Autotrophic carbon sources for heterotrophic bacterioplankton in a floodplain lake of central Amazon

The relative contribution of autotrophic carbon sources (aquatic macrophytes, flooded forest, phytoplankton) for heterotrophic bacterioplankton was evaluated in a floodplain lake of the Central Amazon. Stable carbon isotopes (¹³C) were used as tracers. Values of ¹³C of different autotrophic sources were compared to those of dissolved organic carbon (DOC) and those of bacterially produced CO₂.The percentage of carbon derived from C₄ macrophytes for bacterially produced CO₂ was the highest, on average 89%. The average ¹³C value of CO₂ from bacterial respiration was –18.5 ± 3.3. Considering a fractionation of CO₂ of 3 by bacterial respiration, ¹³C value was –15.5, near C₄ macrophyte ¹³C value (–13.1).The average value of total DOC ¹³C was –26.8 ± 2.4. The percentage of C₄ macrophytes carbon for total DOC was on average 17%. Considering that bacteria consume mainly carbon from macrophytes, the dominance of C₃ plants for total DOC probably reflects a faster consumption of the former source, rather than a major contribution of the latter source.Heterotrophic bacterioplankton in the floodplain may be an important link in the aquatic food web, transferring the carbon from C₄ macrophytes to the consumers.     

Seasonal and diel relationships between the isotopic compositions of dissolved and particulate organic matter in freshwater ecosystems

A study of the isotopic composition of organic matter was conducted in a freshwater marsh over seasonal and diel time scales to determine the sources of dissolved organic matter (DOM) and the processes leading to its formation. Bulk C and N isotopic compositions of the bacterial fraction (0.2–0.7 m) and particulate organic matter (POM; 0.7–10 m) were compared on a seasonal basis with the change in ¹³C of DOM. The bulk isotopic data support the idea that DOM was, in part, derived from the breakdown of larger organic matter fractions. The bacterial fraction and POM were compositionally similar throughout the year, based on a comparison of the ¹³C of individual amino acids in each fraction. Annual variation in the ¹³C of amino acids in DOM was greater relative to the variation in larger fractions indicating that microbial reworking was an important factor determining the proteinaceous component of DOM. The ¹³C enrichment of serine and leucine in each organic matter fraction suggested microbial reworking was an important factor determining organic matter composition during the most productive times of year. Changes in the bulk ¹³C of DOM were more significant over daily, relative to seasonal, time scales where values ranged by 6 and followed changes in chlorophyll a concentrations. Although bulk ¹³C values for POM ranged only from –29 to –28 during the same diel period, the ¹³C of alanine in POM ranged from –30 to –22. Alanine is directly synthesized from pyruvate and is therefore a good metabolic indicator. The ¹³C of individual amino acids in DOM revealed the diel change in the importance of autotrophic versus heterotrophic activity in influencing DOM composition. Diel changes in the ¹³C of phenylalanine, synthesized by common pathways in phytoplankton and bacteria, were similar in both DOM and POM. The diel change in ¹³C of isoleucine and valine, synthesized through different pathways in phytoplankton and bacteria, were distinctly different in DOM versus POM. This disparity indicated a decoupling of the POM and DOM pools, which suggests a greater source of bacterial-derived organic matter at night. The results of this study demonstrate the use of the isotopic composition of individual amino acids in determining the importance of microbial reworking and autotrophic versus heterotrophic contributions to DOM over both diel and seasonal time scales.     

The Use of the [13C]/[12C] Ratio for the Assay of the Microbial Oxidation of Hydrocarbons

The study deals with a comparative analysis of the relative abundances of the carbon isotopes ¹²C and ¹³C in the metabolites and biomass of the Burkholderia sp. BS3702 and Pseudomonas putida BS202-p strains capable of utilizing aliphatic (n-hexadecane) and aromatic (naphthalene) hydrocarbons as sources of carbon and energy. The isotope compositions of the carbon dioxide, biomass, and exometabolites produced during the growth of Burkholderia sp. BS3702 on n-hexadecane (¹³C = –44.6 ± 0.2) were characterized by the values of ¹³C_{CO 2} = –50.2 ± 0.4, ¹³C_biom = –46.6 ± 0.4, and ¹³C_exo = –41.5 ± 0.4, respectively. The isotope compositions of the carbon dioxide, biomass, and exometabolites produced during the growth of the same bacterial strain on naphthalene (¹³C = –21 ± 0.4) were characterized by the isotope effects ¹³C_{CO 2} = –24.1 ± 0.4, ¹³C_biom = –19.2 ± 0.4, and ¹³C_exo = –19.1 ± 0.4, respectively. The possibility of using the isotope composition of metabolic carbon dioxide for the rapid monitoring of the microbial degradation of petroleum hydrocarbons in the environment is discussed.     

Isoform-Specific Membrane Translocation of Protein Kinase C After Ischemic Preconditioning

Mild cerebral anoxic/ischemic/stress insults promote tolerance and thereby protect the brain from subsequent lethal anoxic/ischemic insults. We examined whether specific activation of PKC , , , or isoforms is associated with ischemic preconditioning (IPC) in rat brain. IPC was produced by a 2-minute global cerebral ischemia. Membrane and cytosolic fractions of the hippocampi were immunoblotted using specific antibodies for PKC, , , and . PKC showed a significant translocation to the membrane fraction from 30 min to 4 h and PKC at 4 h following IPC. In contrast, the membrane/cytosol ratio of PKC showed a tendency to decrease at 30 min and 8 h, and the membrane/cytosol ratio of PKC was significantly decreased from 30 min to 24 h following IPC. These findings indicate PKC isoform-specific membrane translocations in the hippocampus after brief global brain ischemia and suggest that activation of PKC and PKC may be associated with IPC-induced tolerance in the rat hippocampus.     

Regulation of cell apoptosis by protein kinase c delta

The isoforms of the PKC family are activated in response to mitogenic stimuli, to inflammatory stimuli, and to stress and play important roles in a variety of cellular functions including apoptosis. PKC a member of the novel PKC subfamily, is actively involved in cell apoptosis in a stimulus and tissue specific manner; it both regulates the expression and function of apoptotic related proteins and is itself a target for caspases. Activation of PKC by various apoptotic stimuli results in the translocation of PKC to distinct cellular compartments such as mitochondria, golgi and nucleus, and the differential translocation contributes to its different effects. In addition, phosphorylation of PKC on distinct tyrosine residues and its association with specific apoptotic related proteins such as c-Abl, DNA-PK, p73 and lamin B are pivotal to its function in cell apoptosis. Recent findings on these aspects of the PKC cascades are the major focus of this review.     

Stimulation of μ- and δ-Opiate Receptors and Tolerance of Isolated Heart to Oxidative Stress: the Role of NO-Synthase

Preliminary intravenous injection of -opiate receptor (OR) agonists DSLET (0.1 mg/kg) or DTLET (0.1 mg/kg) increased tolerance of isolated perfused myocardium to damage by oxidative stress simulated in vivo with FeSO₄ + ascorbic acid. This manifested itself by a decreased level of creatine phosphokinase (CPK) in the perfusate flowing out of the heart during the oxidative exposure. The preliminary systemic injection of -agonists DAMGO (0.1 mg/kg) or DALDA (0.1 mg/kg) failed to affect the release of CPK from the myocardium. The cardioprotective effect of the -agonist DSLET was completely abolished by preliminary intravenous injection of the -OR antagonist ICI 174,864 (2.5 mg/kg). The intravenous injection of the NO-synthase inhibitor L-NAME (50 mg/kg) also completely abolished the cardioprotective effect of -OR stimulation. The preliminary injection of DSLET but not of DAMGO prevented an increase in the level of diene conjugates and a decrease in the activity of superoxide dismutase (SOD) in the isolated myocardium tissue. Thus, the in vivo stimulation of -OR increased the tolerance of the heart to oxidative stress through activation of NO-synthase and SOD.     

Trophic interactions of Antarctic seals as determined by stable isotope signatures

The diets and trophic interactions among Weddell, crabeater, Ross, and leopard seals in the eastern Ross Sea, Antarctica, were investigated by the use of stable isotope techniques during the 1999–2000 summer seasons. The ¹³C and ¹⁵N values in seal serum clearly distinguished the three Antarctic pack-ice seal species at different trophic positions (Weddell>Ross>crabeater). These patterns appeared to reflect a close linkage to their known foraging ecology and diving behaviors, and agreed well with their presumed dietary diversity. The more enriched ¹³C and ¹⁵N values in male Weddell seals than those in females suggested differences in foraging preferences between them. Significant differences in ¹⁵N were also found among different age groups of Weddell seals. A strong correlation between the C:N ratios and serum cholesterol was probably due to extremely high cholesterol levels in phocids. Comparisons of isotope data with harbor seals revealed distinct differences between Antarctic phocids and the northern seal species.     

Affinity Profiles of Novel δ-Receptor Selective Benzofuran Derivatives of Non-Peptide Opioids

Highly selective heterocyclic opioid ligands with potent -antagonist activity have been developed on the basis of the message-address concept. Using this strategy, benzofuran derivatives corresponding to the non-selective opioid antagonist, naloxone, and to the -opioid receptor selective agonists, oxymorphone and oxycodone, were synthesized. In vitro opioid receptor binding profiles and agonist/antagonist character of these compounds were determined in rat brain membrane preparations with highly selective radioligands. All three benzofuran derivatives displayed high affinities for the -opioid receptor, much less potency toward the -binding site, and were the least effective at the -site. The results indicated that the addition of the bezofuran moiety to these fused ring opioids confers -receptor selectivity. The Na⁺ indices suggested a partial agonist character for oxymorphone- and oxycodone-benzofuran, and an antagonist character for naloxone-benzofuran. These compounds were capable of irreversible inhibition of opioid binding sites in a dosedependent.     

Spatial and temporal distribution of carbon isotopes in soil organic matter at the Dinghushan Biosphere Reserve, South China

The spatial and temporal distribution of carbon isotopes (¹³C, ¹⁴C) in soil organic matter (SOM) were studied based on SOM content, SOM ¹⁴C and SOM ¹³C of thinly layered soil samples for six soil profiles with different elevations at the Dinghushan Biosphere Reserve (DHSBR), South China. The results indicate that variations of SOM ¹³C with depth of the soil profiles at different elevations are controlled by soil development, and correlate well with SOM composition in terms of SOM compartments with different turnover rates, and SOM turnover processes at the DHSBR. The effect of carbon isotope fractionation was obvious during transformation of organic matter (OM) from plant debris to SOM in topsoil and SOM turnover processes after the topsoil was buried, which resulted in great increments of OM ¹³C, respectively. Increments of SOM ¹³C of topsoil from ¹³C of plant debris were controlled by SOM turnover rates. Both topsoil SOM ¹³C and plant debris ¹³C increase with elevation, indicating regular changes in vegetation species and composition with elevation, which is consistent with the vertical distribution of vegetation at the DHSBR. The six soil profiles at different elevations had similar characteristics in variations of SOM ¹³C with depth, alterations of SOM contents with depth and that SOM ¹⁴C apparent ages increasing with depth, respectively. These are presumably attributed to the regular distribution of different SOM compartments with depth because of their regular turnover during soil development. Depth with the maximal SOM ¹³C value is different in mechanism and magnitude with penetrating depth of ¹⁴C produced by nuclear explosion into atmosphere from 1952 to 1962, and both indicate controls of topography and vegetation on the distribution of SOM carbon isotopes with depth. Elevation exerts indirect controls on the spatial and temporal distribution of SOM carbon isotopes of the studied mountainous soil profiles at the DHSBR. This study shows that mountainous soil profiles at different elevations and with distinctive aboveground vegetation are presumably ideal sites for studies on soil carbon dynamics in different climatic-vegetation zones.     

The Opioidergic System in the Combined Regulation of Pain and Immunity

Opioidergic mechanisms are involved in responses to nociceptive and antigenic stimuli at all levels and stages (from peripheral nociceptors to the cerebral cortex and from the precursors of immunocompetent cells to mature effector cells). In most experimental and clinical studies, opioid-mediated analgesia proved to be accompanied by immunosuppression. Opioid receptors of , , and types are involved in the mechanisms of combined regulation of pain and immunity, with and receptors suppressing the immune response and receptors enhancing it. By modifying the chemical structure of opioid ligands, it is possible to preserve the analgesic effect and avoid the development of immunosuppression. The opioidergic mechanisms are coupled with nonopioid peptidergic and nonpeptide systems of pain and immunity regulation.     

The structure and genetic control of a new class of disulphide-linked proteins in wheat endosperm

Summary Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of unreduced total protein extracts from the endosperm of hexaploid wheat revealed three high molecular weight protein bands (triplet bands) in a zone of heavy background streaking. Electrophoretic examination of 135 hexaploid cultivars showed at least five different patterns of these triplet bands. Nine durum wheat cultivars showed a single band only. Analysis of nullisomic-tetrasomic and ditelocentric lines of Chinese Spring wheat revealed that the slowest moving band (Tri-1) of the triplet was controlled by gene(s) on chromosome arm 1DS and the fastest moving band (Tri-3) by 1AS. The band with intermediate mobility (Tri-2) was found to be a hybrid aggregate of the subunits controlled by 1DS and 1AS. Using a non-reducing/reducing form of 2-dimensional (2-D) electrophoresis, these triplet bands were shown to be heterotetramers of four subunits designated D (M.W. 58,000), (22,000), A (52,000) and (23,000) where Tri-1=DD, Tri-2 = DA and Tri-3 = AA. With very low concentrations of 2-mercaptoethanol (ME), the tetramers dissociated into dimeric subunit pairs (D, A), the monomers being observed with higher concentrations of ME. The structure of these subunit pairs resembles that of the subunit pairs in the globulin storage proteins of oats and some legumes. The 2-D method employed in this study was useful also for separating low molecular weight (LMW) subunits of glutenin from the monomeric gliadins which have similar electrophoretic mobility in 1-D separation. It was shown that at least four of these LMW glutenin subunits are controlled by genes on 1DS and 1AS and at least one subunit is controlled by gene(s) on 1BS. This electrophoretic separation method has proven useful in understanding the aggregation behaviour of the seed proteins of wheat.     

Stimulation of crystallin synthesis in embryonic brain cells in culture

Summary Expression of -crystallin, a lens-specific protein, in 6-day-old chick embryonic brain cells was examined in situ and in vitro. The presence of minute amounts of -crystallin and its mRNA (-mRNA) in brain cells in situ was demonstrated by immunoblot and Northern blot analysis. In spreading cultures of the brain cells, -crystallin and -mRNA showed a significant increase from their in situ level. Immunohistological staining (peroxidase antiperoxidase) with monospecific anti-serum against -crystallin revealed that -producers were both epithelial cells and dendritic cells. Neither lentoidogenesis nor -crystallin expression was observed. Stimulation of -crystallin synthesis in cultured brain cells differed when compared with transdifferentiating cultures of neural retina cells. In the latter, -crystallin synthesis occurred concomitantly with differentiation of morphologically distinct lens cells containing -crystallin.     

Comparisons of δ13C values in leaves of aquatic macrophytes from different habitats in Britain and Finland; some implications for photosynthetic processes in aquatic plants

Summary The ¹³C values of submerged aquatic plants from contrasting but relatively defined habitats, and the ¹³C values of emergent, floating and submerged leaves of dimorphic aquatic plants, were measured. In many instances the ¹³C values of dissolved inorganic carbon in the water were also measured. Plant ¹³C values in the vicinity of-40 to-50 were found in rapidly flowing spring waters with carbonate ¹³C values of-16 to-21, consistent with the notion that species such as Fontinalis antipyretica almost exclusively assimilate free CO₂ via RuP₂ carboxylase. Plant ¹³C values in the vicinity of-10 to-15 in sluggish water with carbonate ¹³C values of about-5 were observed, consistent with the notion that boundary layer diffusion and/or HCO₃ ^- uptake may determine the ¹³C value of submerged aquatic plants in these circumstances. Comparisons of ¹³C values of the same or related species growing in waters of similar carbonate ¹³C value but different flow rates confirmed this view; more negative ¹³C values were frequently associated with plants in fast moving water. In Britain, but not in Finland, the ¹³C values of submerged leaves of dimorphic plants were almost invariably more negative than in aerial leaves. The ¹³C value of carbonate from chalk streams and in acid springs indicate substantial inputs of respiratory CO₂, as opposed to atmospheric carbon. The contributions of these variations in ¹³C of the carbon source, and of isotope fractionation in diffusion, to the ¹³C value of submerged parts of dimorphic plants is discussed.     

Characterization and expression analysis of CD3ɛ and CD3γ/δ in fugu, Takifugu rubripes

CD3 is an essential component of the CD3-TCR complex. In this report, we describe the cloning, characterization, and expression analysis of the CD3 and CD3/ chain genes from fugu, Takifugu rubripes. Two distinct CD3 homologue cDNAs, designated as CD3-1 and CD3-2, and a CD3/ homologue cDNA were isolated from the fugu thymus. The deduced amino acid sequences of these cDNAs exhibit conserved essential CD3 chain motifs and overall structures. RT-PCR analysis demonstrated that the CD3 and CD3/ genes were expressed in lymphoid organs (e.g. thymus, head kidney, trunk kidney and spleen), mucosal tissues (gill, skin, and intestine), and peripheral blood leucocytes (PBL). The CD3 and TCR genes were expressed only in the surface IgM^– population, which were separated from PBL using an anti-fugu IgM monoclonal antibody. In addition, in situ hybridization confirmed that CD3-expressing cells were distributed randomly in the head kidney, trunk kidney, and spleen, but in the thymus were restricted to the lymphoid outer zone and epithelioid inner zone only. Collectively, these results suggest that CD3 molecules are useful markers for the identification of T cells in teleost fish. The present study thus provides a critical step in identifying T cells in this model organism.Nucleotide sequence data reported in this paper are available in the DBJ/EMBL/GenBank databases and have been assigned the accession numbers AB166798 (CD3-1), AB166799 (CD3-2), and AB166800 (CD3/).     

δ-Toxin,unlike melittin,has only hemolytic activity and no antimicrobial activity: Rationalization of this specific biological activity

The antimicrobial activity of a synthetic peptide corresponding to -hemolysin had been examined. The peptide didnot exhibit antimicrobial activity against gram negative and gram positive micro-organisms unlike other hemolytic peptides like melittin. This lack of antibacterial activity arises due to the inability of -hemolysin to perturb the negatively charged bacterial cell surface and permeabilize the bacterial plasma membrane. However, the red blood cell surface has a structure considerably different from bacteria, and does not act as a barrier to molecules reaching the lipid membrane. Hence -toxin can lyse erythrocytes. Thus, the specificity in biological activity has been rationalized in terms of differences, in the interaction of the toxin with the bacterial and red blood cell surfaces.     

Fugu immunoglobulin D: a highly unusual gene with unprecedented duplications in its constant region

We have isolated and characterized the cDNA that encodes IgD of fugu (Takifugu rubripes). Though the splicing of 1 with the 1 domain was similar to those reported for teleost IgDs, highly unusual and unprecedented domain duplications were found in the constant region of the fugu IgD. The structure of the fugu IgD is like VDJ-1-(1-2-3-4-5-6)₂-7-m1-m2. Genomic sequence analysis of the fugu IgD gene supported the results of cDNA sequencing that the first six domains in the constant region are duplicated. Such a novel duplication pattern has not been reported in any other vertebrates. However, IgD secretory domains could not be identified in this study. The deduced amino acid sequence of the fugu IgD constant region showed high identity (35–55%) to the sequences of previously reported teleost IgDs. Gene expression analyses based on RT-PCR demonstrated that the IgD gene is preferentially expressed in presumptive lymphoid tissues; moreover, in situ hybridization showed that IgD-positive cells are distributed throughout the spleen and head kidney. The expression pattern is similar to that of IgM, corroborating the hypothesis that IgD plays an important role in the humoral immune system of this species.The nucleotide sequence data reported in this paper will appear in the DDBJ/EMBL/GenBank nucleotide sequence databases with the accession numbers AB159481 and AB159482.     

Using stable hydrogen and oxygen isotope measurements of feathers to infer geographical origins of migrating European birds

Successful application of stable-hydrogen isotope measurements (D_f) of feathers to track origins of migratory birds and other wildlife requires a fundamental understanding of the correlation between D_f and deuterium patterns in rainfall (D_p) over continental scales. A strong correlation between D_p and D_f has been confirmed for birds and insects in North America, but not yet for other continents. Here, we compare D_f data from resident European birds to new D_p basemaps for Europe. Three maps, representing growing-season and mean annual D_p estimates from an elevation-explicit, detrended interpolation model and growing-season D_p estimates from simple Kriging, all indicate that strong isotope gradients occur across Europe with a general depletion occurring in a northeast direction. The feather data, representing 141 individuals of 25 avian species from 38 sites, ranged from –131 to –38. Regression analysis showed that strong correlations existed between both mean annual and growing-season D_p estimated by detrended interpolation and D_f of non-aquatic and non-corvid birds (r²=0.66 and 0.65, respectively). We also examined mean annual and growing-season ¹⁸O_p vs. ¹⁸O_f for our samples. Both oxygen regressions were similar (r²=0.56 and 0.57, respectively) but poorer than for deuterium. Our study reveals that D measurements of feathers from migratory birds in Europe may be used to track their origin and movements, and so provide a powerful investigative tool for avian migration research in Europe.     

Solution conformations of proline rings in proteins studied by NMR spectroscopy

Summary Three different conformations of proline rings in a protein in solution, Up, Down and Twist, have been distinguished, and stereospecific assignments of the pyrrolidine -, - and -hydrogens have been made on the basis of ¹H-¹H vicinal coupling constant patterns and intraresidue NOEs. For all three conformations, interhydrogen distances in the pairs -³, ³-³, ²-², ²-², and ³-³ (2.3 Å) are shorter than those in the pairs -², ²-³, ³-², ²-³, and ³-² (2.7–3.0 Å), resulting in stronger NOESY cross peaks. For the Up conformation, the ³-² and ²-³ spin-spin coupling constants are small (<3 Hz), and weak cross peaks are obtained in a short-mixing-time (10 ms) TOCSY spectrum; all other vicinal coupling constants are in the range 5–12 Hz, and result in medium to strong TOCSY cross peaks. For the Down form, the -², ²-³, and ³-² vicinal coupling constants are small, leading to weak TOCSY cross peaks; all other couplings again are in the range 5–12 Hz, and result in medium to strong TOCSY cross peaks. In the case of a Twist conformation, dynamically averaged coupling constants are anticipated. The procedure has been applied to bovine pancreatic trypsin inhibitor and Cucurbita maxima trypsin inhibitor-V, and ring conformations of all prolines in the two proteins have been determined.     

Using stable isotopes to investigate migratory connectivity of the globally threatened aquatic warbler Acrocephalus paludicola

Understanding the links between breeding and wintering areas of migratory species has important ecological and conservation implications. Recently, stable isotope technology has been used to further our understanding. Stable isotope ratios vary geographically with a range of biogeochemical factors and isotope profiles in organisms reflect those in their food and environment. For inert tissues like feathers, isotope profiles reflect the environment in which they were formed. Following large-scale habitat destruction, the globally threatened aquatic warbler Acrocephalus paludicola has a fragmented breeding population across central Europe, largely in Belarus, Poland and Ukraine. The species sub-Saharan African wintering grounds have not yet been discovered, and this significantly hampers conservation efforts. Aquatic warblers grow their flight feathers on their wintering grounds, and we analysed stable isotope ratios (¹⁵N, ¹³C, D) in rectrices of adults from six main breeding sites (subpopulations) across Europe to determine whether different breeding subpopulations formed a single mixed population on the wintering grounds. ¹⁵N varies considerably with dietary trophic level and environmental factors, and D with the D in rainfall; neither varied between aquatic warbler subpopulations. Uniform feather ¹⁵N signatures suggest no major variation in dietary trophic level during feather formation. High variance and inter-annual differences in mean D values hinder interpretation of these data. Significant differences in mean ¹³C ratios existed between subpopulations. We discuss possible interpretations of this result, and consider differences in moulting latitude of different subpopulations to be the most parsimonious. ¹³C in plants and animals decreases with latitude, along a steep gradient in sub-Saharan Africa. Birds from the most north-westerly breeding subpopulation (Karsibor, Poland) had significantly lower variance in ¹³C and ¹⁵N than birds from all other sites, suggesting either that birds from Karsibor are less geographically dispersed during moult, or moult in an area with less isotopic heterogeneity. Mean ¹³C signatures from winter-grown feathers of different subpopulations were positively correlated with the latitude and longitude of breeding sites, suggesting a strong relationship between European breeding and African winter moulting latitudes. The use of stable isotopes provides novel insights into migratory connectivity and migration patterns in this little-known threatened species.     

New organisms—novel genetics

This report describes a method for quantifying -endotoxin contents in spray-dried preparations ofBacillus thuringiensis strain GC-91. -Endotoxin is solubilized in the pro-toxin form and separated from other soluble compounds by ion exchange chromatography. The method does not discriminate between the different -endotoxins produced by strain GC-91. It is suitable for quality control, since -endotoxin concentrations found in different preparations correlate inversely with the LC 50 in bioassays on artificial diet against freshly hatched larvae ofSpodoptera littoralis. A typical batch of spray-dried material contains 1.22%±0.08% -endotoxin. The method is most accurate with preparations containing 0.5 to 2.0% toxin, for which triplicate estimations give standard errors close to±0.2%.