The Emergence of Colistin- and Imipenem-Associated Multidrug Resistance in Escherichia coli Isolates from Retail Meat.

To determine the prevalence of Escherichia coli and their drug resistance profiles in fresh pork sold at two retail outlets (open-air market and closed retail stores) in Alice, South Africa. Retail meat samples (n = 176) collected from four shops (two from open-air markets and two from closed stores) were analyzed by conventional biochemical and PCR-based molecular confirmatory tests. The confirmed isolates were profiled for antimicrobial susceptibility to a panel of 12 commercial antibiotics: tetracycline, ampicillin, sulphamethoxazole trimethoprim, erythromycin, gentamycin, colistin sulphate, cefotaxime, chloramphenicol, norfloxacin, ciprofloxacin, cefuroxime, and imipenem. Colistin, ampicillin, and erythromycin resistance genes were profiled with the gene-specific primers. Multidrug resistance (MDR) and the association of imipenem and colistin in the MDR profile were determined. A total of 68 (39.08%) E. coli isolates were confirmed by PCR analysis. Resistance was most common to erythromycin (100%), followed by cefotaxime (95.58%), ampicillin (88.23%), cefuroxime (88.23%), trimethoprim-sulphamethoxazole (88.23%), and tetracycline (60.29%). Overall, 27/68 (39.70%) were MDR (≥ 3antibiotics classes). MDR E. coli isolates associated with imipenem resistance (50.00%) and colistin resistance (33.82%) were detected. The resistance genes were detected among the isolates though not in all the phenotypically resistant isolates. The detection of colistin resistance among MDR E. coli isolates from retail meat is troubling as the drug is a last resort antibiotic. Overall, the epidemiological implications of the findings are of public health importance.

Socio-demographic and clinical determinants of late presentation among patients newly diagnosed with HIV in the Eastern Cape, South Africa.

South Africa recently implemented the 'test and treat' strategy for all HIV-infected individuals receiving diagnosis at the health facility level. However, the impact of this programme in terms of the prevention of HIV transmission, morbidity and mortality associated with HIV can only be maximized if patients are diagnosed early. This study determines the prevalence of late presentation among newly diagnosed HIV-infected individuals and also examines the socio-demographic and clinical determinants for late presentation in health facilities in the Eastern Cape Province, South Africa.In this cross-sectional study, a total of 335 newly diagnosed patients were recruited consecutively between August 2016 and July 2017. Late presenter for HIV care was defined in accordance with the European Late Presenter Consensus working group as a patient who reports for care when the CD4 count is below 350 cells/µL and/or when there is an established AIDS-defining clinical condition, irrespective of CD4 count. Adjusted and unadjusted logistic regression analysis was used to examine the determinants of late HIV diagnosis.Participants' mean age was 33.6 (SD: 10.6). Almost 96% of the participants believed their route of HIV infection was heterosexual sex. Most newly diagnosed HIV-infected patients (60%) were late presenters (CD4+ count ≤350 cells/µL and/or having an AIDS-defining illness in World Health Organisation (WHO)-defined stage III/IV), with 35% presenting with Acquired Immune Deficiency Syndrome (AIDS)-related complications. In the adjusted model, only male sex (AOR: 2.81; CI: 1.51-5.23), no formal education (AOR: 5.63; CI: 1.68-18.85), and overweight body mass category (AOR: 2.45; CI: 1.04-5.75) were independently associated with late HIV diagnosis.The majority of newly diagnosed HIV-infected individuals were late presenters. To maximize the impact of the 'test and treat' policy aimed at reducing new HIV transmissions and preventing the morbidity and mortality associated with HIV, there is a need for programmes to improve early detection of HIV in the study settings. This programme should target males and individuals with no formal education for maximum impact.

The activity of commercial antimicrobials, and essential oils and ethanolic extracts of Olea europaea on Streptococcus agalactiae isolated from pregnant women.

BACKGROUND: Streptococcus agalactiae also known as Group B Streptococcus (GBS) is a major cause of disease in pregnant women and new born babies where it causes early and late onset disease characterised by sepsis, pneumonia and meningitis. Ten to 37 % of pregnant women in the world are colonised with GBS while intrapartum antibiotic prophylaxis has led to significant reduction in early onset disease. The increase in drug resistant microorganisms has become a major threat. Development of vaccines is still in progress so there is need for new and safer alternatives to treatment. METHODS: Benzyl penicillin, Ampicillin, Cefotaxime, Ceftriaxone, Levofloxacin, Erythromycin, Clindamycin, Linezolid, Vancomycin, Tetracycline and Cotrimoxazole, Olea europaea leaf extracts and essential oil were tested against GBS isolates from South Africa and Namibia. RESULTS: The isolates showed 100% sensitivity to benzyl penicillin, ampicillin, ceftriaxone, levofloxacin, linezolid, vancomycin, O. europaea leaf extracts and essential oils. Only one isolate (0.6%) was resistant to cefotaxime and 23.4 and 10.4% were resistant to clindamycin and erythromycin respectively. CONCLUSION: GBS isolates showed sensitivity to O. europaea extracts at low minimum inhibitory concentrations. Β lactams are still the drugs of choice for treatment of GBS disease but O. europaea extracts potent as an alternative source of antimicrobials.

Prevalence and capsular type distribution of Streptococcus agalactiae isolated from pregnant women in Namibia and South Africa.

BACKGROUND: Streptococcus agalactiae or Group B Streptococcus (GBS) is the leading cause of neonatal morbidity and mortality resulting in septicaemia, bacteraemia and meningitis. Long term problems in children range from loss of hearing to mental retardation. While Intrapartum Antibiotic Prophylaxis (IAP) has reduced the incidence of S. agalactiae infection, it still remains the leading cause of disease in neonates. GBS has ten capsular types whose distribution varies across the world. Therefore, this study sought to determine the prevalence of GBS in Namibia and South Africa amongst pregnant women between 35 and 37 weeks gestation and elucidate the capsular types. METHODS: Lower vaginal and rectal swabs were collected from pregnant women between 35 and 37 weeks gestation. Five hundred and thirty pregnant women were recruited into the study in Windhoek, Namibia while one hundred pregnant women were recruited in the Eastern Cape, South Africa. The swabs were cultured on 5% sheep blood agar (Biomerieux, New Jersey, USA) for isolation of GBS. Presumptive isolates were confirmed using both the Vitek (2) and molecular techniques targeting the scpB gene. Capsular typing was performed in a multiplex PCR with capsular specific primer pairs. RESULTS: The prevalence of GBS in Namibia was 13.6 and 37% in South Africa respectively. In both countries most women were dually colonised with GBS. Capsular types II, III and V were the most prevalent. CONCLUSIONS: The prevalence of GBS in Namibia was lower than in South Africa in this study. The prevalence in both countries was not different from those reported in other African countries and around the world. The predominant capsular types in this study are the ones commonly associated with adverse maternal outcomes.

Global Status of Porcine circovirus Type 2 and Its Associated Diseases in Sub-Saharan Africa.

Globally, Porcine circovirus type 2 (PCV2) is a recognized viral pathogen of great economic value in pig farming. It is the major cause of ravaging postweaning multisystemic wasting syndrome (PMWS) and many other disease syndromes generally regarded as Porcine circovirus associated diseases (PCVAD) in Europe. PCV2 infections, specifically PMWS, had impacted huge economic loss on swine production at different regions of the world. It has been studied and reported at different parts of the globe including: North and South America, Europe, Asia, Oceania, Middle East, and the Caribbean. However, till date, this virus and its associated diseases have been grossly understudied in sub-Sahara African region and the entire continent at large. Two out of forty-nine, representing just about 4% of countries that make up sub-Sahara Africa presently, have limited records on reported cases and occurrence of the viral pathogen despite the ubiquitous nature of the virus. This review presents an overview of the discovery of Porcine circovirus and its associated diseases in global pig herds and emphasizes the latest trends in PCV2 vaccines and antiviral drugs development and the information gaps that exist on the occurrence of this important viral pathogen in swine herds of sub-Saharan Africa countries. This will serve as wake-up call for immediate and relevant actions by stakeholders in the region.

Genetic profiling for Anaplasma and Ehrlichia species in ticks collected in the Eastern Cape Province of South Africa.

BACKGROUND: Anaplasma and Ehrlichia are emerging tick-borne pathogens that cause anaplasmosis and ehrlichiosis in humans and other animals worldwide. Infections caused by these pathogens are deadly if left untreated. There has been relatively no systematic survey of these pathogens among ticks in South Africa, thus necessitating this study. The presence of Anaplasma and Ehrlichia species were demonstrated by PCR in ticks collected from domestic ruminants at some selected communities in the Eastern Cape of South Africa. The ticks were identified by morphological characteristics and thereafter processed to extract bacterial DNA, which was analyzed for the presence of genetic materials of Anaplasma and Ehrlichia. RESULTS: Three genera of ticks comprising five species were identified. The screening yielded 16 positive genetic materials that were phylogenetically related to Ehrlichia sequences obtained from GenBank, while no positive result was obtained for Anaplasma. The obtained Ehrlichia sequences were closely related to E. chaffeensis, E. canis, E. muris and the incompletely described Ehrlichia sp. UFMG-EV and Ehrlichia sp. UFMT. CONCLUSION: The findings showed that ticks in the studied areas were infected with Ehrlichia spp. and that the possibility of transmission to humans who might be tick infested is high.

Macrolide, glycopeptide resistance and virulence genes in Enterococcus species isolates from dairy cattle.

The genus Enterococcus is known to possess the capacity to acquire and disseminate antimicrobial resistant determinants alongside the ability to produce various virulence genes that enables it to establish infections. We assessed the prevalence and antibiogram profiles of Enterococcus spp. in faecal samples of dairy cattle. Faecal swab samples were collected from 400 dairy cattle from two commercial cattle farms in two rural communities in the Eastern Cape, South Africa. Confirmation of enterococci isolates was carried out by PCR targeting of the tuf gene. Species delineation was by species-specific primers targeting the superoxide dismutase (sod A) gene in a multiplex PCR assay. Isolates were screened for the presence of the following virulence genes (ace, gel E, esp, efa A, cyl A and hyl E) and antimicrobial resistance determinants to erythromycin, vancomycin and streptomycin were evaluated molecularly. A total of 340 isolates were confirmed as belonging to the genus Enterococcus . Species distribution among the isolates consisted of Enterococcus faecium (52.94 %) and Enterococcus durans (23.53 %) in preponderance compared to the three other species, namely Enterococcus faecalis (8.8 %), Enterococcus hirae (8.6 %) and Enterococcus casseliflavus (5.9 %). All were resistant to vancomycin, while 99 % showed resistance to aminoglycoside and 94 % to macrolide. Three virulence genes (ace, gel E and esp) were detected in almost all the confirmed isolates. The resistance determinants van B (19.7 %), van C1 (25 %), van C2/3 (26.3 %) erm B (40.29 %) and str A (50.88 %) were detected among the isolates. A high prevalence of multidrug-resistant enterococci isolates was detected in this study and the genetic repertoire to survive in the presence of antimicrobial agents was present in these organisms.

Multiple antibiotic resistances among Shiga toxin producing Escherichia coli O157 in feces of dairy cattle farms in Eastern Cape of South Africa.

BACKGROUND: Shiga toxin-producing Escherichia coli (STEC) O157:H7 is a well-recognized cause of bloody diarrhea and hemolytic-uremic syndrome (HUS). The ability of STEC strains to cause human disease is due to the production of Shiga toxins. The objectives of this study were to determinate the prevalence, serotypes, antibiotic susceptibility patterns and the genetic capability for Shiga toxin production in Escherichia coli (STEC) strains isolated from dairy cattle farms in two rural communities in the Eastern Cape Province of South Africa. METHODS: Fecal samples were collected between March and May 2014, from individual cattle (n=400) in two commercial dairy farms having 800 and 120 cattle each. Three hundred presumptive isolates obtained were subjected to polymerase chain reactions (PCR) for identification of O157 serogroup and Shiga toxin producing genes (stx1, stx2) on genomic DNA extracted by boiling method. Susceptibility of the isolates to 17 antibiotics was carried out in vitro by the standardized agar disc-diffusion method. RESULTS: Based on direct PCR detection, 95 (31.7%) isolates were identified as O157 serogroup. The genetic repertoire for Shiga toxin production was present in 84 (88.42%) isolates distributed as stx1 (37), stx2 (38) and stx1/2 (9) respectively while 11 of the isolates did not harbor Shiga toxin producing genes. Multiple antibiotic resistances were observed among the isolates and genetic profiling of resistance genes identified bla ampC 90%, blaCMY 70%, blaCTX-M 65%, blaTEM 27% and tetA 70% and strA 80% genes among the antimicrobial resistance determinants examined. CONCLUSION: We conclude that dairy cattle farms in the Eastern Cape Province are potential reservoirs of antibiotic resistance determinants in the province.

Virulence and antimicrobial resistance factors of Enterococcusspp. isolated from fecal samples from piggery farms in Eastern Cape, South Africa.

BACKGROUND: Enterococci have emerged as an important opportunistic pathogen causing life-threatening infections in hospitals. The emergence of this pathogen is associated with a remarkable capacity to accumulate resistance to antimicrobials and multidrug-resistance particularly to vancomycin, erythromycin and streptomycin have become a major cause of concern for the infectious diseases community. In this paper, we report the prevalence of Enterococcus in respect to species distribution, their virulence and antibiogram profiles. METHODS: Four hundred fecal samples were collected from two piggery farms in the Eastern Cape Province of South Africa. Enterococcus species were isolated and confirmed with generic specific primers targeting the tuf gene (encoding elongation factor). The confirmed isolates were speciated with enterococci species specific primers that aimed at delineating them into six species that are commonly associated with infections in humans. Antibiotic susceptibility testing was performed by disc diffusion method. Six virulence genes and antimicrobial resistance profiles of the isolates were evaluated molecularly. RESULTS: Molecular identification of the presumptive isolates confirmed 320 isolates as Enterococcus spp. Attempt at speciation of the isolates with primers specific for E. faecalis, E. durans, E. casseliflavus, E. hirae and E. faecium delineated them as follows: E. faecalis (12.5 %), E. hirae (31.25 %), E. durans (18.75 %) and E. faecium (37.5 %) while E. casseliflavus was not detected. All the isolates were resistant to vancomycin, streptomycin and cloxacillin, and to at least two different classes of antibiotics, with 300 (93.8 %) isolates being resistant to five or more antibiotics. Also, three out of the six virulence genes were detected in majority of the isolates and they are Adhesion of collagen in E. faecalis (ace) (96.88 %), gelatinase (gelE) (93.13 %) and surface protein (esp) (67.8 %). CONCLUSION: There was high prevalence of multi-resistant vancomycin Enterococcus spp. (VREs) in the fecal samples of pigs in the farms studied, and this poses health implications as vancomycin is an important drug in human medicine. Further studies are needed to determine the spread of vancomycin resistance among bacteria of human origin in the communities.

Anti-staphylococcal, anti-HIV and cytotoxicity studies of four South African medicinal plants and isolation of bioactive compounds from Cassine transvaalensis (Burtt. Davy) codd.

BACKGROUND: Medicinal plants represent an important opportunity to rural communities in Africa, as a source of affordable medicine and as a source of income. Increased patient awareness about safe usage is important as well as more training with regards to traditional medicine. The aim of this study was to evaluate the ethnomedicinal prowess of some indigenous South African plants commonly used in Eastern Cape Province of South Africa for the treatment of skin and respiratory tract infections, HIV and their toxicity potential. METHODS: Cassine transvaalensis, Vangueria infausta, Croton gratissimus and Vitex ferruginea were tested for antibacterial activities against Staphylococcus aureus and Staphylococcus epidermidis using Kirby-Bauer disk diffusion and minimum inhibition concentration (MIC). Cytotoxic and anti-HIV-1 activities of plants were tested using MTT Assay (3- (Dimethylthiozole-2-yl-2,5-diphenyltetrazolium bromide)) and anti- HIV-1iib assay. In search of bioactive lead compounds, Cassine transvaalensis which was found to be the most active plant extract against the two Staphylocoous bacteria was subjected to various chromatographic. Thin layer chromatography, Column chromatography and Nuclear Magnetic Resonance (NMR), (1H-1H, 13C-13C, in DMSO_d6, Bruker 600 MHz) were used to isolate and characterize 3-Oxo-28-hydroxylbetuli-20(29)-ene and 3,28-dihydroxylbetuli-20(29)-ene bioactive compounds from C. transvaalensis. RESULTS: The four plants studied exhibited bioactive properties against the test isolates. The zones of inhibition ranged between 16 mm to 31 mm for multi-drug resistant staphylococci species. MIC values varied between 0.6 and 0.02 µg/ml. C. gratissimus and C. transvaalensis exhibited the abilities to inhibit HIV-1iib. Two bioactive compounds were isolated from C. transvaalensis. CONCLUSION: Data from this study reveals the use of these plant by traditional healers in the Eastern Cape. Furthermore, C. transvaalensis and C. gratissimus were found to be more active as against HIV-1iib. While C. transvaalensis was most active against the two Staphylococcus bacteria.

Detection of potential risk of wastewater effluents for transmission of antibiotic resistance from Vibrio species as a reservoir in a peri-urban community in South Africa.

We assessed the antibiogram characteristics of some Vibrio species isolated from wastewater final effluents in a typical peri-urban community of South Africa. Marked resistances were noted against erythromycin (100%), chloramphenicol (100%), nitrofurantoin, cefuroxime and cephalothin (90-95%) in V. parahaemolyticus, V. fluvialis and V. vulnificus, respectively. Fourteen antibiotypes were identified, with multiresistance to 8-10 antibiotics being common. The antibiotypes AMP, PEN, STR, SUL, TMP, COT, CHL, ERY, CIP and PB demonstrated by V. fluvialis were the most prevalent (17.24%). Eight putative antibiotic resistance genes were identified with floR being the mostly (100%) detected in all the three species while tet(A) was the least with 65% prevalence in V. vulnificus, 7.14% in V. parahaemolyticus and none in V. fluvialis. These results demonstrate that the treated effluent system are reservoirs for various antibiotic resistance genes which could be disseminated to inhabitants downstream the plant and pose health risk to the communities who are dependent upon the watershed for domestic and recreational purposes.

Teenagers' Experiences of Sexual Health Dialogue in the Rural Villages of the Vhembe District; Limpopo Province

Dialogue with teenagers about sexual health is of global concern; as it is found mostly to be minimal; if not absent. This limitation is influenced by the cultural values; beliefs and norms of teenagers. To a great extent; culture influences which and how sexual health issues can be discussed between teenagers and adults. The purpose of this study was to explore and describe teenagers' experiences of sexual health dialogue in the rural areas of the Vhembe district in Limpopo province; South Africa. A qualitative; explorative; descriptive and contextual research approach was used. Data were collected by means of in-depth; individual interviews and focus-group discussions. A purposive sampling method was used to select the 57 participating teenagers; based on criteria that were set. The findings of the study indicated that there was minimal dialogue between adults and teenagers about sexual health. Most teenagers reported that they learned about issues related to sexual health from friends; the media; initiation schools and relatives who; most often; were aunts. It was also noted with concern that the teenagers always equated sexual health to the act of intercourse and that no attention was paid to other aspects of sexual health. Recommendations to improve the current situation were made

Municipal wastewater effluents as a source of listerial pathogens in the aquatic milieu of the Eastern Cape Province of South Africa: a concern of public health importance.

We evaluated the effluent quality of an urban wastewater treatment facility in South Africa and its impact on the receiving watershed for a period of 12 months. The prevalence and antimicrobial susceptibility of potential Listeria pathogens (L. ivanovii and L. innocua) and the physicochemical quality of the treated wastewater effluent was assessed, with a view to ascertain the potential health and environmental hazards of the discharged effluent. Total listerial density varied between 2.9 x 10(0) and 1.2 x 10(5) cfu/mL; free living Listeria species were more prevalent (84%), compared to Listeria species attached to planktons (59-75%). The treated effluent quality fell short of recommended standards for turbidity, dissolved oxygen, chemical oxygen demand, nitrite, phosphate and Listeria density; while pH, temperature, total dissolved solids and nitrate contents were compliant with target quality limits after treatment. The Listeria isolates (23) were sensitive to three (15%) of the 20 test antibiotics, and showed varying (4.5-91%) levels of resistance to 17 antibiotics. Of seven resistance gene markers assayed, only sulII genes were detected in five (22%) Listeria strains. The study demonstrates a potential negative impact of the wastewater effluent on the receiving environment and suggests a serious public health implication for those who depend on the receiving watershed for drinking and other purposes.

Occurrence of potentially pathogenic vibrios in final effluents of a wastewater treatment facility in a rural community of the Eastern Cape Province of South Africa.

We assessed the occurrence of Vibrio pathogens in the final effluents of a rural wastewater treatment facility in the Eastern Cape Province of South Africa as free or plankton-associated (180 microm, 60 microm and 20 microm plankton sizes) entities using standard culture-based and molecular techniques. The free-living Vibrio densities varied from 0 to 3.45 x 10(1) cfu ml(-1), while the plankton-associated Vibrio densities vary with plankton sizes as follows: 180 microm (0-4.50 x 10(3) cfu ml(-1)); 60 microm (0-4.86 x 10(3) cfu ml(-1)); 20 microm (0-1.9 x 10(5) cfu ml(-1)). The seasonal variations in the Vibrio densities in the 180 and 60 microm plankton size samples were significant (p < 0.05), while the 20 microm plankton size and free-living Vibrio densities were not. Molecular confirmation of the presumptive vibrios isolates revealed fluvialis (36.5%), as the predominant species, followed by Vibrio vulnificus (34.6%), and Vibrio parahaemolyticus (23.1%); only API 20NE was employed to detect Vibrio metschnikovii (5.8%), suggesting a high incidence of pathogenic Vibrio species in the final effluent of the rural wastewater facility. Analysis suggested that the concentration of Vibrio species correlated negatively with salinity and temperature (p < 0.001 and p < 0.002 respectively) as well as with pH and turbidity (p < 0.001) in the final effluent. We conclude that rural wastewater treatment facilities in the Eastern Cape Province of South Africa are potential sources of Vibrio pathogens in the aquatic environment of the communities.

Prevalence and species distribution of E. Histolytica and E. Dispar in the Venda region, Limpopo, South Africa.

The prevalence and species distribution of Entamoeba histolytica and E. dispar in the Venda region were determined in stool samples collected from public hospitals and primary schools by ELISA and a nested polymerase chain reaction (PCR). E. histolytica was detected in 37/197 (18.8%) and 1/47 (2.1%) samples, whereas 50/197 (25.3%) and 4/47 (8.5%) had E. dispar in the hospitals and schools, respectively. The age groups most infected were 0-2 (33%) years followed by 20-29 years (27%). E. histolytica was significantly associated with diarrhea (77.4% versus 22.6%; chi 2 = 39.48, P < 0.05), and with the presence of lactoferrin (85.7% versus 14.2%) in the stools, indicating intestinal inflammation (chi 2 = 29.605, P < 0.05). E. histolytica was found in 5 (16.12%) of the 31 HIV-positive individuals and in 33 (15.5%) of the 213 HIV-negative individuals. E. histolytica infections are common in the Venda region and are associated with diarrhea and intestinal inflammation.

Resistance mutational analysis of HIV type 1 subtype C among rural South African drug-naive patients prior to large-scale availability of antiretrovirals.

Baseline HIV-1 resistance data are important for resistance monitoring purposes especially in regions initiating large-scale antiretroviral treatment programs. We examined 40 protease and 35 reverse transcriptase amino acid sequences of HIV-1 subtype C from drug inexperienced patients from rural settings in South Africa for resistance mutations. Samples were collected between 2001 and 2004 prior to the availability of antiretrovirals through public health institutions. Ninety-five percent of patients had no major mutations in the protease gene, although substitutions M46L (2.5%) and G73S (2.5%), which according to the Stanford Genotypic Resistance Interpretation Algorithm are considered major mutations, were detected. In addition, a high prevalence of minor mutations was observed in the protease, with at least three minor resistance-associated mutations in 37% of the isolates. An isoleucine insertion at codon 37 was detected in one sequence. Most of the RT sequences were wild-type, although V118I (8.5%) and Y318F (5.7%) associated with resistance to lamivudine and nevirapine, respectively, were observed. Our data suggest that major resistance mutations among the drug-inexperienced population in South Africa may be rare, and routine resistance testing before the initiation of therapy in this initial stage of the treatment program may not be necessary.