One of the most challenging aspects of anomalous phenomenon studies is the question of their reproducibility, which is often considered as a condition for a study to be considered scientific. Another critical aspect in such a study is the validity of human testimonies, which is the object of some specific branches of human sciences and has led to a number of famous controversies in terms of what is science (Abelson, 1974; Bauer, 1979) or what the value of testimonies is (Loftus 1979). [See: Sturrock, 1987, for review in a similar area.]

Nevertheless, there are several scientific domains, undoubtedly accepted as full sciences, that do not actually need any experimental reproducibility. For instance, in paleontology, no one can say when and where the next discovery of an Australopithecus skeleton will occur, although this is never depicted in a joking manner as, for instance, observations (even including material evidence, such as photographs or sonar recordings) concerning the Loch Ness Monster (see Bauer, 1987, for review).

Now the major problem is to record indisputable traces of something that is presently interpreted as an unknown or anomalous event, in view of further classification after more knowledge has been received by the scientific community.

The aim of this paper is to give an example of how to study the effects of a phenomenon of unknown origin (of the UFO-type) on the biochemistry of living nonhuman organisms (i.e., on facts that cannot be suspected of lacking objectivity). The question of comparison with controls arises, and will also be dealt with in this study; despite the fact that one cannot know where and when such phenomena will occur, so that no experimental protocol and planning can be actually organized in view of the comparison of "treated" organisms with untreated ones in as exactly similar conditions as possible.

Principles of Sampling Procedure

The first point to be clarified is the exact area where the unknown event (UE) has landed or been in the closest contact with the environment. In the present case, this was the object of a police report referred to as P.V. nr. 28, 9-1-8 1, relating a visible circular trace on the ground.

Then, an ecological axis should be chosen, along which, a series of plants or sedentary animals belonging to the same species can be found at intervals. This axis should go across the "contact area" of the UE and preferably join the epicentre.

The landing area, visible on the ground, was about 2.5 to 3 m in diameter and plants of a wild strain of alfalfa, Medicago minima, were found inside, on the trace, and throughout the surrounding area. This species was thus chosen as the biological model.

The first samples were collected by the local police on the border of the trace (point A) and at a point situated at 20 m (point B) for controls by four days after the observation of the UE.

The second batch of samples (points C to G) were collected by 40 days after the day of observation by a team of technicians of the National Space Research Center. It should be noted that nobody other than the author was aware in advance of when, where, and what was to be collected. This decreases the risk that artifacts could be produced by hoaxers.

A last batch of samples (points H to L) were then collected along the same axis, but two years later, (i.e.,in February 1983) the same plant species were growing on the site, but, of course, samples could not be collected at exactly the same distances. Table 1 indicates the position of the various samples along the axis.

Living plants were taken with a large clod of earth and immediately driven to the laboratory and frozen, except sample (A, B), which was transported by policemen in paper sacks.

In sample (A, B), the plants looked rather dry, but without any sign of burning. In all other samples, the alfalfa leaves, of various size, were quite similar in aspect. No visible morphological alteration was discernible after examination under a Meopta DM23 binocular microscope.

Biochemical Procedure

Samples of 100 mg (fresh weight or equivalent) of young leaves (2 to 3 mm with 7.0 ± 3.6 mg average weight by leaf) were ground in Potter homogenizers with chloroform. Older leaves, which were present in samples A and B, were also analyzed. After 5 mn centrifugation at 5,000 g, the lipid phase was recuperated and concentrated under low pressure to a final volume of 5 µl per mg. These extracts were spotted on thin layer plates, and the various pigments (chlorophylls and derivatives, carotenoïds, quinons, and chromenols) separated according to the previously described techniques. Chromatograms were recorded at 425 nm using a CS920 densitometer.

The pellets were resuspended and homogenized in a mixture of water-ethanol-pyridine-acetic acid (80-10-5-5 v/v) for extraction of carbohydrates and free amino acids. Volumes were adjusted to 0. 5 µl per mg. Quantitative thin layer chromatographies were performed as previously described for carbohydrates (Bounias, 1976, 1980a) and amino acids (Bounias, 1980b).

The pH of the soil was determined after homogenization of 5 g of the earth clod in 100 ml water.

Statistical Methods

Means and SD calculated from (N) determinations were used in student's t test for comparisons. Variances were compared using Fisher's F test. The probabilities of significances corresponding to these comparisons and to the correlation and regression (least square method) calculations, were determined from the equations of distribution of t and F. For correlation coefficients (p), the "t" value was calculated from: t = [gamma· p²/(1 - p²)]^1/2 where v = degree of freedom = N - 2. The standard deviations of the regression slopes (b) was calculated from: sigma_b = [(b/p)² - b²)/v]^1/2. The slopes corresponding to two aleatory variables plotted together are given byd= b/p.

Additionally, in (D+ 370) control samples ("H" to "L"), considered as likely representative of the natural biological variability on the site, correlations with distances to the epicenter were artificially increased by switching the values of the two parameters situated at the extreme parts (i.e., H and L) to their upper and lower possible values, (or reversally), taking into account the range of their standard deviation. Then, correlation calculations will give an estimation of what can be considered as the strongest "fortuitous" correlation, in conditions where no particular correlation is expected. The following notations will be used in the text for the regression slopes: b_E for exposed to the event (A to G); b₀ for controls (H to L); b_r for reconstituted theoretical extreme values in controls.

Only the major features will be represented here.

Acknowledgements. This work was partly supported by grants from the Centre National d'Études Spatiales, Toulouse, France (CNES). Thanks are due to Dr. A. Esterle and to Ing. J-J. Velasco (GEPAN) for their assistance in samples collection, to Mrs. M. M. Daurade for technical assistance and to Miss D. Fernandez for typing the manuscript.