Spatial and temporal variation in malaria transmission in a low endemicity area in northern Tanzania

Background

Current detection or screening for malaria infection necessitates drawing blood by fingerprick or venipuncture, which poses risks and limitations for repeated measurement. This study presents PCR detection of Plasmodium falciparum in human urine and saliva samples, and illustrates this potential application in genotyping malaria infections.

Methods

Urine and saliva were obtained from 47 thick film positive and 4 negative individuals one day after collection of blood slides and filter paper blood spots. P. falciparum DNA was extracted from blood, urine and saliva, in separate groups, using the Chelex method or Qiagen DNEasy^® kit (urine and saliva only). Blood, urine and saliva extracts were subjected to PCR in separate batches. Amplicons from the various sample types were examined for MSP2 polymorphisms and restriction fragment patterns on DHFR amino acid codon 59.

Results and discussion

Malaria infections exhibited primarily low-grade parasite densities, with a geometric mean of 775 asexual parasites/μl. Regularly matching polymorphic MSP2 genotypes were found between the corresponding urine, saliva and peripheral blood amplicons of each individual, with different inter-individual polymorphic genotypes. Amplicon yields were significantly dependent on DNA extraction method, parasite density and primer set (p < 0.001). A Qiagen^® kit extraction had more than 2× higher amplicon yield than the Chelex method, for both urine and saliva. Amplicon yields were 1.6 fold higher from saliva than urine. For each unit increase in log parasite density, the probability of amplicon enhanced 1.8 fold. Highest amplicon yields were obtained from the primer set with the shortest PCR product.

Conclusion

P. falciparum infection is detectable by PCR on human urine and saliva samples. Subject to further refinement of extraction technique and amplicon yields, large-scale malaria parasite screening and epidemiological surveys could be possible without the need to collect blood and use of needles or sharps.     

Existing in plenty: abundance, biomass and diversity of ciliates and meiofauna in small streams

1. The ciliate and metazoan meiofaunal assemblages of two contrasting lowland streams in south‐east England were examined over the period of a year, using a high taxonomic resolution. Monthly samples were taken from an oligotrophic, acid stream (Lone Oak) and a circumneutral, nutrient‐rich stream (Pant) between March 2003 and February 2004. 2. We assessed the relative importance of ciliates and rotifers within the small‐sized benthic assemblage with respect to their abundance, biomass and species richness. In addition, we examined the influence of abiotic and biotic parameters and season on the assemblage composition at two levels of taxonomic resolution (species and groups). 3. Ciliates dominated the assemblages numerically, with maximum densities of over 900 000 and 6 000 000 ind. m^?2 in Lone Oak and Pant respectively. Rotifers and nematodes dominated meiofaunal densities, although their contribution to total meiofaunal biomass (maxima of 71.9 mgC m^?2 in Lone Oak and of 646.8 mgC m^?2 in the Pant) was low and rotifer biomass equalled that of ciliates. 4. Although the two streams differed in terms of total abundance of ciliates and meiofauna and shared only 7% of species, the relative proportion of groups was similar. Sediment grain size distribution (the percentile representing the 0.5–1 mm fraction) was correlated with assemblage structure at the species level, revealing the tight coupling between these small organisms and their physical environment. Seasonal changes in the relative abundance of groups followed similar patterns in both streams, and were correlated with the abundance of cyclopoid copepods and temperature. 5. Information on these highly abundant but often overlooked faunal groups is essential for estimates of overall abundance, biomass, species richness and productivity in the benthos, and as such has important implications for several areas of aquatic research, e.g. for those dealing with trophic dynamics.     

Salinity stress and cytoplasmic factors. A comparison of cell permeability and lipid partiality in salt sensitive and salt resistant cultivars and lines of Triticum aestivum and Hordeum vulgare

Salinity effects on the cell membranes of four lines of wheat ( Triticum aestivum L.). and two cultivars of barley ( Hordeum vulgare L.), differing in salt resistance were investigated. Plants were grown for 10 days in 1/4-strength Hoagland solution and then for 5 more days in 1/4-strength Hoagland with and without NaCl (100 m M ) or (for Hordeum only) polyethylene glycol (PEG). Permeability to three non-electrolytes (urea, methylurea and ethylurea) of subepidermal cells of leaf sheaths ( Triticum ) and coleoptiles ( Hordeum ) was determined and membrane partiality calculated, a parameter which numerically indicates the degree of lipophilicity of a membrane. Non-electrolyte permeability significantly increased and membrane partiality decreased in the salt sensitive cultivars or lines under salt stress. Neither parameter changed significantly in the salt resistant lines and cultivar in a saline environment. Osmotic stress in Hordeum by PEG 10000 had no significant effect on permeability and thus membrane partiality neither in sensitive nor in resistant cultivars.
The osmotic component of salinity stress did not seem to be a major factor causing injury, rather ion toxicity may be a cause of cell damage. The results indicate differences in the membrane between salt sensitive and salt resistant genotypes. Salt resistance seems to be controlled by genetic factors independent of external salinity levels.