478 THE COCCUS OF MEDITERRANEAN FEVER 



cutaneous inoculations of heated agar cultures found that he could then 

 inoculate them with virulent cultures without producing a typical attack of 

 Mediterranean fever ; but one of these monkeys showed no immunity to a 

 second test inoculation. 



By repeatedly inoculating horses and goats sub-cutaneously with living 

 cultures Shaw and Eyre obtained powerfully agglutinating serums (1 in 3000 

 and 1 in 5000). These serums have no therapeutic properties when tested 

 on man and animals. In a case of laboratory infection in man Nicolle found 

 that recovery coincided with the inoculation of 10 c.c. of serum from an 

 hyper-immunized ass. 



Bassett-Smith attempted the treatment of Mediterranean fever with a 

 vaccine prepared by heating emulsions of ten-day old agar cultures in distilled 

 water for half an hour to 60 C. The dose of vaccine used was O'5-l c.c. 

 In acute cases the inoculation aggravated the symptoms but in chronic cases 

 it appeared to stimulate the destruction of the micro-organisms and certainly 

 shortened the duration of the disease. 



5. Agglutination. 



Wright, Birt and Lamb have shown that the serum of persons suffering 

 from Mediterranean fever, like the serum of immunized animals, agglutinates 

 the micrococcus. 



Generally speaking the agglutination reaction is poorly developed in the blood 

 of patients (1 in 15 to 1 in 50), though Lamb and Kesava have obtained agglutina- 

 sion in dilutions of 1 in 160 and even 1 in 280. For purposes of clinical diagnosis 

 the reaction of a 1 in 10 or 1 in 15 dilution of the serum should be determined and 

 if this gives a positive result higher dilutions may be tested. The agglutination 

 reaction always appears at the end of the first week of the disease, and may still 

 be present years after recovery. In artificially infected monkeys Birt and Lamb 

 have found it present as early as the fifth day. 



The blood and the milk of infected goats agglutinate the coccus. 



For carrying out the agglutination reaction Nicolle advises using broth emulsions 

 of agar cultures 3-5 days old, and mixing the serum and emulsion in small straight 

 tubes. The reaction can be observed with the naked eye. 



Pollaci and Ceranlo have shown that blister fluid and the saliva of infected persons 

 agglutinate the organism. (Dilute a loopful of an agar culture in 5-20 drops of 

 filtered saliva : the agglutination can be seen under the microscope in 3060 minutes.) 

 The reaction with the saliva is said to be always present in persons suffering from 

 Mediterranean fever and absent in healthy individuals. 



6. Immune body. 



Sicre has demonstrated the presence of an immune body in the blood of 

 inoculated animals and of persons suffering from Mediterranean fever. 



SECTION IV, DETECTION, ISOLATION AND IDENTIFICATION 

 OF THE ORGANISM. 



Post mortem, the spleen, liver and kidneys should be examined for the 

 Micrococcus melitensis. During life it may be recovered by puncture of the 

 spleen (p. 198) or by sowing cultures with the blood, milk or urine. 



Scrapings of the internal organs should be sown on ordinary or litmus- 

 nutrose agar, incubated at 37 C. for about a week and then examined and 

 tested. 



The number of organisms in the blood is always small : it is best to examine 

 the blood during the height of the fever and to sow at least 2-4 c.c. in 250 c.c. 

 of broth. Pollaci recommends the addition of bile to the broth. 



