The Association between Tumour Markers and Meta-iodobenzylguanidine Scans in South African Children with High-risk Neuroblastoma.

AIMS: Diagnostic and post-induction 123I-meta-iodobenzylguanidine (123I-mIBG) scans have prognostic significance in the treatment of neuroblastoma, but data from low- and middle-income countries are limited due to resource constraints. The aim of this study was to determine the association between neuroblastoma-associated tumour markers (lactate dehydrogenase [LDH], ferritin and MYCN amplification) and 123I-mIBG scans (modified Curie scores and metastatic disease patterns) in predicting complete metastatic response rates (mCR) and overall survival. MATERIALS AND METHODS: Two hundred and ninety patients diagnosed with high-risk neuroblastoma in South Africa between January 2000 and May 2018 and a subanalysis of 78 patients with diagnostic 123I-mIBG scans were included. Data collection included LDH, ferritin and MYCN amplification at diagnosis. Two nuclear physicians independently determined the modified Curie scores and pattern of distribution for each diagnostic and post-induction 123I-mIBG scans with high inter-rater agreement (r = 0.952) and reliability (K = 0.805). The cut-off values for the diagnostic and post-induction modified Curie scores of ≥7.0 (P = 0.026) and 3 (P = 0.009), respectively, were generated. The association between the tumour markers and the modified Curie score of the 123I-mIBG scans was determined using post-induction mCR and 2-year overall survival. RESULTS: Diagnostic LDH (P < 0.001), ferritin (P < 0.001) and the diagnostic modified Curie scores (P = 0.019) significantly predicted mCR. Only ferritin correlated with diagnostic modified Curie scores (P = 0.003) but had a low correlation coefficient of 0.353. On multivariable analysis, the only significant covariate for 2-year overall survival at diagnosis was LDH <750 U/l (P = 0.024). A post-induction chemotherapy modified Curie score ≤3.0 had a 2-year overall survival of 46.2% compared with 30.8% for a score >3.0 (P = 0.484). CONCLUSION: LDH, ferritin and the diagnostic 123I-mIBG scans significantly predicted mCR, but only LDH predicted 2-year overall survival. Ferritin and the modified Curie scores correlated with each other. MYCN amplification neither correlated with any aspect of the 123I-mIBG scans nor significantly predicted mCR or 2-year overall survival. LDH and ferritin are therefore appropriate neuroblastoma tumour markers to be used in low- and middle-income countries with limited or no access to mIBG scans and/or MYCN amplification studies.

Geographic range of vector-borne infections and their vectors: the role of African wildlife.

The role of African wildlife in the occurrence of vector-borne infections in domestic animals has gained renewed interest as emerging and re-emerging infections occur worldwide at an increasing rate. In Africa, biodiversity conservation and the expansion of livestock production have increased the risk of transmitting vector-borne infections between wildlife and livestock. The indigenous African pathogens with transboundary potential, such as Rift Valley fever virus, African horse sickness virus, bluetongue virus, lumpy skin disease virus, African swine fever virus, and blood-borne parasites have received the most attention. There is no evidence for persistent vector-borne viral infections in African wildlife. For some viral infections, wildlife may act as a reservoir through the inter-epidemic circulation of viruses with mild or subclinical manifestations. Wildlife may also act as introductory or transporting hosts when moved to new regions, e.g. for lumpy skin disease virus, Rift Valley fever virus and West Nile virus. Wildlife may also act as amplifying hosts when exposed to viruses in the early part of the warm season when vectors are active, with spillover to domestic animals later in the season, e.g. with bluetongue and African horse sickness. Some tick species found on domestic animals are more abundant on wildlife hosts; some depend on wildlife hosts to complete their life cycle. Since the endemic stability of a disease depends on a sufficiently large tick population to ensure that domestic animals become infected at an early age, the presence of wildlife hosts that augment tick numbers may be beneficial. Many wild ungulate species are reservoirs of Anaplasma spp., while the role of wildlife in the epidemiology of heartwater (Ehrlichia ruminantium infection) has not been elucidated. Wild ungulates are not usually reservoirs of piroplasms that affect livestock; however, there are two exceptions: zebra, which are reservoirs of Babesia caballi and Theileria equi, and buffalo, which are reservoirs of Theileria parva. The latter causes Corridor disease when transmitted from buffaloto cattle, butthis appearsto be a self-limiting condition, at least in southern Africa. Wild animals are important reservoirs of tsetse-transmitted Trypanosoma spp. infection. The distribution and abundance of some tsetse species, e.g. Glossina morsitans and G. pallidipes, are closely related to the occurrence of their preferred wildlife hosts.

Psychosocial presentation of revisional LAGB patients: a qualitative study.

This qualitative study offers insight into the experiences, expectations, perceptions and beliefs that may lead to laparoscopic adjustable gastric band patients' failure to achieve expected weight loss and seek revisional bariatric surgery. The 23 participants from two sites were interviewed and data were analysed from a grounded theory methodology in order to build a causal model. Analysis of participants' reports identified 'unrealistic expectations of the LAGB' as the core category. Additionally, the restriction of the band had a negative impact on participants' social interactions, leading to feelings of deprivation and, thus, to a desire for reward from food choices and consequently an increase of consumption of high-calorie-dense foods. These foods were chosen because of their specific texture or ability to provide reward. The resulting increase in weight or failure to achieve excess weight loss, led to feelings of shame and loneliness and emotional eating resulting in increased the consumption of rewarding foods. Thus, identifying unrealistic expectations of laparoscopic adjustable gastric band (LAGB) and emotional eating behaviours are important in those who are present initially for primary bariatric and revisional bariatric surgery, as they may contribute specifically to these patients' weight regain and consequent failure to achieve excess weight loss.

Malignant Catarrhal Fever: An Emerging Disease in the African Buffalo (Syncerus caffer).

Within the tribe Bovini in the subfamily Bovinae, the water buffalo (Bubalus Bubalis), American bison (Bison bison), European bison (Bubalus bonasus) and yak (Bos grunniens) are recognized as species highly susceptible to malignant catarrhal fever (MCF). In contrast, the lack of reports describing clinical MCF in the African buffalo (Syncerus caffer) whether free ranging or captive has led to a perception that African buffaloes are resistant to MCF. During the last decade, several cases of MCF in African buffaloes were confirmed in South Africa and experience with seven of these cases is described in this report. Detection of viral nucleic acid in blood or tissues was successful in six African buffaloes that suffered from clinical signs compatible with MCF. Four were positive for infection with ovine herpesvirus type 2 (the causative virus of sheep-associated MCF), and two were positive for alcelaphine herpesvirus type 1 (causative virus of wildebeest-associated MCF). Histopathological examination of tissue samples from all the animals yielded typical lesions that were consistent with those described for MCF in domestic cattle. Developments in the management of African buffaloes translocated from their traditional habitats have likely contributed to the identification of another susceptible host in the subfamily Bovinae.

Development of a real time polymerase chain reaction assay for equine encephalosis virus.

Equine encephalosis virus (EEV) is the cause of equine encephalosis. The disease is similar to mild forms of African horse sickness (AHS) and the two diseases are easily confused. Laboratory identification and serotyping of EEV is based on viral isolation in BHK-21 cells and a viral plaque inhibition neutralisation test. These procedures are time-consuming and therefore a more rapid diagnostic assay for EEV that can distinguish EEV from African horse sickness virus (AHSV) infections was developed. The S7 (VP7) gene from 38 EEV isolates representing all seven serotypes was amplified and sequenced. A conserved region at the 5' end of the gene was identified and used to design group-specific EEV primers and a TaqMan(®) MGB™ hydrolysis probe. The efficiency of the EEV real-time RT-PCR assay was 81%. The assay was specific, as it did not detect any of the nine serotypes of AHSV, nor 24 serotypes of bluetongue virus (BTV) and sensitive, with a 95% limit of detection of 10(2.9) TCID50/ml blood (95% confidence interval: 10(2.7) to 10(3.3)). The real-time format was selected because of its convenience, sensitivity and ability to produce results rapidly.

Genetic characterization of bovine viral diarrhoea (BVD) viruses: confirmation of the presence of BVD genotype 2 in Africa.

Bovine viral diarrhoea virus (BVDV) has emerged as one of the economically important pathogens in cattle populations, with a worldwide distribution and causing a complex of disease syndromes. Two genotypes, BVDV 1 and 2, exist and are discriminated on the basis of the sequence of the 5' non-coding region (5' NCR) using real-time PCR. Real-time PCR is more sensitive, specific, and less time-consuming than conventional PCR, and it has less risk of cross-contamination of samples. Limited information exists on BVDV genetic subtypes in South Africa. The aim of this study was to determine the genotypes of BVDV currently circulating in South African feedlots. A total of 279 specimens (219 tissue samples, 59 trans-tracheal aspirates and 1 blood sample) were collected from dead and living cattle with lesions or clinical signs compatible with BVDV infection. Pooled homogenates from the same animals were prepared, and total RNA was extracted. A screening test was performed on the pooled samples, and positive pools were investigated individually. A Cador BVDV Type 1/2 RT-PCR Kit (QIAGEN, Hilden, Germany) was used for the real-time PCR assay on a LightCycler(®) V2.0 real-time PCR machine (Roche Diagnostics, Mannheim, Germany). The results were read at 530 and 640 nm for BVDV 1 and 2, respectively. Bovine viral diarrhoea virus was detected in a total of 103 samples that included 91 tissue samples, 1 blood sample and 11 trans-tracheal aspirates. Eighty-five (82.5 %) of the strains were genotype 1 and 18 (17.5 %) were genotype 2. Comparing the sequencing data, genotypes 1 and 2 from the field strains did not cluster with vaccine strains currently used in feedlots in South Africa. The present study revealed the presence of BVDV genotype 2 in cattle in South Africa based on the high sequence similarity between genotype 2 field strains and strain 890 from North America. The presence of genotype 2 viruses that phylogenetically belong to different clusters and coexist in feedlots is consistent with the possibility of multiple virus introductions. These results represent the first documented evidence for the presence of BVDV genotype 2 in African cattle.

Evaluation of the effects of long-term storage of bovine ear notch samples on the ability of 2 diagnostic assays to identify calves persistently infected with bovine viral diarrhoea virus.

Research aimed at optimising diagnostic laboratory procedures is central to the development of effective bovine viral diarrhoea virus (BVDV) control programmes. BVDV is a single-stranded RNA virus that crosses the placenta to infect foetuses, resulting in reproductive losses due to foetal death or persistently infected calves that die early in life. Persistently infected animals are widely accepted to be the primary reservoir of BVDV and the largest source of infection. This poses important challenges to overall animal/herd health and can cause major losses to the cattle industry. Long-term storage of bovine ear notch samples from calves persistently infected with BVDV may adversely affect the ability of diagnostic assays to detect the virus efficiently. In order to test this hypothesis, ear notch samples from 7 animals were divided into 2 groups. One set was subjected to prompt formalin fixation and the other set stored either as fresh samples without preservatives at -2 degrees C, or soaked overnight in phosphate buffered saline followed by freezing of the supernatant fluid at -2 degrees C. Frozen ear notches and ear notch supernatant yielded positive results with an antigen-capture, enzyme linked immunosorbent assay (AC-ELISA) for the duration of the study (6 months) and optical density (OD) values remained significantly within range. There was no significant difference between storing fresh ear notch samples or PBS at -2 degrees C. However, positive immunohistochemistry (IHC) staining on formalin fixed ear notches started to fade between Day 17 and Day 29 when stored at room temperature. It was concluded that fresh ear notches could safely be stored at -2 degrees C for a period of 6 months prior to testing for BVD viral antigens.

A competitive ELISA for the detection of group-specific antibody to equine encephalosis virus.

A polyclonal antibody-based, group-specific, competitive ELISA (C-ELISA) for the detection of antibodies to equine encephalosis virus (EEV) was developed. The assay measures the competition between a specific guinea pig antiserum and a test serum, for a pre-titrated EEV antigen. The C-ELISA detected antibodies to the seven known EEV serotypes. Reference antisera raised against other arboviruses did not cross react with EEV antigen. Negative sera from horses in the United Kingdom were used to establish the baseline for a negative population. Negative and positive populations of South African horses, selected on the basis of virus neutralisation were assayed subsequently. Optimal test parameters, where sensitivity≅specificity≅100%, were calculated by two-graph receiver operator characteristic (TG-ROC) analysis to be at a cut-off value of 29.5% inhibition. Results show the EEV C-ELISA described to be sensitive, specific and reliable. Used in conjunction with ELISAs available for African horse sickness virus (AHSV), differential serological diagnosis between EEV and AHSV can be achieved.

Efficacy of vaccination at 4 and 6 weeks in the control of canine parvovirus.

Seroconversion after early vaccination at four weeks against canine parvovirus (CPV) using a high antigen titre vaccine was evaluated in 121 puppies from three breeds of dogs housed in kennels representative of the private practitioner's environment. The trial included 52 German shepherd pups, 25 Rottweiler pups and 44 Boerboel pups. From each group 11, 4, and 18 puppies acted as control dogs, respectively. Depending on the different groups, puppies were vaccinated at 4, 6, 9 and 12 weeks. The experimental group differed from the control group in that they received the high titre vaccine at 4 weeks of age, whereas the control group was not vaccinated at 4 weeks. Blood was collected from all pups prior to vaccination to measure maternally derived colostral antibody. The results indicated that vaccination at 4 weeks of age in pups with high maternally derived antibody levels, results in seroconversion rates that may lead to a reduction in the window of susceptibility with respect to CPV infection. The implications of the findings with respect to dogs in heavily contaminated environments are discussed.

A literature review and investigation of staphylococcal necrotic dermatitis in sheep.

An outbreak of necrotic dermatitis in sheep was intensively investigated. Initially 19 of 147 Letelle (Merino-type) ewes were identified but closer inspection revealed 57 sheep that had skin lesions, some very slight, and that the majority (46 or 80%) had lesions only above the lips. A small number of them had multiple lesions on the legs or vulvae apart from lip lesions. Seven had only vulvar lesions and 2 only leg lesions. Among the sheep with lip lesions, twice as many had lesions on the right as on the left. Electron micrographs did not reveal any virus particles from the lesions, but all bacterial swabs yielded pure cultures of beta-haemolytic, Gram-positive cocci that were catalase, coagulase and DNase positive. The organism was identified as Staphylococcus aureus. Histopathology was consistent with a dermotoxic insult. A review of available literature indicated that this outbreak was consistent with a diagnosis of ovine necrotic (staphylococcal) dermatitis, supported by data on signalment, lesions, distribution, size, number, epidemiology as well as specific tests. The range of differential diagnoses and possible confusers are discussed.

Monoclonal gammopathy and other serum protein electrophoresis patterns in patients with HIV infection in South Africa.

BACKGROUND: Monoclonal gammopathy has been noted with increased frequency in patients with HIV infection compared with the general population. The prevalence in patients on antiretroviral therapy (ART) has not been widely investigated. The present study aims to describe the prevalence and characteristics of monoclonal proteins in a population of HIV-infected patients in the Western Cape region of South Africa. METHODS: Clinical information and blood and urine samples were collected from patients with proven HIV infection attending two clinics in the Western Cape. Serum protein electrophoresis was performed on all patients using agarose gel. Those with visible mono- or oligoclonal bands or increased gamma fractions were subjected to immunofixation electrophoresis. RESULTS: Three hundred and sixty-eight patients were included in the study with a mean age of 37.7 y. Of these patients, 326 were on ART for a mean duration of 19.6 months. Monoclonal bands were found in 12 patients (3.2%) and oligoclonal bands in 14 (3.8%). These bands were associated with a shorter duration of ART, increased total protein, acute on chronic phase pattern on electrophoresis and an increased gamma fraction, but not associated with CD4 count and viral load. The majority were of low concentration and of the IgG isotype. CONCLUSION: Our study confirms that the prevalence of monoclonal and oligoclonal bands is higher in this HIV-positive cohort on ART compared with that of the general population. However, the prevalence is lower than previously reported in other studies.

Intradermal tuberculin testing of wild African lions (Panthera leo) naturally exposed to infection with Mycobacterium bovis.

African lions in the southern half of Kruger National Park (KNP) are infected with Mycobacterium bovis. Historically, reliable detection of mycobacteriosis in lions was limited to necropsy and microbiological analysis of lesion material collected from emaciated and ailing or repeat-offender lions. We report on a method of cervical intradermal tuberculin testing of lions and its interpretation capable of identifying natural exposure to M. bovis. Infected lions (n=52/95) were identified by detailed necropsy and mycobacterial culture. A large proportion of these confirmed infected lions (45/52) showed distinct responses to bovine tuberculin purified protein derivative (PPD) while responses to avian tuberculin PPD were variable and smaller. Confirmed uninfected lions from non-infected areas (n=11) responded variably to avian tuberculin PPD only. Various non-tuberculous mycobacteria (NTM) were cultured from 45/95 lions examined, of which 21/45 were co-infected with M. bovis. Co-infection with M. bovis and NTM did not influence skin reactions to bovine tuberculin PPD. Avian tuberculin PPD skin reactions were larger in M. bovis-infected lions compared to uninfected ones. Since NTM co-infections are likely to influence the outcome of skin testing, stricter test interpretation criteria were applied. When test data of bovine tuberculin PPD tests were considered on their own, as for a single skin test, sensitivity increased (80.8-86.5%) but false positive rate for true negatives (18.75%) remained unchanged. Finally, the adapted skin test procedure was shown not to be impeded by persistent Feline Immunodeficiency Virus(Ple) co-infection.

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 vaccine-induced, 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.

Multi-host pathogens and carnivore management in southern Africa.

A retrospective serosurvey of multi-host feline and canine viruses among carnivore species in southern Africa (n=1018) identified widespread pathogen exposure even in remote protected areas. In contrast to mortality experienced in East African predators, canine distemper virus (CDV) infection among African wild dogs (Lycaon pictus) in Botswana was not associated with identifiable change in pup survivorship or disease related mortality of adults. A disease outbreak of unknown aetiology occurred in the same population over 4 weeks in 1996. Outbreak boundaries coincided with ecotones, not the spatial distribution of contiguous packs, highlighting the potential importance of landscape heterogeneities in these processes. Direct management of pathogens in domestic animal reservoirs is complicated by the apparent complexity of pathogen maintenance and transmission in these large systems. Conservation effort should be focused at securing large metapopulations able to compensate for expected episodic generalist pathogen invasion and attention directed to addressing underlying causes of population depression such as habitat loss and wildlife conflict.

The epidemiology of lion lentivirus infection among a population of free-ranging lions (Panthera leo) in the Kruger National Park, South Africa.

Feline immunodeficiency virus is a lentivirus of domestic cats that causes significant lifelong infection. Infection with this or similar lentiviruses has been detected in several nondomestic feline species, including African lions (Panthera leo). Although lion lentivirus (FIVple) infection is endemic in certain lion populations in eastern and southern Africa, little is known about its pathogenic effects or its epidemiological impact in free-ranging lions. This report describes the epidemiological investigation of lentivirus positivity of free-ranging lions in the Kruger National Park, South Africa. A nested polymerase chain reaction assay for virus detection was performed on all whole blood samples collected. In addition, serum samples were tested for cross-reactive antibodies to domestic feline lentivirus antigens and to puma lentivirus synthetic envelope peptide antigen. The results were analysed in conjunction with epidemiological data to provide a descriptive epidemiological study on lion lentivirus infection in a free-ranging population of lions. The overall prevalence of lentivirus infection was 69%, with a prevalence of 41% in the north of the park, and 80% in the south. Adult males had the highest prevalence when combining the factors of sex and age: 94%. The lowest prevalences were found among juveniles, with male juveniles at 29%. Adults were 5.58 times more likely to test positive for FIVple than juveniles, with adult males being 35 times more likely to be test positive for FIVple compared with juvenile males. This research represents the 1st epidemiological study of the lion lentivirus among free-ranging lions in the Kruger National Park.

Gammaherpesvirus carrier status of black wildebeest (Connochaetes gnou) in South Africa.

Malignant catarrhal fever (MCF) is an economically important disease primarily of domestic cattle with a high case fatality rate. It is caused by either alcelaphine herpesvirus type 1 (AlHV-1) or ovine herpesvirus type 2 (OvHV-2). The major reservoir host of AlHV-1 is the blue wildebeest (Connochaetes taurinus), but it is generally accepted that the black wildebeest (Connochaetes gnou) is also a reservoir host. No viral studies in the black wildebeest have been reported and the carrier status of black wildebeest has not been documented. Specimens were collected from several game farms and conservation areas in central South Africa representing the geographical area historically linked to the natural habitat of the black wildebeest. Specimens were obtained from 304 black wildebeest of different ages and sex, as well as 51 black wildebeest foetuses at different stages of gestation. Virus was isolated from a black wildebeest calf. Morphological features and antigenic characteristics suggested it to be a gammaherpesvirus closely related to AlHV-1. All serum samples tested positive with a competitive inhibition enzyme-linked immunosorbent assay (CI-ELISA) for group-specific malignant catarrhal fever virus antibody. A SYBR Green real-time PCR assay was developed for the detection of gammaherpesviral DNA. Only 15.8 % of the animals tested positive with the real-time PCR assay whereas 90 % of the foetuses tested positive. This finding suggests that, unlike OvHV-2 infection in lambs in which the infection takes place after weaning, the virus in black wildebeest is mainly transmitted in utero or soon after birth. The results suggest that black wildebeest are latent carriers of a gammaherpesvirus similar or closely related to AlHV-1 present in blue wildebeest and that it is likely that all black wildebeest are persistently infected.

The use of a probiotic in captive cheetahs (Acinonyx jubatus).

Juvenile captive cheetahs (Acinonyx jubatus) often present with diarrhoea that is commonly associated with bacterial infections. A species-specific probiotic containing Lactobacillus Group 2 and Enterococcus faecium was prepared from healthy adult cheetahs. Juvenile cheetahs (n = 27) between 8 and 13 months of age were included in the probiotic trial. The animals were observed prior to and after feeding of the probiotic which was made available for 28 days. Feeding of the probiotic resulted in a significantly increased body weight in the treatment group (P = 0.026), while there was no increase in the control group. A relative improvement in the faecal quality in the probiotic group during the treatment period compared with the pre-treatment (P = 0.0363) and post-treatment (P = 0.004) period was observed. This was accompanied by an absence of blood and mucus in the faeces during the treatment period in the probiotic group.

Ulcerative balanitis and vulvitis of Dorper sheep in South Africa: a study on its aetiology and clinical features.

Ovine ulcerative balanitis and vulvitis in sheep of the Dorper breed has been observed in South Africa since 1979. Its aetiology has not been conclusively resolved, and there is some discrepancy in descriptions of its clinical features. In order to identify the pathogenic micro-organism/s that contribute to the occurrence of the disease, the microflora in the genital tracts of both clinically healthy and affected sheep were isolated and compared. Bacteriological examination of materials from affected and unaffected sheep resulted in the isolation of Arcanobacterium pyogenes from 44.2% and 17.2% of them respectively. This difference is statistically significant (P < 0.01). Seventy-four per cent of the isolates originated from severe clinical cases. Mycoplasmas were isolated from 49.3% of 116 clinically normal sheep and 78.2% of 104 affected sheep. There were significant differences in their rates of isolation in clinical groups (P < 0.05). Of all the mycoplasma isolates, Mycoplasma mycoides mycoides large colony variant (MmmLC) was isolated from 61.5% of clinically diseased sheep while 6.0% of the isolates were from apparently healthy animals (P < 0.05). The study threw light on the prevalence of mycoplasmas in the genital tract of apparently healthy sheep and, at the same time the identity of the mycoplasma pathogen associated with ulcerative balanitis and vulvitis was revealed. The findings of this investigation therefore confirmed the involvement of mycoplasma, particularly that of MmmLC large colony, in the disease in Dorper sheep in South Africa, and it was concluded that this microorganism is an important pathogen of balanitis and vulvitis in them. The study furthermore demonstrated a probable synergism between A. pyogenes and MmmLC. Finding these 2 organisms together occurred 53.4 times more frequently in the affected sheep than in the unaffected, which emphasises the probable multifactorial nature of the disease. The association between age and the presence of clinical signs was statistically significant. It was found that young sheep were more likely to have lesions than adult sheep. Clinical observations showed that the typical ulceration appears to be confined to the glans penis and lips of the vulva; no ulceration was observed on the shaft of the penis and prepuce or vaginal vestibule. In uncomplicated cases inflammation of the prepuce and vaginal vestibule is not a regular feature of the disease. Therefore the names ulcerative balanitis and vulvitis most accurately describe the nature of the disease in South Africa.