ELISA absorbance cut-off method affects malaria sporozoite rate determination in wild Afrotropical Anopheles

ABSTRACT. Malaria sporozoite infection rates in a mixed species group of 244 Anopheles gambiae Giles sensu lato and 115 An. funestus Giles wild female mosquitoes were compared using three methods to determine cutoff absorbance values for positivity of a Plasmodium falciparum Welch enzyme-linked immunosorbent assay (ELISA). Positive controls were based on P. falciparum circumsporozoite protein. As negative controls, four wild male Anopheles were included on each microtitre plate; tests were repeated on four consecutive days for each plate.
Infection rates were estimated at 13.1–22.8% using the mean absorbance value of negative controls plus three standard deviations, 11.7–12.8% using double the mean and 12.5–13.6% using the fixed cut-off value of 0.20 (allowing for 20% variation in negative control absorbance values).
Observed agreement for positivity or negativity among samples tested four times was 98.6% for the 2× mean method, 97.2% for the fixed cut-off 0.20 value, but only 82.7% for the mean +3 SD method. It was concluded that the 2× mean cut-off method is most reliable for field studies. P. falciparum sporozoite rates of 12.2% in An. funestus and 11.9% in An. gambiae s. l . were thus determined on the basis of the 2× mean cut-off method.
This comparative evaluation demonstrates that vector infectivity rates can be seriously over-estimated from sporozoite ELISA tests, by as much as 87% in one case considered here, depending on the absorbance cut-off method applied for negative controls.     

The Fine Structure of the Scopula-Stalk Region of Vorticella convallaria

ABSTRACT. We examined by SEM and TEM the stalk-scopular junction, the stalk, and stalk formation in Vorticella convallaria Linnaeus, 1767. The stalk sheath is anchored to the walls of the scopular lip and to the scopular cilia by thin fibrils. Experimental extraction of these fibrils weakens this junction enough to separate the stalk from the cell body. Telotrochs escape from the stalk by means of violent contractions of the cell body, accelerated beating of the trochal band cilia, and twisting of the cell body against the stalk. The edges of the scopular lip spread over the scopular cilia after escape and, in some cases, fuse to enclose the entire, aboral scopular surface in a cupola-like structure. The sessile cells contain fewer and smaller scopular granules than telotrochs. The presence of disintegrating scopular granules in the stalk matrix of some sessile cells suggests that they contain material which is secreted over a period of time to form the stalk. Eruptive formation of the initial adhesive pad and quick elongation of the distal part of the stalk suggests a rapid exocytosis of the larger, more numerous granules of the telotroch. The stalk sheath is formed of fibrils making up complete and incomplete compartments peripherally arranged along the major stalk axis.     

Isolation of highly polymorphic microsatellite markers from the intertidal barnacle Chthamalus montagui Southward

This study reports the isolation and characterization of seven highly polymorphic microsatellite loci in Chthamalus montagui (Crustacea, Cirripedia). The loci were isolated from a library constructed from genomic DNA enriched for CA repeats. The markers yielded three to 43 alleles per locus (mean 16.7) in samples averaging 49 individuals. Observed heterozygosity ranged from 0.08 to 0.58 (mean 0.39). These microsatellite loci will be valuable tools for population genetic studies of this species.