Dodonaea viscosa

The virulence factors of Candida albicans are germ tube and biofilm formation, adherence to host tissues, and production of hydrolytic enzymes. This study investigated the effect of Dodonaea viscosa var. angustifolia extract on the germ tube and biofilm formation of C. albicans. Serum containing the three subinhibitory concentrations of leaf extract was inoculated with C. albicans, incubated, and viewed under a light microscope. Number of cells with germ tube was recorded and the results were analysed using Scheffe test for pairwise comparison. Biofilms were grown on coverslips in the presence of plant extracts and processed for scanning electron microscopy (SEM). Planktonic cells were grown in the presence of plant extract for 6 h and processed for electron microscopy (TEM). The crude plant extract significantly (P<0.01) reduced the germ tube formation of C. albicans at 3.125 (85.36%), 1.56 (61.91%), and 0.78 mg/mL (26.27%) showing a concentration dependent effect. SEM results showed concentration dependent reduction in biofilm and hyphae formation. TEM results showed that the plant extract caused damage to the cell wall and cell membrane. DVA extract has ability to reduce virulence of C. albicans by inhibiting germ tube and biofilm formation through damage to the cell wall. Therefore, it has therapeutic potential

An outbreak of lymphocutaneous sporotrichosis among mine-workers in South Africa

BACKGROUND The largest outbreak of sporotrichosis occurred between 1938 and 1947 in the gold mines of Witwatersrand in South Africa. Here, we describe an outbreak of lymphocutaneous sporotrichosis that was investigated in a South African gold mine in 2011. METHODOLOGY Employees working at a reopened section of the mine were recruited for a descriptive cross-sectional study. Informed consent was sought for interview, clinical examination and medical record review. Specimens were collected from participants with active or partiallyhealed lymphocutaneous lesions. Environmental samples were collected from underground mine levels. Sporothrix isolates were identified by sequencing of the internal transcribed spacer region of the ribosomal gene and the nuclear calmodulin gene. PRINCIPAL FINDINGS Of 87 male miners, 81 (93%) were interviewed and examined, of whom 29 (36%) had skin lesions; specimens were collected from 17 (59%). Sporotrichosis was laboratory-confirmed among 10 patients and seven had clinically-compatible lesions. Of 42miners with known HIV status, 11 (26%) were HIV-infected. No cases of disseminated disease were detected. Participants with 3 years’ mining experience had a four times greater odds of developing sporotrichosis than those who had been employed for >3 years (adjusted OR 4.0, 95%CI 1.2–13.1). Isolates from 8 patients were identified as Sporothrix schenckii sensu stricto by calmodulin gene sequencing while environmental isolates were identified as Sporothrix mexicana. CONCLUSIONS/SIGNIFICANCE S. schenckii sensu stricto was identified as the causative pathogen. Although genetically distinct species were isolated from clinical and environmental sources, it is likely that the source was contaminated soil and untreated wood underground. No cases occurred following recommendations to close sections of the mine, treat timber and encourage consistent use of personal protective equipment. Sporotrichosis is a potentially re-emerging disease where traditional, rather than heavily mechanised, mining techniques are used. Surveillance should be instituted at sentinel locations.http://www.plosntds.orgam201

South African Ebola diagnostic response in Sierra Leone : a modular high biosafety field laboratory

BACKGROUND : In August 2014, the National Institute for Communicable Diseases (NICD) in South Africa established a modular high-biosafety field Ebola diagnostic laboratory (SA FEDL) near Freetown, Sierra Leone in response to the rapidly increasing number of Ebola virus disease (EVD) cases. METHODS AND FINDINGS : The SA FEDL operated in the Western Area of Sierra Leone, which remained a ªhotspotº of the EVD epidemic for months. The FEDL was the only diagnostic capacity available to respond to the overwhelming demand for rapid EVD laboratory diagnosis for several weeks in the initial stages of the EVD crisis in the capital of Sierra Leone. Furthermore, the NICD set out to establish local capacity amongst Sierra Leonean nationals in all aspects of the FEDL functions from the outset. This led to the successful hand-over of the FEDL to the Sierra Leone Ministry of Health and Sanitation in March 2015. Between 25 August 2014 and 22 June 2016, the laboratory tested 11,250 specimens mostly from the Western Urban and Western Rural regions of Sierra Leone, of which 2,379 (21.14%) tested positive for Ebola virus RNA. CONCLUSIONS : he bio-safety standards and the portability of the SA FEDL, offered a cost-effective and practical alternative for the rapid deployment of a field-operated high biocontainment facility. The SA FEDL teams demonstrated that it is highly beneficial to train the national staff in the course of formidable disease outbreak and accomplished their full integration into all operational and diagnostic aspects of the laboratory. This initiative contributed to the international efforts in bringing the EVD outbreak under control in Sierra Leone, as well as capacitating local African scientists and technologists to respond to diagnostic needs that might be required in future outbreaks of highly contagious pathogens.S1 Video. ªHotº processing of Ebola clinical specimens, PPE and decontamination procedures in South African modular, field-operated biocontainment facility in Sierra Leone.Janusz T Paweska was supported by funding from National Research Foundation and the Global Disease Detection Programmehttp://www.plosntds.orgam2017Microbiology and Plant Patholog

The effect of Dodonaea viscosa var. Angustifolia (L.F.) on the ultrastructure of Candida albicans cell wall and biofilm formation

Dissertation submitted to the Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, in fulfillment of the requirements for the degree of Master of Science in Medicine. Johannesburg, 2012Oral candidiasis is an infection prevalent in immunocompromised individuals. The main causative agent is Candida albicans. Many antifungal agents are available and are effectively used. However, due to the development of drug resistance, toxicity and poor solubility resulting in poor absorption; medicinal plants have been investigated. Dodonaea viscosa var. angustifolia, an indigenous South African plant has shown to have an antifungal effect including inhibition of adherence of C. albicans to oral epithelial cells; which is the crucial first step of infection. This study investigated the effect of the crude extract on the ultrastructure of C. albicans cell wall, which might be responsible for the reduced adherence to oral epithelial cells. The effect of the plant extract on C. albicans germ tube and biofilm formation was also studied since biofilm structure allows for high resistance to antifungal agents and host defense mechanisms. Crude plant extracts were prepared using dried leaves and acetone. Three C. albicans strains were used throughout the study. Minimal fungicidal concentrations of plant extract were determined using a microdilution technique. Three subinhibitory concentrations 3.125, 1.562 and 0.781 mg/ml were selected for further studies. The effect of these subinhibitory concentrations of plant extract on the C. albicans cell wall structure, cell membrane, germ tube formation, biofilm formation and cell wall proteins were studied using transmission electron microscopy, light microscopy, scanning electron microscopy and SDS-PAGE respectively. v The subinhibitory concentrations of crude plant extract rendered C. albicans cell wall thinner and at some places caused cell wall breakage and disruption. This effect increased with a decrease in plant extract concentration. The cell membrane was also damaged by the plant extract showing increased undulation. This effect was not concentration dependent. The subinhibitory concentrations decreased C. albicans germ tube formation and the effect increased with an increase in concentration. Biofilm formation was reduced by the plant extract and in addition, hyphal formation by cells within the biofilm was also reduced. However, SDS-PAGE showed that on a molecular level, the plant extract did not remove any specific adhesin proteins from the cell wall. The crude plant extract of D. viscosa var. angustifolia at high concentrations, kills C. albicans and at low concentrations, renders the surviving cells avirulent. Therefore it has the potential to be developed into an effective therapeutic agent to treat and prevent oral candidiasis. However, further research is required to identify the mode of action of the extract, the specific chemicals responsible for the effect, and the cytotoxicity

Candida auris Clinical Isolates Associated with Outbreak in Neonatal Unit of Tertiary Academic Hospital, South Africa

Candida auris was first detected at a university-affiliated hospital in Johannesburg, South Africa, in 2009. We used whole-genome sequencing to describe the molecular epidemiology of C. auris in the same hospital during 2016–2020; the neonatal unit had a persistent outbreak beginning in June 2019. Of 287 cases with culture-confirmed C. auris infection identified through laboratory surveillance, 207 (72%) had viable isolates and 188 (66%) were processed for whole-genome sequencing. Clade III (118/188, 63%) and IV (70/188, 37%) isolates co-circulated in the hospital. All 181/188 isolates that had a fluconazole MIC >32 µg/mL had ERG11 mutations; clade III isolates had VF125AL substitutions, and clade IV isolates had K177R/N335S/E343D substitutions. Dominated by clade III, the neonatal unit outbreak accounted for 32% (91/287) of all cases during the study period. The outbreak may have originated through transmission from infected or colonized patients, colonized healthcare workers, or contaminated equipment/environment

Two echinocandin-resistant Candida glabrata FKS mutants from South Africa

Echinocandins are recommended as first-line agents to treat invasive infections caused by Candida glabrata since this organism is inherently less susceptible to azoles. However, resistance to echinocandins has been described in C. glabrata due to amino acid changes in the hotspot regions of the FKS1 and FKS2 genes. In this report, we describe the first two South African C. glabrata isolates with echinocandin resistance mediated by mutations in the FKS2 gene. Both isolates were cultured from urine specimens from private-sector patients

High Prevalence of Candida auris Colonization during Protracted Neonatal Unit Outbreak, South Africa

One third of patients were colonized by Candida auris during a point-prevalence survey in a neonatal unit during an outbreak in South Africa. The sensitivity of a direct PCR for rapid colonization detection was 44% compared with culture. The infection incidence rate decreased by 85% after the survey and implementation of isolation/cohorting

Molecular type distribution and fluconazole susceptibility of clinical Cryptococcus gattii isolates from South African laboratory-based surveillance, 2005–2013

Como es el caso a nivel mundial, Cryptococcus gattii es una causa menos frecuente de criptococosis que Cryptococcus neoformans en Sudáfrica. Realizamos tipificación de secuencias multilocus (MLST) y pruebas de susceptibilidad a fluconazol de 146 aislamientos seleccionados al azar de 750 Pacientes sudafricanos con enfermedad por C. gattii identificados mediante vigilancia de laboratorio mejorada, de 2005 a 2013. El tipo molecular dominante fue VGIV (101/146, 70 %), seguido de VGI (40/146, 27%), VGII (3/146, 2%) y VGIII (2/146, 1%). Entre los 146 aislamientos de C. gattii, se identificaron 99 tipos de secuencia (ST) diferentes, con ST294 (14/146, 10 %) y ST155 (10/146, 7%) siendo el más comúnmente observado. Los valores de CIM50 y CIM90 de fluconazol de 105 (de 146) aislados de C. gattii seleccionados al azar fueron de 4 μg/ml y 16 μg/ml, respectivamente. Los aislamientos VGIV tenían un valor MIC50 más bajo en comparación con los aislamientos no VGIV, pero estos los valores estaban dentro de una dilución doble uno del otro. Los pacientes seropositivos para el VIH tenían una probabilidad ajustada diez veces mayor de una infección por VGIV en comparación con los pacientes seronegativos para el VIH. aunque con números pequeños (99/136; 73% vs. 2/10; 20%), el intervalo de confianza (IC) fue ancho (IC 95%: 1,93-55,31, p = 0,006). La filogenia del genoma completo de 98 aislamientos del tipo molecular más prevalente de Sudáfrica, VGIV, identificó que este tipo molecular es altamente diversos, con dos grupos interesantes de diez y seis aislamientos estrechamente relacionados identificados respectivamente. Uno de estos grupos consistía únicamente en pacientes de la provincia de Mpumalanga en Sudáfrica, lo que sugiere una fuente ambiental similar. Este estudio aportó nuevos conocimientos sobre la estructura de la población mundial de este importante patógeno humano.As is the case globally, Cryptococcus gattii is a less frequent cause of cryptococcosis than Cryptococcus neoformans in South Africa. We performed multilocus sequence typing (MLST) and fluconazole susceptibility testing of 146 isolates randomly selected from 750 South African patients with C. gattii disease identified through enhanced laboratory surveil lance, 2005 to 2013. The dominant molecular type was VGIV (101/146, 70%), followed by VGI (40/146, 27%), VGII (3/146, 2%) and VGIII (2/146, 1%). Among the 146 C. gattii iso lates, 99 different sequence types (STs) were identified, with ST294 (14/146, 10%) and ST155 (10/146, 7%) being most commonly observed. The fluconazole MIC50 and MIC90 val ues of 105 (of 146) randomly selected C. gattii isolates were 4 μg/ml and 16 μg/ml, respec tively. VGIV isolates had a lower MIC50 value compared to non-VGIV isolates, but these values were within one double-dilution of each other. HIV-seropositive patients had a ten fold increased adjusted odds of a VGIV infection compared to HIV-seronegative patients, though with small numbers (99/136; 73% vs. 2/10; 20%), the confidence interval (CI) was wide (95% CI: 1.93–55.31, p = 0.006). Whole genome phylogeny of 98 isolates of South Afri ca’s most prevalent molecular type, VGIV, identified that this molecular type is highly diverse, with two interesting clusters of ten and six closely related isolates being identified respectively. One of these clusters consisted only of patients from the Mpumalanga Prov ince in South Africa, suggesting a similar environmental source. This study contributed new insights into the global population structure of this important human pathogen

Molecular type distribution and fluconazole susceptibility of clinical Cryptococcus gattii isolates from South African laboratory-based surveillance, 2005-2013.

As is the case globally, Cryptococcus gattii is a less frequent cause of cryptococcosis than Cryptococcus neoformans in South Africa. We performed multilocus sequence typing (MLST) and fluconazole susceptibility testing of 146 isolates randomly selected from 750 South African patients with C. gattii disease identified through enhanced laboratory surveillance, 2005 to 2013. The dominant molecular type was VGIV (101/146, 70%), followed by VGI (40/146, 27%), VGII (3/146, 2%) and VGIII (2/146, 1%). Among the 146 C. gattii isolates, 99 different sequence types (STs) were identified, with ST294 (14/146, 10%) and ST155 (10/146, 7%) being most commonly observed. The fluconazole MIC50 and MIC90 values of 105 (of 146) randomly selected C. gattii isolates were 4 μg/ml and 16 μg/ml, respectively. VGIV isolates had a lower MIC50 value compared to non-VGIV isolates, but these values were within one double-dilution of each other. HIV-seropositive patients had a ten-fold increased adjusted odds of a VGIV infection compared to HIV-seronegative patients, though with small numbers (99/136; 73% vs. 2/10; 20%), the confidence interval (CI) was wide (95% CI: 1.93-55.31, p = 0.006). Whole genome phylogeny of 98 isolates of South Africa's most prevalent molecular type, VGIV, identified that this molecular type is highly diverse, with two interesting clusters of ten and six closely related isolates being identified, respectively. One of these clusters consisted only of patients from the Mpumalanga Province in South Africa, suggesting a similar environmental source. This study contributed new insights into the global population structure of this important human pathogen