Biological cost of tolerance to heavy metals in the mosquito Anopheles gambiae.

The global rate of heavy metal pollution is rapidly increasing in various habitats. Anopheles malaria vector species (Diptera: Culicidae) appear to tolerate many aquatic habitats with metal pollutants, despite their normal proclivity for 'clean' water (i.e. low levels of organic matter). Investigations were conducted to establish whether there are biological costs for tolerance to heavy metals in Anopheles gambiae Giles sensu stricto and to assess the potential impact of heavy metal pollution on mosquito ecology. Anopheles gambiae s.s. were selected for cadmium, copper or lead tolerance through chronic exposure of immature stages to solutions of the metals for three successive generations. Biological costs were assessed in the fourth generation by horizontal life table analysis. Tolerance in larvae to cadmium (as cadmium chloride, CdCl(2)), copper [as copper II nitrate hydrate, Cu(NO(3))(2) 2.5 H(2)O] and lead [as lead II nitrate, Pb(NO(3))(2)], monitored by changes in LC(50) concentrations of the metals, changed from 6.07 microg/L, 12.42 microg/L and 493.32 microg/L to 4.45 microg/L, 25.02 microg/L and 516.69 microg/L, respectively, after three generations of exposure. The metal-selected strains had a significantly lower magnitude of egg viability, larval and pupal survivorship, adult emergence, fecundity and net reproductive rate than the control strain. The population doubling times were significantly longer and the instantaneous birth rates lower in most metal-selected strains relative to the control strain. Our results suggest that although An. gambiae s.s. displays the potential to develop tolerance to heavy metals, particularly copper, this may occur at a significant biological cost, which can adversely affect its ecological fitness.

Visual assessment of sporozoite and bloodmeal ELISA samples in malaria field studies.

The accuracy of visually assessing positivity for samples of field-collected Anopheles tested by enzyme-linked immunosorbent assays (ELISA) for Plasmodium falciparum sporozoites and human bloodmeals was determined during malaria field studies in Kenya. Six observers familiar with ELISAs evaluated 5,344 sporozoite ELISA samples and four observers evaluated 360 bloodmeal samples as either positive or negative based on the presence and strength of green-colored peroxidase reactions relative to controls on each microtiter plate. Interobserver agreement ranged from 97.9 to 99.8% for sporozoite samples and from 90.3 to 96.1% for bloodmeal samples. For both assays, the mean sensitivity and specificity of visual readings, compared with spectrophotometric readings, exceeded 98% when absorbance values were greater than or equal to 0.4 (on a scale of 0.0 to 2.0). Most incorrect visual readings occurred for samples with absorbance values between 0.2 and 0.4. The total percentage of samples classified correctly by visual examination ranged from 97.7 to 99.5% for the sporozoite ELISA and from 95.0 to 96.7% for the bloodmeal ELISA. Thus, there was minimal error associated with visually determining positive reactions for the ELISA assays used in malaria field studies.