RESPIRATION IN SOUTHERN WHALES 393 



spherical, and their diameter varied from 0-5 to 2-0/^. They were motile, insomuch that 

 they moved with an irregular motion distinct from Brownian movement. They were 

 white or sometimes pale blue and highly refractive. Unsuccessful attempts were made 

 to count these particles by haemocytometer. The best way of assessing the number 

 present was by comparison with the number of erythrocytes. A great variety of numbers 

 was found, ranging from 10 to 30 million per c.mm. These particles retained their 

 activity at all room temperatures below 40° C. 



Cultures. In the absence of more elaborate facilities than are afforded on board a 

 floating factory, it was impossible to make as comprehensive an investigation of these 

 organisms as was desired. They were cultured in a crude manner by placing the whole 

 blood in a sterile Petri dish, diluting it with freshly boiled 07 per cent solution of 

 sodium chloride, and keeping it at 30° C. No figures are available to show the numbers 

 of organisms present from day to day, because of the difficulty of counting mentioned 

 above, but superficial examination of smears from the culture was ample to show that 

 the organisms were increasing at a rapid rate. A sample of fresh blood treated in this way 

 became so congested with X organisms in 4 days that the surface of the culture was 

 covered with a grey scum which was composed exclusively of the organisms. After some 

 days, of course, the other formed elements of the blood became disrupted, the haemo- 

 globin turned to methaemoglobin, and the blood became unrecognizable as such. As 

 far as could be seen the cultures consisted entirely of the X organisms. 



Cultures showed no signs of infection until some days had elapsed, when, owing to 

 frequent lifting of the lid in an insanitary laboratory, foreign bodies found their way in 

 and multiplied. An infected culture usually cleared itself if kept covered and undisturbed 

 for a week. For comparison, a sample of blood was taken from a rotten whale and was 

 found to be teeming with many kinds of bacteria, including streptococci, in addition to 

 the X organisms. This sample was treated in the same way as the fresh samples, and in 

 one week there were no bacteria visible in the blood ; the only sign of life was in the X 

 organisms, which had increased substantially. The aggregation of organisms at the 

 surface of the culture suggested that they were aerobic. One portion of a culture was 

 aerated continually for a week, another was covered with paraffin oil, and the remainder 

 was left standing open to the air. The X organisms in the aerated portion diminished 

 steadily, in the covered portion they remained almost stationary, while in the third they 

 showed a steady increase. It appeared that too much air was harmful. 



Experiments were performed on samples of blood which were kept in Petri dishes 

 until all the corpuscles had disappeared to determine the gaseous condition of the 

 culture. The methods were exactly the same as for fresh blood. The blood of a Blue 

 whale (taken 2. i. 33), v/hich had been kept in a Petri dish for 23 days, was subjected to 

 one of these experiments. The sample had been open to the air under the lid of the dish. 



Gas content of the culture: Nitrogen o-66 vol. per cent n.t.p. 



Oxygen 0-22 ,, ,, 



Carbon dioxide 18-50 ,, ,, 



