BILIARY CALCULI IN' THE SQUETEAGUE. 
133 
calcium, and magnesium, smaller quantities of sulphuric acid, and a slight amount of 
iron. The residue (6) was extracted with hot chloroform, and the extract (7) upon 
evaporation gave but little bilirubin. The residue (8) was boiled with alcohol. The 
alcohol extract (9) when evaporated gave a small amount of bilirubin. Thinking that 
there might be yet. a bilirubinate which had not been decomposed by the dilute HC1, 
the residue (10) was extracted with hot dilute I TCI alcohol for three hours. The 
acid-alcohol extract (11) upon evaporation yielded a considerable quantity of pigment 
which had the properties of bilirubin. A portion of the residue (12) gave strong- 
reactions with Mil Ion’s reagent and by the xanthoproteic test. The remainder was 
divided into two portions. The smaller (11) was fused with KN () 3 and HNaC0 3 in 
order to determine the presence of P and S in the organic molecule. The product 
dissolved in hot dilute nitric acid gave good reactions for phosphoric and sulphuric 
acids. The larger portion (13) was washed, dried to constant weight at 105°C., and 
the per cent of N estimated by the Dumas method. An accident at the close of the 
determination prevented an accurate estimation. There was at this time 13+ per 
cent N. The estimation, being of necessity low, can not be taken as an absolute 
indication of the nature of the substance, yet it would seem to suggest a nucleo- 
albumin rather than a mucin. (See Table II.) 
The quantitative analysis of these calculi dried to constant weight at 100° C. 
yielded: 
Per cent. 
Cholesterin (and fat) 0.47 
Bilirubin 22.39 
Nucleo-albumin 70.69 
Mineral ,. 5.10 
68.65 
The difference in per cent of fat and cholesterin in the two analyses is certainly 
striking. The amount of cholesterin in the two cases varied but little as far as could 
be determined by qualitative reactions, but the fat in the latter case was evidently 
very much less. 
The gallstones in the third or lower group, taken from the third squeteague 
(a female), were much smaller than the preceding. The largest, dried over sulphuric 
acid, weighed 0. 1 1 gram and the entire 2d weighed only 0.935 gram. The appearance, 
except size, did not differ from No. 2. The calculi were also found in the gall bladder, 
gall duct, intestines, and embedded in the tissues surrounding the intestines and 
liver. The gall bladder contained a quantity of bile. The qualitative analysis did 
not differ from the two preceding, and a quantitative determination was not made. 
The three fish from which the above calculi were taken were apparently in 
normal condition as far as could be determined when taken from the water. The 
livers were perfectly normal in appearance. It is a difficult matter to collect the 
urine from these animals, as it is generally eliminated as soon as they are taken from 
the water; but the small quantity of urine that was collected showed the presence of 
no bile pigments. Observations of this character would certainly be of interest in 
the light of comparative physiology. 
Perhaps the most interesting feature in the analyses described above is the high 
per cent of nucleo-albumin, differing in this respect from the gallstones reported 
from other animals. Further observations on the formation of these calculi might 
prove to be of value in explaining the causes of such deposits. 
