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.     

Quantitation of malaria sporozoites transmitted in vitro during salivation by wild Afrotropical Anopheles

Abstract. The malaria transmission potential of wild, infective Anopheles from western Kenya was evaluated by determining the number of sporozoites transmitted in vitro by salivation when their mouthparts were inserted into capillary tubes containing either sucrose or blood. With sucrose, 86.6% of 102 infective Anopheles transmitted a geometric mean (GM) of 3.84 sporozoites (range 1–34). With blood, 23.1% of 104 infective Anopheles , tested on the day of collection, transmitted a GM of 2.30 sporozoites (range 1–117). For Anopheles held 5 days postcapture before testing with blood, 53.6% of 56 transmitted a GM of 6.04 sporozoites (range 1–420). Transmitting Anopheles contained significantly more salivary gland sporozoites than non-transmitters. No significant differences were detected between Anopheles gambiae Giles sensu lato and Anopheles funestus Giles in sporozoite transmission by individuals with sporozoites in their salivary glands.
Sporozoites were detected microscopically in the salivary duct from heads in 80.3% of 117 infective Anopheles (GM=11.2, range 1–71). Sporozoite detection in mosquito heads by ELISA was 25% less efficient than microscopic detection.
Over 98% of the infective Anopheles transmitted less than twenty-five sporozoites. Transmitted sporozoites represented only about 3% of the total sporozoites in the salivary glands suggesting that sporozoite transmission may be restricted to sporozoites in the salivary duct at the time of feeding. Results are discussed in relation to anti-sporozoite vaccine development.     

The Movement of Sugars into the Sieve Elements of Bark Strips of Willow: I. METABOLISM DURING TRANSPORT

¹⁴C-labelled sugars (sucrose, glucose, and fructose) were suppliedto the cambial surface of bark strips of willow, and in conjunctionwith the aphid stylet technique (Weatherley, Peel, and Hill,1959), the movement into and distribution of activity in thesieve tube sap was examined. As well as sucrose, free hexosesand sugar phosphates were found to contain the ¹⁴C-label, andrapid interconversions occurred between these compounds somewherealong the entry route. Sucrose entry into the sieve elementswas accompanied by at least a partial breakdown into its componenthexoses, and the involvement of a sugar phosphate pool was alsodemonstrated. Possible transformation sites on the entry routebetween the solution bathing the cambial surface and the sieveelement vacuole are discussed.     

Neurosecretion and Salt and Water Balance in the Annelida and Crustacea

The possible role of the neurosecretory system in regulationof salt and water has been studied in the annelids and crustaceans.In the earthworm, Lumbricus terrestris, a brain factor influences'the salt and osmotic concentration of the blood and coelomicfluid. Removal of the brain results in the increase of waterinflux with a decrease in the salt and osmotic concentrationsof the body fluids. The decreases in salt and osmotic concentrationscan be prevented by the implantation of the brain or the injectionof brain homogenates. In the freshwater crayfish, Procambarus clarkii, a factor, presumablysecreted in the brain and released in the eyestalk, seems tomaintain the normal permeability of the body surfaces to water.Eyestalk removal, which eliminates the release site, resultsin the increased influx of water with a decrease in the saltconcentration of the blood. A brain factor also seems to beinvolved in maintaining the sodium and osmotic concentrationsof the blood. In the semi-terrestrial grapsid crab, Metopograpsus messor,the thoracic ganglion, under the control of an eyestalk element,secretes a factor involved in increasing the permeability ofthe body surfaces to water. The removal of the eyestalks, theimplantation of the thoracic ganglion, or the injection of extractsof thoracic ganglia, results in changes in the osmotic concentrationof the blood tending toward that of the medium. In all threespecies studied, the neuroendocrine factors seem to be involvedprimarily in the regulation of the permeability of the bodysurfaces to water.     

In Vitro Culture of Phytomonas Sp. Isolated from Euphorbia characias. Metabolic Studies by 1H NMR

ABSTRACT. We describe the in vitro culture of Phytomonas species isolated from Euphorbia characias . The best choice between tested media was SDM-79, in which promastigotes, after 6 days of culture, reached cell densities as high as 4 × 10⁷ cells/ml. Cells growing in LIT or MTL medium showed longer division times and lower cell densities. We succeeded in obtaining Phytomonas sp. amastigote and spheromastigote forms in modified GRACE's medium, yielding transformation rates of up to 70%. Electron microscopy studies were performed in order to characterize the ultrastructural features of these forms obtained in vitro. On the other hand, metabolic studies based on qualitative (nuclear magnetic resonance spectroscopy) and quantitative metabolic methods (enzymatic assays) showed that promastigote forms secreted mainly ethanol, acetate, glycine, glycerol, piruvate and succinate in SDM-79 medium, whereas the major metabolites found after transformation in modified Grace's medium were ethanol, acetate, glycine, piruvate and smaller amounts of glycerol.     

Treatment of alcoholism; problems arising from the substitution of other drugs in therapy

Of the 139 patients admitted to hospital for chronic alcoholism, 32 had been taking other drugs also, and 17 were addicted to the drugs. Of the 32 patients, 16 used barbiturates, and 8 were addicted. Five took large amounts in suicidal attempt. Ten patients combined still other drugs with alcohol and barbiturates; and seven of them were addicted to barbiturates. Of the six patients combining alcohol with drugs other than barbiturates, two were addicted to the use. Of the 16 patients who used drugs other than barbiturates, eight used one or more opiates such as meperidine, morphine, codeine or dihydromorphinone. Four used stimulants such as benzedrine or dexedrine, alone or in combination. Still other drugs were used in some combination by 32 patients.     

Inter-organ Synergism and the Control of Chlorophyll Accumulation in Sunflower (Helianthus annuus) Cotyledons

Prior to illumination, the embryonic axis of sunflower seedlingsmakes a contribution to the cotyledons which enables them togreen more rapidly when subsequently irradiated with white light.The contributions made to this stimulation by the hypocotylhook, hypocotyl, and roots have been investigated. Greeningin sunflower cotyledons reaches an optimum rate when they are6 d old. The developing embryonic axis not only promotes theattainment of this optimum rate but also aids in the maintenanceof this optimum rate as the cotyledons continue to age. Thisstimulatory effect appears to arise primarily from the rootsand can be duplicated by the exogenous application of benzyladenine. In contrast to previous reports we find that the hypocotyl hookplays no role in the control of greening when present duringillumination.     

Territorial behaviour in an Australian nectar-feeding bird

The red wattlebird, a large Australian honey eater, defends feeding territories from other nectar-feeding birds. One such territory decreased in size and experienced fewer intruders as the density of flowers increased. Daily energy production from the nectar in the territory closely approximated the energy requirements of the territory holder. The bird was probably minimizing energetic costs rather than maximizing energy intake.     

Effects of variation in ear temperature on growth and yield of spring wheat

Wheat plants were kept in a growth room at 15 ^oC from 7 days after an-thesis until maturity and their ears were warmed to 20 or 25 ^oC for various periods. Continuous warming initially increased the growth rate of the ear and decreased that of the stem, but hastened senescence of the ear and decreased final grain yield by decreasing dry weight per grain. Warming the ears increased the movement of ¹⁴C and nitrogen to them from the leaves and stem during the early stages of grain growth but decreased it later. Warming the ears for only the first 10 or 20 days also hastened ear senescence; grain yield decreased progressively with increase in duration of the warm period. All effects were greater at 25 than at 20 ^oC. The effects of changing ear temperature from 15 to 20 ^oC were independent of the temperature of the rest of the plant. Altering the humidity of the air around the ears by 4–7 mb at constant temperature had no effect on ear growth or senescence. Warming all of the plant except the ears from 15 to 20 ^oC increased ear growth slightly during early grain growth and decreased it later, irrespective of ear temperature: stem dry weight, leaf area and net photosynthetic rate of flag leaves and green stems were decreased and dark respiration rate of stems was increased.