1975 Onderstepoort Journal of Veterinary Research, Volume 42, 1975

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CONTENTS

*Volume 42: Number 1*

Electron microscopy of Cowdria ruminantium infected reticulo-endothelial cells of the mammalian host	Du Plessis, JL	1

Inhibition of macrophage migration in Salmonella immunity	Cameron, CM & Van Rensburg, JJ	15

The effect of Clostridium perfringens epsilon toxin on the blood brain barrier of mice	Worthington, RW & Mulders, MSG	25

Helminths recovered from the bontbok, Damaliscus dorcas dorcas (Pallas, 1776)	Verster, A, Imes, GD Jr & Smit, JPJ	29

Ixodes (Afrixodes) drakensbergensis n. sp. from domestic and wild animals in Natal, Republic of South Africa	Clifford, C, Theiler, G & Baker, M	33

Preliminary report on the transmission of Parafilaria bovicola in South Africa	Nevill, EM	41

An outbreak of ringworm in karakul sheep caused by a physiological variant of Trichophyton verrucosum Bodin	Scott, DB	49

A histological study of the effect of cortisol and some sex steroids on the immune response to sheep erythrocytes by the mouse	Hellig, HR & Gerneke, WH	53

Some observations on the use of D-glucose-2-³H as a tracer in turnover studies in Merino sheep	Van der Walt, JG	63

Research note:

Some protozoan parasites of wild birds from the vicinity of Onderstepoort	Thomas, SE & Dobson, LD	67


*Volume 42: Number 2*

Vitamin A deficiency in the captive African lion cub Panthera Leo (Linnaeus, 1758)	Bartsch, RC, Imes, GD Jr & Smit, JPJ	43

Blood parasites of some wild Bovidae in Botswana	Carmichael, IH & Hobday, E	55

Immunization of mice and guinea-pigs against Salmonella dublin infection with live and inactivated vaccine	Cameron, CM & Fuls, WJP	63

Studies on schistosomiasis. 7. A comparison of various methods for the infestation of sheep with Schistosoma mattheei	Van Wyk, JA, Heitmann, LP & Van Rensburg, JL	71

Research note:

An apparatus for the restraint of sheep during infestation with schistosomes	Van Wyk, JA	75


*Volume 42: Number 3*

The occurrence and pathology of chlamydiosis in domestic and laboratory animals : a review	Schutte, AP & Pienaar, JG	77

The partial purification of Clostridium perfringens Beta toxin	Worthington, RW & Mulders, MSG	91

Some toxic, physical and chemical properties of the oral secretion of the sand tampan, Ornithodoros savignyi Audouin (1827)	Howell, CJ, Neitz, AWH & Potgieter, DJJ	99

A rapid spectrophotometric method for the monitoring of embryonic development in ticks (Acarina : Ixodoidea)	Londt, JGH	103


*Volume 42: Number 4*

Gross and microscopic observations of ovarian abnormalities from five Burchell's zebra, Equus Burchelli antiquorum, Smith 1841	Imes, GD & Smuts, GL	109

Observations on naturally acquired hepatozoonosis of wild carnivores and dogs in the Republic of South Africa	Young, E, De Vos, V, Bigalke, RD, Basson, PA & McCully, RM	117

Blindness and encephalopathy caused by Helichrysum argyrosphaerum D.C. (Compositae) in sheep and cattle	Basson, PA, Kellerman, TS, Albl, P, Von Maltitz, LJF, Miller ES & Welman, WG	135

Author index		149

Subject index		151

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Subject index
(Pretoria : The Government Printer, 1975) Anonymous; Bigalke, R.D.; Cameron, Colin McKenzie; Verster, Anna J,M.; Walker, Jane B.; De Kock, V.E.
Author index
(Pretoria : The Government Printer, 1975) Anonymous; Bigalke, R.D.; Cameron, Colin McKenzie; Verster, Anna J,M.; Walker, Jane B.; De Kock, V.E.
Ixodes (Afrixodes) drakensbergensis n. sp. from domestic and wild animals in Natal, Republic of South Africa
(Published by The Government Printer, Pretoria, 1975) Clifford, C.M.; Baker, M.; Theiler, Gertrud; Bigalke, R.D.; Cameron, Colin McKenzie; Verster, Anna J,M.; Walker, Jane B.; De Kock, V.E.
Ixodes (Afrixodes) drakensbergensis n. sp., is described from females, males, nymphs and larvae collected on a drag at Giant's Castle Nature Reserve, Natal, Republic of South Africa; it was also taken from an eland in the same area and from goats and a cow in the adjacent Tank Area 118. The occurrence of I. (A.) drakensbergensis on domestic animals suggests that it may be of economic importance in this area. Information is provided to separate the new species from other closely related Ixodes species that occur or may occur in this region.
Some observations on the use of D-glucose-2-³H as a tracer in turnover studies in Merino sheep
(Published by The Government Printer, Pretoria, 1975) Van der Walt, J.G.; Bigalke, R.D.; Cameron, Colin McKenzie; Verster, Anna J,M.; Walker, Jane B.; De Kock, V.E.
Bolus Injection of a mixture of D-g1ucose-2-³H and -(U)-¹⁴C into each sheep enabled a turnover rate to be calculated for each isotope and to be compared in the same animal. No statistically significant differences were found in pool size, or glucose space. The total entry rates calculated from the D-glucose-2-³H results were found to be significantly higher (P>0,95 paired t test) than those calculated from the D-glucose -(U)-¹⁴C results, the average difference being 18%. This difference indicated that the extent of glucose recycling was somewhat less than that reported for monogastric mammals (30%-Katz & Dunn, 1967) and similar to that found by Judson & Leng (1972) in sheep. Glycogen was isolated from skeletal, cardiac and smooth muscle as well as from the liver and the ³H:¹⁴C ratios determined. These ratios were found to be significantly higher than those reported for monogastric mammals such as the rat or dog.
Helminths recovered from the bontbok, Damaliscus dorcas dorcas (Pallas, 1766)
(Published by The Government Printer, Pretoria, 1975) Smit, J.P.J.; Verster, Anna Johanna Maria.; Imes, G.D.; Bigalke, R.D.; Cameron, Colin McKenzie; Verster, Anna Johanna Maria; Walker, Jane B.; De Kock, V.E.
A report is given on the helminths collected from 4 bontbok, Damaliscus dorcas dorcas, which died following capture at the Bontebok National Park, Swellendam, and transfer to the National Zoological Gardens, Pretoria. Seven of these helminths are new host records. Lungworms submitted to the institute for identification over the last 2 years are also reported; these were all Dictyocaulus magna. The various scientific and common names that have been applied to the bontbok in the past are reviewed.
A histological study of the effect of cortisol and some sex steroids on the immune response to sheep erythrocytes by the mouse
(Published by The Government Printer, Pretoria, 1975) Hellig, H.R.; Gerneke, W.H.; Bigalke, R.D.; Cameron, Colin McKenzie; Verster, Anna J,M.; Walker, Jane B.; De Kock, V.E.
Sections of the spleen, thymus, lymph nodes and liver, collected from mice at various time intervals after injection with either steroid hormones only, sheep red blood cells (SRBC) only, or SRBC plus steroid hormones, were compared by histological examination. A regimen of 3 injections of 4 mg of cortisol given at 24 hourly intervals was shown to have a more severe effect than 3 injections of 1 mg given at the same times irrespective of whether SRBC were injected or not. The thymic cortex showed rapid and extensive depletion of lymphocytes very soon after corticosteroid treatment and did not recover until about the 8th day. The medulla was affected to a lesser extent. Spleens and lymph nodes showed early lymphocyte destruction, active ingestion of debris by macrophages, and germinal centres were considerably decreased in number and less clearly demarcated in corticosteroid-treated animals than in SRBC immunized controls. Spleens and lymph nodes of mice that received SRBC only exhibited the characteristic morphology of active germinal centre development associated with the immune response. Corticosteroid treatment of mice sensitized with SRBC caused an increase in neutrophilic promyelocytes in bone marrow smears to the 4th day, whereafter their numbers returned to normal. The normoblasts were decreased on the 2nd and 3rd days’ whereafter they increased to normal. Plasma cells were increased in SRBC injected animals in bone marrow smears. Of the effects of the sex steroids studied the most notable was a drastic effect of estradiol on the thymus; both the cortex and medulla were completely depleted of lymphocytes and could hardly be distinguished.
The effect of Clostridium perfringens epsilon toxin on the blood-brain barrier of mice
(Published by The Government Printer, Pretoria, 1975) Worthington, R.W.; Mulders, M.S.G.; Bigalke, R.D.; Cameron, Colin McKenzie; Verster, Anna J,M.; Walker, Jane B.; De Kock, V.E.
It was shown that Clostridium perfringens epsilon toxin has the effect of allowing the passage of ¹²⁵I polyvinyl-pyrrolidone and ¹²⁵I human serum albumin into mouse brain. These substances did not enter the brains of normal control mice. The passage of albumin into the brains of mice poisoned with epsilon toxin was extremely rapid. When large doses of toxin (± 4 000 MLD) were given death ensued within 2-3 min at which stage 1,5% of the injected albumin had already entered the brain. In cases where smaller doses were given and the time interval between injection and death was longer the figure was increased to 2-2½% of the injected plasma albumin.
An outbreak of ringworm in Karakul sheep caused by a physiological variant of Trichophyton verrucosum Bodin
(Published by The Government Printer, Pretoria, 1975) Scott, D.B.; Bigalke, R.D.; Cameron, Colin McKenzie; Verster, Anna J,M.; Walker, Jane B.; De Kock, V.E.
An outbreak of ringworm in Karakul sheep occurred during 1973 among the flocks of at least 14 farmers in South West Africa and the Northern Cape Province. Infections spread from one farm to another as a result of introducing infected stud rams. On one farm a few cattle, a goat and a horse developed lesions after being in contact with infected sheep. Some of the attendants handling infected sheep also contracted the disease. Strains recovered from infected animal’s resembled Trichophyton verrucosum morphologically, but when grown on laboratory media they did not require an exogenous source of vitamins, which was found to be indispensable for strains of this species studied previously.
Preliminary report on the transmission of Parafilaria bovicola in South Africa
(Published by The Government Printer, Pretoria, 1975) Nevill, E.M.; Bigalke, R.D.; Cameron, Colin McKenzie; Verster, Anna J,M.; Walker, Jane B.; De Kock, V.E.
The filarial worm Parafilaria bovicola causes streaks of blood on the skins of live cattle and slimy bruise-like lesions on the subcutaneous surfaces of their carcases. To determine the vectors of this worm a field survey was conducted in November 1972 and during the summer of 1973/74 on 6 farms in the Transvaal. A total of 10 093 flies was collected off cattle and examined for the infective 3rd stage Parafilaria larvae. Of the 12 fly species collected, Muscalusoria, Musca domesrica, Musca xanthomelas, Musca n. sp., Musca sorbens and Musca fasciata were the commonest. Third stage larvae were found in 3 species all belonging to the subgenus Eumusca, viz. M. fusaria, M. xanthomelas and a new Musca species. The infection rate in these flies was usually less than 1% and most of the worms were recovered from the heads of female flies. The same 3 species were successfully infected artificially in the laboratory, the infection rate ranging from 40, 0%-53, 8%. The measurements of larvae from these flies agreed closely with those of larvae recovered from field-collected flies. Female P. bovicola worms perforate the skin of cattle and deposit their eggs and/or microfilariae into the blood which trickles down from the holes. Most of these bleeding marks were noticed between July and December 1973. As 3rd stage larvae were recovered from flies from August 1973 to February 1974 the main period for transmission is likely to be between July and February. Since the 3 flies suspected of being vectors feed mainly on eye secretions it is believed that most transmission takes place via the orbital route. Thereafter development of the worms in cattle to the adult stage will take approximately 7-10 months.
Inhibition of macrophage migration in Salmonella immunity
(Published by The Government Printer, Pretoria, 1975) Cameron, Colin McKenzie; Van Rensburg, J.J.; Bigalke, R.D.; Cameron, Colin McKenzie; Verster, Anna J,M.; Walker, Jane B.; De Kock, V.E.
Protein antigens were prepared from rough strains of Salmonella typhimurium and S. dublin by phenol and veronal-buffer extraction. It was shown that the in vitro migration of peritoneal exudate cells from guinea pigs that were immunized with rough avirulent mutants could be inhibited effectively with these antigens. The cells obtained from S. typhimurium-immunized guinea pigs were also sensitive to S. dublin antigens and vice versa. A degree of sensitivity and inhibition could be demonstrated consistently in a group of immunized guinea pigs. However, the variation in samples, even from among individual animals that had survived challenge, was so great that it precludes the use of the macrophage migration technique as a routine standard assay procedure for immunity.
Blindness and encephalopathy caused by Helichrysum argyrosphaerum DC (Compositae) in sheep and cattle
(Published by The Government Printer, Pretoria, 1975) Basson, P.A.; Miller, E.S.; Kellerman, T. Stephanus; Albl, P.; Von Maltitz, L.J.F.; Welman, Wilhelmina G.; Bigalke, R.D.; Cameron, Colin McKenzie; Verster, Anna J,M.; Walker, Jane B.; De Kock, V.E.
An outbreak of amaurosis and paresis was observed amongst sheep, and occasionally cattle, which had been grazing on pastures consisting mainly of Helichrysum argyrosphaerum DC. A similar syndrome was produced by feeding the suspect plant to sheep, thereby providing proof of its toxicity. Although none of the sheep became blind and only one developed paralysis, typical spongy lesions were detected in the brains and optic fasciculi of all the experimental animals. The lesion had specific predilection sites, such as the white matter around the lateral ventricles, the optic tracts plus chiasm, the pyramidal tracts and the brachium pontis. Enlargement of the optic fasciculi furthermore gave rise to malacia, papilloedema and retinal changes. Some aged sheep developed cataracts approximately 2-3 months after the initial outbreaks of amaurosis had occurred. Circumstantial and histopathological evidence suggests that the cataracts may be due to chronic Helichrysum poisoning, but this could not be confirmed in the present investigation.
Observations on naturally acquired hepatozoonosis of wild carnivores and dogs in the Republic of South Africa
(Published by The Government Printer, Pretoria, 1975) Young, E.; De Vos, Valerius; Bigalke, R.D.; Basson, P.A.; McCully, R.M.; Bigalke, R.D.; Cameron, Colin McKenzie; Verster, Anna J,M.; Walker, Jane B.; De Kock, V.E.
Hepatozoonosis was studied in hyaenas, lions, jackals, cheetahs and one leopard in the Kruger National Park and compared with the condition seen in dogs in the Republic of South Africa. Hepatozoon schizonts were found in the wild carnivores. The genesis of microschizonts was followed and is illustrated. The schizonts were sometimes very plentiful in the lung, myocardium and skeletal muscle, and were also encountered in the spleen, liver and lymph nodes. Gametocytes were present in leucocytes. The host response was usually very mild. Sporogenous development in ticks was observed in Rhipicephalus simus females removed from an infected hyaena and R. sanguineus adults fed on an infected jackal in the nymphal stage. Attempts to transmit Hepatozoon from a jackal to dogs by means of ticks gave inconclusive results. The developmental stages of Hepatozoon seen in dogs were very similar to those encountered in wild carnivores, except for the presence of more prominent residual material in the mature microschizonts. Schizonts were found in the spleen, liver, lungs and lymph nodes. In most cases the host response to the schizonts was mild and the parasite appeared to be of little consequence to the animal. Nevertheless, severe lesions were sometimes found in uncomplicated cases as well as those complicated by intercurrent diseases such as babesiosis and viral infections. In dogs with severe infections there was considerable necrosis and a marked reticulo-endothelial response with granuloma formation in the spleen, liver and lymph nodes.
Gross and microscopic observations of ovarian abnormalities from five Burchell's zebra, Equus burchelli antiquorum, Smith 1841
(Published by The Government Printer, Pretoria, 1975) Imes, G.D.; Smuts, G.L.; Bigalke, R.D.; Cameron, Colin McKenzie; Verster, Anna J,M.; Walker, Jane B.; De Kock, V.E.
Five pair of zebra ovaries were submitted to the Onderstepoort Veterinary Research Institute for examination because of unusual uni- and/or bilateral gross appearances. Microscopic findings revealed an accessory spleen encapsulated by the ovarian peritoneum, a case of tuba-ovarian cysts, one example of ovarian aplasia, an example of germinal inclusion cysts and an abnormal follicular haemorrhage related to pressure necrosis of a nearby corpus luteum. Similarities to the ovary of the domestic mare were also seen and some are briefly described.
Electron microscopy of Cowdria ruminantium infected reticulo-endothelial cells of the mammalian host
(Published by The Government Printer, Pretoria, 1975) Du Plessis, J.L.; Bigalke, R.D.; Cameron, Colin McKenzie; Verster, Anna J,M.; Walker, Jane B.; De Kock, V.E.
An electron microscopic study of structures suspected to represent a possible developmental cycle of Cowdria ruminantium in reticulo-endothelial cells of mice and ruminants is reported. After infection dense bodies increase in size and undergo division to form fragmented dense bodies. These in turn apparently sub-divide and become organized to give rise to mature "organisms". In none of these structures do limiting membranes separate the parasitic inclusions from the host cell cytoplasm. Present observations suggest that growth of the organism in reticulo-endothelial cells differs from that of chlamydial and rickettsial agents and somewhat resembles the replication of some viruses. Developmental stages observed after infection of ruminants with the Ball 3 strain of the heartwater agent are indistinguishable from those seen with the mouse adapted strain. These observations support the hypothesis that C. ruminantium released from reticulo-endothelial cells subsequently penetrates endothelial cells where further multiplication by binary fission occurs.
A rapid spectrophotometric method for the monitoring of embryonic development in ticks (Acarina : Ixodoidea)
(Published by The Government Printer, Pretoria, 1975) Londt, J.G.H.; Bigalke, R.D.; Cameron, Colin McKenzie; Verster, Anna J,M.; Walker, Jane B.; De Kock, V.E.
A rapid spectrophotometric method for the monitoring of embryonic development in Boophilus decoloratus (Koch, 1844) is described. The method is based on a quantitative assessment of guanine, the principal end-product of nitrogenous metabolism in ticks, which is gradually built up and stored in the rectal sac during embryonic development of the larvae. A study of the growth of tick embryos under constant temperature conditions and 6 different humidity conditions demonstrated that embryonic development was dependent on the water content of the eggs at the time of oviposition. When eggs lost more than 35% of their initial mass through evaporation, nitrogenous metabolism (as indicated by guanine production) was seriously affected and embryos died.
Some toxic, physical and chemical properties of the oral secretion of the sand tampan, Ornithodoros savignyi Audouin (1827)
(Published by The Government Printer, Pretoria, 1975) Howell, C.J.; Neitz, Albert Walter Herman; Potgieter, D.J.J.; Bigalke, R.D.; Cameron, Colin McKenzie; Verster, Anna J,M.; Walker, Jane B.; De Kock, V.E.
Mice, exposed to the effects of the oral secretion of Ornithodorus savignyi by injection, revealed that a potent toxin is present in the secretion. Electrophoretic and chromatographic separations of the secretion showed a heterogeneous pattern of protein-like substances. Toxicity trials on these fractions showed that the toxic principle is protein in nature and fairly heat stable
The partial purification of Clostridium perfringens Beta toxin
(Published by The Government Printer, Pretoria, 1975) Mulders, M.S.G.; Worthington, R.W.; Bigalke, R.D.; Cameron, Colin McKenzie; Verster, Anna J,M.; Walker, Jane B.; De Kock, V.E.
An attempt was made to purify Clostridium perfringens Beta toxin. Crude toxin prepared by ammonium sulphate precipitation of culture supernatants was purified by chromatography on Sephadex G50, Sephadex G100 and DEAE cellulose. This material, although highly purified, was not homogeneous on polyacrylamide gel electrophoresis. It had a toxicity of 800 000 mouse MLDs/mg N, a typical protein absorption spectrum in the UV region, an iso-electric point of 5, 6 and the main component had a molecular mass of 42 000 ± 2 000 (estimated by electrophoresis in sodium dodecyl sulphate containing polyacrylamide gels)
The occurrence and pathology of chlamydiosis in domestic and laboratory animals : a review
(Published by The Government Printer, Pretoria, 1975) Pienaar, J.G.; Schutte, A.P.; Bigalke, R.D.; Cameron, Colin McKenzie; Verster, Anna J,M.; Walker, Jane B.; De Kock, V.E.
The literature on the various disease syndromes caused by chlamydia in domestic and laboratory animals is summarized. A review of the pathological lesions which characterize these diseases is presented and the pathogenesis of chlamydiosis is briefly discussed. Some aspects of unpublished findings on the occurrence of intestinal, kidney and genital lesions in natural and experimental cases of chlamydiosis in cattle and sheep, abortions and conjunctivitis in horses in South Africa are recorded.
An apparatus for the restraint of sheep during infestation with schistosomes
(Published by The Government Printer, Pretoria, 1975) Van Wyk, Jan Aucamp; Bigalke, R.D.; Cameron, Colin McKenzie; Walker, Jane B.; Verster, Anna J.M.; De Kock, V.E.
With this adjustable apparatus sheep aged from five months to five years and varying in mass from 16-95 kg could be restrained effectively. Cercarial suspension was spilt by only 2/44 sheep during infestation with Schistosoma mattheei. Using a battery of seven cages, it was possible to infest 23 sheep, each for 30 minutes, in one day. No cercariae were more than 4 and a half hours old at the end of exposure.
Studies on schistosomiasis. 7. A comparison of various methods for the infestation of sheep with Schistosoma mattheei
(Published by The Government Printer, Pretoria, 1975) Van Rensburg, L. J.; Heitmann, L.P.; Van Wyk, Jan Aucamp; Bigalke, R.D.; Cameron, Colin McKenzie; Verster, Anna J,M.; Walker, Jane B.; De Kock, V.E.
The percutaneous (leg and thorax and abdomen) and subcutaneous routes of infestation with Schistosoma mattheei were compared in 29 sheep. Larger percentages of cercariae developed after percutaneous than subcutaneous infestation and the difference was highly significant (P<0,001). Furthermore, if the leg was used for percutaneous infestation worm development was significantly higher (P < 0,02) when the skin was washed thoroughly in water before exposing it to cercariae, than when it was left unwashed. Washing was apparently not necessary if the thorax and abdomen served as the route of infestation.

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