No deviations from normal were seen in the erythrocyte sedimentation rate of any of the animals studied (Appendix 7). Potassium levels were generally within the extreme normal range in the various chronic cases of the disease. Extensive changes, mainly in the middle part of the lobule, were found in the liver.
The same could be said for the plasma iron level found in the studied cases.
BROWN Plasma iron levels were not studied in the case of Sheep 1541, but remained
Icterogenin appears to have a definite deleterious effect on the erythrocytes of animals administered with it. The symptomatology of the various forms of selenosis has been reviewed in a previous paper (Brown & de Wet, 1962). The generally higher levels of selenium in the livers of cases of enzootic jaundice are consistent with its often more acute and rapidly fatal nature of the two syndromes.
The lamb's liver contained more selenium than is found in the average case of geldikkop, and both livers contained the amounts usual in typical cases of enzootic icterus. Composition of Karoo vegetation, types of vegetation represented and plant associations. However, the selenium content of the Karoo vegetation is much lower than that in the seleniferous areas of the Americas.
A preliminary classification of the Karoo vegetation according to the apparent uptake of selenium from the soil was attempted. There appears to be a definite correlation between the selenium content of the vegetation and the incidence of yellow thick head and enzootic jaundice. A definite correlation was found between mismanaged field types, the selenium content of the vegetation and the occurrence of enzootic icterus and yellow thick head.
CHAPTER 14
This aspect of the disease and the stress factors in question are the subject of the next chapter. Some of these cases are undoubtedly due to negligence and careless use of the vaccine, but many of them have been shown to be true. The calculation of the absolute heemiltological indices in these cases showed that the anemia was normocytic hypocytemic and normochromic.
Three of the Karoo animals responded to the infection and all three showed a similar transient increase in urinary coproporphyrin excretion to that found in the three Onderstepoort sheep just mentioned. However, all were tested for photosensitivity by solar radiation for approximately two hours each day for the two-week period after the onset of the febrile reaction, in the case of the Onderstepoort animals. 15 shows these symptoms in one of the Karoo animals at the time when his urinary coproporphyrin excretion was maximal.
No changes of any particular significance were found in the total plasma protein levels of any of the sheep that developed a febrile reaction. Plasma iron levels were studied in five of the Onderstepoort sheep and in all animals sourced from Karoo farms. Swelling of the coronary band and surrounding tissues in the right forelimb of the same animal.
Glob
CHAPTER 15
The previous chapter of this thesis highlighted many features of epizootiology, symptomatology, histopathology, chemical pathology and general biochemistry common to both conditions. The further course of events then largely depends on the nature of the syndrome itself, e.g. Investigation into the occurrence of potentially harmful levels of selenium in Karoo vegetation.
The histological evidence of liver damage from pyrrolizidine alkaloids: Megalocytosis of hepatocytes and inclusion globules. 34;Diseases of the kidney and genito-urinary tract" in Biochemical Disorders in Human Disease, pp. Effects of the pyrrolizidine alkaloid, lasiocarpine N-oxide, on nuclear and cell division in rat liver.
The sensitized Merino skin response to radiation in different areas of the spectrum. Normal values for the activity of some enzymes of the Embden-Meyerhof pathway and tricarboxylic acid cycle occurring in sheep plasma. DETAILS OF THE SHEEP USED JN THE STUDIES ON GEELDIKKOP A'S HAEMATOLOGY, GENERAL CHEMICAL PATHOLOGY, BIOCHEMISTRY AND HISTOPATHOLOGY.
APPENDIX 2
MCH Examination of blood and bone marrow smear-Remarks Blood smear: Very marked anisocytosis, large number of macrocytes. VH-V Not cloned I Not done Not done I 5·12 0 Blood counts: Fair degree of anisocytosis, mild anemia Blood staining: Severe anisocytosis with numerous large macrocytes. Numerous cells present with Jolly bodies Blood staining: Fair degree of anisocytosis with numerous large macrocytes.
Numerous normoblasts and cells with Jolly bodies Blood staining: Fair degree of anisocytosis with large number of macrocytes. I Blood and Bone Marrow Stain Examination-Remarks Blood smear: Fair degree of anisocytosis Blood stain: Mild anisocytosis I Blood stain: Nothing unusual Blood stain: Nothing unusual I I Blood stain: Nothing unusual. Blood and Bone Marrow Stain Examination-Remarks I ~mear Blood: Anisocytosis, polychromase Blood Appearance: Nothing unusual Blood Staining: Nothing unusual erythrocyte and granulocyte precursors.
No normoblasts seen Blood smear: Nothing unusual 1 Bone marrow smear: Marked reduction of all cellular elements Blood smear: Nothing unusual Blood smear: Nothing unusual Blood smear: Nothing unusual Blood smear: Marked anisocytosis I Blood smear: Nothing unusual Blood smear: Nothing unusual Smear bone marrow: Very active hemopoiesis Blood smear: Nothing unusual Blood smear: Nothing unusual. Blood smear: Nothing unusual Blood smear: Nothing unusual Blood smear: Nothing unusual I Blood smear: Nothing unusual I Blood smear: Nothing unusual I Blood smear: Nothing unusual Blood smear: Nothing unusual Blood and bone marrow smear Examination-notes - I Blood smear: Nothing unusual 5·87 I Blood smear: Nothing unusual Blood smear: Marked anisocytosis with numerous macrocytes and cells containing Jolly bodies I. Examination-notes of blood smear and bone marrow Blood smear: Nothing unusual j Blood smear: Nothing unusual Blood smear: Nothing unusual Blood smear: Nothing unusual Blood smear: Nothing unusual Blood smear: Nothing unusual Blood smear: Nothing unusual Blood smear: Nothing unusual Blood smear: Nothing unusual.
APPENDIX 4
APPENDIX 5
DETAILS OF THE SHEEP USED IN THE STUDIES ON HAEMATOLOGY, GENERAL CHEMICAL PATHOLOGY, BIOCHEMISTRY AND HISTOPATHOLOGY OF ENZOOTIC ICTERUS. Principle of the methods: Copper forms a deep pink complex with bis-acetaldehyde-oxalyldihydrazone at pH 8 ·4-9 ·I. The formation of the pink-purple complex takes place best in the presence of ammonium ions (Peterson & Bollier, 1955).
Add to the control tube 0·25 ml of water and to the rest of the tubes 0·25 ml of oxalyl dihydrazide solution. In three clean tubes labeled as above, place 2 ml of the blank and standard mixtures and 2 ml of the clear centrifuge. The last step includes heating strongly until the evolution of white fumes after adding 5 ml of water and reducing the volume of the digestion to about 5 ml.
After measuring the pH of the contents of each tube, 0.25 ml of oxalyldihydrazide and acetaldehyde and ammonia solutions were added, the contents of the tubes were mixed and allowed to stand for one hour before the optical densities were read against the reagent blank. The color develops in fifteen minutes and is maximal one hour after the addition of the color reagents. Dissolve one gram of tissue, 1 ml of standard solution and 1 ml of water in a 100 ml Kjeldahl flask as described under (f) above.
APPENDIX 10
Most of the mercury sulfate formed during fermentation will settle out at this stage. In the outlined method, all residual hydrogen peroxide is removed during the final stage, when the volume of the digestate is reduced to 10 ml. The variation is greatest at the low concentrations of selenium and is directly due to the color of the blanks used in each batch of determinations.
Ferrous sulfate present as 0.5 mg Fe prevented the normal color development of 50 meg selenium. In this regard, the elements tested and the amounts in which each was added to the contents of the test flasks are listed in the table below. This is visible in the case of raw materials, where various shades of light yellow can appear.
In the first place, considerable and laborious foaming is invariably encountered in the initial stages of digestion. It should be left overnight in sulfuric acid, with the addition of mercuric oxide and only 5 ml of water at this stage. Quantitative adaptation of the codeine test to the colorimetric determination of selenium in plant materials.
APPENDIX 11
Changes in Official and Experimental Methods of Analysis at the Fifty-Fourth Annual Meeting. Excretion of volatile selenium compounds after administration of sodium selenite to white rats. The dry residues of this process are represented by fractions called "dried bile extracts".
The large spots appearing above the designation "BSP" were obtained from samples of the material injected into the animals in question which were chromatographed as reference standards at the same time as samples of the bile extracts. It is clear from these chromatograms that conjugate B is present in the bile in the greatest amounts. Thin Layer Chromatography: Samples of each conjugate were prepared for subsequent chemical work by chromatography of the bulk material on a large number of thin layers prepared from a mixture of 35 g of Kieselgel G (Merck's "nach Stahl" containing 13 percent CaSO., and particle size 0.10 to 401 -') and 55 ml of water are mixed immediately before use.
The same three conjugates mentioned above were present together with traces of two or three other slower purple-reacting compounds in the alkaline developing medium. After the chromatograms had become almost dry, the colored spots were scooped off with small stainless steel spatulas into suitable labeled containers and stored in the dry state at - 10°C until required for later work, when elution of the conjugates from the Kieselgel leftovers were. achieved with minimal amounts of acetone 3: water l made alkaline with a drop or two of concentrated ammonia to visualize the completeness of elution. Preliminary identification of the conjugates: The pure pigments eluted from thin layer chromatograms as described above were dissolved in 0·1 N NH4OH and the spectral absorption of the solutions was determined in the ultraviolet and visible regions of the spectrum.
