RESPONSES OF THE ALLIGATOR 693 



Phase 4 was designed to study the treatment of infected animals 

 that had been thermally stressed. An antibiotic ointment containing 

 (1) polymyxin B sulfate, 5,000 units/gram, (2) zinc bacitracin, 400 

 units/gram, and (3) neomycin sulfate, 5 mg (equivalent to 3.5 mg 

 neomycin base), was applied topically to lesions. Kanamycin sulfate 

 (10 mg/kg body weight) was given intramuscularly to some of the 

 infected animals daily for 5 days. Injections were stopped for 2 days 

 and then resumed for 5 days with an increased dose, up to 50 mg/kg 

 body weight. Fifty milliliters of oxy tetracycline hydrochloride (25 

 mg/ml) was added to the infected water in the 600-liter tank 

 containing a third set of animals. A fourth tank of animals remained 

 untreated as controls. 



Normal values for animals were established beginning with initial 

 capture in their natural habitats off the South Carolina coast. The 

 procedures in studying the alligator blood were: 



1. Blood was collected from the dorsal vein of the tail with 

 1.5-ml ethylenediaminetetraacetic acid tubes to provide an anti- 

 coagulated specimen. Slides were prepared, and microhematocrits 

 were determined. 



2. Red blood cells were counted in an automated unit that gives 

 total red count, total white count, hemoglobin, hematocrit, and 

 mean corpuscular hemoglobin concentration. Hemoglobin was mea- 

 sured by a cyanomethemoglobin method on a 1 : 500 dilution in an 

 automated hemoglobinometer. 



3. Platelet counts and white blood cell counts were performed by 

 adding 20 /jl of well-mixed anticoagulated blood to 1.98 ml of 1% 

 ammonium oxalate to make a 1 : 100 dilution. An estimated platelet 

 count was performed using a Miller disk and 5 Ox oil objective (Dacie 

 and Lewis, 1968). 



Cytochemical stains; periodic-acid— Schiff (PAS), as described by 

 Lillie (1965); peroxidase, by the Kaplow method (Kaplow, 1965); 

 and leukocyte alkaline phosphatase (LAP) (Kaplow, 1968) were used 

 to determine the identity of certain white blood cells from the 

 chemical content. 



The methods for protein studies on serum were as follows: 



1. Total protein was determined by the biuret method. 



2. Electrophoresis was performed on cellulose acetate gels with a 

 sodium barbital buffer at a pH of 8.6. Migration was performed at 

 180 V, 11.5 mA for 13 min. The plates were then stained for 3 min 

 in Ponceau S, a general protein stain. The stained gels were scanned 

 and counted by an automated, digital, integrated scanner. 



