Performance of local light microscopy and the ParaScreen Pan/Pf rapid diagnostic test to detect malaria in health centers in Northwest Ethiopia

Background

Diagnostic tests are recommended for suspected malaria cases before treatment, but comparative performance of microscopy and rapid diagnostic tests (RDTs) at rural health centers has rarely been studied compared to independent expert microscopy.

Methods

Participants (N = 1997) with presumptive malaria were recruited from ten health centers with a range of transmission intensities in Amhara Regional State, Northwest Ethiopia during October to December 2007. Microscopy and ParaScreen Pan/Pf® RDT were done immediately by health center technicians. Blood slides were re-examined later at a central laboratory by independent expert microscopists.

Results

Of 1,997 febrile patients, 475 (23.8%) were positive by expert microscopists, with 57.7% P.falciparum, 24.6% P.vivax and 17.7% mixed infections. Sensitivity of health center microscopists for any malaria species was >90% in five health centers (four of which had the highest prevalence), >70% in nine centers and 44% in one site with lowest prevalence. Specificity for health center microscopy was very good (>95%) in all centers. For ParaScreen RDT, sensitivity was ≥90% in three centers, ≥70% in six and <60% in four centers. Specificity was ≥90% in all centers except one where it was 85%.

Conclusions

Health center microscopists performed well in nine of the ten health centers; while for ParaScreen RDT they performed well in only six centers. Overall the accuracy of local microscopy exceeded that of RDT for all outcomes. This study supports the introduction of RDTs only if accompanied by appropriate training, frequent supervision and quality control at all levels. Deficiencies in RDT use at some health centers must be rectified before universal replacement of good routine microscopy with RDTs. Maintenance and strengthening of good quality microscopy remains a priority at health center level.     

Phosphatidic acid regulation of PIPKI is critical for actin cytoskeletal reorganization

Type I phosphatidylinositol-4-phosphate 5-kinase (PIPKI) is the main enzyme generating the lipid second messenger phosphatidylinositol-4,5-bisphosphate [PI(4,5)P2], which has critical functions in many cellular processes, such as cytoskeletal reorganization, membrane trafficking, and signal transduction. All three members of the PIPKI family are activated by phosphatidic acid (PA). However, how PA regulates the activity and functions of PIPKI have not been fully elucidated. In this study, we identify a PA-binding site on PIPKIγ. Mutation of this site inhibited the PA-stimulated activity and membrane localization of PIPKIγ as well as the formation of actin comets and foci induced by PIPKIγ. We also demonstrate that phospholipase D (PLD) generates a pool of PA involved in PIPKIγ regulation by showing that PLD inhibitors blocked the membrane localization of PIPKIγ and its ability to induce actin cytoskeletal reorganization. Targeting the PIPKIγ PA-binding-deficient mutant to membranes by a membrane localization sequence failed to restore the actin reorganization activity of PIPKIγ, suggesting that PA binding is not only involved in recruiting PIPKIγ to membranes but also may induce a conformational change. Taken together, these results reveal a new molecular mechanism through which PA regulates PIPKI and provides direct evidence that PA is important for the localization and functions of PIPKI in intact cells.     

Water-membrane partition thermodynamics of an amphiphilic lipopeptide: an enthalpy-driven hydrophobic effect

To shed light on the driving force for the hydrophobic effect that partitions amphiphilic lipoproteins between water and membrane, we carried out an atomically detailed thermodynamic analysis of a triply lipid modified H-ras heptapeptide anchor (ANCH) in water and in a DMPC (1,2-dimyristoyl-sn-glycero-3-phosphocholine) bilayer. Combining molecular mechanical and continuum solvent approaches with an improved technique for solute entropy calculation, we obtained an overall transfer free energy of ~−13 kcal mol⁻¹. This value is in qualitative agreement with free energy changes derived from a potential of mean force calculation and indirect experimental observations. Changes in free energies of solvation and ANCH conformational reorganization are unfavorable, whereas ANCH-DMPC interactions—especially van der Waals—favor insertion. These results are consistent with an enthalpy-driven hydrophobic effect, in accord with earlier calorimetric data on the membrane partition of other amphiphiles. Furthermore, structural and entropic analysis of molecular dynamics-generated ensembles suggests that conformational selection may play a hitherto unappreciated role in membrane insertion of lipid-modified peptides and proteins.     

Genome of a virulent bacteriophage Lb338-1 that lyses the probiotic Lactobacillus paracasei cheese strain

There is a lack of fundamental knowledge about the influence of bacteriophage on probiotic bacteria and other commensals in the gut. Here, we present the isolation and morphological and genetic characterization of a virulent narrow-host-range bacteriophage, φLb338-1. This phage was isolated from fresh sewage and was shown to infect the probiotic cheese strain Lactobacillus paracasei NFBC 338. Electron microscopy studies revealed that φLb338-1 is a member of the Myoviridae family, with an isometric head, a medium-sized contractile tail, and a complex base plate. Genome sequencing revealed a 142-kb genome with 199 open reading frames. Putative functions could be assigned to 22% of the open reading frames; these had significant homology to genes found in the broad-host-range SPO1-like group of phages which includes the Enterococcus faecalis phage φEF24C, Listeria phage A511, and Lactobacillus plantarum phage LP65. Interestingly, no significant genomic similarity was observed between the phage and the probiotic host strain. Future studies will determine if the presence of bacteriophage φLb338-1 or others in the human or animal gut plays an antagonistic role against the probiotic effect of beneficial bacteria.     

Importance of Coverage and Endemicity on the Return of Infectious Trachoma after a Single Mass Antibiotic Distribution

Background

As part of the SAFE strategy, mass antibiotic treatments are useful in controlling the ocular strains of chlamydia that cause trachoma. The World Health Organization recommends treating at least 80% of individuals per community. However, the role of antibiotic coverage for trachoma control has been poorly characterized.

Methodology/Principal Findings

In a collection of cluster-randomized clinical trials, mass oral azithromycin was administered to 40 villages in Ethiopia. The village prevalence of ocular chlamydia was determined before treatment, and at two and six months post-treatment. The mean prevalence of ocular chlamydia was 48.9% (95% CI 42.8 to 55.0%) before mass treatments, decreased to 5.4% (95% CI 3.9 to 7.0%) at two months after treatments (p<0.0001), and returned to 7.9% (95% CI 5.4 to 10.4%) by six months after treatment (p = 0.03). Antibiotic coverage ranged from 73.9% to 100%, with a mean of 90.6%. In multivariate regression models, chlamydial prevalence two months after treatment was associated with baseline infection (p<0.0001) and antibiotic coverage (p = 0.007). However, by six months after treatment, chlamydial prevalence was associated only with baseline infection (p<0.0001), but not coverage (p = 0.31).

Conclusions/Significance

In post-hoc analyses of a large clinical trial, the amount of endemic chlamydial infection was a strong predictor of chlamydial infection after mass antibiotic treatments. Antibiotic coverage was an important short-term predictor of chlamydial infection, but no longer predicted infection by six months after mass antibiotic treatments. A wider range of antibiotic coverage than found in this study might allow an assessment of a more subtle association.     

Landscape genetic structure of <Emphasis Type="Italic">Olea europaea</Emphasis> subsp. <Emphasis Type="Italic">cuspidata</Emphasis> in Ethiopian highland forest fragments

Understanding patterns of genetic diversity at the landscape scale will enhance conservation and management of natural populations. Here we analyzed the genetic diversity, population connectivity, and spatial genetic structure among subpopulations and age groups of Olea europaea subsp. cuspidata, a cornerstone species of the Afromontane highlands. The study was conducted at the landscape level within a radius of approximately 4 km, as well as on a fine scale (intensive study plot) of less than 300 m radius. In total 542 samples from four natural subpopulations in northwestern Ethiopia were analyzed using ten nuclear microsatellite markers. Inbreeding was higher in smaller populations. No genetic difference was detected among cohorts of different tree sizes in the intensive studied plot. Average population differentiation was low but significant (F _ST ?=?0.016). Landscape genetic analysis inferred two groups: the most distant subpopulation WE located less than 4 kms from the other three subpopulations formed a separate group. Sixty-four percent of the total migrants were shared among the three latter subpopulations, which are spatially clustered. Immigrants were non-randomly distributed inside of the intensive study plot. Significant spatial genetic structure (SGS) was found both at the landscape scale and in the intensive study plot, and adults showed stronger SGS than young trees. An indirect estimate of 220 m as mean gene dispersal distance was obtained. We conclude that even under fragmentation migration is not disrupted in wild olive trees and that large protected populations at church forests are very important to conserve genetic resources. However, the higher level of inbreeding and evidence for population bottlenecks in the small populations, as well as the persisting heavy pressure on most remaining populations, warrants quick action to maintain genetic diversity of wild olive in the Ethiopian highlands.     

Stone walls and sacred forest conservation in Ethiopia

Many tropical forests worldwide are protected due to their sacredness to religious communities. In the south Gondar region of Ethiopia, most of the remaining native forests are tiny fragments (5.42 ha ± 0.34) surrounded by pasture and agriculture that are protected because they encompass churches of the Ethiopian Orthodox Tewahido Church (EOTC). A small number of communities have erected stone walls around the perimeter of the forests to demarcate the boundary, and/or protect the interior of the forest. We evaluated the effectiveness of these walls at protecting ecological conditions by examining tree and seedling communities among sacred forests with and without walls in Montane and Upper Montane sites. We found the wall to be an effective conservation tool as regeneration potential was higher in forests with a wall. The density and species richness of seedlings were significantly higher in forests with a wall and these effects were more pronounced in Upper Montane forests. Forests with a wall also had seedlings of many native tree species that were not found in forests without walls. As expected, there were few differences in tree communities in forests with and without a wall. Although rare on the landscape, the presence of a stone wall around these forests was effective at protecting the seedling community because it likely reduced access to the forests by grazers and directed human visitors to trails. The use of a stone wall may protect seedling communities in other sacred forest fragments, particularly for those that are small, isolated, surrounded by agriculture, and have a depleted seed bank.     

Removal of fluoride, arsenic and coliform bacteria by modified homemade filter media from drinking water

An attempt was made to investigate the removal of fluoride, arsenic and coliform bacteria from drinking water using modified homemade filter media. Batch mode experimental study was conducted to test the efficiency of modified homemade filter for reduction of impurities under the operating condition of treatment time. The physico-chemical and biological analysis of water samples had been done before and after the treatment with filter media, using standard methods. Optimum operating treatment time was determined for maximum removal of these impurities by running the experiment for 2, 4, 6, 8, 10 and 12h, respectively. The maximum reduction of fluoride, arsenic and coliform bacteria in percentage was 85.60%, 93.07% and 100% and their residual values were 0.72 mg/l, 0.009 mg/l and 0 coliform cells/100ml, respectively after a treatment time of 10h. These residual values were under the permissible limits prescribed by WHO. Hence this could be a cheap, easy and an efficient technique for removal of fluoride, arsenic and coliform bacteria from drinking water.