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.     

A Multi-Unit Sampling System and Its Use in the Characterization of Ultradian and Infradian Growth in Tetrahymena*

SYNOPSIS. A multi-unit automatic sampling device for investigation of microbial growth under a wide variety of conditions is described. The kinetics of asynchronous population growth for batch cultures of Tetrahymena pyriformis (W) at several temperatures show that there are 2 distinct growth phases: an exponential ultradian growth phase that is strongly temperature dependent and a non-stationary growth phase, the infradian phase, that shows little or no temperature dependence over the range from 15–27 C.     

Phylogenetic Position of Amblyospora Hazard & Oldacre (Microspora: Amblyosporidae) Based on Small Subunit rRNA Data and Its Implication for the Evolution of the Microsporidia

ABSTRACT. Sequences of the small subunit rRNA genes of Amblyospora californica and an Amblyospora sp. from Culex salinarius were determined. These sequences were compared phylogenetically with 16 other microsporidia. The results suggest Amblyospora forms a sister taxon to the rest of the microsporidia examined. The basal position of Amblyospora is discussed with respect to the evolution of microsporidian life cycles. These sequences represent the longest microsporidian small subunit rRNA genes sequenced to date, 1,359 and 1,358 bp, respectively. Structural features and GC content (49% for both) are comparable to those of other microsporidia which have been sequenced.     

Tropical zooplankton: phenotypic and spatial analyses of polymorphic Ceriodaphnia cornuta (Crustacea: Cladocera)

SUMMARY. 1. The predation-balanced polymorphism of Ceriodaphnia cornuta (a small-eyed horned form and a large-eyed unhorned form) in Lake Gatun, Panama, is re-examined approximately 12 years after the original study (Zaret, 1972a). We consider morphological variability, distribution and fecundity of the population inhabiting Laboratory Bay.
2. The two morphological characters emphasized by Zaret, eye size and horn length, are studied in detail. The C. cornuta population exhibits wide, yet continuous variation; coefficients of variation are 28% or eye diameter and range from 114% to 136% for horn and spine lengths. Phenotypic variation in the population is characterized by: (1) negative correlations between spine lengths and eye size; (2) positive correlations among spines from different body regions (anterior horn, lateral spines, and tail spine); and (3) weak correlations between body length and spine lengths.
3. While the horned and unhorned forms continue to coexist in Laboratory Bay. the frequencies of horned forms have increased in the open water areas since the time of the original study. The increase in horned form frequencies may be due to changes in fish predation intensity. Did vertical distribution patterns of the forms remain very similar to each other.
4. In contrast to previous laboratory findings, egg ratio calculations from field samples revealed similar clutch sizes for the horned and unhorned forms. The egg development time of the horned form, however, was found to be approximately 30% longer than that of the unhorned form, perhaps because of increased moulting difficulties produced by the presence of spines.     

Uptake, translocation and metabolism of fluazifop-butyl in Setaria viridis

The uptake and translocation of fluazifop-butyl was investigated in Setaria viridis. Young plants (three to four leaves) with a portion of the second, third or fourth leaf covered, were sprayed with a dose equivalent to 0.25 kg a.i. ha-¹. ¹⁴C-fluazifop-butyl was subsequently applied to the unsprayed area and the treatment resulted in plant death within 2 wk. Uptake by leaf 3 was rapid, with less than 5% of the applied dose remaining on the leaf surface after 24 h. The highest proportion of ^14C-activity was retained in the treated portion of the leaf. Only 2% of the applied dose was translocated from leaf 3 and 0. 76% accumulated in the apical meristem. Uptake by the younger leaf 4 was more rapid and the pattern of translocation differed in that more ¹⁴-^-activity accumulated in apical meristematic tissue. 6–30% of the applied dose was undetected and this was greatest when foliar uptake was slow. Artificial leaf surface experiments indicated that this undetected activity may have been due to volatility of fluazifop-butyl or degradation to volatile products. ^14C-activity extracted from treated leaves was identified as fluazifop-butyl, fluazifop acid and polar conjugates. The major ¹⁴C-activity extracted from the apical meristem was fluazifop acid and no fluazifop-butyl was detected in this extract.     

Potassium-Ammonium Uptake Interactions in Tobacco Seedlings

Short-term (< 12 h) uptake experiments were conducted with6–7-week-old tobacco (Nicotiana tabacum L. cv. Ky 14)seedlings to determine absorption interactions between K⁺ andNH₄⁺. At equal solution concentrations (0.5 mol m^–3) netK⁺ uptake was inhibited 30–35% by NH₄⁺ and NH₄⁺ uptakewas decreased 9–24%. Removal of NH₄⁺ resulted in completerecovery in K⁺ uptake rate, but NH⁴⁺ uptake rate did not recoverwhen K⁺ was removed. In both cases, inhibition of the uptakerate of one cation saturated as the concentration of the othercation was increased up to 0.5 mol m^–3. The relative effectof K⁺-NH₄⁺ interactions was not altered when Cl- was replacedwith SO₄^2–, but the magnitudes of the uptake rates wereless in the absence of Cl-. The V_max for NH₄⁺ uptake was reducedfrom 128 to 105 µmol g^–1 dry wt. h^–1 in thepresence of 0.5 mol m^–3 K⁺ and the K_m for NH₄⁺ doubledfrom 12 to 27 mmol m^–3 in the presence of K⁺. The resultsof these K⁺-NH₄⁺ experiments are interpreted as mixed-noncompetitiveinteractions. However, an enhanced efflux of K⁺ coupled to NH₄⁺influx via an antiporter cannot be ruled out as contributingto the decrease in net K⁺ uptake. Key words: Nicotiana tabacum, K⁺, NH₄⁺, Uptake interactions     

NEUROBIOLOGY OF ECHINODERMATA

During the past ten years much information has been added to our knowledge of nerve and muscle systems of echinoderms. 1. Electron-microscopy has shown that all the main nerve trunks consist of large numbers of small, parallel-running unmyelinated axons which are packed tightly together. Glial cells are generally absent. Discrete regions of neuropile are recognizable by the interweaving of axons, and the presence of vesicles. It has not yet been found possible to locate synapses with certainty in the nervous system, but it appears that they are chiefly confined to neuropile. The obvious nerve cords are massive accumulations of neurons which do not appear to interact locally. 2. Peripheral axons are difficult to distinguish because both interstitial and muscle cells have processes which often resemble axons. Ultrastructural analysis of this problem is aggravated by difficulties in fixation. However, the electron-microscope has shown that much of the echinoderm body wall contains a thick subepithelial plexus of processes from epithelial cells. Epithelial cells may thus act as sensory cells and supply axons to the plexus. 3. With the exception of striated muscles in some pedicellariae, all echinoderm muscles so far examined are of the smooth type. These muscles characteristically contain large filaments, and in this way do not resemble vertebrate smooth muscle. Some muscles are innervated by simple axonal contact, in others the muscles themselves send processes towards the nervous tissue. 4. Physiological studies of electrical activity in nerve and muscle systems have not added significantly to our knowledge of function. Several authors have demonstrated that massed electrical activity is conducted decrementally along the radial nerve cords, but this does not explain any known aspect of coordination. The only records of electrical activity from single neurons (Takahashi, 1964) have not been repeated. 5. There is strong evidence for two types of neurons in the central nervous system of echinoderms. One of these contains acetylcholine, the other dopamine and/or noradrenaline. Electron-microscopical histochemistry has given good indication that catecholamines are bound in echinoderm nerve tissue to particles similar to those reported in other invertebrate nervous tissues, and there is good evidence that acetylcholine is bound to synaptic vesicles which are morphologically identical to those present in the mammalian brain. The available data further indicate that acetylcholine is a transmitter in sensory and motor neurons, while dopamine and/or noradrenaline are transmitters in interneurons. Such interneurons may be involved in the coordination of the movement of the tube-feet. Other substances which have been implicated in neuro-effector mechanisms in other animal groups have not been found or are present in very small quantities. 6. Studies on the reproductive physiology of starfish have shown that several substances in the radial cords play important roles in its control. Such substances cannot at present be called neurosecretions because it is not known if they are derived from neurons. 7. Pharmacological studies on isolated muscle tissues have not added significantly to our knowledge of their control. The potency of ACh in causing contraction is well documented, and anticholinesterases are similar in effect. Catecholamines, although clearly very important in the nervous system, do not produce clear-cut effects. The published reports of relaxation to noradrenaline may well be due to direct effects on the muscle. No definite information has been obtained on the role of the adrenergic parts of the nervous systems of echinoderms, other than showing that they are not involved in motor responses. Extensive studies with a wide variety of drugs have produced inconsistent and largely negative results.