80 487 6.24 5.84 1 019 13.66 5.30 393 8.68 5.40 336 9.76 5.70 290 14.16 5.56 LOW DOSES HIGH DOSES A 30 a >°20 10 0 B 30 ~20(|,I X , 0 0 10 20 30 0 10 20 30 Diameter (mm) Diameter (mm) 341062 Sensitin RT 23 RS 10 or RS 30 S.D. RT 23 RS 10 RS 30 RS 64 659 L. A. SIMEONOV & Z. JAREC LOW DOSES A HIGH DOSES B C D 102D03(0 Diameter (mm) 10 20 30 Diameter (mm) WHO 01063 Fig. 2. Frequency distributions (%) of reaction sizes in BCG-vacci- nated persons, after testing with (A) human sensitin RT 23, (B) atypical sensitin RS 10, (C) atypical RS 30, (D) atypical sensitin RS 64. 30 X 20a) - 00 30 L- 10 0 30 aa lon L 0 30 20 a) 10 _ 0 30 - 2000m15 cL 10 1- 0 0 660 SKIN SENSITIVITY TO DIFFERENT MYCOBACTERIAL SENSITINS Table 3. Number of reactors to non-mammalian cinated with BCG. Because of this procedure any sensitins who had reactions more than 8 mm larger than persons infected with atypical mycobacteria would their reaction to human sensitin have been considered as reactors and should be Reactors to Reactors to found among the non-vaccinated tuberculin-positive BCG status Sensitin low doses high doses population; such persons are unlikely to have been No. % No. % BCG-vaccinated. If the prevalence of atypical infec- tion has been high, the distribution of reactions toUnvaccinated RS 10 - - - - 1 TU of RT 23 with 0.005% Tween 80 would have RS 30 1 1.7 - - shown a large number of small and medium-size RS 64 - - - - reactions; no such result was found in this study, however (see Fig. IA). Vaccinated RS 10 2 0.4 3 0.7 As the numbers of unvaccinated persons testde RS 30 2 0.4 1 0.3 with atypical, sensitins was small, the reactions to RS 64 13 2.6 12 4.1 RS 10 and RS 30 were pooled together. These two groups were pooled in this way because of the similarity of the sensitivity induced by these two ferences between them in reaction size were not large. sensitins in BCG-vaccinated persons. There was The distribution of the reactions to low doses of poor correlation between the reactions to RT 23 and RT 23 is typical for a BCG-vaccinated population the pooled reactions to RS 10 and RS 30, both to tested a long time after BCG vaccination-a bimodal the low and to the high doses (Table 4); the reac- shape with no clear separation between the first tions to RT 23 were much stronger than reactions mode of non-reactors and the second mode of to RS 10 and RS 30 (Table 2 and Fig. 1). There reactors (Fig. 2A). Between the reactors and the was no separate group of reactors with larger reac- non-reactors are those with small and medium-size tions to RS 10 or RS 30 than to RT 23, thus there reactions representing the BCG-vaccinated children was no evidence of the existence of reactors with whose sensitivity has already waned. The distribu- higher sensitivity to atypical sensitins among per- tion of reactions to the low dose of RS 64 is similar sons not vaccinated with BCG. The same conclu- (Fig. 2D). The distribution patterns of reactions to sions were reached after previous investigations with low doses of RS 10 and RS 30 are skewed and do not the avian sensitin RS 10, and the Weybridge avian show any typical shape (Fig. 2B and 2C). tuberculin (Simeonov, 1964, 1967; Bleiker, 1969). The distribution patterns of reactions to high Whereas there was some variation in the way BCG- doses are more or less bimodal, symmetrical in the vaccinated children reacted to RT 23 and to the cases of RT 23 and RS 64 and skewed with RS 10 other sensitins given in low doses, the similarity of and RS 30; the modes of the distribution patterns their reactions to high doses was remarkable, especi- of the two latter sensitins are at lower values than ally the similarity in reactions to RT 23 and to with RT 23 and RS 64. RS 64. In spite of the fact that all 4 sensitins had Table 3 shows the numllbers of children in whom shown definite specificity in experimental animals the reaction to the atypical sensitin was at least (Magnusson, 1961), they showed a large number of 8 mm larger than the reaction to RT 23. This dif- cross-reactions when given in high doses to BCG- ference was chosen arbitrarily, based on the experi- vaccinated children. Very few children showed larger ence of other investigators (Hart et al., 1962). reactions to the atypical sensitins than to RT 23 and these results may have been the result of experimental DISCUSSION Data from investigations in cattle and poultry indicate that there is a potential risk, for the popula- tion of Yugoslavia, of infection with non-mammalian mycobacteria. At the beginning of the BCG vaccina- tion programme it was decided that all persons eligible for BCG vaccination or revaccination were to be tuberculin-tested. This has been done since 1960 with 3 TU of RT 23 with 0.0050% Tween 80. Those who show reactions of less than 5-6 mm are revac- error. In conclusion, it is clear that there is no evidence that infection with mycobacteria antigenically akin to the atypical mycobacteria belonging to groups of non-chromogens, photochromogens and scotochro- mogens is prevalent in Bosnia. Atypical sensitins elicited cross-reactions with the human sensitin RT 23 that were more pronounced in BCG-vaccinated per- sons, in persons tested with the scotochromogen sensitin RS 64, and in persons tested with high doses. 661 662 L. A. SIMEONOV & Z. JAREC Table 4. Distribution of reactions by size: persons not vaccinated with BCG who were tested with low doses (above) and high doses (below) of RT 23 and RS 10 or RS 30 Reaction Reaction to RT 23 (mm) to RS 10 Total or RS 30 (mm) 0-1 2-3 4-5 6-7 8-9 10-11 12-13 14-15 16-17 18-19 20-21 22-23 :24 0-1 61 2 - - 1 2 1 - 1 - - 1 - 69 2-3 1 1 - - - 2 - 1 - - 1 - - 6 4-5 - - 1 - - - - 2 1 - 3 2 - 9 6-7 - - - 1 - - - 1 - - - - 2 8-9 - - - - - - 1 - - - 1 - 2 10-11 - - - - - - - 1 - 1 1 - 1 4 12-13 - - - - - - 1 2 1 3 1 - - 8 14-15 - - - - - - 2 2 - 4 - - 1 9 16-17 - - - - - - - - 4 2 - 2 - 8 18-19 - - - - - - - - - - - - 2 2 20-21 - - - - - - - - - 1 1 - - 2 22-23 _- __ _ >24 - - - - - - - - - - - - 1 1 Total 62 3 1 1 1 4 4 10 7 11 7 6 5 122 Reaction Reaction to RT 23 (mm) to RS 10 Total or RS 30 (mm) 0-1 2-3 4-5 6-7 8-9 10-11 12-13 14-15 16-17 18-19 20-21 22-23 >24 0-1 41 2 - 2 - - 1 - - - - - - 46 2-3 2 - - - 1 - 1 - - - - - 4 4-5 - - - 1 1 - - - - - - - - 2 6-7 - - - 1 - 1 - - 2 - - - - 4 8-9 - - - - 1 - 2 1 4 10-11 - 1 - - 1 1 2 - - - 1 - - 6 12-13 - - - - - - - 2 - - - - 2 14-15 - - - - - 1 - 1 1 - - 3 16-17 - - - 1 - - -1 1 Total 43 3 4 3 4 4 4 4 1 2 - - 72 RESUME SENSIBILITE CUTANEE A DIFFERENTES SENSITINES MYCOBACTtRIENNES CHEZ DES ECOLIERS DE YOUGOSLAVIE Pour la presente enquete, 1735 ecoliers de Yougoslavie tines ci-apres: a) une sensitine de mycobacterie aviaire ont e repartis sur une base aleatoire en trois groupes qui non chromogene (RS 10) avec Tween 80; b) une sensitine ont subi simultanement une epreuve intradermique avec de mycobacterie photochromogene (RS 30) avec Tween une faible dose de tuberculine humaine RT 23 avec 80; et c) une sensitine de mycobacterie scotochromog&ne Tween 80 et avec une faible dose de l'une des trois sensi- (RS 64) avec Tween 80. Les sujets qui ont reagi par une SKIN SENSITIVITY TO DIFFERENT MYCOBACTERIAL SENSITINS 663 induration de diametre inferieur a 12 mm ont subi un nouveau test avec une dose 10 fois plus elevee des memes sensitines. En raison de la generalisation de la vaccination par le BCG au cours des ann&es anterieures, la proportion des enfants non porteurs de cicatrices de BCG dans les groupes etudies n'etait que de 10% environ. Les resultats des epreuves n'ont fait ressortir aucune prevalence de l'infection par des mycobacteries atypiques appartenant aux groupes des mycobacteries non chromo- genes, photochromogenes et scotochromogenes. On a note un certain nombre de reactions croisees a la tuber- culine RT 23 et aux sensitines de mycobacteries atypiques. Ces reactions etaient particulierement frequentes chez les sujets vaccines par le BCG, chez ceux qui ont subi une epreuve cutanee avec la sensitine RS 64, et chez les enfants qui ont e testes avec de fortes doses. REFERENCES Bleiker, M. A. (1969) Bull. int. Un. Tuberc., 42, 65 Guld, J., Bentzon, M. W., Bleiker, M. A., Griep, W. A., Magnusson, M. & Waaler, J. (1958) Bull. Wld Hlth Org., 19, 845 Hart, P. D'Arcy, Miller, C. L., Sutherland, I. & Lesslie, 1. W. (1962) Tubercle (Edinb.), 43, 268 Kozamernik, M. & Gasi, A. (1965) In: Report of the VIII Conference of Microbiologists, Ljubljana, Yugo- slavia Magnusson, M. (1961) Amer. Rev. resp. Dis., 83, 57 Simeonov, L. (1964) Acta tuberc. pneumol. scand., 44, 39 Simeonov, L. (1967) Z. Tuberk., 127, 89 Simeonov, L., Vinik-Kos, L., Jarec, Z. & Jovanovic, D. (1969) In: Report of the International Symposium on Atypical Mycobacteria, Berlin-Buch Teofanovic, S. & Simeonov, L. (1968) Probl. Tuberk., No. 9, p. 12 Todorovic, L. (1963) Tuberkuloza, 15, 293 Vlajinic, S. (1967) Tuberkuloza, 21, 497 Wayne, L. C., Runyon, E. H. & Kubica, C. P. (1969) Amer. Rev. resp. Dis., 100, 733 WHO Tuberculosis Research Office (1952) Mass BCG vaccination in Yugoslavia, Copenhagen, International Tuberculosis Campaign 664 L. A. SIMEONOV & Z. JAREC Appendix table 1. Distribution of reactions by size: BCG-vaccinated persons tested with low doses (above) and high doses (below) of RT 23 and RS 10 Reaction Reaction to RT 23 (mm) to RS 10 Total (mm) 0-1 2-3 4-5 6-7 8-9 10-11 12-13 14-15 16-17 18-19 20-21 22-23 >24 0-1 188 22 19 23 15 9 20 19 4 4 1 - - 324 2-3 5 7 7 6 9 7 8 12 7 1 2 - - 71 4-5 4 2 3 4 6 4 15 12 5 4 - - - 59 6-7 1 2 1 1 2 8 6 10 3 3 - - - 37 8-9 1 - - 2 3 3 4 7 7 4 1 - - 32 10-11 - - - - - - 1 2 - 7 1 1 - 12 12-13 - - - - - - 1 2 2 3 2 1 - 11 14-15 - - - 1 - - 2 1 1 - 1 - - 6 16-17 - - - - 1 - 1 18-19 - - - - - - - - - - 1 - - 1 Total 199 33 30 37 35 31 57 65 30 26 9 2 - 554 Reaction Reaction to RT 23 (mm) to RS 10Toa(mm) 0-1 2-3 4-5 6-7 8-9 10-11 12-13 14-15 16-17 18-19 20-21 22-23 >24 T 0-1 24 1 3 2 10 8 8 6 2 2 - 1 - 67 2-3 2 - 1 4 2 3 4 4 1 - - - 1 22 4-5 - - 2 5 3 4 7 5 3 3 1 - - 33 6-7 - - - 1 1 5 6 11 2 3 1 - 1 31 8-9 - 1 - 1 - 11 9 13 9 3 2 - 1 50 10-11 1 - 1 - 1 4 11 19 10 4 - - 3 54 12-13 - 1 - - 2 2 8 14 17 9 2 - - 55 14-15 - - - - 1 - 4 6 15 11 6 4 2 49 16-17 - - - - 1 - - 4 7 3 1 1 1 18 18-19 - - - - - - - - 4 1 4 - - 9 20-21 - - - - - - _ - - - - - 1 1 22-23 - - - - - - - 1 - - - 1 1 3 >24 - - - - 1 - 1 Total 27 3 7 13 21 37 57 83 70 39 17 8 11 393 SKIN SENSITIVITY TO DIFFERENT MYCOBACTERIAL SENSITINS 665 Appendix table 2. Distribution of reactions by size: BCG-vaccinated persons tested with low doses (above) and high doses (below) of RT 23 and RS 30 Reaction to RS 30 - (mm) Reaction to RT 23 (mm) 0-1 2-3 4-5 6-7 8-9 10-11 12-13 14-15 16-17 18-19 20-21 22-23 >24 Total 165 15 18 16 21 14 4 4 5 5 3 8 4 - 8 3 4 3 2 - 2 1 4 3 - - 2 1 2 3 - 1 - 1 2 2 - - 1 - - 1 1 18 23 7 8 6 2 7 5 5 8 9 4 6 4 3 1 8 4 1 2 3 2 5 2 - 2 3 _ 1 1 1 - - - 298 2 1 - - 48 2 - - - 39 3 - _ - 36 2 - _ - 23 7 - - 1 27 5 - _ 1 2 4 - - 1 16 6 1 - 2 14 - - 1 1 3 - 1 - - 2 1 1 Total 175 20 34 27 37 34 51 65 34 32 3 1 6 519 Reaction Reaction to RT 23 (mm) to RS 30 Total (mm) 0-1 2-3 4-5 6-7 8-9 10-11 12-13 14-15 16-17 18-19 20-21 22-23 >24 0-1 24 3 1 5 5 5 7 3 3 4 1 1 - 62 2-3 - - 1 1 3 - 2 - - - - - - 7 4-5 - - 1 2 1 2 3 3 2 - - - - 14 6-7 - - - - 6 2 7 5 2 1 - - - 23 8-9 - - - - 3 1 9 10 6 - 1 1 - 31 10-11 - - - - - 4 4 14 10 7 1 1 - 41 12-13 - - - - 1 - 5 22 13 8 2 - - 51 14-15 - - - - - 2 5 14 19 16 3 - 1 60 16-17 - - - - 1 - 1 7 12 5 1 - 1 28 18-19 - - - - - - 2 3 3 4 4 1 - 17 20-21 - - - - - - - - - - - - 2 2 Total 24 3 3 8 20 16 45 81 70 45 13 4 4 336 0-1 2-3 4-5 6-7 8-9 10-11 12-13 14-15 16-17 18-19 20-21 22-23 666 L. A. SIMEONOV & Z. JAREC Appendix table 3. Distribution of reactions by size: BCG-vaccinated persons tested with low doses (above) and high doses (below) RT 23 and RS 64 Reaction to RS 64 (mm) 0-1 2-3 4-5 6-7 Reaction to RT 23 (mm) 8-9 10-11 12-13 14-15 16-17 18-19 20-21 22-23 >24 131 12 5 6 11 7 5 4 5 1 7 7 4 3 4 3 2 - - 4 3 1 1 2 - 2 - 1 - 1 - 1 2 - 6 3 2 6 3 2 4 5 4 4 5 2 4 3 3 5 2 6 8 4 9 10 5 11 16 1 7 10 1 2 12 - 1 4 - 2 4 2 3 2 6 5 6 9 3 3 2 1 1 - _ _ 1 - 1 - - 1 1 - 1 - - - 1 1 1 - 7 1 - - 2 2 1 1 2 - 1 2 3 - - 1 Total 158 27 22 28 26 26 47 77 43 20 5 4 4 487 Reaction Reaction to RT 23 (mm) to RS 64 Total(mm) 0-1 2-3 4-5 6-7 8-9 10-11 12-13 14-15 16-17 18-19 20-21 22-23 >24 0-1 15 - 2 1 1 - - - - 1 - - - 20 2-3 1 1 - - 1 1 - - - - 4 4-5 - - - - - - - - - - - - - - 6-7 - - - - 1 1 1 1 1 - - - - 5 8-9 - - 2 2 1 3 2 1 1 - - - - 12 10-11 - - 2 1 2 1 6 10 1 3 - - - 26 12-13 - 1 - 5 3 5 10 13 3 3 - - - 43 14-15 - - - - 4 3 17 19 8 7 - - - 58 16-17 - - - 1 2 2 2 16 13 9 - - 1 46 18-19 - - - - - 1 4 11 9 11 4 - - 40 20-21 - - - - - - 2 2 3 3 3 4 1 18 22-23 1 - - - - - - - 4 2 1 - 1 9 24-25 - - - - - - 1 1 1 1 2 - 1 7 26-27 - - - - - - - - - - 1 1 28-29 _- - - - - - - >30 - - - - - - - - - 1 - - - 1 Total 17 2 6 10 15 16 46 74 44 41 10 4 5 290 Total 0-1 2-3 4-5 6-7 8-9 10-11 12-13 14-15 16-17 18-19 20-21 22-23 >- 24 176 43 37 39 40 47 34 34 14 14 8 1