THE EFFECT OF VARYING RELATIVE DENSITY AND VARYING FOOD SUPPLY ON INTERSPECIFIC COMPETITION BETWEEN HELISOMA DURYI AND BULINUS TRUNCATUS (GASTROPODA, PLANORBIDAE)

Interspecific competition between Helisoma duryi and the intermediatehost of schistosomes, Bulinus truncatus results in reduced growthand reproduction of B. truncatus and the effects are more pronouncedas the density of H. duryi is increased. The drastic reductionin reproduction of B. trunauus is the result of a predationby H. duryi on egg masses. It appears that increasing the foodsupply cannot completely eliminate the effects of competition.Although the growth and reproduction of B. truncatus in competitionwith H. duryi increases significantly with the increased foodsupply, both growth and reproduction are still reduced comparedto control snails given the same amount of food. The effectsof food supply and competition on the allometric size-weightrelationships of the two species are slight. (Received 23 February 1987; accepted 4 May 1988)     

The effect of temperature,darkness, starvation and various food types on growth,survival and reproduction of Helisoma duryi,Biomphalaria alexandrina and Bulinus truncatus (Gastropoda: Planorbidae)

Helisoma duryi has been proposed as a biological control agent in schistosomiasis due to its superiority in laboratory competition experiments with various species of the intermediate host snails. Therefore it was considered important to evaluate the response of this snail species and the intermediate host species, Biomphalaria alexandrina and Bulinus truncatus, to various physical, chemical and biological factors under laboratory conditions in order to obtain information on the similarities in the ecological niches of these species. The factors considered in the present paper are: temperature, darkness, starvation and food. All three species had optimal growth and egg laying at 26–28 °C. Only H. duryi survived for a longer period at 33°C and it was capable of starting egg laying at this temperature although the onset was delayed. However, low temperature (18°C) caused a relatively larger decrease in egg laying of H. duryi than in the other two species. Growth and egg laying was reduced for H. duryi and B. truncatus kept under darkness and B. alexandrina could not tolerate maintenance under darkness. A few days of starvation of juvenile snails had no effect on later growth and egg laying capacity of the survivors, although mortality in B. truncatus was increased. B. alexandrina had a lower tolerance to starvation than the other two species. Egg laying of snails fed only one of the three laboratory food types decreased for all three species in the order: Vov-vov (dog food in dry pellets), Tetramin (fish food) and lettuce. Combinations of lettuce and one or more proteinaceous food types gave optimal growth and egg laying for all three species.     

INTRASPECIFIC AND INTERSPECIFIC VARIATION IN SHELL MORPHOLOGY OF BIOMPHALARIA PFEIFFERI (KRAUSS, 1848) AND HELISOMA DURYI (WETHERBY, 1879) FROM MOSHI, TANZANIA, ANALYSED BY MORPHOMETRIC STUDIES

Helisoma duryi, a planorbid snail species, which is a potentialbiological control agent of the intermediate hosts of schistosomiasis,was introduced in an irrigation scheme (T.P.C.) in northernTanzania in 1972. The species has been present at T.P.C. since,and during a snail survey in January 1981, a great variationin the shell morphology of H. duryi, ranging from typical H.duryi forms to forms resembling Biomphalaria pfeifferi, theintermediate host of Schistosoma mansoni in the area, was observed.The Biomphalaria- Wkc forms of H. duryi could constitute anentire population but also occasionally occurred in populationsof typical H. duryi. This study was undertaken to determine whether simple shellmeasurements, or ratios between some of these could be usefulin discriminating between H. duryi and B. pfeifferi. Followingparameters were considered: shell height, shell diameter, diameterof the umbilicus, ratio between shell height and diameter ofthe umbilicus, and the number of whorls. (Received 29 July 1983;     

ALLOZYME COMPARISONS AND PLOIDY LEVELS AMONG SPECIES OF THE BULINUS TRUNCATUS/TROPICUS COMPLEX (GASTROPODA: PLANORBIDAE) IN CAMEROON

Allozyme electrophoresis, ploidy level, and, to a lesser extent,anatomy and shell morphology were used to characterize snailsof the Bulinus truncatus/tropicus complex in Cameroon. Starchgel electrophoresis revealed no intrapopulation variation within20 populations examined for 14 allozyme loci in 12 enzyme systems.Interpopulation differences in allozyme patterns were used todivide the populations studied into five groups. Each groupis distinguished by a unique combination of zymograms for hydroxybutyratedehydrogenase and isocitrate dehydrogenase. Four of these groupsare tetraploids and are identified as B. truncatus. Populationsof the most commonly found B. truncatus group are generallyaphallic, are frequently associated with transmission of S.haematobium, and generally occur at altitudes below 800 m abovesea-level. Populations of B. truncatus belonging to the otherthree groups are usually euphallic, not associated with S. haematobiumtransmision, and generally occur at altitudes above 1000 m.These four groups may represent distinct biotypes of B. truncatus.Populations belonging to the fifth group are diploid, euphallic,not associated with S. haematobium transmission, and occur inhabitats above 1000 m. This is the first confirmed report ofdiploid B. truncatus/tropicus-complex snails from this regionof Africa. They are tentatively identified as B. natalensis,a species previously thought to be limited to East and SouthernAfrica. (Received 31 October 1988; accepted 20 February 1989)     

FRESHWATER SNAILS OF THE BULINUS TRUNCATUS/TROPICUS COMPLEX IN KENYA: TETRAPLOID SPECIES

Studies in recent years on chromosome number in freshwater snailsbelonging to the Bulinus truncatus/tropicus complex demonstratedthe occurrence in Kenya of one diploid species, B. tropicus,and 2 tetraploid species, B. truncatus and B. permembranaceus.Further observations are now described on the morphologies ofthe tetraploid species, and their distributions. Observations were made on B. truncatus from 26 localities (11newly reported) and B. permembranaceus from 26 localities (7newly reported), in respect of chromosome number, egg proteins,enzymes (5 systems), shell (10 variables measured), male genitalsystem (normal or aphallic) and radula (1st lateral tooth, sizeand mesocone shape). B. permembranaceus differs most clearly from B. truncatus inegg proteins, enzyme-loci and in lacking aphallic individuals.Mesocone shape is generally less angular in B. permembranaceus.The shell of B. permembranaceus grows larger, has the columellamore commonly concave and its spire is proportionally higherand more acute. To distinguish these tetraploid species fromthe partly sympatric B. tropicus, observations on chromosomenumber and biochemical characters are necessary. The tetraploids are allopatric, B. truncatus being found rarelyas high as 1900 m altitude, whereas B. permembranaceus occupiesthe altitude range 1940–2760 m. B. truncatus may be expandingits distribution in Kenya in man-made waterbodies, while B.permembranaceus possibly is restricted by adaptation to coolconditions and by interaction with B. tropicus. Differencesbetween B. truncatus and B. permembranaceus, both morphologicaland biochemical, indicate their origins lie in independent episodesof tetraploidy. ^*Member of External Scientific staff, Medical Research Council (Received 31 October 1988; accepted 6 December 1988)     

THE EFFECT OF WATER QUALITY ON OVIPOSITION IN BIOMPHALARIA GLABRATA (SAY, 1818) (PLANORBIDAE), AND A DESCRIPTION OF THE STAGES OF THE EGG-LAYING PROCESS

With the overall goal of developing a method to reliably induce ovipositionin the freshwater pulmonate Biomphalaria glabrata, the effectsof water quality on female reproductive physiology were examined.Groups of snails were subjected to controlled experimental conditionsconsisting of a daily regimen of feeding and water change. Aftera period of acclimatiz-ation, egg mass (EM) output under theseconditions was relatively stable, and snails laid a majority(82.5%) of their EM during the initial 4 h following daily waterchange. When this regimen was perturbed by halting water changefor 24 h (dirty-water treatment), EM output was significantly inhibited.When water change was resumed, EM output returned to previouslevels within 4 h post-water change (PWC). This dirty-water treatmentfollowed by water change also resulted in a significant increasein mean EM size during the 4 h PWC when compared to controls.To better describe the events preceding egg-laying in B. glabrata,we then used these experimental manipulations to induce ovipositionin groups of snails, and dissected them during the 4 h followingwater change. Observations of the reproductive tracts of stimulatedsnails allowed us to divide the egg-laying process, from ovulationto oviposition, into discrete stages, after de Jong-Brink, Koop,Roos & Bergamin-Sassen (1982). Stage I was characterized bythe presence of ova in the hermaphroditic duct and carrefour,and fertilized, packaged eggs in the oviduct and muciparousgland. Stage II was characterized by the presence of packagedeggs in the othecal gland embedded in a mucous layer, constitutingthe egg mass to be laid on the substratum. No packaging eventswere occurring in the carrefour/albumen gland region duringthis stage. When snails were dissected immediately after oviposition(Stage III), unpackaged ova were observed in the hermaphroditicduct, carrefour, and oviduct. The mean time it took for snailsto reach Stage III was 120 6 49 min (SD), and this value wasstatistically different from the mean time to Stages I and II,showing that our induction protocol results in a temporal progressionthrough the egg- laying process. Gonadal oocyte density (oocytes/mm²of ovotestis) was quantified as a function of these stages ofthe reproductive cycle, and was found to be significantly lowerduring Stage II (fully formed egg mass in othecal gland) thanall other stages examined. Taken together, these results showthat female reproductive activity can be experimentally controlledthrough the manipulation of water quality, and that such a protocolis a valuable tool for addressing specific questions regardingthe reproductive physiology of B. glabrata. The implicationsof these results as they pertain to the regulation of femalereproductive activity in B. glabrata are discussed. (Received 15 January 1999; accepted 17 May 1999)     

SEASONAL FLUCTUATIONS IN POPULATION DENSITES OF BULINUS SENEGALENSIS AND B. TRUNCATUS (PLANORBIDAE) IN TEMPORARY POOLS IN A FOCUS OF SCHISTOSOMA HAEMATOBIUM IN NIGER: IMPLICATIONS FOR CONTROL

Annual and seasonal changes in population densities of Bulinussenegalensis Müller 1781 and B. truncatus (Audouin, 1827)were studied in temporary pools of the west sahelian zone inNiger in relation to environmental factors. B. senegalensisis present in the temporary pools whatever their duration, whereasB. truncatus inhabits only the more persistent pools. B. senegalensispopulations develop mostly during the rainy season when thewater temperature is warm. B. truncatus is most abundant afterthe rainy season when the water temperature is cooler, but thissnail may increase when the water temperature is warmer duringunusual rainy seasons with low and intermittent rainfall. Fieldobservations suggest that interspecific competition is exceptionallyoccurring in the pool colonized by both B. senegalensis andB. truncatus. The implications of these observations on treatmentand retreatment schedules for schistosome infection in peopleand on snail control in temporary pools are discussed. (Received 4 May 1993; accepted 22 July 1994)     

EFFECT OF CONTROLLED TEMPERATURES ON GAMETOGENESIS IN THE GASTROPODS PHYSA ACUTA (PHYSIDAE) AND BULINUS TROPICUS (PLANORBIDAE)

The effects of a range of constant temperatures on gametogenesiswere investigated in two freshwater snail species which occursympatrically in the Umsin-dusi River, Natal, South Africa,Physa acuta (invasive) and Bulinus tropicus (endemic). Bothwere shown to be simultaneous hermaphrodites with slight protandry.An increase in temperature accelerated the process of gametogenesisbut P. acuta responded more rapidly to these changes than B.tropicus. This, coupled with the greater fecundity and shorterincubation period of P. acuta, gives it a reproductive advantageover the endemic species, providing a possible explanation forthe invasive's success in rapidly repopulating disturbed riverinehabitats. (Received 16 November 1990; accepted 1 May 1991)     

MATING SYSTEM ANALYSIS USING PROTEIN ELECTROPHORESIS IN THE SELF-FERTILE HERMAPHRODITE SPECIES BULINUS TRUNCATUS (GASTROPODA: PLANORBIDAE)

Uniparental reproduction in the tetraploid hermaphrodite speciesBulinus truncatus has been suggested to occur via self-fertilization,on the basis of cytological and genetic studies. However, recentanalyses of population genetic structure by protein electrophoresisindicated the occurrence of multi-banded phenotypes referredto as ‘fixed hetero-zygosity’. Although intrapopulationand geographical variation of multibanded phenotypes occur,many populations bear only one type of such patterns. This ledto the suggestion that parthenogenesis may well be the matingsyste in B. truncatus. However, such fixed heterozygosity patternsare expected in some tetraploid species in which double disomicinheritance of alleles occurs. It is therefore not possibleto determine from population genetic structure analysis alonewhether segregation occurs or not. Here, we investigate thestability of such patterns over one generation of uniparentalreproduction among four populations. We also present resultsof crossing experiments between individuals from two populations,using two diagnostic loci to analyse their offspring. Our resultsclearly indicate Mendelian segregation of alleles, and confirmsexual reproduction by self-fertilization and cross-fertilization.We interpret the multibanded patterns observed in populationsas the product of both diploid loci of the tetraploid genomewhen they are monomorphic for different alleles. Our study alsoallows us to suggest that partial selfing may be the regularmating system in B. truncatus. (Received 12 February 1992; accepted 7 September 1992)     

Fate of organic compounds in mountain streams of the Marmot basin: A process reactor model

Helisoma duryi is a planorbid snail that has been proposed as biological control agent against intermediate hosts of schistosomiasis. When the population density of this species is increased from 200 to 25 ml per snail in containers of constant volume (i.e. 200 ml) growth decreases. Whereas the single snails (i.e. 200 ml/snail) laid no egg masses, egg-laying in the other groups decreased with increasing density. When four snails were kept in various volumes of water giving the same population densities as above both growth and egg-laying decreased with increasing density. Cross-breeding experiments between an albino and a pigmented strain of H. duryi showed that the albino character is inherited as a recessive autosomal character, cross-fertilization appears to be the only mode of reproduction in H. duryi, stored foreign sperm remains viable for up to five months and stored sperm can survive when snails are aestivating or starving. The implications of the present findings for the use of H. duryi in biological control are discussed.     

A COMPARISON OF THE LIFE CYCLES OF THE SLUGS DEROCERAS RETICULATUM (MULLER) AND ARION INTERMEDIUS NORMAND ON PERMANENT PASTURE

Life cycles of the slugs Deroceras reticulatum and Arion intermediuswere compared over a four-year period on permanent pasture nearOvingham, Northumberland. Both species were semelparous, theirlife cycles taking about one year. D. reticulatum, unlike A.intermedius and other species, had two overlapping generationsthat laid their eggs in late spring and autumn respectivelyand also had one instead of two immature phases in its lifecycle. The pattern and rates of growth for the immature phasesin the life cycle of A. intermedius differed markedly from thosedescribed under laboratory conditions. Growth in the infantilephase was delayed in the field by low winter temperatures whilesecond stage growth was usually delayed in late spring by dryconditions. The life cycle of A. intermedius appears to be synchronisedby seasonal changes in photoperiod, unlike that of D. reticulatum.Growth in D. reticulatum continued throughout the winter monthsexcept under exceptionally cold conditions and was not usuallydelayed by dry conditions in spring. The size of hermaphroditegland relative to body weight in D. reticulatum reached a maximumin mature-unmated slugs and then became progressively smalleras the slug approached the post-reproductive stage which wasrelatively brief. Exceptionally dry conditions in 1962 delayedthe development of the hermaphrodite gland, the maximum sizereached was significantly reduced and fewer eggs were laid. (Received 17 September 1987; accepted 2 January 1988)     

PATTERNS OF REPRODUCTION IN THREE SANDY BEACH WHELKS OF THE GENUS BULLIA GRIFFITH

The breeding cycles of two species of sandy beach whelk (Bulliadigitalis and B. pura) are presented and compared with a thirdspecies (B. rhodostoma). In all three species, egg maturationand copulation occur in spring while summer marks the depositionand spawning of egg capsules. B. digitalis and B. pura migrateoffshore to lay their eggs, the juveniles of both species restrictingthemselves to beyond the breaker zone. Newly hatched snailsof B. rhodostoma appear in the intertidal towards late summer.The discovery of a penis-like structure on females of B. rhodostomais investigated. The pseudopenis was also found on B. pura butnever on B. digitalis. The possibility of a sex-change was eliminatedon the histological investigation of the gonads of a range ofsnails and the examination of sex-ratios over a period of fivemonths. General trends in reproductive behaviour are also discussed. (Received 18 March 1984;     

A COMPARATIVE STUDY OF THE MATURE SPERMATOZOA OF BULINUS AFRICANUS (KRAUSS, 1848) AND B. GLOBOSUS (MORELET, 1866) (GASTROPODA: PLANORBIDAE)

The morphology of the mature spermatozoa of the closely-relatedfreshwater pulmonates, Bulinus africanus and Bulinus globosus,was examined using scanning and transmission electron microscopy.A comparison showed no useful differences that could be usedto distinguish the species though they did differ in one respectfrom Bulinus tropicus, the only other bulinid for which spermmorphology is known. (Received 12 August 1996; accepted 6 December 1996)     

MORPHOLOGICAL DIVERGENCE AND EVIDENCE FOR REPRODUCTIVE ISOLATION IN LITTORINA SAXATILIS(OLIVI) IN NORTHEAST ENGLAND

Current work at Old Peak (Ravenscar) on the northeast coastof England indicates that boulder-dwelling Lutonna saxatilis(Olvi) comprise at least two distinct forms, inhabiting high-and mid-shore boulders Using multivariate morphometrics we showthat these forms are significantly different in shape. Rareintermediates between these two distinct forms were occasionallyfound, usually with the low shore animals The two forms showdifferent reproductive strategies. High shore females (L. saxatilisH) mature at a smaller size and produce a smaller number oflarge eggs giving rise to large crawlaways The mid-shore form(L. saxatilis M) matures at a larger size and produces a largenumber of small eggs which give rise to small crawlaways. Anovel observation is that intermediate females (L. saxatilisI) contain eggs of both sizes, giving rise to a bimodal distributionof egg sizes within the brood pouch. The usual incidence ofabortion (observed in H and M forms) is Very low (<1%) However,in the intermediates the large eggs showed a high incidenceof abortion (about 60%) and the incidence of abortion in thesmall eggs was also increased (about 30%) Thus there appearsto be a partial reproductive barrier between the two forms,presumably resulting in reduced fitness due in the intermediatesto reproductive dysfunction. These findings extend to othershores in the region, over total distance of about 66 km. Thefacts are suggestive of a hitherto unrecognised reproductivebarrier with L. saxatilis, the nature and strength of whichis under investigation ^*Present address: University College Scarborough. Filey Road.Scarborough Yo11 3AZ. (Received 12 June 1995; accepted 15 September 1995)     

THE GROWTH AND BREEDING CYCLE OF LITTORINA OBTUSATA (GASTROPODA : PROSOBRANCHIATA) FROM CARDIGAN BAY

The population structure of Littorina obtusata was for mostof the year bimodal, with a stable adult component and an activelygrowing juvenile component. Growth rate of the juvenile componentwas similar at different beach levels and there was a retardationof growth as the snails approached sexual maturity. There was little variation in the histology and external appearanceof the reproductive organs throughout the year and the breedingcycle has been characterized by the amount of spawn laid andthe presence of mature eggs in the oviduct of the female. ^*Present address: Portway School, Penpole Lane, Shirehampton,Bristol. (Received 7 September 1977;     

OBSERVATIONS ON THE ECOLOGY OF BULINUS (PHYSOPSIS) GLOBOSUS (MORELET), B. (BULINUS) FORSKALII (EHRENBERG) AND LYMNAEA NATALENSIS (KRAUSS) IN RICE SWAMPS IN SIERRA LEONE.

Changes in population of Bulinus globosus, B. forskalii andLymnaea natalensis were studied in four different locationsnamed Sites A, B, C. and D in Bo town Sierra Leone. The snailpopulations and oxygen concentrations of water increased inthe rainy season. The biological oxygen demand did not havesignificant correlation with changes in snail populations inSites B and D. B. globosus was absent from Site A while B. forskaliiwas the predominant snail in Site C. Site D with the highestmean monthly conductivity of 176 µmhos contained 54% whileSite A with a similar reading of 90µmhos contained 1%of all snails found. (Received 22 May 1980;     

Salt Tolerance in Suaeda maritima (L.) Dum: THE EFFECT OF SODIUM CHLORIDE ON GROWTH, RESPIRATION, AND SOLUBLE ENZYMES IN A COMPARATIVE STUDY WITH PISUM SATIVUM L

The effect of sodium, chloride on the growth of a halophyte,Suaeda maritima (L.) Dum., was compared with its effect on Pisumsativum L. cv. Alaska under controlled environmental conditions.The salt stimulated the growth of Suaeda maximally at concentrationsof 170 to 340 mM while the growth of Pisum was inhibited evenby 100 mM. Both species accumulated ions in the tops and themaximum concentrations of Na⁺ and Cl^– rose in Suaeda to860 mM (based on the water content) and 730 mM and in Pisumto 170 mM and 300 mM respectively. Respiration in both specieswas inhibited as the NaCl level in the culture solution wasraised. Four supernatant enzymes (malic dehydrogenase, glucose-6-phosphatedehydrogenase, peroxidase, and acid phosphatase) prepared fromPisum and from Suaeda (grown either in the absence of addedNaCl or in the presence of 340 mM NaCl) were assayed in variouslevels of sodium chloride. The dehydrogenases were markedlyinhibited by increasing salt concentrations while there wasa smaller effect on the peroxidase and acid phosphatase. Therewas no difference in the effect of salt on the enzymes preparedfrom the two species although one is halophilic and the otherhalophobic.     

Phosphate Absorption, Fluxes, and Symplasmic Transport in Osmotically-Shocked Zea mays Roots

Osmotic shock with sequential 30 min treatments in ice-coldsaline solutions and distilled water inhibited both the subsequentuptake of orthophosphate (Pi) and its transport into the xylemof excised corn (Zea mays L.) roots. Measurements of Pi fluxeswith ³²P indicated that the decrease in net Pi uptake over a24 h period caused by osmotic shock was due primarily to delayedrecovery of Pi influx rather than to increasing efflux. Despitecomplete recovery of Pi absorption within 2–6 h aftershocking with 150–200 mM NaCl, transport to the xylemduring the subsequent 24 h only partially recovered. Leucineuptake and incorporation into protein was also markedly inhibitedby osmotic shock but both almost completely resumed controlrates within 24 h after shocking with up to 150 mM NaCl. Tetracyclineinhibited recovery of Pi uptake after NaCl treatment whereaspuromycin did not. These results with corn roots are consistentwith the hypothesis that recovery of Pi uptake activity aftermoderate osmotic shock requires de novo synthesis of membraneproteins. Incomplete recovery of Pi transport to the xylem suggeststhat osmotic shock may damage plasmodesmata. Key words: Corn, Ion uptake, Leucine uptake, NaCl, Puromycin, Tetracycline     

Genetic variation within and between populations of hermaphroditic <Emphasis Type="Italic">Bulinus truncatus</Emphasis> tetraploid freshwater snails of the Albertine Rift,East Africa

Genetic variability within and among Bulinus truncatus of the Albertine Rift freshwater bodies were assessed to investigate the degree of inbreeding and gene flow in the snail populations. The effect of ploidy on the genetic structuring of B. truncatus is also described. We characterized the genetic structure of seven B. truncatus populations from Lake Albert, Lake Kivu, and Katosho swamp in Tanzania using five polymorphic microsatellite loci. Genetic differentiation was quantified using pairwise FST values and Nei’s standard genetic distances. Different alleles were observed across all loci and genetic diversity was low although it varied greatly across populations; observed heterozygosity was, however, higher than the expected heterozygosity in three of the populations studied. Significant heterozygote deficiencies were observed coupled with significant linkage disequilibria in five populations for all the five loci examined in this study. We found significant genetic differentiation among the seven freshwater bodies; private alleles were observed across all loci indicating restricted or absence of gene flow between populations. Limited snail dispersal and the reproductive biology of B. truncatus are the major forces shaping the genetic variation observed. Low genetic variation within B. truncatus populations exposes them to a high parasite infection risk as predicted in the Red Queen hypothesis.     

OBSERVATIONS ON THE POPULATION DYNAMICS OF SNAIL HOSTS FOR SCHISTOSOMES IN THE DELTA OF THE SENEGAL RIVER BASIN

Between 1987 and 1990, snail surveys were carried out in theSenegal River Basin at Lampsar, in a natural ‘marigot’,and at Richard Toll, in an irrigation canal. Lymnaea natalensis.Bulinus guernet^* and B.forskalii, were regularly found at boththe sites. Bulinus globosus/jousseaumei and B. umbilicatus wereonly present at Lampsar. A decrease of the snail density wasobserved during the rainy season at Lampsar. The recent appearanceof Biomphalaria pfeifferi at Richard Toll is discussed. Thedata show that adult B. pfeiffert are present in the main irrigationcanal all through the year. Consequently parasite transmissionwill be possible throughout the year as well. (Received 23 October 1991; accepted 20 May 1992)