Development of a Monoclonal Antibody for the Detection and Quantification of Predation on Slugs within the Arion hortensis agg. (Mollusca: Pulmonata)

The Arion hortensis agg. (Mollusca: Pulmonata) are major slug pests of agriculture and horticulture throughout much of the temperate World. A monoclonal antibody was created that could specifically identify and quantify the remains of these slugs within the carabid predator Pterostichus melanarius Illiger, a polyphagous species found in arable crops in Europe and North America. The antibody was shown to be specific for the two species of pest slug within this species aggregate, Arion distinctus Mabille and A. hortensis Férussac, and did not cross-react with any other invertebrates tested, including other slugs in the same genus. It proved to be highly sensitive in an enzyme-linked immunosorbent assay. The mean detection period for the remains of A. distinctus in predator foregut samples was 3.28 days at 16°C. However, A. distinctus remains could be identified in male carabids for 30% longer than in females, emphasizing the importance of separate analyses. The value of the antigen half-life as a measure of the detection period was questioned because analysis of covariance showed that the regression slopes for antigen decay were not significantly different between the two sexes, giving very similar half-lives (approx 18.5 h for females and 19 h for males). Calculated quantities of soluble slug proteins in predator foreguts declined more rapidly over time than concentrations, allowing separate assessment of the importance of these two variables on detection periods. It was concluded that a valuable new antibody had been created and characterized, which can be used for the analysis of predation in the field.     

THE INFLUENCE OF AN ALTERNATIVE FOOD ON THE EFFECTIVENESS OF PROPRIETARY MOLLUSCICIDAL PELLETS AGAINST TWO SPECIES OF SLUGS

The effectiveness of three types of molluscicidal pellets againstDeroceras reticulatum (Müller) and Arion hortensis (Férussac)was assessed in the presence and absence of potato leaf discs.Pellets containing methi-ocarb were significantly more effectiveagainst both of the slug species than either of the metaldehydeformulations, in the absence of leaf discs. When leaf discswere present, methiocarb was no more effective against D. reticulatumthan were the metaldehyde formulations. Leaf discs did not interferewith the effectiveness of methiocarb against A. hortensis. Theresults are discussed in relation to past work and more recentfield studies. (Received 24 March 1986;     

THE IMPORTANCE OF MUCUS AS A DEFENCE AGAINST CARABID BEETLES BY THE SLUGS ARION FASCIATUS AND DEROCERAS RETICULATUM

The effect of mucus exudation on the survival of Arion fasciatusand Deroceras reticulatum was studied by exposing mechanicallystimulated and control slugs to Carabidae beetles for 24 hours.A light stimulation, lasting three minutes, exhausted the copiousflow of mucus for one day. A generalist, Pterostichus niger,significantly preferred stressed D. reticulatum over controlones. Similarly, P. niger exclusively ate stressed individualsof A. fasciatus. Two beetles which specialize in gastropods,Cychrus caraboides and Carabus violaceus, consumed an equalnumber of stressed and control D. reticulatum and A. fasciatus.The susceptibility of the slug species to predation was different:for each beetle species, the proportion of available D. reticulatumpreyed upon was significantly higher than that of A. fasciatus.The differences in the behaviour of A. fasciatus and D. reticulatumin defending themselves against attacks is described. The mainreason why specialist beetles are able to hunt slugs successfullyis their ability to prevent the slugs from exuding large amountsof mucus. This may succeed by different means: C. violaceustargeted their killing strokes against the posterior part ofthe mantle while C. caraboides hit the head of the slug. Inboth case the strokes seemed to paralyze the slugs. (Received 31 March 1993; accepted 1 October 1993)     

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)     

Daily variation in the number of slugs under refuge traps

Daily variation in the number of slugs under and around 25 cm × 25 cm pad‐traps was recorded in plots of crop cultures. Grey field slugs Deroceras reticulatum and garden slugs (Arion distinctus and Arion hortensis) left traps at dusk and came back at the end of the night. The time of day at which the number of trapped slugs was maximal did not occur in the middle of the day, when slugs were inactive, but in the early morning and in the evening, a few hours before dusk. This was due to a rise in temperature under the traps in the middle of the day. However, the grey field slugs stayed under traps throughout the morning, when temperatures under the traps reached values more favourable for rest.     

LABORATORY STUDIES ON DAMAGE TO POTATO TUBERS BY SLUGS

The ability of eight species of slugs to damage intact potatotubers c .v. Maris Piper was investigated. Four species, previouslyimplicated as pests of field crops, caused primary damage totubers. Interactions between three of the species Milax budapestensis(Hazay), Arion hortensis (Fèrussac) and Deroceras reticulatum(Müller) were studied using potato tubers of five cultivars.Single species groups caused most damage to tubers and interactionbetween species reduced damage irrespective of cultivar. Theresults are discussed in relation to past work. ^*Present address: 59 Valley Park Drive, Clanfield, Hants. PO80PL. (Received 19 August 1986;     

Phylogeny of slug species of the genus Arion: evidence of monophyly of Iberian endemics and of the existence of relict species in Pyrenean refuges

The Iberian Peninsula contains the majority of the Paleartic land slug species of the genus Arion, which exhibits diverse taxonomic problems. The present study investigated Arion taxonomy on the basis of analyses of the mitochondrial ND1 gene and nuclear internal transcribed spacer 1 (ITS1) sequences. The Iberian endemic species were monophyletically clustered in two divergent sister clades. The topotype specimens of Arion lusitanicus and the closely related species Arion nobrei and Arion fuligineus, as well as Arion hispanicus and Arion flagellus, were grouped into an ‘Atlantic’ clade, whereas Arion baeticus, Arion gilvus, Arion anguloi, Arion wiktori and Arion paularensis were included in a ‘Continental–Mediterranean’ clade. Calibration of mutation rate in the ND1 gene suggested that the divergence of these two clades occurred around the Pliocene–Pleistocene boundary, with subsequent speciation events during the Pleistocene. A group of ancestral and divergent endemic species with distribution centred in the Pyrenean mountain range (Arion molinae, Arion lizarrusti, Arion antrhacius and Arion iratii) arose in the Pliocene and survived through the Pleistocene in geographically confined small populations. Arion lusitanicus showed up to be polyphyletic: specimens, sampled outside the geographic range of the topotype in the north‐western Iberian Peninsula, were included in a non‐monophyletic clade together with the widely distributed species Arion ater and Arion rufus. The divergent species with a wide European distribution (Arion subfuscus, Arion hortensis, Arion fagophilus and Arion intermedius) were located in basal positions in all topologies. The evolutionary history of these slug species (highly sensitive to climatic factors, with capacity for both outcrossing and selfing, and with low dispersal ability) appears to have been moulded by Pliocene–Pleistocene climate events and by the rugged topography of southern Europe, giving rise to repeated cycles of population isolation during periods of glaciation alternating with interglacial expansions limited by geographic barriers.     

SLUG DAMAGE AND NUMBERS OF SLUGS IN OILSEED RAPE BORDERING ON GRASS STRIPS

In 1995, slug damage and numbers of slugs were estimated intwo grass strips and adjacent rape fields. Investigations beganas soon as rape seedlings emerged and lasted for five weeks.Slug damage to rape plants 1 m from the grass strips was significantlyhigher than at greater distances from the strips. Derocerasreticulatum was the most abundant slug species recorded in bothgrass strips and adjacent rape fields. Arion lusitanicus andArion fasciatus were much less abundant than D. reticulatum.In one field, D. reticulatum declined steadily with increasingdistance from the grass strips and therefore appeared to havecaused the majority of severe damage to rape plants close tothe strips. This finding was surprising because until now severeslug damage in oilseed rape beside semi-natural habitats hasbeen observed only where A. lusitanicus was abundant. (Received 12 November 1997; accepted 26 January 1998)     

DOES TANDONIA BUDAPESTENIS (MOLLUSCA: PULMONATA) CONTAIN TOXINS? EVIDENCE FROM FEEDING TRIALS WITH THE SLUG PREDATOR PTEROSTICHUS MELANARIUS (COLEOPTERA: CARABIDAE)

Slugs are known to be killed and consumed by a range of invertebrateand vertebrate predators in the field. Carabid beetles (Coleoptera:Carabidae) in particular are major natural enemies of slugs,and have been shown to be capable of controlling certain speciesin a crop environment. This paper reports experiments to investigatethe effects, on the carabid Pterostichus melanarius, of feedingon the milacid slug Tandonia budapestensis. The slugs provedto be toxic, 50% of the beetles dying within two days. Mortalityof beetles fed on two other species of slug, Deroceras reticulatumand Arion distinctus, was not significantly different from thatfor beetles fed upon a control prey (Calliphora vomitoria).These results represent the only known case of a European slugproving to be toxic to potential predators, and is one of avery small number of reported instances of possible toxicityamongst terrestrial gastropods. The possibility that the orangeline down the dorsal keel of T. budapestensis may act as warningcoloration to birds is discussed. (Received 4 December 1996; accepted 5 March 1997)     

FOOD-INDUCED BODY PIGMENTATION QUESTIONS THE TAXONOMIC VALUE OF COLOUR IN THE SELF-FERTILIZING SLUG CARINARION SPP

Body pigmentation is a popular taxonomic marker in slugs to discriminateclosely related species. However, the genetic background ofbody pigmentation is known only for a few species, while inmany others body pigmentation is influenced by age, food and/orclimate. In this study, we investigated the effects of differentfood items on body pigmentation expression in two selfing pulmonategastropods, Arion (Carinarion) silvaticus and Arion (Carinarion) fasciatus.Both species mainly differ in the distribution of yellow-orangegranules on the body, which in A. fasciatus are concentratedin lateral bands, and in A. silvaticus are evenly scattered.Animals were raised individually under the same conditions, whilethey laid eggs as a consequence of selfing. This F₁ generation wasafterwards divided into two groups, which were fed with different fooditems. A diet of carrot, lettuce or paper had no effect on the distributionof the yellow-orange granules in A. silvaticus, but provokeda loss of the yellow-orange lateral bands in A. fasciatusso that externally these F₁ specimens became similar to A. silvaticus.In both species, a diet of nettle resulted in a strong yellow-orangepigmentation, which often formed yellow-orange lateral bands.These results indicate that food can probably influence the'species-specific' body pigmentation in Carinarion, and thusquestion the reliability of colour traits to distinguish A.silvaticus and A. fasciatus. (Received 5 May 2000; accepted 29 September 2000)     

A HISTOLOGICAL SURVEY OF GONADAL DEVELOPMENT IN ARION ATER L. (MOLLUSCA, PULMONATA)

A method of breeding the slug Arion ater L. in the laboratorywas established. Measurements were made at monthly intervalsof body weight, weight and volume of the hermaphrodite gland,diameter and number of acini, oocytes and other cell types insidethe acini in field and laboratory-reared animals. Male and femalegametes grow simultaneously in the hermaphrodite gland of fieldanimals during the year. High humidity and low temperature causedinhibition of spermatogenesis and increase in the number offemale gametes in laboratory-reared animals. The average rateof increase in diameter of the oocytes of field samples was10 µm per month and in laboratory samples 9 µm permonth. ^*Present address: Department of Biology, School of Sciences,Ferdowsi University, Mashhad, Iran. (Received 30 September 1977;     

VARIATIONS IN INFESTATIONS OF SIPHONARIA CAPENSIS AND S. CONCINNA(PULMONATA) BY LARVAL TREMATODES IN SOUTHERN AFRICA

The prevalence of digenean trematode parasites in Siphonariacapensis (Pulmonata) was examined in populations from 24 sites,and in S. concinna from 10 sites along the coast of southernAfrica from 1989 to 1991. Sporocysts containing cercariae werefound in the digestive gland of both species. To date, two typesof cercaria have been isolated from S. capensis and one typefrom S. concinna, these types possibly representing three distinctspecies of trematode. In approximately 30% of parasitized S.capensis and 10% of parasitized S. concinna the digestive glandwas completely destroyed. In both species of Siphonaria therewas considerable temporal and spatial variation in parasiteprevalence. At some locations up to 40% of S. capensis wereinfected whereas at others, no parasites were found. Prevalenceof trema-todes in S. concinna rarely exceeded 12%. At severalsites the prevalence of infestation within the limpets increasedwith increasing host size. (Received 8 June 1992; accepted 6 July 1992)     

The influence of weather upon the activity of slugs

Summary The activity of slugs was measured by a catch per unit effort sampling system based upon night searching, and was related to the microclimatological conditions in the habitat by regression analysis. The activity of Arion hortensis Fér., Arion subfuscus (Drap.), Arion lusitanicus Mab., Milax budapestensis (Hazay), and of all slugs irrespective of species, was found to be best related to temperature and vapour pressure deficit. The relevance of these results to the application of effective methods for the control of slugs is discussed.     

Immunocytochemical Localisation of Glycosaminoglycan-Like Activity in the Mucus and Associated Tissues of Terrestrial Slug Species (Gastropoda: Pulmonata)

Indirect immunofluorescence assays were conducted on cryotome sections of four terrestrial slug species from three distinct phylogenetic groups, Arion ater (L.), Arion hortensis (Férussac), Tandonia (Milax) budapestensis (Hazay), and Deroceras reticulatum (Müller) using monoclonal antibodies for two glycosaminoglycans (GAGs), heparan sulphate, and chondroitin sulphate. Specific staining for a heparan sulphate-like component was demonstrated in the foot and tail regions of each species and was particularly intense in A. ater and A. hortensis, notably in the epidermis and associated mucus-like material, and in mucus-like material from the pedal gland region of the latter species. Subsequent studies with A. ater confirmed the presence of heparan-sulphate–like activity in the caudal gland duct region. No evidence of specific staining for chondroitin sulphate-like activity was found in any of the slug species. This work suggests that a specific GAG, or a group of closely related GAGs, is a common component of mucus in a range of slug species and of different types of mucus, including trail (pedal) mucus and the more viscous mucus produced by the caudal gland.     

Assimilation efficiencies of slugs on different food materials

Summary The relative assimilation efficiencies of two common British species of slug, Limax flavus (L.) and Arion hortensis (Fér.), were studied in the laboratory using a gravimetric method. Results are given for adult slugs feeding on carrot root and potato tuber at 5°, 10°, 15°, and 20° C, and for L. flavus juveniles of known age feeding on potato at 10° and 15° C. A few results are also provided for Agriolimax reticulatus (Müller) feeding on both potato and carrot at 15° C.High mean assimilation efficiencies of 76.94 and 76.87% were obtained for L. flavus feeding on carrot and potato respectively. The equivalent results for A. hortensis were 88.60 and 89.42%. For A. reticulatus at 15° C the figures were 76.1 and 71.9%. Ingestion rates and assimilation rates are given for the first two species at all temperatures. Assimilation efficiency was found to be independent of temperature for L. flavus but not for A. hortensis. Ingestion rate and assimilation rate were temperature dependent in all cases except for A. hortensis feeding on potato. A. hortensis was more efficient at consuming both carrot and potato per gram live weight. All species consumed more potato than carrot per gram live weight.Juvenile L. flavus were found to have higher assimilation efficiencies and ingestion rates than the adults per gram live weight.Linear regression equations were determined from the relationship between dry weight of food consumed, and dry weight of faeces produced from that food for L. flavus.It is concluded that food quality and age of slug may be the factors most strongly influencing assimilation efficiency in this group.     

ESTERASE VARIABILITY IN THE GASTROPODS LITTORINA SAXATILIS (OLIVI) AND L. ARCANA ELLIS

The esterase system has been studied using isoelectric focusingin two closely related rough periwinkles, Littorina saxatilisand L. arcana, in an investigation of possible differences betweenthe two species at this level of discrimination. Collectionsof both species were obtained from Yorkshire, north Cornwalland south Devon; L. saxatilis was also obtained from sites insouth Cornwall (in and near the estuary of the Tamar), Dorsetand Scotland, where L. arcana was not present. There is considerablevariation both within and between sites which tends to maskany overall differences between the two species. However, ingeneral there is a much greater degree of heterogeneity withinsamples of L. saxatilis than within samples of L: arcana whenboth species are found together; this is reflected in the ShannonWiener diversity index for bands. Also the number of esterasebands is greater for L. saxatilis than for L. arcana at eachof the five sites where both occurred. A dendrogram derivedfrom the percentage occurrence of each band present in eachsample produced a reasonable degree of separation of the twospecies (Received 8 January 1988; accepted 24 February 1988)     

GROWTH AND MORTALITY OF THE MUSSELS CHOROMYTILUS MERIDIONALIS (KRAUSS) AND AULACOMYA ATER(MOLINA) AS INDICATORS OF BIOTIC CONDITIONS

Two mussels, Choromytilus meridionalis (Krauss) and Aulacomyaater, (Molina) were used as indicators of biotic factors thatapparently result in radically different sublittoral communitiesat two closely situated islands on the South African west coast.Specimens of both species were attached to submerged plates,which were either unprotected or protected from predation bycages. C. meridionalis grew faster and tolerated silting andsmothering better than A. ater. At Malgas Island adults of C.meridionalis are completely absent and those of A. ater scarcedue to intense predation by rock-lobsters. When protected againstpredation, growth of both species was significantly faster andsurvivership higher at Malgas than at Marcus Island, where C.meridionalis is abundant but A. ater absent. When the musselswere unprotected at Malgas, they were eaten in a few hours.At Marcus Island where rock-lobsters are virtually absent, cagedmussels grow faster than unprotected individuals and survivedlonger. Drilling whelks (mostly Nucella squamosa (Lamarck))were major predators at Marcus. Relative intensity of rock-lobsterpredation and competition between the mussels appear to controlthe structure and diversity of the sublittoral communities atthese sites, rather than physical factors. Growth rate, shellthickness and tolerance of smothering by other species determinewhich species of mussel predominates. (Received 5 April 1988; accepted 11 July 1988)     

ORGAN CULTURE STUDIES ON CELL DIFFERENTIATION IN THE HERMAPHRODITE GLAND OF ARION ATER L. (MOLLUSCA, PULMONATA)

Organ culture experiments were set up to investigate cell differentiationin the hermaphrodite gland of Arion ater L. They were carriedout in the absence and presence of oestradiol, oestriol, testosteroneand follicle stimulating hormones. In the absence of hormonesexplants of young animals showed mitotic divisions in spermatogonia;newly-differentiated spermatogonia were also observed in explantsof the hermaphrodite gland of young and old animals. In thepresence of oestriol hormone new germinal cells differentiatedfrom the neck region of the hermaphrodite glands of mature animals.Other hormones did not affect the explants. (Received 31 August 1981;     

Molluscicidal and slug-repellent properties of anaerobically digested organic matter

Slug problems in arable crops and vegetables have increased drastically during the past few decades. Observations on slug damage to oilseed rape suggested that fresh, anaerobically digested organic material from a biogas production plant is molluscicidal. To find out whether digested matter can be used for the control of agricultural pest slugs, a series of exeriments were carried out. The laboratory experiments demonstrated strong mollusc repellent and molluscicidal effects of digested organic matter against the three most important pest slugs of Switzerland, Arion lusitanicus, A. distinctus and Deroceras reticulatum. The effects were restricted to fresh digested matter and were rapidly lost when the material was stored, and also after application in the field. In the field experiment, fresh digested matter greatly reduced slug damage to lettuce in comparison with the untreated plots. At present, the chemical nature of the molluscicidal compound(s) in digested matter is unknown, but environmental pollutants such as heavy metals can be ruled out. Current research aims at a new formulation which is easier to apply and has a longer-lasting molluscicidal or slug-repellent effect, and at optimising the dosage and number of applications.