The incidence of infection with mycobacterium tuberculosis is an index of the risk of infection to which a community is exposed. An accurate estimation of incidence rate is of considerable importance in understanding the epidemiology of tuberculosis in organising control measures. A new method of estimating incidence of infection is discussed. The material from 3 studies of National TB Institute has been utilized. Study I: is a part of a survey of a random sample of 134 villages. No previous tuberculin testing or BCG vaccination had been carried out in the area, but each person was examined for BCG scar in order to exclude persons vaccinated probably from other areas. After a complete census, a Mantoux test with 1 TU of PPD RT 23 with Tween 80 given on two occasions (Round I and II). Those with reaction of 13mm or less at Round I were offered a test with 20 TU with Tween 80 within a week of 1 TU test. The interval between the rounds was about 18 months. From the analysis of the data from the first 50 villages for which complete information for both rounds was available, it was seen that there was a general increase in the size of reactions elicited in the second round. Study 2: tuberculin testing was carried out with 1 TU and 20 TU among selected ‘control’ groups which provided the data regarding the “enhancing of tuberculin allergy” seen in repeat tuberculin tests. Study 3: in the course of the longitudinal “survey reader assessments” were carried out periodically to judge the standards of the tuberculin test readers. Inter & intra-reader comparisons were made. The findings have been used to estimate the magnitude of reader variation. The data was also used to study variations in the technique of testing and reading.

It was estimated that on an average inter & intra-reader variations between the rounds were unlikely to exceed 6mm or more in more than 5% of the observations. The reading errors have an equal chance of being positive or negative except at extreme ends of the distribution where zero readings of Round I can only show an increase, and the very large reactions had a greater chance of showing only a decrease at a subsequent round. The study mainly concerns with the problems of estimating the incidence of tuberculous infection in a community. Calculations based on age-specific prevalence rates or on rates of tuberculin conversion or both subject to gross error, leading to unreliable epidemiological conclusions. For estimating the newly infected, a new approach has been suggested based on the drawing of a curve for the distribution of differences in reaction size from one round of tuberculin testing to another. It is assumed that if new infection causes a distinct rise in the degree of tuberculin sensitivity which is greater than the combined rise due to enhancement and reader variation, the distribution of differences between the rounds should indicate the newly infected. It is shown that the newly infected probably constitute a homogeneous group with an increase in mean reaction size of about 24mm and standard deviation of 4mm. Accordingly, 98% of the newly infected show an increase in reaction size of 16mm or more.

Tuberculin tests repeated after an interval of time, at a different site have been reported to elicit reactions larger than the first test. A study was undertaken where reactors of 13mm or less to 1 TU have been tested with 20TU for the study of low grade reactions. Study was carried out in a previously untested and unvaccinated rural population (Longitudinal Survey), where only about 25% of the population showed 14mm or more to 1 TU and the remaining about 60% showed 10mm or larger reactions to 20 TU. These results confirm the high prevalence of non-specific allergy in the area.

It was found that a tuberculin test does enhance the allergy elicited by a subsequent test. The enhancing effect is associated with the initial allergy i,e., 8-13mm to 1 TU tuberculin, especially those elicited by a 20 TU test, increase being almost confined to those with 10mm and larger reactions to 20 TU. The enhancing effect increases with increase in age especially among those with 10mm or bigger reactions to 20 TU. It is possible that the enhancing effect is more in communities with high prevalence of non-specific allergy.

Studies on the distribution of tuberculous infection and disease in households have mostly been restricted to the examination of contacts of known cases. Clinical experience has lead to a strong belief that tuberculosis is a family disease and contact examination is a “must” for case-finding programmes. A representative picture of the distribution of infection and disease in households can be obtained only from a tuberculosis prevalence survey.

This paper reports an investigation, based on a prevalence survey in a rural community in south India. The survey techniques and study population have been described in an earlier report. Briefly, the defacto population was given a tuberculin test with 1 TU of PPD RT 23 with Tween 80 and those aged 10 years and above were examined by 70mm photofluorography. All the X-ray pictures were read by two independent readers. Those with any abnormal shadows by either of the two readers were eligible for examination of a single spot specimen of sputum by direct smear and culture. The defacto population numbered 29,813 and tuberculin test results were available for 27,115. After excluding BCG scars, the study population of 24,474 was distributed over 5,266 households which were further classified as “bacillary case household” with atleast one bacteriologically confirmed case, “X-ray case household” with atleast one radiologically active case but with no bacillary cases and ‘non-case household’ with neither a bacillary nor an X-ray case. Total bacillary cases were 77 and were distributed in 75 household. 74 households had one case each and one household had 3 bacillary cases.

The findings of the study have thrown considerable doubt on the usefulness of contact examination in tuberculosis control; (1) over 80% of the total number of infected persons, in any age group, occurred in households without cases, (2) cases of tuberculosis occurred mostly singly in households, and the chance of finding an additional case by contact examination in the same household is extremely small, (3) a common belief has been that prevalence of infection in children in 0-4 age group is a good index of disease in households, but in this study about 32% of households with cases of tuberculosis had no children in this age group, (4) in houses with bacteriologically confirmed case only 12% of the children in 0-4 age group showed evidence of infection, a possible explanation of such a low intensity of infection could be that there is resistance to infection. It is well known that some children even after repeated BCG vaccination do not become tuberculin positive. It is felt that a large number of children do inhale tubercle bacilli, but a primary complex does not develop or even if it develops, the children remain tuberculin negative. A hypothesis has been made that in addition to resistance to infection, there is something known as “resistance to disease”. Otherwise, it is difficult to explain why under conditions of heavy exposure in infection, only some individuals develop evidence of infection and very few develop disease thereafter.

The report describes the changes that occurred during second survey carried out after an interval of one and half years in the cases diagnosed at the first survey done during 1961-62 from among a total population of about 62,000 in 119 villages in Bangalore District. It was observed that (1) Of the 62 sputum smear positive cases also having suggestive chest X-ray shadows, 34% had died, 35% were sputum positive and 31% had become culture negative after 1½ years. Of the 10 smear positive cases who were X-ray normal, non-e was culture positive at the start and 7 were negative by culture and smear after 1½ years. Of the 67 scanty smear positive cases (1 to 3 bacilli seen), only 3 were sputum positive, 10 were having X-ray shadows and half were tuberculin negative after 1½ years. (2) Of the 88 culture only positive cases (20 or more colonies and with X-ray evidence of disease) 31% had died and 47% continued to be sputum positive after 1½ years. A much smaller proportion of these changes occurred among culture positive cases with less than 20 colonies. (3) There were 457 persons having radiologically active tuberculosis on the basis of interpretation of a single X-ray picture by two independent readers but whose sputum were negative for AFB (suspect cases). Of these, 38% were tuberculin negative also. Of those suspect cases who were tuberculin positive, 9% become sputum positive after 1½ years, while only 2% of the tuberculin negative suspect cases became sputum positive.

It is concluded that there is a lot of variation in fate among the different categories of cases of pulmonary tuberculosis. Further, attention has been drawn to the possibility of self healing in about 30% of the bacillary cases after 1½ years.

Generally there is no acceptable definition of the term “case of pulmonary tuberculosis”, although such a definition is of fundamental importance both in clinical medicine where results of various chemotherapeutic regimens are compared, as well as for the comparison of different epidemiological data. The main purpose of this paper is to focus attention on the difficulties of defining a case on the basis of bacteriological examination, X-ray examination and tuberculin test. Data from two successive prevalence surveys in a random sample of 134 villages in Bangalore district with a population 70,000 have been utilized to illustrate some of the difficulties in defining a “case” of pulmonary tuberculosis for reporting the prevalence or incidence of the diseases. The entire population was tuberculin tested with 1 TU RT 23 with Tween 80 at both rounds and those 5 years of age and older were examined by 70mm photofluorogram. The sputum specimens (spot and overnight) were collected from those with any abnormality on X-ray as recorded by either of the two independent readers. Both the specimens were examined by fluorescent microscopy and Ziehl-Neelsen technique and by culture.

Analysis of data has shown that the term “a case of pulmonary tuberculosis” does not represent a single uniform entity, but embraces cases of several types, differing considerably in their tuberculin sensitivity, results of X-ray and sputum examination, in the reliability of their diagnosis and mortality experience. The status of cases found at initial and subsequent surveys showed changes with time, and such changes show considerable differences for the various types of cases. It was felt that a single straight-forward definition of a case was not possible to suit all situations. One has to use more than one definition. Although theoretically, finding a single bacillus in sputum should be adequate proof of pulmonary tuberculosis, it was shown that finding of a few bacilli (3 or less) was very often due to artifacts and should not be the basis for a diagnosis. It has also been found that positive radiological findings, in the absence of bacteriological confirmation, indicate only a high risk of the disease and not necessarily pulmonary tuberculosis. Direct microscopy appears to be a consistent index of disease but in community surveys has the limitation of missing a substantial proportion of cases and of adding some false ones.

In view of the difficulty of providing a single definition of a case of tuberculosis, four indices have been suggested. (1) Cases definitely positive by direct smear; (2) Cases definitely positive by culture; (3) All cases positive by culture (including less than twenty colonies); (4) Sputum positive cases which are radiologically active. Each of these could be used for different situations. However, it was concluded that, there seems to be no option but to use more than one definition for assessing the prevalence and incidence of disease.

Prevalence surveys are useful for estimating the tuberculosis problem in different countries. Three techniques are commonly used in surveys, tuberculin test, mass miniature radiography and sputum examination. Each has its own limitations. A limitation of sputum examination is that all the sputum positive cases in the community cannot be diagnosed when only one sample of sputum is examined from each eligible person. Multiple sputum examinations are not often possible under field conditions of surveys covering the whole community. It would be worthwhile to have some idea of the extent of under-diagnosis in sputum examination. For this purpose, during an epidemiological survey, four specimens of sputum were collected within seven days of X-ray examination from each person with an abnormal chest X-ray in 30 villages of a district of south India. Each specimen was examined by Fluorescent Microscopy (FM), Ziehl Neelson (ZN) technique and culture.

There were 34 culture positive cases among 2,164 persons for whom all the four culture examination results were available. Of them, 21 (62%) were found positive on one specimen. The second specimen increased the positivity to 32 (95%). Thus, for detecting both smear and culture positive cases two specimens are adequate. A third specimen is helpful for detecting cases positive by culture alone. An estimate of prevalence obtained from one sputum specimen can be estimated for the prevalence obtained from many specimens by applying correction factor of 1.67 and estimates based on two specimens by applying 1.26. Of the remaining 37 smear positive cases detected by one specimen, 20 were smear positive and culture negative. Of the remaining 17 smear positive and culture positive, 14(82%) were detected by one smear examination only.

ZN positives not confirmed by culture (mostly with less than four bacilli reported in the smear) increased from 7 from the first specimen to 18 from all four specimens, while positives confirmed by culture method showed only a marginal increase from 13 to 15. FM did not have this disadvantage as only two were culture negative among the 18 smear positive results by FM method. Examination of two specimens by FM detected about 95% of cases demonstrable by this method. But with the ZN technique additional specimens may add more “false positives”. Thus, for detecting cases both smear and culture-positive two specimens appear adequate. A third specimen is helpful for detecting cases positive on culture only.

Data from 62 randomly selected villages in a district of south India, which formed part of a prevalence survey carried out by the National Tuberculosis Institute, Bangalore, during 1960-61, has been made use of. The survey covered 29,813 persons in 5,266 households. There were 70 cases with bacilli demonstrable either in smear or culture and 300 suspect cases. Using the village map (prepared by survey staff), ‘case clusters’ were formed first, with each case household as nucleus and adjacent households within a maximum distance of about 20 meters on either side of the case households. Households closest to the nucleus household on either side have been called as 1st neighbourhood and those coming next in proximity on either side as a 2nd neighbourhood and so on. The case household and its four neighbourhood together was called a cluster. If another case household was found within 4th neighbourhood of the first case the cluster was extended by including the 4th neighbourhood of the new case also. Such clusters were called composite case clusters and clusters with only one case household as simple case clusters. Similarly, suspect case clusters were formed and differentiated as simple suspect clusters or composite suspect clusters. Further, to serve as a control group, non-case clusters were constituted from a systematic sample of 10% households that were not included in case or suspect case clusters.

Out of 60 case clusters formed, only 7 have multiple cases showing that there was no evidence of high concentration of disease in case clusters. While the percentage of child contacts (0-14 years) infected was considerably higher in case clusters (25.8%), there was not much difference between suspect case clusters (14.9%) and non-case clusters (9.8%). Similarly, there was not much difference between simple and composite clusters. Infection among child contacts was higher in case households as compared to their neighbourhoods. To get some idea of the zone of influence of a case or suspect case, prevalence of infection was studied for 10 neighbourhoods, in simple clusters to avoid the influence of multiple cases. It appeared that the zone of influence of a case may extend at least upto the 10th neighbourhood. It was also noted that there was very little difference between zones of influence of suspect cases and non-cases. Case clusters in which the nucleus case had shown activity of lung lesion (evident on X-ray reading) or had cough showed significantly higher infection among child contacts. Clusters around cases positive on both smear and culture did not show higher infection than those around cases positive on culture only. (This may be due to sputum examination of single specimen only).

Out of the total infected persons in the community, only 2% were in case households and 7% in suspect case households, over 90% being in non-case households. The zone of influence of a case extending at least upto the 10th neighbourhood and the overlapping of such zones of influence of cases, present and past, seems to be the most probable explanation for the wide scatter of infection in the community. Prevalence of infection among child contacts was definitely higher in case clusters. But, the significance of this could be understood only from a study of the incidence of disease during subsequent years in different types of clusters. It is significant that only 10% of the total infected persons in the community were found in case clusters. The case yield in general population, cluster contacts, household contacts and symptomatics attending general health institutions have been also compared. The case yield in the last group (10%) is much higher than the case yield from both types of contacts (0.7% and 0.6%) which where only slightly higher than the case yield from the general population (0.4%).

This report is based on the study of 735 cultures of tubercle bacilli identified as human type. Sputum specimens were collected from patients attending the Lady Willingdon Tuberculosis Demonstration and Training Centre (LWTDTC), Bangalore, and from the mass Case-finding studies in semi-urban areas. Drug sensitivity tests for streptomycin, isoniazid, PAS and thioacetazone with different drug concentrations, different size of inoculum and for various length of incubation were carried out.

No difference was observed in the duration of growth between sensitive and resitant cultures in their first appearance on primary diagnostic cultures or sub-cultures on drug free slopes when innoculated with standard suspension. The primary cultures took about 3 weeks and sub-cultures 2 weeks to grow on drug free media. Large sensitive bacillary population required higher concentration of thioacetazone to inhibit the growth, suggesting standardization of inoculum size for sensitivity tests. Prolonged incubation period on drug slopes showed profound influence on the level of drug inhibiting concentration of thioacetazone; with the increase in incubation period, fall in growth of sensitive culture was not observed even on high drug concentration. The reproducibility of this observation on duplicate specimens from the same patients after shorter intervals excluded the possibility of experimental error. A reduction in the inhibition of growth of sensitive organisms on drug media with time is presumed to be due to either deterioration of the drugs in the media or due to adaptation by the micro-organisms. Because of the decrease in inhibition of growth, even sensitive organisms may be classified as resistant if reading of culture for drug sensitivity is prolonged beyond 3 weeks of the inoculation period. It is suggested that a standard inoculum size and a maximum limit of 3 weeks incubation period should be adopted for finding out sensitivity to thioacetazone. Cultures classified as sensitive to the three first line drugs or resistant to one or more, showed no difference in the pattern of sensitivity to thioacetazone.