Journal of the South African Veterinary Association

Research on animal blood groups and biochemical polymorphisms at Onderstepoort (1956-1990)

The introduction and wide use of artificial insemination in cattle in the 1950s led to a need for accurate parentage identification. Blood group determination by means of the newly emerging scientific discipline called immunogenetics provided the answer. A blood group laboratory was consequently established at Onderstepoort in 1956, initially concentrating on the production of blood typing reagents. Once established the technology was also applied to studies on a variety of problems in various animals as summarised in this paper. Investigations include zygosity in cattle twins, blood transfusion in domestic animals, breed relationships, genetic polymorphisms and the identification of useful genetic markers for production and disease parameters in breeding programmes.

Effect of heterologous seminal plasma and semen extenders on motility of frozen-thawed ram spermatozoa

Ram seminal plasma increases the fertility of frozen-thawed ram spermatozoa deposited into the cervix. The aim of the current study was to compare the effect of ram seminal plasma to that of bull seminal plasma, dog prostatic fluid, protein-free TALP, TrilEq (Triladyl with 0.5 mℓ of Equex STM paste added to each 100 mℓ) and heat-treated skim milk on longevity and percentages of progressively motile and aberrantly motile frozen-thawed ram spermatozoa. Three ejaculates from each of 6 rams were extended in TrilEq, pooled and frozen in straws as a single batch per ram. One hundred and eight straws (3 straws from each ram for each fluid) were thawed in random order. Once thawed, a straw was emptied into a tube with 0.85 mℓ of the appropriate fluid at 37 ºC and kept at that temperature for 6 h. Motility was assessed at ×200 magnification immediately (time zero) and 2, 4 and 6 h after thawing. Progressive motility decreased from each time to the next (P < 0.05) and was 39.0 % (0 h), 26.0 % (2 h), 19.6 % (4 h) and 12.6 % (6 h); SEM 1.24, n = 108 for each group. Ram seminal plasma resulted in higher progressive motility than bull seminal plasma, lower than milk, and similar to the other fluids. Ram seminal plasma resulted in lower aberrant motility than protein-free TALP and similar aberrant motility to other fluids. The effect of ram seminal plasma and dog prostatic fluid was very similar. The effect of ram seminal plasma on the fertility of frozen-thawed ram spermatozoa deposited into the cervix is not due an exceptionally beneficial effect on the motility of spermatozoa.

Cardiovascular effects of epidural morphine or ropivacaine in isoflurane-anaesthetised pigs during surgical devascularisation of the liver

The cardiovascular effects of non-abdominal and abdominal surgery during isoflurane anaesthesia (A-group) or isoflurane anaesthesia supplemented with either epidural ropivacaine (AR-group; 0.75 % solution, 0.2 mℓ/kg) or morphine (AM-group; 0.1 mg/kg diluted in saline to 0.2 mℓ /kg) were evaluated in 28 healthy pigs with a mean body weight of 30.3 kg SD ± 4.1 during surgical devascularisation of the liver. Anaesthesia was induced with the intramuscular injection of midazolam (0.3 mg/kg) and ketamine (10 mg/kg). Anaesthesia was deepened with intravenous propofol to enable tracheal intubation and maintained with isoflurane on a circle rebreathing circuit. The vaporiser was set at 2.5 % for the A-group and 1.5 % for the AR-and AM-groups. Differences between treatment groups were not statistically significant (P > 0.05) for any of the variables. Differences between AMand AR-groups were marginally significant heart rate (HR) (P = 0.06) and mean arterial blood pressure (MAP) (P = 0.08). Within treatment groups, differences for the A-group were statistically significant (P < 0.05) between non-abdominal and abdominal surgery for HR, systolic blood pressure, diastolic blood pressure (DIA) and MAP. Within the AM-group differences were statistically significant (P < 0.05) for DIA and MAP, and within the AR group differences for all variables were not statistically significant (P > 0.05). It was concluded that in isoflurane-anaesthetised pigs, the epidural administration of ropivacaine decreased heart rate and improved arterial blood pressure during surgery.

**Diversity and seasonal occurrence of Eimeria species in a mixed flock of communally reared sheep and goats in Mafikeng in the North West Province, South Africa**

Diversity and seasonal occurrence of coccidia in a communally reared mixed flock of sheep and goats at Mafikeng, North West Province, South Africa, was determined between March 2008 and February 2009. Faecal specimens were collected directly from the rectum of the animals and the number of oocysts per gram of faeces (opg) determined. The mean monthly opg for goats was significantly higher than that for sheep. Higher oocyst counts were observed during the hot, rainy season than during the cold, dry season. The highest mean values for both the sheep (862.5 opg) and goats (1200 opg) were recorded during March. Six species (Eimeria crandallis, E. bakuensis, E. weybridgensis, E. ahsata, E. intricata, and E. ovinoidalis) were recovered from sheep, with E. crandallis and E. bakuensis occurring most frequently. The last 2 species, together with E. ahsata, are considered among the most pathogenic species in sheep. In goats, 7 species (E. arloingi, E. jolchijevi, E. caprina, E. alijevi, E. caprovina, E. christenseni and E. hirci) were recovered, with E. arloingi and E. jolchijevi occurring most frequently. Up to 5 Eimeria species were recovered from individual specimens in goats while up to 4 were recovered in sheep. No cross-infections between goats and sheep were recorded and no clinical coccidiosis was noted during the study. It is increasingly becoming evident that the pathogenic E. arloingi is one of the most commonly occurring Eimeria species in goats in South Africa.

Prevalent organisms on ostrich carcasses found in a commercial abattoir

The prevalent microbial growth on carcasses before and after overnight cooling in an ostrich abattoir and de-boning plant was investigated. The effect of warm or cold trimming of the carcasses was examined together with possible causes of contamination along the processing line. An attempt was made to link the prevalent microorganisms that were identified from carcasses to those from specific external contamination sources. Samples of carcasses and possible contaminants were collected in the plant, plated out and selected organisms were typed using a commercial rapid identification system. It was indicated that the cold trim (mainly of bruises) of carcasses was advantageous in terms of microbiological meat quality. Results indicated pooled water in the abattoir as the most hazardous vector for carcass contamination and that contaminants from this source are mostly Gram-negative pathogens. Pseudomonas and Shigella were frequently isolated from surface and air samples and indicated that the control of total plant hygiene is a requirement for producing ostrich meat that is safe to consume and has an acceptable shelf-life.

**A serological survey of antibodies to Leptospira species in dogs in South Africa**

Leptospirosis, a disease more common in the tropics, can cause a life-threatening multisystemic syndrome in humans and animals. Immunity, whether natural or vaccineinduced, is serogroup-specific with the infecting serovars varying according to geographical locality. In South Africa, in spite of the fact that the bacterin vaccine for some Leptospira serovars is often used, there is no recent information on the incidence of canine leptospirosis as well as the infecting serovar/s. The aim of this study, which was undertaken on sera collected in 2008 and 2009 from both strays and owned dogs predominantly in the coastal regions of South Africa, was to determine the presence of leptospiral antibodies to 15 serovars known to infect dogs. Of the 530 samples tested, 25 tested positive to 7 different serovars with the microscopic agglutination test (MAT). Nine of the 25 samples tested positive to more than one serovar. The 2 serovars most frequently represented were Canicola, which reacted to 17 sera, and Pyrogenes, which reacted to 10 sera. Currently the only vaccines available in South Africa in different combinations contain serovars Canicola, Icterohaemorrhagiae, Pomona and Grippotyphosa. The results showed that the use of vaccines containing serovar Canicola is still justifiable in certain regions of the country. However, the presence of antibodies to serovar Pyrogenes in several dogs, pending a broader investigation, indicates that this serovar should also be included in the range of Leptospira vaccines for use in South Africa.

Veterinary extension on sampling techniques related to heartwater research

Heartwater, a tick-borne disease caused by Ehrlichia ruminantium, is considered to be a significant cause of mortality amongst domestic and wild ruminants in South Africa. The main vector is Amblyomma hebraeum and although previous epidemiological studies have outlined endemic areas based on mortalities, these have been limited by diagnostic methods which relied mainly on positive brain smears. The indirect fluorescent antibody test (IFA) has a low specificity for heartwater organisms as it cross-reacts with some other species. Since the advent of biotechnology and genomics, molecular epidemiology has evolved using the methodology of traditional epidemiology coupled with the new molecular techniques. A new quantitative real-time polymerase chain reaction (qPCR) test has been developed for rapid and accurate diagnosis of heartwater in the live animal. This method can also be used to survey populations of A. hebraeum ticks for heartwater. Sampling whole blood and ticks for this qPCR differs from routine serum sampling, which is used for many serological tests. Veterinary field staff, particularly animal health technicians, are involved in surveillance and monitoring of controlled and other diseases of animals in South Africa. However, it was found that the sampling of whole blood was not done correctly, probably because it is a new sampling technique specific for new technology, where the heartwater organism is much more labile than the serum antibodies required for other tests. This qPCR technique is highly sensitive and can diagnose heartwater in the living animal within 2 hours, in time to treat it. Poor sampling techniques that decrease the sensitivity of the test will, however, result in a false negative diagnosis. This paper describes the development of a skills training programme for para-veterinary field staff, to facilitate research into the molecular epidemiology of heartwater in ruminants and eliminate any sampling bias due to collection errors. Humane handling techniques were also included in the training, in line with the current focus on improved livestock welfare.

**An investigation of the effects of secondary processing on Mycobacterium spp. in naturally infected game meat and organs**

The risk for humans to contract bovine tuberculosis through the consumption of undercooked game meat as well as biltong (traditionally dried game meat) is a concern. The survival potential of Mycobacterium bovis during the cooking and drying processes was researched in a preceding study on beef and the positive results compelled the authors to investigate the results with a similar preliminary study on game meat. Muscular, lymphatic and visceral tissues from skin test positive African buffalo (Syncerus caffer) and greater kudu (Tragelaphus strepsiceros) with tuberculous lesions were collected from the Hluhluwe iMfolozi Park during the park's culling programme. The different tissues were exposed to cooking and the muscular tissue to the drying process prior to culture. All acid-fast isolates were analysed by polymerase chain reaction for the presence of Mycobacterium bovis. All tissues were found negative for Mycobacterium bovis but non-tuberculous mycobacteria were isolated from kidney, liver, heart and lymph nodes. The results showed that these processes will kill Mycobacterium bovis but the unexpected recovery of non-tuberculous mycobacteria suggests possible survival and resistance characteristics of these strains which might be of veterinary public health interest.

**Gousiekte in African buffalo (Syncerus caffer)**

Three African buffalo (Syncerus caffer) that died after capture and translocation from Mutirikwe Recreational Park in southern Zimbabwe showed macroscopic and microscopic lesions of cardiomyopathy compatible with a diagnosis of gousiekte. The buffalo had had access to Pavetta schumanniana, a plant that is known to cause gousiekte. Death was attributed to cardiac failure as a result of previous consumption of the plant, exacerbated by the stress of translocation.

Surgical uterine drainage and lavage as treatment for canine pyometra

Pyometra is a common post-oestral syndrome in bitches. Classical treatment consists of either ovariohystorectomy or medical intervention. Surgical uterine drainage and lavage via direct trans-cervical catheterisation using a 5 % povidone-iodine in saline solution was performed successfully in 8 bitches with pyometra. All bitches conceived and whelped without complications subsequent to this treatment. It is concluded that this method offers an effective alternative treatment for canine pyometra with shorter recovery times as well as good clinical recovery and pregnancy rates in bitches destined for further breeding.

Foal in Mare: insights inside the foaling mare

Medicinal plants of South Africa, 2nd edition

Parasitological Society of Southern Africa: abstracts