Chromate tolerance in strains of Rhodosporidium toruloides modulated by thiosulfate and sulfur amino acids

Cr(VI) tolerance was studied in four strains of Rhodosporidium toruloides and compared with that of a fifth strain, DBVPG 6662, isolated from metallurgical wastes and known to be Cr(VI) resistant. Tolerance was studied in relation to different species of sulfur (sulfates, thiosulfates, methionine, cysteine) at different concentrations. Djenkolic acid, a poor source of sulfur and an activator of sulfate transport, was also considered. In synthetic medium all strains except the Cr(VI)-resistant one started to be inhibited by 10 g ml^– (0.2 mm) Cr(VI) as K₂Cr₂O₇. DBVPG 6662 was inhibited by 100 g ml^– (2.0 mm) Cr(VI). In Yeast Nitrogen Base without amino acids (minimal medium), supplemented with varying concentrations of chromate, all Cr(VI)-sensitive strains accumulated concentrations of total chromium (from 0.8 to 1.0 g mg^– cell dry wt) after 18 h of incubation at 28 °C. In minimal medium supplemented with 10 g ml^– Cr(VI), the addition of sulfate did not significantly improve the yeast growth. Cysteine at m levels increased tolerance up to 10 g ml^–, whereas methionine only reduced the Cr(VI) toxicity in the strain DBVPG 6739. Additions of djenkolic acid resulted in increased Cr(VI) sensitivity in all strains. The best inorganic sulfur species for conferring high tolerance was thiosulfate at concentrations up to 1 mm. In all cases increased Cr(VI) tolerance was due to a significantly reduced uptake in the oxyanion by the cells and not to the chemical reduction of Cr(VI) to Cr(III) by sulfur compounds.     

The Biogeochemical Cycle of Methane in the Coastal Zone and Littoral of the Kandalaksha Bay of the White Sea

Microbiological and biogeochemical investigations of the processes of methane production (MP) and methane oxidation (MO) in the coastal waters and littoral of the Kandalaksha Bay of the White Sea were carried out. The studies were conducted in the coastal zones and in the water areas of the Kandalaksha Preserve, Moscow State University White Sea Biological Station, and the Zoological Institute (RAS) biological station in August 1999, 2000, and 2001 and in March 2001. The rate of CO₂ assimilation in the shallow and littoral sediments was 35–27800 g C/(dm³ day) in summer and 32.8–88.9 g C/(dm³ day) in winter. The maximal rates of MP were observed in the littoral sediments in the zone of macrophyte decomposition, in local depressions, and in the estuary of a freshwater creek (up to 113 l/(dm³ day)). The maximal level of MO was observed in the shallow estuarine sediments (up to 2450 l/(dm³ day)). During the winter season, at the temperature of –0.5 to 0.5°C, the MP rate in the littoral sediments was 0.02–0.3 l/(dm³ day), while the MO rate was 0.06–0.7 l/(dm³ day). The isotopic data obtained indicate that the C_org of the mats and of the upper sediment layers is enriched with the heavy ¹³C isotope by 1–4 as compared to the C_org of the suspension. A striking difference was found between the levels of methane emission by the typical littoral microlandscapes. In fine sediments, the average emission was 675 l CH₄/(m² day); in stormy discharge stretch sediments, it was 1670 l CH₄/(m² day); and under stones and in silted pits, 1370 l CH₄/(m²day). The calculation, performed with consideration of the microlandscape areas with a high production, allowed the CH₄ production of 1 km² of the littoral to be estimated as 192–300 l CH₄/(km² day).     

Ecotoxicity of lead to the zebra musselDreissena Polymorpha,Pallas

The effect of lead on the filtration rate of the zebra musselDreissena polymorpha was investigated, together with the accumulation of Pb in the soft tissues of the mussels. The NOEC-filtration was 116 g.l^–1 (0,56 mol.l^–1) and the EC₅₀-filtration was 370 g.l^–1 (1.79 mol.l^–1). The NOEC-accumulation was the concentration found in the control water (1.4g.l^–1). These experiments show that the EC₅₀-filtration for Pb is similar to that for Cd, higher than that for Cu and lower than that for Zn. The water quality criteria for lead allow 25 g Pb.l^–1 in surface water. This will not cause short-term effects. Long-term effects may, however, occur, since an accumulation of Pb as low as 16 g.l^–1 was recorded in this study.     

Asymmetric diffusion into the postsynaptic neuron: An extremely efficient mechanism for removing excess GABA from synaptic clefts on the Deiters' neurone plasma membrane

Microdissected Deiters' neuron plasma membranes have been used for studying the passage of GABA through the membrane both in the inward and outward direction. Working with 0.2 mM GABA in the compartment simulating the outside of the neurone and with 2.0 mM GABA in the one simulating the inside we found a net transport of GABA towards the inside. This mechanism does not require a Na⁺ ion gradient across the membrane. The nature of the transport process involved was studied by determining the rate of [³H]-GABA inward passage as a function of GABA concentration (1 nM–800 M) on the outward side of the membrane. The results have shown that until 50 M a diffusion process (v=D₁×C, where D₁=3.1×10^–11 1/m²×sec) is the sole mechanism involved. Above 50 M a second diffusion process is activated v=D₂×(C–50×10^–6), where D₂=2.8×10^–11 1/m²×sec. Taking in account both inward and outward directed diffusion, one can calculate 16 M as the equilibrium concentration of GABA on the outward side of the membrane. From a kinetic point of view, these diffusion processes are able to reduce GABA concentration in a synaptic cleft from 3 mM to 20 M within 3 sec. These diffusion systems are discussed as extremely efficient in removing the excess of released GABA in the synaptic cleft.     

Effects of GNA and other mannose binding lectins on development and fecundity of the peach-potato aphid Myzus persicae

Three mannose-binding lectins were assayed in artificial diets for their toxic and growth-inhibitory effects on nymphal development of the peach-potato aphid Myzus persicae. The snowdrop (Galanthus nivalis) lectin GNA was the most toxic, with an induced nymphal mortality of 42% at 1500 g ml^–1 (30 M) and an IC₅₀ (50% growth inhibition) of 630 g ml^–1 (13 M). The daffodil (Narcissus pseudonarcissus) lectin NPA and a garlic (Allium sativum) lectin ASA induced no significant mortality in the range 10–1500 g ml^–1, but did result in growth inhibition of 59% (NPA) and 26% (ASA) at 1500 g ml^–1 (40 M for NPA, 63 M for ASA). All three lectins were responsible for a slight but significant growth stimulation when ingested at 10 g ml^–1, reaching +26%, +18% and +11% over the control values for the garlic lectin, the daffodil lectin and the snowdrop lectin, respectively. GNA, as well as the glucose/mannose binding lectin Concanavalin A, were also provided at sublethal doses throughout the life cycle of the aphids, and effects on adult performance were monitored. Adult survival was not significantly altered, but both lectins adversely affected total fecundity and the dynamics of reproduction, resulting in significant reduction in calculated r _ms (population intrinsic rate of natural increase) on lectin-containing diets. These effects are discussed in relation to the use of transgenic plants expressing these toxic lectins for potential control of aphid populations.     

Effect of Naloxone on the Activity of Cl–-Activated Mg2+-ATPase from Plasma Membrane Fraction of Bream Brain (Abramis brama L.) in the Presence of GABAa-ergic Substances

We studied the effect of naloxone—an antagonist of the opioid receptors—on sensitivity of Cl^–-activated Mg²⁺-ATPase from the plasma membrane fraction of bream brain (Abramis brama L.) to GABA_a-ergic substances. Preincubation of the plasma membranes with 1–100 M naloxone increased the basal Mg²⁺-ATPase activity and suppressed its activation by chloride ions. The same effects were observed in the presence of the agonists of GABA_a/benzodiazepine receptors: 0.1–100 M GABA, 1–500 M pentobarbital, and 0.1–100 M phenazepam. Naloxone (10 M) inhibited activation of the basal Mg²⁺-ATPase by the studied ligands and restored the enzyme sensitivity to Cl^–. However, the effect of naloxone was not observed in the presence of high concentrations of pentobarbital (500 M) and phenazepam (100 M). The obtained data show that naloxone modulates the activity of Cl^–-activated Mg²⁺-ATPase from the plasma membranes of bream brain and antagonizes the GABA_a receptor ligands.     

The effects of aflatoxin B1 on immature germinating maize (Zea mays) embryos

Immature maize (Zea mays L.) embryos were treated with aflatoxin B₁ concentrations, ranging from 0.1 g ml^–1 to 25 g ml^–1. Below 5 g ml^–1 aflatoxin B₁, root and shoot elongation was not significantly inhibited. Ultrastructurally, root tip cells showed little deterioration, except a possible diffused clearing in mitochondria and plastids. As the toxin concentration was increased above 5 gml^–1, shoot, and particularly root elongation, was progressively inhibited. Associated with this, there was an apparent decrease in the ribosome population. Furthermore, membranes, particularly the vacuolar membrane, became abnormal and vacuolar distension occurred. At 20 and 25 g ml^–1, these effects were exacerbated, and mitochondria and plastid structure was disrupted. At these concentrations, there was evidence of a disruption in lipid metabolism. The results are discussed in the context of known aflatoxin effects on cellular control mechanisms and ultrastructure in animal systems.     

Phenotypic expression of Vogesella indigofera upon exposure to hexavalent chromium,Cr6+

A eubacterium producing a blue pigment was isolated from a drinking water filter, and subsequently identified as Vogesella indigofera. This bacterium was further investigated for its morphological and biochemical characteristics after exposure to hexavalent chromium, Cr⁶⁺. The threshold Cr⁶⁺ concentration inhibiting the pigment production by V. indigofera was 200–300 g ml^–1 in liquid cultures of nutrient broth and 100–150 g ml^–1 on nutrient agar plates. The Cr⁶⁺ concentration preventing V. indigofera growth was 300–400 g ml^–1 in liquid cultures, but greater than 150 g ml^–1 on agar plates. Moreover, rugose colonies without the blue pigmentation were observed on agar plates amended with 150 (g Cr⁶⁺) ml^–1. The biochemical utilization profiles of the colonies without pigmentation did not differ from the original pigment-producing ones, indicating phenotypic plasticity of this bacterium. The difference of phenotypic expression of V. indigofera under various Cr⁶⁺ concentrations might have potential application as a pollution bioindicator for heavy metals.     

Tissue culture ofKarwinskia humboldtiana—a plant producing toxins with antitumoural effects

Isolated embryos ofKarwinskia humboldtiana were cultured in vitro. The growth of embryos and development to plantlets on woody plant medium supplemented with indole-3-acetic acid 6.10^-2 mol l^–1, gibberellic acid (GA₃) 3.10^-2 mol l^–1, and 6-benzylaminopurine (BA) 2 mol l^–1 was obtained. Multiplication of shoots and rooting of excised shoots has been achieved. Callus formation on modified Murashige-Skoog medium supplemented with 1-naphthaleneacetic acid 10 mol l^–1, GA₃ 14 mol l^–1, and kinetin 5 mol l^–1 on hypocotyls, or on root cultures on medium supplemented with 2.4-dichlorophenoxyacetic acid 10 mol l^–1 and BA 10 mol l^–1 was induced.Abbreviations BA 6-benzylaminopurine - 2,4-d 2,4-dichlorophenoxyacetic acid - GA₃ gibberellic acid - IAA indole-3-acetic acid - NAA 1-naphthaleneacetic acid - TEM transmission electron microscopy     

Pattern of cadmium accumulation and essential cations during growth of cadmium-tolerant fungi

The present study evaluates the growth response of two strains of filamentous fungi; a Fusarium sp. and Alternaria tenuis, grown on both solid and liquid Czapek Dox medium amended with different concentrations of CdCl₂. Colony extension and the mycelial dry weight of both fungi were significantly inhibited by high concentrations of cadmium. Extended lag phases and low growth rates resulted from cadmium administration. Cadmium drastically affected fungal morphogenesis by the production of stunted sterile thick mycelial filaments of the Fusarium sp. and chains of uncharacterized swellings instead of conidia in A. tenuis. Experiments showed that cadmium accumulation by the Fusarium sp. grown in liquid medium was a concentration dependent, and over the incubation time it displayed a plateau pattern. The cells grown on medium containing 0.25 mmol l^–1 CdCl₂ accumulated up to 89 ± 12 mol Cd (gm dw)^–1 after two days, falling to 29 ± 10 mol Cd (gm dw)^–1 after five days. At 0.5 mmol l^–1 CdCl₂ treatment the maximum cellular cadmium content was 132 ± 14 mol (gm dw)^–1, attained after 3 days, and decreased to 98 ± 9 mol (gm dw)^–1 at the end of the incubation time. There was a simultaneous marked drop in cadmium content and pH of the growth medium during the first few days. The presence of cadmium markedly altered the cellular essential cations; K⁺ and Mg²⁺ being decreased while Na⁺ increased during the growth period. Such findings resulted a reverse pattern of cellular Na⁺/K⁺ ratio for cells grown on cadmium-containing medium in respect to the control treatment. The results are discussed in relation to a further dimension of cadmium effects that might reflect its toxicity, as well as the implication of cadmium extrusion for tolerance during fungal growth.     

Nitrous oxide production by nitrogen-fixing,fast-growing Rhizobia

Rhizobium trifolii, R. leguminosarum, andR. hedysarum, grownex planta under anoxic conditions in a chemically defined medium, evolve N₂O from NO₃ ^–, NO₂ ^–, and (NH₄)₂NO₃. The amount of nitrous oxide formed after 96 hours is about 0.2M×mg^–1 cells d.w. Large availability of organic matter enhances the production of N₂O from nitrate by free-livingR. trifolii in peat/sand mixtures. Denitrification of the above species andR. meliloti was detected also in planta. Nitrous oxide production increases almost linearly from 10–45M×mg^–1 nodules d.w. when nitrogen-fixing plants are exposed to increasing concentrations of nitrate (1–12M).     

Influence of inorganic microelements on the production of camptothecin with suspension cultures of <Emphasis Type="Italic">Camptotheca acuminata</Emphasis>

The effects of eight microelements (I^–, BO₃ ^3–, MoO₄ ^2–, Co²⁺, Cu²⁺, Mn²⁺, Fe²⁺, Zn²⁺) on the biosynthesis of camptothecin and the growth of suspension cultures of Camptotheca acuminata were studied. The increase of I^– to 25 M l^–1, Cu²⁺ to 1 M l^–1, Co²⁺ to 2 M l^–1 and MoO₄ ^2– to 10 M l^–1 in Murashige and Skoog (MS) medium resulted in 1.66, 2.84, 2.53 and 2.04 times higher of camptothecin yield than that in standard MS medium respectively. Combined treatment of I^– (25 M l^–1), Cu²⁺ (1 M l^–1), Co²⁺ (2 M l^–1) and MoO₄ ^2– (10 M l^–1) lead to improve cell dry weight, camptothecin content, and camptothecin yield to 30.56 g l^–1, 0.0299%, and 9.15 mg l^–1, respectively, which were 20.2, 208.9 and 273.8% increment respectively when compared with those of control.     

Seasonal changes in inorganic and organic phosphorus in the soil of a riparian forest

This study addresses the temporal distribution of forms of phosphorus in the soil of a temporarily flooded riparian forest of the valley of the river Garonne (Southwest of France). A sequential extraction for forms of phosphorus of increasing chemical stability was used. During the study period (13 months), the forest was flooded a few days during March and May. In winter, resin-Pi concentration was high (26 g g^–1) in comparison to spring values (<9 g g^–1). NaHCO₃-Po, NaHCO₃-Pi or NaOH-Pi concentrations increased during winter (up to 74, 124 and 78 g g^–1 respectively) and decreased significantly during spring (32, 44 and 32 g g^–1 respectively). This pattern was attributed to simultaneous mineralization and plant uptake during the growing season and to the flood events (erosional processes and P-release). During summer and fall, resin-Pi concentration increased significantly (up to 26 g g^–1 in October). NaHCO₃-Po concentrations remained low during spring and summer (<33 g g^–1), and increased significantly in fall (>45 g g^–1 NaHCO₃-Pi or NaOH-Pi increased in late spring or summer (90 g g^–1 and 68 g g^–1 respectively). Increasing concentrations of the labile forms during late spring or summer were ascribed to the warm temperature and soil dryness that limited plant growth. HCl-Pi increased regularly after the floods (174 g g^–1 before the flood events to 254 g g^–1 after the floods). Residual P presented a similar pattern i.e. 214 g g^–1 and 279 g g^–1 respectively before and after the flood events. This pattern was attributed to a progressive incorporation of flood deposits to the soil.     

Diurnal variation in physical-chemical properties and primary production in the interconnected marine,mangrove and freshwater biotopes of Kakinada coast,Andhra Pradesh,India

Diurnal variation in hydrological variables and dissolved inorganic nutrients such as PO _inf4 ^sup3– -P, N O _inf2 ^sup– -N, NO _inf3 ^sup– -N and NH _inf4 ^sup+ -N were studied in three interconnected biotopes including freshwater, marine and mangrove brackish water of the Kakinada coastal zone, Andhra Pradesh. Samples were collected at intervals of 3 hours, for a period of 24 hours. In the marine environment salinity varied from 26 to 32 whereas in the mangrove waters it fluctuated from 12 to 20 and in both biotopes salinity showed bimodal type of oscillation. Dissolved oxygen content was high in the mangrove waters during day time but decreased rapidly during the night hours. In the marine environment PO_f4 ^p3–-P concentration varied from 0.345 to 1.195 g at l^–1, NO _inf3 ^sup– -N from 1.03 to 6.62 g at l^–1 and NO _inf2 ^sup– -N from 0.086 to 0.506 g at l^–1. The highest and the lowest concentrations of PO _inf4 ^sup3– -P, NO _inf3 ^sup– -N, NO _inf2 ^sup– -N recorded in the mangrove waters were 0.790 and 0.325 g at l^–1, 7.10 and 1.60 g at l^–1 and 0.278 and 0.060 g at l^–1, respectively. The concentration of PO _inf4 ^sup3– -P, NO _inf3 ^sup– -N and NO _inf2 ^sup– -N were high in the freshwater canal, the maximum and minimum values being 1.110 and 0.730 g at l^–1, 26.40 and 9.98 g at l^–1 and 0.520 and 0.252 g at l^–1 respectively. The concentration of ammonia was relatively high in the mangrove water. Gross and net primary production in the mangrove water was 4 times higher than in the marine biotope. There was no export of dissolved nutrients from the mangrove environment to the adjacent marine waters.     

Influence of nitrogen on the behaviour of nickel in wheat

A glasshouse experiment was conducted to study the effect of Ni on the growth and nutrients concentration in wheat (Triticum aestivum Cv. WH 291) in the presence and absence of applied N as urea. Responses to N application were observed up to 120 g N g^–1 soil. No response to Ni was observed in the dry matter yield of wheat tops (leaves + stem) in the absence of applied N while in the presence of applied N, significant yield increases were obtained at 12.5g Ni g^–1 soil. Nickel was not toxic to wheat up to 50g Ni g^–1 soil in the presence of 120g N g^–1 soil. Nitrogen and Ni concentration in wheat tops and roots increased with increasing levels of applied N and Ni, respectively. Applied Ni had an antagonistic effect on N concentration. Similarly, N reduced the Ni concentration in the wheat tissues. Positive growth responses to Ni were associated with 22 and 15g Ni g^–1 in wheat tops, in the presence of applied N at 60 and 120g N g^–1 soil, while Ni toxicity was associated with 63, 92.5 and 112.5g Ni g^–1 in wheat tops, in the absence and presence of applied N at 60 and 120g N g^–1 soil, respectively.     

Accumulation of cadmium by immobilizedZoogloea ramigera 115

Summary Zoogloea ramigera 115 was immobilized into beads of calcium-alginate and placed into batch air-bubbled column reactors. In the absence of any added nutrients the immobilized bacterium adsorbed Cd from solutions containing levels of 2 and 20 g ml^–1 per day, over a period of 21 and 20 days, respectively. Adsorption of Cd from solutions containing 20 g ml^–1 Cd was better than 90% for 16 days. Beads treated with Cd at 2 g ml^–1 never adsorbed less than 95% of the metal. Alginate adsorbed Cd as well, but inclusion of cells changed the effectiveness of adsorption. Of a 250 g ml^–1 Cd solution, alginate adsorbed 70.4% Cd in 60 min whereas alginate plus cells adsorbed 90.5% in the same time span. Temperature had no effect on adsorption by immobilized cells at levels of 2 and 10 g ml^–1 Cd. However at higher concentrations, binding was enhanced as temperature increased.Z. ramigera beads were stable during all treatments and for prolonged periods of time (21 days).     

Linear growth in microbial cultures limited by accumulated substrate

Summary The linear growth phase in cultures limited by intracellular (conservative) substrate is represented by a flat exponential curve. Within the range of experimental errors, the presented model fits well the data from both batch and continuous cultures ofEscherichia coli, whose growth is limited in that way.List of symbols D dilution rate, h^–1 - K_S saturation constant, g.L^–1 - S concentration of the limiting substrate, g.L^–1 - S_i concentration of the limiting substrate accumulated in the cells, g.g^–1 - S_o initial concentration of the limiting substrate, g.L^–1 - t time of cultivation, h - t₁ time of exhaustion of the limiting substrate from medium, h - t_o beginning of exponential phase, h - X biomass concentration, g.L^–1 - X₁ biomass concentration at the time of exhaustion of the limiting substrate from the medium, g.L^–1 - X_o biomass concn. at the beginning of exponential phase, g.L^–1 - biomass concn. at steady-state, g.L^–1 - Y growth yield coefficient (biomass/substrate) - specific growth rate, h^–1 - _m maximum specific growth rate, h^–1     

Procedures for the axenic isolation of conchocelis and monospores from the red seaweed Porphyra yezoensis

In order to maintain axenic seedstock cultures axenically of thecommercially important red seaweed, Porphyra yezoensis, aprocedure was developed for axenic isolation and culture of conchocelis andmonospores. For axenic isolation of the conchocelis, contaminated microalgaewere most effectively removed by filtering contaminated samples through a100-m mesh after sonication. Removal of bacteria and otheralgaewas accomplished using a mixture of 5 agents (0.02% chitosan, 100 gml^–1 GeO₂, 10 gml^–1 ampicillin, 40 gml^–1 kanamycin and 200 gml^–1 streptomycin). Axenic single colonies wereisolatedfrom a semi-solid medium prepared from 1% transfer gel. After collectingmonospores from the 40–50% density layer on a percoll-gradient, removalofbacteria and fungi from the monospores was accomplished using a mixture of 5antibiotics (3.5 g ml^–1 nystatin, 2 mgml^–1 ampicillin, 400 gml^–1 kanamycin, 50 gml^–1 neomycin and 800 gml^–1 streptomycin). Axenic single juvenile blades wereisolated from a semi-solid medium prepared from 0.5% transfer gel.     

Insecticidal activity of avidin and streptavidin against four species of pest Lepidoptera

Two biotin-binding proteins, avidin and streptavidin, were found to be insecticidal to the larvae of four species of Lepidoptera – light brown apple moth, Epiphyas postvittana (Walker) (fam. Tortricidae), green-headed leaf-roller, Planotortrix octo (Dugdale) (fam. Tortricidae), brown-headed leaf-roller, Ctenopseustis obliquana (Walker) (fam. Tortricidae) and potato tuber moth, Phthorimaea operculella (Zeller) (fam. Gelechiidae). Mortality occurred in all species, but there was a wide range in susceptibility. P. operculella larvae were the most susceptible with an LC₅₀ of respectively, 2.3 g ml^–1 for avidin and 1.4 g ml^–1 for streptavidin after 9 days. E. postvittana larvae had an LC₅₀ of 43.4 g ml^–1 for avidin after 21 days, and C. obliquana larvae of 45.7 g ml^–1 for avidin after 28 days. Although significant mortality occurred in P.octo at the highest doses of avidin, there was no sufficient dose-mortality response to calculate an LC₅₀ for this species. For all species mortality curves were steep over a close range of doses followed by a plateau where mortality did not increase significantly with dose and did not reach 100%. Mortality was significantly affected by the amount of biotin in the diet on which the parental generation had been reared. Where this was rich in biotin, significant mortality of the offspring was much lower: larval offspring of a colony of E. postvittana, reared for five generations on a biotin-free diet had an LC₅₀ of 5.1 g ml^–1 after 14 days compared with 76.7 g ml^–1 for larvae from a colony reared on general purpose diet. The implications for use of these proteins to confer insect resistance on transgenic plants are discussed.     

Isolation and characterization of a toxic intracellular protein from Vibrio parahaemolyticus

A toxic factor released from disrupted cells of Vibrio parahaemolyticus was partially purified by gel filtration after precipitation with (NH₄)₂SO₄ at 40% saturation. The factor, which was a thermostable protein of 63 kDa, lysed human erythrocytes at a concentration of 0.15 g ml^-1. Its LD₅₀ by intravenous injection into mice was 6.4 g. Fluid accumulated in suckling mice force-fed with the toxic material (1 to 25 g). Haemolytic activity, which occurred maximall at 37°C and pH 7.0 was enhanced by Ca²⁺, Cu²⁺ and Zn²⁺, each at 1 mm. Anti-toxic-factor serum agglutinated V. parahaemolyticus cells. The factor may play a role in the pathogenesis of V. parahaemolyticus infections and in the host's defence mechanisms against infection by the microorganism.