Objective: The main objective of this study was to compare the performance of nested PCR with expert microscopy as a means of detecting Plasmodium parasites during active malaria surveillance in western Thailand. Methods:The study was performed from May 2000 to April 2002 in the village of Kong Mong Tha, located in western Thailand. Plasmodium vivax (PV) and Plasmodium falciparum (PF) are the predominant parasite species in this village, followed by Plasmodium malariae (PM) and Plasmodium ovale (PO). Each month, fingerprick blood samples were taken from each participating individual and used to prepare thick and thin blood films and for PCR analysis.Results: PCR was sensitive (96%) and specific (98%) for malaria at parasite densities ≥ 500/µl; however, only 18% (47/269) of P. falciparum-and 5% (20/390) of P. vivax-positive films had parasite densities this high. Performance of PCR decreased markedly at parasite densities <500/µl, with sensitivity of only 20% for P. falciparum and 24% for P. vivax at densities <100 parasites/µl. Conclusion:Although PCR performance appeared poor when compared to microscopy, data indicated that the discrepancy between the two methods resulted from poor performance of microscopy at low parasite densities rather than poor performance of PCR. These data are not unusual when the diagnostic method being evaluated is more sensitive than the reference method. PCR appears to be a useful method for detecting Plasmodium parasites during active malaria surveillance in Thailand.
Abstract. In Zimbabwe, high‐resolution (10 Hz to an accuracy of ± 0.1 p.p.m.) measurements were made of atmospheric and host‐produced CO2 in tsetse habitats during the dry season. The diel structure of atmospheric CO2 concentrations is bimodal, with a minimum at approximately 16.00 hours and maxima at approximately 05.00 hours and 20.00 hours, respectively. The background CO2 noise is greater in densely vegetated riverine woodland than in leafless, deciduous (mopane) woodland. Variation in atmospheric CO2 concentrations is correlated with decreasing wind speed and increasing thermal stability. Consequently, the background noise during the day is greatest in riverine woodland during early morning and late afternoon, when winds are typically light and stable, and thermal inversion conditions are developing. Measurements were made of CO2 at 8–64 m downwind from natural (two cattle) or synthetic sources (4–20 L min−1 CO2). The signal from the sources appears as fluctuations above threshold (approximately 355–362 p.p.m.), in the form of intermittent ‘bursts’ of CO2. The strength, duration and intermittency of the signals attributable to these sources declines with source strength and distance from the source. In riverine woodland, approximately 50% of all bursts are 0.1 s long, and 10% are > 2.0 s long. Carbon dioxide signals from equivalent sources are stronger in riverine woodland than mopane. Carbon dioxide dispensed at rates of 4–20 L min−1 is detected up to 64 m downwind of the source but peaks are typically < 10 p.p.m. above threshold. Consequently, host‐CO2 signals are obscured during periods of large fluctuations in atmospheric CO2. These results suggest that CO2 is detectable, at least in some circumstances, at tens of metres downwind and hence dispels the notion that its action is limited to that of a short‐range attractant.
BackgroundThe population dynamics of Plasmodium sporogony within mosquitoes consists of an early phase where parasite abundance decreases during the transition from gametocyte to oocyst, an intermediate phase where parasite abundance remains static as oocysts, and a later phase where parasite abundance increases during the release of progeny sporozoites from oocysts. Sporogonic development is complete when sporozoites invade the mosquito salivary glands. The dynamics and efficiency of this developmental sequence were determined in laboratory strains of Anopheles dirus, Anopheles minimus and Anopheles sawadwongporni mosquitoes for Plasmodium vivax parasites circulating naturally in western Thailand.MethodsMosquitoes were fed blood from 20 symptomatic Thai adults via membrane feeders. Absolute densities were estimated for macrogametocytes, round stages (= female gametes/zygotes), ookinetes, oocysts, haemolymph sporozoites and salivary gland sporozoites. From these census data, five aspects of population dynamics were analysed; 1) changes in life-stage prevalence during early sporogony, 2) kinetics of life-stage formation, 3) efficiency of life-stage transitions, 4) density relationships between successive life-stages, and 5) parasite aggregation patterns.ResultsThere was no difference among the three mosquito species tested in total losses incurred by P. vivax populations during early sporogony. Averaged across all infections, parasite populations incurred a 68-fold loss in abundance, with losses of ca. 19-fold, 2-fold and 2-fold at the first (= gametogenesis/fertilization), second (= round stage transformation), and third (= ookinete migration) life-stage transitions, respectively. However, total losses varied widely among infections, ranging from 6-fold to over 2,000-fold loss. Losses during gametogenesis/fertilization accounted for most of this variability, indicating that gametocytes originating from some volunteers were more fertile than those from other volunteers. Although reasons for such variability were not determined, gametocyte fertility was not correlated with blood haematocrit, asexual parasitaemia, gametocyte density or gametocyte sex ratio. Round stages and ookinetes were present in mosquito midguts for up to 48 hours and development was asynchronous. Parasite losses during fertilization and round stage differentiation were more influenced by factors intrinsic to the parasite and/or factors in the blood, whereas ookinete losses were more strongly influenced by mosquito factors. Oocysts released sporozoites on days 12 to 14, but even by day 22 many oocysts were still present on the midgut. The per capita production was estimated to be approximately 500 sporozoites per oocyst and approximately 75% of the sporozoites released into the haemocoel successfully invaded the salivary glands.ConclusionThe major developmental bottleneck in early sporogony occurred during the transition from macrogametocyte to round stage. Sporozoite invasion into the salivary glands was very efficient. Information on the natur...
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