Investigating selective media for optimal isolation of Brucella spp. in South Africa.

Bovine brucellosis in South Africa is caused mainly by Brucella abortus biovar (bv.) 1 and less frequently by B. abortus bv. 2. Bacterial isolation is regarded as the gold standard for diagnosis of Brucella species; however, it is not very sensitive. The aim of this study was to determine the selective medium with optimum antibiotic composition that will allow the growth of Brucella species (spp.) while inhibiting moulds, yeast and most, if not all, Gram-negative contaminants in South Africa. In the controlled experiment, modified Agrifood Research and Technology Center of Aragon (CITA) medium (mCITA) seemed to be the optimum selective medium for isolation of Brucella spp. as compared with Farrell's medium (FM) and modified Thayer Martin (mTM), while FM inhibited the growth of most fungal and bacterial contaminants. Mean comparison between the three media used to culture B. abortus resulted in lower mean difference ranging from 0 to 2.33. In case of Brucella ovis, high mean difference was obtained when comparing FM with mCITA (10.33) and mTM (12). However, the mean differences of 0.67 and 1.67 were obtained when comparing mCITA and mTM media used to, respectively, culture pasteurised and raw milk spiked with B. ovis. Further optimisation at the Agricultural Research Council - Onderstepoort Veterinary Research Institute resulted in a comparable performance between FM and mCITA; however, mCITA allowed optimal growth of the fastidious B. ovis, which is generally inhibited on FM. Generally, mCITA seemed to be the optimum selective medium for isolation of Brucella spp., while FM inhibits the growth of most fungal and bacterial contaminants. Thus, veterinary laboratories can use mCITA and/or FM but should take into consideration the detection of factious Brucella isolated in the country or region.

Seroprevalence and characterization of Brucella species in cattle slaughtered at Gauteng abattoirs, South Africa.

BACKGROUND: Brucellosis is an infectious and contagious zoonotic bacterial disease of both humans and animals. In developing countries where brucellosis is endemic, baseline data on the prevalence of brucellosis, using abattoir facilities, is important. OBJECTIVES: The aim of this study was to determine the seroprevalence of antibodies against Brucella in slaughter cattle at Gauteng province, South Africa and to characterize isolates of Brucella spp. METHODS: In this cross-sectional study, un-clotted blood samples with corresponding organ tissue samples were collected from slaughtered cattle. Serological [Rose Bengal test (RBT), complement fixation test (CFT) and indirect ELISA (iELISA)], molecular (PCR) and bacteriological methods were used to detect Brucella antibodies and Brucella spp. from 200 slaughtered cattle in 14 abattoirs. RESULTS: The RBT revealed a seroprevalence of brucellosis as 11.0% (22 of 200) and iELISA confirmed 5.5% (11 of 200). The estimated seroprevalence from RBT and iELISA was 5.5% while RBT and CFT was 2.0% (4 of 200). Brucella melitensis (n = 6) and B. abortus (n = 5) were isolated from 11 cattle tissues (5.5%) as confirmed to species level with AMOS PCR and differentiated from vaccine strains with Bruce-ladder PCR. Seven of the 11 isolates originated from seropositive cattle of which five were biotyped as B. abortus bv 1 (n = 2) and B. melitensis bv 2 (n = 1) and B. melitensis bv 3 (n = 2). CONCLUSIONS: This is the first documentation of B. melitensis in cattle in South Africa. The zoonotic risk of brucellosis posed by Brucella-infected slaughter cattle to abattoir workers and consumers of improperly cooked beef cannot be ignored.

Detection of Salmonella from animal sources in South Africa between 2007 and 2014.

Retrospective laboratory-based surveillance was conducted on Salmonella serotypes isolated from various animal species from 2007 to 2014 at the Agricultural Research Council, Onderstepoort Veterinary Research Institute, South Africa. During the surveillance period, 1229 salmonellae isolations were recorded. Around 108 different serotypes were recovered from nine different food and non-food animal host species. The three most common serotypes were Salmonella entericasubspecies enterica serotype Heidelberg (n = 200), Salmonella enterica subspecies enterica serotype Enteritidis (n = 170) and Salmonella enterica subspecies enterica serotype Typhimurium (n = 146). These were followed by Salmonella enterica subspecies enterica serotype Anatum (n = 62) and Salmonella entericasubspecies enterica serotype Infantis (n = 57). Salmonella enterica subspecies enterica serotype Schwarzengrund and Salmonella enterica subspecies entericaserotype Muenchen were recovered in 50 and 48 cases, respectively. Of the total number of isolations recorded during the period under review, 871 (70.8%) occurred in poultry and other birds, 162 (13.2%) in horses, 116 (9.4%) in cattle, 26 (2.1%) in sheep and goats, 22 (1.8%) in rhinoceroses, 16 (1.3%) in pigs, 8 (0.6%) in crocodiles, 6 (0.5%) in cats and 6 (0.5%) in leopards. Food animals accounted for 83.5% of the total isolations, with cattle and poultry representing approximately 72.7%. Forty-two (3.4 %) isolates were found from non-food animals that include rhinoceroses (n = 22), crocodiles (n = 8), leopards (n = 6) and cats (n = 6). Salmonella Heidelberg was the most frequently isolated serotype, whereas S. Typhimurium had the widest zoological distribution. Clinical laboratory isolation of different Salmonella serotypes from various hosts may aid in recognising the threat to livestock, public and environmental health. Moreover, it may also highlight the potential zoonotic and food safety risk implications of the detected Salmonella serotypes.

Epidemiology and characterization of Escherichia coli outbreak on a pig farm in South Africa.

An investigation of mortality of piglets through clinical signs, post-mortem, histopathology and bacteriological analyses revealed the causal organism to be Escherichia coli, mainly O149:K91:K88 which belongs to the enterotoxigenic biotypes. Molecular characterization and epidemiologic analysis elucidated it as shiga-toxin (ST) E. coli resistant to ampicillin, cefotaxime, tetracycline, trimethoprim-sulfamethoxazole, tylosin and neomycin. Conventional PCR results detected genes for ST-2, adhesin involved in diffuse adherence (AIDA-1) and F18 fimbriae virulence factors. Survival analyses and logistic regression of piglet mortality patterns showed that season of weaning, weaning weight and age of dam had significant influence on survival rate of piglets. Factors affecting pathogenicity of bowel edema and survival of affected piglets on a farm with persistent infection were reported for the first time. An association of E. coli O149:K91:K88 (F4) with clinical edema disease was made even though it has been reported in the past that this serotype does not produce ST. It was concluded that more stringent measures to mitigate the impact of the disease need to be targeted for spring and in older sows.

Salmonella contamination, serovars and antimicrobial resistance profiles of cattle slaughtered in South Africa.

Antimicrobial resistant Salmonella are among the leading causes of foodborne infections. Our aim was to determine Salmonella contamination during cattle slaughter in South African rural abattoirs (n = 23) and environmental samples. Furthermore, antimicrobial resistance patterns of the Salmonella isolates were determined. Samples of cattle faeces (n = 400), carcass sponges (n = 100), intestinal contents (n = 62), hides (n = 67), and water from the abattoirs (n = 75) were investigated for Salmonella species using microbiological techniques and species-specific polymerase chain reaction targeting the invA gene. In total 92 Salmonella species isolates were recovered. The Salmonella mean frequency of occurrence on hides, carcasses, and intestinal contents was 35.37% (n = 81). Eleven faecal samples (2.75%) tested positive for Salmonella. The predominant serovar was Salmonella Enteritidis. Diverse serovars that were identified on carcasses were not necessarily found on the hides and intestinal contents. The inconsistent occurrence of the diverse Salmonella serovars on hides, carcasses, and intestinal contents implies that in addition to carriage on hides and in intestinal contents, other external factors also play an important role regarding carcass contamination. The 92 Salmonella were serotyped and tested for susceptibility towards the following antimicrobials: ampicillin, cefotaxime, enrofloxacin, kanamycin, and oxytetracycline using the disk diffusion method. Most Salmonella (n = 66; 71.7%) isolates were resistant to at least one antimicrobial with highest resistance observed towards oxytetracycline (51.90%), which highlights the need for strict hygiene during slaughter and prudent antimicrobial use during animal production. In conclusion, cattle slaughtered in South African rural abattoirs harbour diverse Salmonella serovars that are resistant to antimicrobials, which could be a public health risk. The findings should assist policymakers with improving implementation of hygienic slaughter of cattle in rural abattoirs, which is paramount from socioeconomic, public health, and epidemiological standpoints.