Fecundity, longevity and establishment of Otiorhynchus sulcatus (Fabricius) and Otiorhynchus ovatus (Linnaeus) (Coleoptera: Curculionidae) from the Pacific North-west of the United States of America on selected host plants

1 The fecundity, longevity and establishment of Otiorhynchus sulcatus and Otiorhynchus ovatus from the Pacific North‐west U.S.A. was studied on five selected host plants: Picea abies‘Nidiformis’, Picea glauca‘Conica’, Taxus baccata, Rhododendron catawbiense‘Boursault’ and Fragaria×ananassa‘Totem’. 2 Teneral adults were used to study adult longevity and reproductive success. Leaves of these host plants were used for sustenance for 9 months. Larval establishment was studied by infesting potted host plants with eggs. 3 Fragaria×ananassa‘Totem’ produced the longest survival, shortest preoviposition time, the greatest number of eggs, and the highest fertility for adults of both species. Picea spp. were not good adult hosts for O. sulcatus. Taxus was a good adult host for O. sulcatus, but was a nonhost for adults and larvae of O. ovatus. 4 Adult hosts did not affect preoviposition time or egg viability with O. ovatus adults. With O. sulcatus, preoviposition time was greatly increased and egg viability was < 50% on Picea spp. 5 The best larval host was F.×ananassa‘Totem’ for O. sulcatus and P. glauca‘Conica’ for O. ovatus. Rhododendron was a poor larval host for both species. 6 When all of the studies on these two pests are considered, O. sulcatus appears to have varying host preferences from among its many geographical areas of occurrence whereas O. ovatus has a more universal host selection.     

Host influences on the pathogenicity of Heterorhabditis megidis

The infectivity of infective juveniles(IJs) of Heterorhabditis megidis (strain NLH-E87.3) produced on small, medium and large larvae ofGalleria mellonella, and on medium and largelarvae of Otiorhynchus sulcatus was tested underlaboratory conditions against G. mellonella andO. sulcatus larvae. Infective juvenilesoriginating from small G. mellonella exposed toan initial dose of one IJ were more infectious thanthose from small cadavers exposed to a dose of 30 IJs.Independent of the initial inoculum size, IJs fromsmall cadavers of G. mellonella were moreinfectious than those from medium and large cadavers.At a dose of one IJ per larva, IJs originating frommedium size O. sulcatus cadavers were moreinfective against G. mellonella than againstO. sulcatus larvae. Large G. mellonellalarvae were less susceptible to all IJ batches thanmedium and small sized larvae.     

Physiological and behavioural responses to seasonal changes in environmental temperature in the Australian spiny crayfish Euastacus sulcatus

The strategies used by ectotherms to minimise the detrimental effects of suboptimal thermal environments on physiological performance are often related to whether they inhabit a terrestrial or aquatic environment. Most terrestrial ectotherms use thermoregulatory strategies to maintain body temperature within an optimal range, while many aquatic ectotherms utilise thermal acclimation to maintain performance over varying seasonal temperatures. This study aimed to elucidate the relative contributions of acclimation and behavioural thermoregulation to maintaining whole-animal performance over varying seasonal temperatures in the semi-terrestrial Lamington spiny crayfish (Euastacus sulcatus). Crayfish activity and surface temperatures were determined by means of radio tracking and behavioural observations. Field studies demonstrated that E. sulcatus is exposed to stable daily temperatures, varying only between seasons from 10°C in late winter to over 20°C in summer. Also, terrestrial behaviour corresponded to a small portion of crayfish time (1.13%), much lower than predicted, indicating that E. sulcatus has limited opportunity for behavioural thermoregulation. We also tested the effect of acclimation to either 10 or 20°C on chela strength and stamina. We found acclimation had a more marked effect on chela stamina than maximum strength measures; however, overall acclimatory capacity was limited in E. sulcatus. Thus, we found that the semi-terrestrial crayfish E. sulcatus used neither thermoregulatory behaviours nor physiological strategies to deal with seasonal changes in environmental temperature.     

Use of a nematode, Heterorhabditis heliothidis, to control black vine weevil, Otiorhynchus sulcatus, in potted plants

Application of aqueous suspensions of infective juvenile Heterorhabditis heliothidis, isolate T327, to the soil resulted in up to 100% parasitisation of larvae of the black vine weevil, Otiorhynchus sulcatus, in potted yew, raspberries and grapes in nurseries, and over 87% parasitisation on potted cyclamens and strawberries. Pupae and newly emerged adults on grapevines were also parasitised. Another isolate, T310, produced 92.5 to 98.5% parasitism of O. sulcatus larvae on potted cyclamens in glasshouse, but was less effective on strawberries. Neoaplectana bibionis was found to be less effective than H. heliothidis T327 strain. The use of these nematodes provides an economical and effective method for controlling O. sulcatus on potted plants in glasshouses and nurseries.     

A review of the biology and control of the vine weevil, Otiorhynchus sulcatus (Coleoptera: Curculionidae)

At the turn of the century, damage by Otiorhynchus sulcatus was sporadic and limited to small areas. Increasing horticultural intensification and the adoption of husbandry techniques favourable to the weevil, such as the use of polythene mulches, increased its pest status. The development of the early inorganic pesticides reduced the number of serious outbreaks of this pest and weevil control was further improved by the development of the persistent organochlorine insecticides in the 1940's. The banning of a number of the more persistent insecticides over recent years has now left the horticultural industry in a very vulnerable position. O. sulcatus is now a pest on a range of horticultural crops throughout the temperate regions of the world. Infestations are most common in Europe (where it originated) and the USA, and nearly 150 plants species have been identified as potential hosts to O. sulcatus. Damage is most frequently caused by the root feeding larval stage. Populations as low as one larva plant can kill sensitive species such as Cyclamen. Severe damage by the leaf feeding adults is less common, although low levels of damage or contamination by adults may be unacceptable in certain situations. There is one generation a year. Oviposition by the flightless parthenogenetic females occurs over the summer months with oviposition rates of c. 500 and 1200 eggs adult^-1for outdoor and laboratory populations, respectively. O. sulcatus mainly overwinters as larvae, although significant numbers of adults may survive in areas where winter temperatures are not too severe. A number of natural enemies, such as hedgehogs, frogs and predatory beetles, help to maintain O. sulcatus populations at a low level in natural environments, but they are less successful in intensive horticultural systems where persistent chemicals have been heavily relied on to maintain the population below the economic threshold level. Increasing environmental concern is now forcing growers to consider new pest control strategies. Controlled release formulations of non-persistent products, such as fonofos and chlorpyrifos, have shown potential as control agents for O. sulcatus larvae. Biological control agents, such as insect parasitic nematodes, have been developed commercially and new microbial control agents are in the process of development. Most of the new control products are directed towards control of O. sulcatus larvae. Adult vine weevils are nocturnal and a much more difficult target for the new control agents. It is likely that an integrated approach to pest control will be required to maintain O. sulcatus populations below their economic threshold level.     

Clarification of the Rhipicephalus sanguineus group (Acari,Ixodoidea, Ixodidae). I. R. sulcatus Neumann, 1908 and R. turanicus Pomerantsev, 1936

For many years the biosystematic status of several species in the Rhipicephalus sanguineus group has been confused, with the result that they have often been misidentified. Over the years some 18 of these species have been synonymised with R. sanguineus (Latreille, 1806) itself. More recently four new species in the group have been described: R. guilhoni Morel & Vassiliades, 1963; R. moucheti Morel, 1964; R. bergeoni Morel & Balis, 1976; and R. camicasi Morel, Mouchet & Rodhain, 1976.The literature on this group is critically reviewed and the main systematic problems analysed. R. sulcatus Neumann, 1908 and R. turanicus Pomerantsev, 1936 are two species that were confused both morphologically and ecologically. This has resulted in erroneous conclusions regarding their host relationships and distributions. Integrated biological, morphological and ecological studies on these two species have been conducted. Cross-breeding experiments have proved that both are distinct taxonomic entities. Interbreedings between African and Cypriot strains of R. turanicus demonstrated marked heterosis.A scanning electron microscope was used to determine the main diagnostic morphological differences between the larvae, nymphae and adults of R. sulcatus and R. turanicus, and the morphological similarities between African and Cypriot strains of R. turanicus. Rhipicephalus sulcatus occurs widely in the Afrotropical region in wetter ecological habitats and most frequently parasitises hares, dogs and jackals. R. turanicus occurs more-or-less throughout the Afrotropical region in a wide range of climatic biotopes, as well as in parts of southern Europe, Arabia and Asia, and is most abundant in the late rainy/early dry seasons. It occurs on a wide range of domesticated and wildlife hosts, including ground-feeding birds.     

Nitrous oxide emissions from multiple combined applications of fertiliser and cattle slurry to grassland

Identifying forage species that are productive in saline environments is an important research priority in many areas of the world affected by salinity. The salt and waterlogging tolerances of 19 species of Melilotus were evaluated in a series of glasshouse experiments. Measurements taken on each species included: dry matter (DM) production, root growth and development, shoot ion (Na⁺, K⁺ and Cl⁻) concentrations, root porosity, and in vitro estimates of nutritive value. Research on several species was restricted because of their potential as weed risks. Of the remaining species, M. siculus (syn. M. messanensis), an annual species, showed high relative salt and waterlogging tolerances, good DM production under non-stressed and stressed (saline and hypoxic) conditions, a high level of root porosity under stagnant conditions, low tissue ion (Na⁺, Cl⁻) concentrations, and a reasonable dry matter digestibility content (range 66–69%) under highly saline conditions. M. sulcatus ssp. segetalis and M. indicus were also identified as species with good DM production and tolerance to salinity and waterlogging stresses. Further weed risk assessments and field trials on these species are required before they can be promoted for use as pasture forages on saline areas.     

Two new species of <Emphasis Type="Italic">Parspina</Emphasis> Pearse, 1920 (Digenea: Cryptogonimidae) from freshwater fishes (Gymnotiformes) of the Paraná River basin in Argentina

Two new species of the cryptogonimid genus Parspina Pearse, 1920 are described from gymnotiform fishes of the Paraná River basin, P. carapo n. sp. from the banded knifefish Gymnotus carapo Linnaeus and P. virescens n. sp. from the glass knifefish Eigenmannia virescens (Valenciennes). Parspina carapo differs from P. virescens in the number of oral spines (32–39 vs 30–33) and their length (28–47 vs 16–28 μm), the distribution of tegumental spines and their anchorage, the types of sensory papillae on the body surface (three vs two types), the extent of body length posterior to the caeca (5 vs 13% of the total body length), the dimensions of the pars prostatica (52 × 34 vs 24 × 10 μm), and in the absence of a gonotyl (vs presence). Both P. carapo and P. virescens differ from P. bagre Pearse, 1920 and P. argentinensis (Szidat, 1954) in the number of oral spines (20–21 and 21–28 in the latter pair) and their length (28–32 and 35–60 μm), and in total body length. Additionally, the two new species differ from P. argentinensis in the arrangement of the vitelline follicles (one continuous band vs two groups on each side of the body) and in having a smaller pars prostatica (149 × 49 μm in the latter). Parspina carapo is the fifth intestinal helminth found in G. carapo, and P. virescens is the first found in E. virescens.     

A new species of monorchiid digenean from marine fishes in the Southwestern Atlantic Ocean off Patagonia

Proctotrema bartolii n. sp. (Digenea: Monorchiidae) is described based on naturally and experimental obtained adults from the marine fishes Odontesthes smitti (Lahille), O. nigricans (Richardson) (Atherinopsidae) and Eleginops maclovinus (Cuvier) (Eleginopidae) off Patagonia, Argentina, in the Southwestern Atlantic Ocean. Its generic identification is based on the presence of a unipartite terminal organ with the metraterm uniting with its distal region, an unarmed genital atrium, a single testis, a vitellarium follicular lateral to the ovary and ventral sucker, and uterine coils occupying most of hindbody. The new species differs from P. bacilliovatum Odhner, 1911, P. amphitruncatum Fischthal & Thomas, 1969 and P. guptai Ahmad & Dhar, 1987 in having a smaller body (305–650 vs 1,600–3,080, 1,500–1,800 and 2,150–2,670 μm, respectively), a round vs funnel-shaped oral sucker, a smooth vs lobed ovary, a saccular rather than tubular excretory vesicle, the number of vitelline follicles (12–16 vs 8–9, 9 and 6–8, respectively), and wider eggs (25–31 × 15–20 vs 28–37 × 9–12, 24–28 × 7–10, and 24–30 × 8–10 μm, respectively). Moreover, the new species differs from P. bacilliovatum and P. amphitruncatum in having a saccular rather than a coiled seminal vesicle, and from P. bacilliovatum and P. guptai in having its tegument completely vs partly spined. Proctotrema Odhner, 1911 is considered to be restricted to these four species. This is the first report of a species of this genus from South American waters.     

Analysis of beta-tubulin cDNAs from taxol-resistant Pestalotiopsis microspora and taxol-sensitive Pythium ultimum and comparison of the taxol-binding properties of their products

The anti-cancer drug taxol binds to β-tubulin in assembled microtubules and causes cell cycle arrest in animal cells; in contrast, in fungi, the effect of taxol varies. For instance, the taxol-producer Pestalotiopsis microspora Ne32, an ascomycete, is resistant to taxol (IC₅₀ greater than 11.7 μM), whereas Pythium ultimum, an oomycete, is sensitive to taxol (IC₅₀ 0.1 μM). In order to understand the differential fungal response to taxol, we isolated cDNAs encoding β-tubulin from both P. microspora and P. ultimum. The deduced amino acid sequence of β-tubulin from P. microspora is very similar to those from other Ascomycetes, many of which are resistant to taxol. The sequence of β-tubulin from P. ultimum is very similar to those from Oomycetes and non-fungal organisms, many of which are sensitive to taxol. To examine the interaction between taxol and fungal microtubules, binding studies were performed with fungal cells, using [³H]taxol. The labeled taxol was found to bind specifically to P. ultimum, but not to P. microspora. In addition, the amount of [³H]taxol specifically bound to P. ultimum was reduced by the microtubule-depolymerizing drug thiabendazole, in a dose-dependent manner. These results suggest efficient binding of taxol to microtubules in P. ultimum, but not in P. microspora, and are consistent with the differential taxol sensitivity of these two organisms. Finally a comparison of previously characterized taxol binding sites in various β-tubulin sequences showed that β-tubulins of taxol-sensitive organisms, including P. ultimum, contain Thr219, but β-tubulins of resistant organisms, including P. microspora, contain Asn or Gln at this position, suggesting an important role for residue 219 in the interaction between taxol and β-tubulin. Received: 16 March 1999 / Accepted: 21 August 1999     

Responses of Acilius sulcatus (Coleoptera: Dytiscidae) to chemical cues from perch (Perca fluviatilis)

In this study we tested the hypothesis that the presence of chemical stimuli from a hungry predator would initiate anti-predator responses, while stimuli from a satiated predator would not. We used chemical stimuli released from starved perch (Perca fluviatilis) and from satiated perch (predator). As prey we used adult Acilius sulcatus (Coleoptera: Dytiscidae). The reaction of the beetles to different predator conditions was tested during daytime. We also tested the reaction to starved perch during the night. A. sulcatus activity decreased when it was exposed to stimuli released from starved perch during daytime when visibility was poor, due to the presence of artificial vegetation. There was, however, no reaction to satiated perch under the same experimental conditions. These results indicate that A. sulcatus can discriminate between chemical cues from hungry and satiated fish predators. When visibility was good and the concentration of chemical cues was constant, the beetles did not react to starved perch in the daytime, but their activity decreased at night in response to stimuli released from starved perch. Visual as well as chemical cues seem to be important for detecting a potential predator. When visibility is good, beetles seem to rely on visual stimuli, while in darkness they seem to use chemical stimuli to detect the presence of predators. Received: 4 October 1996 / Accepted: 7 February 1997     

Comparative pollen morphology of Turkish species of <Emphasis Type="Italic">Petrorhagia</Emphasis> (Caryophyllaceae) and its systematic implications

Pollen morphology of 10 taxa from Turkey, four of them endemic, belonging to the genus Petrorhagia (Caryophyllaceae), P. alpina subsp. alpina, P. alpina subsp. olympica, P. cretica, P. dubia, P. hispidula, P. lycica, P. pamphylica, P. peroninii, P. prolifera and P. saxifraga, has been investigated using light (LM) and scanning electron (SEM) microscopy. Pollen morphology differences among these taxa have been determined. The pollen type of investigated taxa is polyporate (12–22-porate), sphaeroidal, the exine exhibits a tectate structure and a microechinate ornamentation. Petrorhagia alpina subsp. alpina has the smallest pollen grain diameter (18.57 μm) and P. dubia the largest one (37.80 μm). The number of pores ranges from 12 (in P. dubia and P. saxifraga) to 22 (in P. alpina subsp. alpina and P. alpina subsp. olympica and P. hispidula) with a minimum pore diameter of 2.37 μm in P. alpina subsp. alpina and a maximum pore diameter of 4.23 μm in P. peroninii. The exine thickness ranges from 1.73 μm in P. saxifraga to 3.78 μm in P. pamphylica. In this study, the systematic implications are discussed in the light of palynological results.     

Molecular phylogeny and systematics of Neotropical toucanets in the genus Aulacorhynchus (Aves,Ramphastidae)

Bonaccorso, E., Guayasamin, J. M., Peterson, A. T. & Navarro‐Sigüenza, A. G. (2011). Molecular phylogeny and systematics of Neotropical toucanets in the genus Aulacorhynchus (Aves, Ramphastidae). —Zoologica Scripta, 40, 336–349. We studied the phylogenetic relationships in the genus Aulacorhynchus, an assemblage of Neotropical toucanets distributed from Mexico south to Bolivia. Based on mitochondrial and nuclear DNA characters, we obtained a robust hypothesis of relationships for all recognized species, including good representation of distinct geographic populations. Our results support the monophyly of the genus Aulacorhynchus, but contradict previous taxonomic arrangements. The genus is made up of three major clades: the Aulacorhynchus prasinus complex, Aulacorhynchus huallagae + Aulacorhynchus coeruleicinctis, and Aulacorhynchus haematopygus + Aulacorhynchus sulcatus + Aulacorhynchus derbianus. Andean populations of A. derbianus are more closely related to A. sulcatus than to Pantepuian populations of A. derbianus, rendering A. derbianus paraphyletic. Based on the molecular phylogeny, and information on geographic distributions and morphological and behavioural characters, we review the specific status of these taxa and propose a new taxonomic arrangement within Aulacorhynchus.     

A revision of the <Emphasis Type="Italic">Paraulopus oblongus</Emphasis> group (Aulopiformes: Paraulopidae) with description of a new species

 Paraulopus brevirostris, P. filamentosus, P. japonicus, P. legandi, P. maculatus, and P. oblongus are redescribed. All species are included in the Paraulopus oblongus group, characterized by having no supraocular ridge, 2.5–3.5 scales above the lateral line, and small adult body size (70–150 mm SL). In addition, P. atripes, from the Indian Ocean, is described as a new species of the P. oblongus group, being defined by the following combination of characters: small antrorse dentary process on chin; eye directed laterally; pelvic fin black in males; caudal fin white; 32–34 gill rakers; 45–46 pored lateral-line scales; and 3.5 scales below lateral line. Paraulopus albimaculatus is regarded as a junior synonym of P. brevirostris, based on examination of type specimens. A key to species in the P. oblongus group is included. Received: September 2, 2002 / Revised: January 31, 2003 / Accepted: February 17, 2003     

Behavioural response of Heterorhabditis megidis towards plant roots and insect larvae

The behavioural response of infective juveniles (IJs) of Heterorhabditis megidis (strain NLH-E87.3) to cues from roots of strawberry (Fragaria x ananassa Duch.), thuja (Thuja occidentalis L.) and to larvae of the black vine weevil, Otiorhynchus sulcatus, was studied. Choice assays were conducted in an Y-tube olfactometer filled with moist sand. Infective juveniles were activated by the presence of intact roots of both strawberry and thuja plants. Some nematodes aggregated in the compartments with roots but most moved away from the roots to the opposite side. Given a choice, IJs showed a preference for strawberry roots above O. sulcatus larvae. No difference in preference was observed between thuja roots and O. sulcatus larvae. The combination of strawberry roots with vine weevil larvae was preferred above roots alone. In the assays with thuja roots and larvae versus thuja roots alone, however, IJs were stimulated to move but showed preference for the opposite compartment away from the arms with roots and larvae. Nematodes responded differently to mechanically damaged roots as opposed to roots damaged by vine weevil larvae. In assays with damaged thuja roots, IJs were most attracted by the roots damaged by larvae, whereas in the strawberry assays IJs showed a clear preference for the mechanically damaged roots. When challenged with a choice between strawberry and thuja roots, IJs moved preferentially to strawberry than to thuja roots. A preference for the combination of strawberry roots plus larvae over the thuja roots plus larvae was also observed.     

A new species of freshwater threadfin, <Emphasis Type="Italic">Polynemus aquilonaris</Emphasis>, from Indochina,and redescription of <Emphasis Type="Italic">Polynemus dubius</Emphasis> Bleeker, 1853 (Perciformes: Polynemidae)

 The taxonomic status of two nominal species of Polynemus, viz. P. dubius Bleeker, 1853 and P. longipectoralis Weber and de Beaufort, 1922, is revised. Although regarded as separate taxa up to the present time, examination of the holotype of P. longipectoralis revealed its close similarity to the type series of P. dubius, in the synonymy of which the former is now included. Polynemus dubius is redescribed as a valid species and a lectotype of the species is designated. In addition, a new species, P. aquilonaris, previously identified as P. dubius or P. longipectoralis, is described from Indochina on the basis of 28 specimens. Polynemus aquilonaris differs from P. dubius in having higher counts of pored lateral-line scales [80–86 (mode 81) vs. 69–79 (78) in the latter] and scale rows below the lateral line [14–17 (mode 14, rarely 13 or 17) vs. 13 (rarely 12)], and lower counts of gill rakers [25–29 (mode 27) vs. 29–33 (30), respectively]. The former is known from Indochina (Chao Phraya and Mekong River systems including Lake Tonle Sap), whereas the latter is currently known from the Malay Peninsula, Sumatra, and Kalimantan. Received: March 29, 2002 / Revised: February 2, 2003 / Accepted: February 10, 2003     

Lagomorphs from the Miocene of Sandelzhausen (southern Germany)

Three genera of lagomorphs, Prolagus, Lagopsis, and “Amphilagus,” were identified during a revision of the lagomorph material from Sandelzhausen (MN5, Early/Middle Miocene boundary, southern Germany). Evidence of two morphological and dimensional classes were observed at some tooth positions in Prolagus (some p3 show an unmistakable P. oeningensis morphology, others closely resemble P. crusafonti), but not at other tooth positions (e.g., M1–2). Insufficient data from Sandelzhausen precludes identification of two different species of Prolagus from this locality, and to define the characteristics of the possible P. crusafonti-like species. Thus, all Prolagus specimens have been classified as P. aff. oeningensis. The genus Lagopsis is represented by L. cf. penai, whose presence is compatible with a MN5 age. The relative abundance of Lagopsis to Prolagus may indicate relatively cool and wet palaeoclimatic conditions. The largest primitive lagomorph species from continental Europe is present at Sandelzhausen. Morphological and dimensional comparisons with other European primitive lagomorphs exclude any affinity with the genera Eurolagus and Titanomys and with the species included in “Amphilagus ulmensis”. Some common features with “Amphilagus antiquus” were observed, although they are not sufficient for the attribution to this taxon. Until there is a general revision of European primitive lagomorphs, the Sandelzhausen giant lagomorph is classified as “Amphilagus” sp. Its origins, whether from evolution within Europe or migration from Asia, remain unknown.      

Cardiac responses to salinity variations in two differently zoned Mediterranean limpets

Cardiac activity of two Mediterranean limpets was tested at different salinities. Patella caerulea inhabits the lower midlittoral where it is exposed to variations in salinity, while P. aspera experiences more stable salinity conditions in the infralittoral fringe. When exposed to moderate hypo- and hypersalinity (23 g l⁻¹ and 43 g l⁻¹) for 24 min, P. caerulea showed no significant variation in heart rate with respect to the control salinity (33 g l⁻¹), while P. aspera exhibited a significant increase in heart rate in both conditions. This suggests a rise in metabolic rate due to activation of behavioural responses or physiological regulation. When exposed to extremely low salinity (3 g l⁻¹) for 24 min, heart contractions ceased in most specimens of P. caerulea. A smaller number of specimens also displayed cessation of heart beat when exposed to extremely high salinity (63 g l⁻¹). The heart beat resumed quickly in all specimens when they were returned to control salinity conditions. In contrast, cardiac activity was not interrupted in any of the P. aspera specimens at the 3 g l⁻¹ and 63 g l⁻¹ salinity levels, but strong bradycardia was evident. Contractile activity of the heart ceased in all specimens of P. caerulea and P. aspera when they were exposed to prolonged hypo-osmotic stress (3 g l⁻¹ for 24 h). This acardia was largely reversible in P. caerulea, but most specimens of P. aspera did not recover from the treatment. Accepted: 3 July 1999