Entomopathogenic nematodes from north-eastern South Africa and their virulence against false codling moth,Thaumatotibia leucotreta (Lepidoptera: Tortricidae)

A survey was undertaken to determine the diversity and frequency of endemic entomopathogenic nematodes (EPN) in subtropical regions in the Mpumalanga, Limpopo and KwaZulu-Natal provinces of South Africa. A total of 136 soil samples were randomly taken from cultivated and uncultivated habitats, including subtropical fruit tree orchards (avocado, litchi, macadamia, mango and guava) and adjoining natural vegetation. EPNs were isolated from 14 samples (10.3%) by means of baiting with Tenebrio molitor larvae. Heterorhabditis was the most common genus isolated from 12 samples, whereas only two Steinernema species were isolated. The most common Heterorhabditis isolated were Heterorhabditis noenieputensis and H. zealandica, which were both isolated from four samples each. The present report is the first occurrence of H. taysearae and H. baujardi in South Africa. The two Steinernema species are both unknown species. Laboratory bioassays were conducted to determine the potential of seven native EPN isolates to control the false codling moth (Thaumatotibia leucotreta) (FCM). Last instar FCM larvae were found to be most susceptible to S. yirgalemense (157-C), H. zealandica (WS 23) and Steinernema litchii (WS9), when exposed to 50 IJs/50?µ? water for 48?h, causing 100%, 94.2% and 93.5% mortality, respectively.     

Comparative secretome analysis of <Emphasis Type="Italic">Trichoderma asperellum</Emphasis> S4F8 and <Emphasis Type="Italic">Trichoderma reesei</Emphasis> Rut C30 during solid-state fermentation on sugarcane bagasse

Background

The lignocellulosic enzymes of Trichoderma species have received particular attention with regard to biomass conversion to biofuels, but the production cost of these enzymes remains a significant hurdle for their commercial application. In this study, we quantitatively compared the lignocellulolytic enzyme profile of a newly isolated Trichoderma asperellum S4F8 strain with that of Trichoderma reesei Rut C30, cultured on sugarcane bagasse (SCB) using solid-state fermentation (SSF).

Results

Comparison of the lignocellulolytic enzyme profiles of S4F8 and Rut C30 showed that S4F8 had significantly higher hemicellulase and β-glucosidase enzyme activities. Liquid chromatography tandem mass spectrometry analysis of the two fungal secretomes enabled the detection of 815 proteins in total, with 418 and 397 proteins being specific for S4F8 and Rut C30, respectively, and 174 proteins being common to both strains. In-depth analysis of the associated biological functions and the representation of glycoside hydrolase family members within the two secretomes indicated that the S4F8 secretome contained a higher diversity of main and side chain hemicellulases and β-glucosidases, and an increased abundance of some of these proteins compared with the Rut C30 secretome.

Conclusions

In SCB SSF, T. asperellum S4F8 produced a more complex lignocellulolytic cocktail, with enhanced hemicellulose and cellobiose hydrolysis potential, compared with T. reesei Rut C30. This bodes well for the development of a more cost-effective and efficient lignocellulolytic enzyme cocktail from T. asperellum for lignocellulosic feedstock hydrolysis.

     

A Risk Prediction Model for the Assessment and Triage of Women with Hypertensive Disorders of Pregnancy in Low-Resourced Settings: The miniPIERS (Pre-eclampsia Integrated Estimate of RiSk) Multi-country Prospective Cohort Study

Background

Pre-eclampsia/eclampsia are leading causes of maternal mortality and morbidity, particularly in low- and middle- income countries (LMICs). We developed the miniPIERS risk prediction model to provide a simple, evidence-based tool to identify pregnant women in LMICs at increased risk of death or major hypertensive-related complications.

Methods and Findings

From 1 July 2008 to 31 March 2012, in five LMICs, data were collected prospectively on 2,081 women with any hypertensive disorder of pregnancy admitted to a participating centre. Candidate predictors collected within 24 hours of admission were entered into a step-wise backward elimination logistic regression model to predict a composite adverse maternal outcome within 48 hours of admission. Model internal validation was accomplished by bootstrapping and external validation was completed using data from 1,300 women in the Pre-eclampsia Integrated Estimate of RiSk (fullPIERS) dataset. Predictive performance was assessed for calibration, discrimination, and stratification capacity. The final miniPIERS model included: parity (nulliparous versus multiparous); gestational age on admission; headache/visual disturbances; chest pain/dyspnoea; vaginal bleeding with abdominal pain; systolic blood pressure; and dipstick proteinuria. The miniPIERS model was well-calibrated and had an area under the receiver operating characteristic curve (AUC ROC) of 0.768 (95% CI 0.735–0.801) with an average optimism of 0.037. External validation AUC ROC was 0.713 (95% CI 0.658–0.768). A predicted probability ≥25% to define a positive test classified women with 85.5% accuracy. Limitations of this study include the composite outcome and the broad inclusion criteria of any hypertensive disorder of pregnancy. This broad approach was used to optimize model generalizability.

Conclusions

The miniPIERS model shows reasonable ability to identify women at increased risk of adverse maternal outcomes associated with the hypertensive disorders of pregnancy. It could be used in LMICs to identify women who would benefit most from interventions such as magnesium sulphate, antihypertensives, or transportation to a higher level of care. Please see later in the article for the Editors'' Summary     

Beta thalassemic mutations recognized by DNA mapping with Hph I and Rsa I in the Algerian population

By using Hph I and Rsa I restriction enzymes and beta globin large intervening sequence as a probe, we have investigated the DNA of 20 Algerian patients with beta(0) or beta(+) thalassemia. In any of them, we detected the nucleotide change which is known to generate an additional Hph I site at the 5' splice junction of the beta globin large intervening sequence and which yields a beta(0) phenotype. In one of them, we detected the nucleotide change which is known to generate an additional Rsa I site within the beta globin large intervening sequence and which is supposed to yield a beta(+) phenotype. These results indicate that these two types of mutation are relatively rare in the Algerian population.     

Electrochemical assay of the nitrate and nitrite reductase activities of Rhizobium japonicum

This work describes an electrochemical method for the determination of the nitrate and nitrite reductase activities of Rhizobium japonicum. The advantage of the method lies in the use of whole cells for the analysis and we earlier developed this protocol for the assay of NO. The results obtained are comparable to the spectrophotometric Griess assay. As the method is based on electrochemical reduction, the commonly interfering biological components like ascorbic acid, uric acid, dopamine, etc., will not interfere with the analysis. This method can be extended to the fabrication of biosensors for nitrate and nitrite using the same principle.     

Four Models of HIV Counseling and Testing: Utilization and Test Results in South Africa

Background

HIV Counseling and Testing (HCT) is the point-of-entry for pathways of HIV care and prevention. However, HCT is not reaching many who are HIV infected and this may be related to the HCT provision model. We describe HCT utilization and HIV diagnosis using four models of HCT delivery: clinic-based, urban mobile, rural mobile, and stand-alone.

Methods

Using cross-sectional data from routine HCT provided in South Africa, we described client characteristics and HIV test results from information collected during service delivery between January 2009 and June 2012.

Results

118,358 clients received services at clinic-based units, 18,597; stand-alone, 28,937; urban mobile, 38,840; and rural mobile, 31,984. By unit, clients were similar in terms of median age (range 28–31), but differed in sex distribution, employment status, prior testing, and perceived HIV risk. Urban mobile units had the highest proportion of male clients (52%). Rural mobile units reached the highest proportion of clients with no prior HCT (61%) and reporting no perceived HIV risk (64%). Overall, 10,862 clients (9.3%) tested HIV-positive.

Conclusions

Client characteristics varied by HCT model. Importantly, rural and urban mobile units reached more men, first-time testers, and clients who considered themselves to be at low risk for HIV.     

Soil microbial counts in relation to site characteristics at a subantarctic Island

Enumerations of colony-forming units of aerobic bacteria and fungi at Marion Island (46°54S, 37°45E) showed that the mire and bog peats investigated possessed greater numbers of microorganisms on a dry soil weight basis than did the slope fernbrake soils but that counts from both sites were similar per unit soil volume. Afjaeldmark soil of an exposed rocky ridge contained very low microbial populations. Manuring by sea-birds and seals caused an increase in levels of soil N and P and in the numbers of bacteria and fungi. Regression analyses indicated that inorganic N and soil pH explained more of the variation in bacterial and fungal counts respectively than all other soil parameters included in the regression. Variation in botanical composition between the sites was examined using multivariate analysis and the resulting pattern compared with the soil microorganism and nutrient levels. This analysis emphasized the importance of manuring influences in elevating NH₄-N and bacterial levels in the soil and allowing the occurrence of a vegetation cover dominated by copriphilous species. No correlation occurred between site botanical factors and soil fungi counts.     

Ureolytic bacteria in sheep rumen.

Estimates were made of the numbers of viable bacteria in the rumens of sheep receiving different rations. Representative colonies were isolated and tested for urease production. Some urease-positive isolates were characterized and identified. The ureolytic activities of the urease-producing isolates were determined and compared with the activity of rumen fluid. The rations fed to the sheep did not exert a significant influence on the relative numbers of the urease-producting organisms in the rumen. No obligately anaerobic ureolytic bacteria were found. All urease-positive isolates were facultatively anaerobic, Gram-positive, catalase-positive cocci. Out of ten isolates, nine were identified as Staphylococcus saprophyticus and one as Micrococcus varians. The total urease activity of the different isolates based on the lowest numbers in which they were present in the rumen, compared favourably with the urease activity of rumen fluid. The facultatively anaerobic Gram-positive cocci were probably responsible for a large proportion of the urease activity of the rumen fluid. Conditions prevailing in the rumen were found to be conducive to urease production by the isolates tested.     

Biosynthesis of cyclopiazonic acids in Penicillium cyclopium: The isolation of dimethylallylpyrophosphate: Cyclo-acetoacetyltryptophanyl dimethylallyltransferase

During the production of β-cyclopiazonic acid (βCA) by Penicillium cyclopium, the following points are noted: (a) Dimethylallylpyrophosphate (DMAPP), which is incorporated into α-cyclopiazonic acid (αCA) in vivo, stimulates overall CA synthesis, whereas tryptophan, although incorporated into αCA, inhibits overall CA synthesis. (b) A previously suggested substrate, γ,γ-dimethylallyltryptophan, is not a precursor of αCA. (c) The accumulation of cyclo-acetoacetyl-l-tryptophanyl (cAATrp) is described in both the culture medium and mycelium with increasing growth. (d) A cell-free extract of mycelium will catalyze the conversion of exogenous cAATrp and exogenous DMAPP into βCA in 1:1 stoichiometry, the βCA being bound to a protein.