LEAD INHIBITION OF CHLOROPHYCEAN MICROALGAE1

The effects of lead on the growth of 9 microalgae representing 4 chlorophycean genera were investigated in axenic laboratory cultures. Variations in sensitivity to lead existed among the algae tested with availability of phosphate often limiting growth. Under conditions in which phosphate was growth limiting at pH 6.2, Chlorella pyrenoidosa Chick, C. ellipsoidea Gerneck, C. vulgaris Beij. Scenedesmus sp. (IUCC 1591).Sc. Obtusiusculus Chodat and Selenastrum capricornutum Printz first showed toxicity at 0.5 mg·l^?1 pb. Two planktonic Scenedesmus spp. And an Ankistrodesmus sp. Were more tolerant, developing toxicity at 1 mg· l^?1 Pb. The relationship of phosphate supply to Pb toxicity was investigated with S. capricornutum and Sc. Obtusiusculus. Al pH 8.0, Selenastrum developed toxicity at 3 mg·l^?1 pb either with or without phosphate. When phosphate supply was growth-limiting, 6 times as much pb (i.e., 3 vs. 0.5 mg l^?1 was required to demonstrate toxicity with Selenastrum at pH 8.0 as opposed to pH 6.2 Sc. Obtusiusculus was inhibited at 0.5 mg· l^?1 pb in phosphate-limiting medium and developed toxicity at 1 mg·l^?1 in media containing phosphate. Control of colonial morphology with Sc. Obtusiusculus was inhibited. Phosphate availability as well as pH would seem to be important factors determining lead toxicity with some algae.     

Intermittent lead‐induced stress on antioxidant enzyme activity and subcellular distribution of Pb in Pogonatherum crinitum seedlings

• Pogonatherum crinitum is a promising lead (Pb) hyperaccumulator due to its high Pb tolerance and accumulation ability. However, the mechanisms that support Pb accumulation and tolerance in P. crinitum are not yet clearly understood.
• An indoor hydroponic experiment was conducted by cultivating P. crinitum seedlings exposed to intermittent Pb stress for 60 days, divided into four stages (T1, T2, T3 and T4), with a 15‐day duration per stage. The following concentrations of Pb were used: 0, 500, 0, 500 mg·l^?1 and 0, 1000, 0, 1000 mg·l^?1). Antioxidant enzyme activity, Pb concentration and subcellular distribution of Pb were measured at each of the above stages.
• The results showed that superoxide dismutase (SOD) activity in shoots, and SOD, peroxidase (POD) and malondialdehyde (MDA) activity in shoots and roots significantly increased from T1 (no Pb stress) to T2 (Pb stress) in both 500 mg·l^?1 and 1000 mg·l^?1 treatments; however, no significant difference was noted between stages T3 (no Pb stress) and T4 (Pb stress). There was no obvious effect of Pb stress on catalase (CAT) activity in shoots and roots among different stages. The Pb concentration in shoots was up to 5090.90 mg·kg^?1 and 7573.57 mg·kg^?1, and the bioconcentration factor (BFC) was 10.18 and 7.57 for the 500 mg·l^?1 and 1000 mg·l^?1 treatments, respectively, which confirmed the Pb hyperaccumulator characteristics of P. crinitum. For plants under Pb stress, most of the Pb was fixed in the cell walls, with a smaller amount in leaves and root vacuoles.
• Both SOD and POD scavenging of reactive oxygen radicals and fixing and compartmentalisation of Pb in the cell wall might play important roles in detoxification of P. crinitum seedlings in response to Pb stress. There was no phased response of P. crinitum to intermittent Pb stress and the physiological response to Pb stress may be contiguous.

     

Expression of phenylalanine ammonia-lyase gene during tracheary-element differentiation from cultured mesophyll cells ofZinnia elegans L.

Expression of the phenylalanine ammonialyase (PAL) gene during tracheary-element differentiation was studied in mesophyll cell-suspension cultures ofZinnia elegans. Dose-response curves of benzyladenine (BA) and α-naphthaleneacetic acid (NAA) were obtained for the cultures in order to achieve the highest percentage of differentiation. The optimal concentrations of BA and NAA were 0.1 mg·l^-1 and 0.06 mg·l^-1, respectively, which normally stimulated about 40% differentiation by 96 h of culture. The effects of the same ratio but different amounts of BA and NAA on tracheary-element formation have been tested and the results indicate that the absolute amounts of BA and NAA rather than the ratio of them were important for tracheary-element formation in theZinnia cultures. The cells when cultured in the presence of 0.001 mg·l^-1 of BA and 0.06 mg·l^-1 of NAA expanded and divided but did not differentiate. The level of PAL activity, synthesis of PAL protein and the level of PAL mRNA peaked during 72 to 96 h when lignin was actively deposited. This indicated that the PAL gene was temporally and preferentially expressed in association with the lignification during tracheary-element differentiation and thus it can be regarded as a molecular marker for the process. We thank Bejing Agricultural University, the Lundgren Research fund (University of Cambridge) and an ORS award for financial support.     

Effect of biotic and abiotic elicitors on production of betulin and betulinic acid in the hairy root culture of Betula pendula Roth

Abstract

Betulin (B) and betulinic acid (BA) are two triterpenoids with a wide range of biological and medicinal activities in different organs of Betula pendula. This research aimed to increase the accumulation of B and BA in the hairy root culture of B. pendula by seven biotic and abiotic elicitors. Hairy root was induced in the stem’s inner bark of B. pendula using the C58C1 strain in the WPM (Woody Plant Medium). The effects of different concentrations of elicitors and different time of root harvest in hairy root culture of B. pendula showed that highest level of growth index (GI), B, and BA was acquired in treated hairy roots with chitosan (CTS), chlorocholine chloride (CCC) and chitosan nano-fiber (CTS NF). Highest GI of B. pendula hairy roots was 13 that was obtained in the roots treated with CTS 150?mg l^?1 on the 8th day. The highest content of BA was 1.3?mg g^?1 DW after treatment with 1?mg l^?1CCC on the 4th and 6th days and 200?mg l^?1CTS NF on the 10th day. The highest B content (0.94?mg g^?1DW) was obtained in the treated hairy root by 2?mg l^?1 CCC after 4 and 6?days.     

Ventilatory and behavioural responses of the marbled sole Pseudopleuronectes yokohamae to progressive hypoxia

This study identified ventilatory and behavioural responses in the marbled sole Pseudopleuronectes yokohamae under experimentally induced progressive decreases in dissolved oxygen (DO) levels. Ventilation frequency showed an increase with decreasing DO levels from normoxia to 2·75 mg O₂ l^?1, followed by a decrease in ventilation frequency at decreased DO levels from 2·00 to 0·75 mg O₂ l^?1. At DO levels below 2·00 mg l^?1, behaviours at the bottom were suppressed, whereas avoidance behaviours increased. A decrease in avoidance behaviours was observed from 1·00 to 0·75 mg O₂ l^?1. Upside‐down reversal and incapacitation at DO levels of 1·00–0·75 mg O₂ l^?1 suggested that sublethal effects on P. yokohamae were induced. The responses observed before the sublethal DO level could be interpreted as an effort to maintain oxygen uptake, reduce routine activities and facilitate avoidance. The observed DO level thresholds that induce behavioural responses, in addition to sublethal effects, indicate hypoxia‐tolerance that is important for understanding the effects of hypoxia on coastal ecosystems.     

Assessment of EDDS and vermicompost for the phytoextraction of Cd and Pb by sunflower (Helianthus annuus L.)

The effects of Ethylenediamine disuccinic acid (EDDS) (0 and 5?mmol·kg^?1) as a synthetic chemical amendment, vermicompost (0 and 5%w/w) as an organic amendment and their combined application were evaluated for the phytoextraction by sunflower (Helianthus annuus L.) of cadmium (Cd) and lead (Pb) at three artificial contamination levels in soils (0, 50, and 100?mg·kg^?1 for Cd and 0, 100, and 200?mg·kg^?1 for Pb). The results showed that the application of EDDS was the most effective method to increase Pb and Cd concentrations in both parts of the plant. The results also showed that the application of EDDS increased 9.27% shoot Pb content at 200?mg·kg^?1 but decreased 15.95% shoot Cd content at 100?mg·kg^?1 contamination level with respect to the respective controls. The bioavailable concentrations of Cd at 100?mg·kg^?1 and Pb at 200?mg·kg^?1 contamination level in the soil at the end of experiment increased 25% and 26%, respectively after the application of EDDS but vermicompost decreased 43.28% the bioavailable Pb concentration relative to their controls. Vermicompost increased the remediation factor index of Cd, thus making it the best treatment for the phytoextraction of Cd. The combined application of EDDS and vermicompost was the best amendment for Pb phytoextraction.     

Physiological responses,tolerance efficiency,and phytoextraction potential of Hylotelephium spectabile (Boreau) H. Ohba under Cd stress in hydroponic condition

Abstract

A sand hydroponic experiment with different concentrations of 0, 5, 10, 20, 40?mg L^?1 Cd was used to study the growth and physiological response of Hylotelephium spectabile (Boreau) H. Ohba. and its phytoextraction potential for Cd. The results showed that total plant biomass under 5?mg L^?1 Cd treatment was slightly affected. The content of malondialdehyde (MDA) in leaf exposed to Cd was higher, and the POD and CAT activity exhibited a positive response to the low level of Cd addition (5?mg·L^?1). The photosynthesis pigments were slightly inhibited, and the ultrastructure of chloroplast remained intact after treatment with 10?mg L^?1 Cd. The maximum leaf Cd content (603?mg·kg^?1) was found in 5?mg L^?1 Cd treatment, then decreased with the Cd level increased. The maximum Cd content in the shoots far exceeds the threshold level (100?mg kg^?1) for a Cd-hyperaccumulator plant with the value of translocation factor (TF_shoot/root) for Cd reaching up to 5.62. In conclusion, H. spectabile showed normal growth and physiological response and high shoot Cd accumulation under 5?mg L^?1 Cd stress, which made it to be a good candidate for phytoextraction of low-level Cd polluted environment.     

Enhanced biological denitrification of high concentration of nitrite with supplementary carbon source

Nitrite accumulates during biological denitrification processes when carbon sources are insufficient. Acetate, methanol, and ethanol were investigated as supplementary carbon sources in the nitrite denitrification process using biogranules. Without supplementary external electron donors (control), the biogranules degraded 200 mg l^?1 nitrite at a rate of 0.27 mg NO₂–N g^?1?VSS h^?1. Notably, 1,500 mg l^?1 acetate and 700 mg l^?1 methanol or ethanol enhanced denitrification rates for 200 mg l^?1 nitrite at 2.07, 1.20, and 1.60 mg NO₂–N g^?1?VSS h^?1, respectively; these rates were significantly higher than that of the control. The sodium dodecyl sulfate polyacrylamide gel electrophoresis of the nitrite reductase (NiR) enzyme identified three prominent bands with molecular weights of 37–41 kDa. A linear correlation existed between incremental denitrification rates and incremental activity of the NiR enzyme. The NiR enzyme activity was enhanced by the supplementary carbon sources, thereby increasing the nitrite denitrification rate. The capacity of supplementary carbon source on enhancing NiR enzyme activity follows: methanol?>?acetate?>?ethanol on molar basis or acetate?>?ethanol?>?methanol on an added weight basis.     

Isolation and characterization of halotolerant phosphate solubilizing bacteria naturally colonizing the peanut rhizosphere in salt-affected soil

Abstract

In search of efficient salt-tolerant phosphate solubilizing bacteria (PSB), we conducted a survey of PSB that naturally colonize the rhizosphere of peanuts in Xinjiang, China, a typical inland saline area. A total of 23 PSB were isolated, which included 12 Bacillus strains, three Acinetobacter strains, two Pseudomonas strains, two Brevibacillus, and one strain of Gordonia terrae, Chryseobacterium lathyri, Ensifer sesbaniae, and Paenibacillus illinoisensis. All PSB have high potential for dissolving calcium phosphate [Ca₃(PO₄)₂] within the range of 65–496?mg·L^?1, but relatively weak ability to dissolve AlPO₄ in amounts ranging from 0.4 to 11.4?mg·L^?1, FePO₄ ranging from 0.1 to 5.5?mg·L^?1, and lecithin ranging from 2.1 to 10.3?mg·L^?1. The maximum tolerance for pH is 10, and the maximum tolerance for Na₂CO₃ is 50?mm. Six isolates that can grow under the condition of 1.5?M NaCl (YMX5, YMX11, TPMX5, TPMX16, TPMX18, and TPMX19) were selected to determine their ability to dissolve Ca₃(PO₄)₂ and tolerate pH changes. This study characterized salt-tolerant PSB isolates that can be used as bioinoculants to protect plants against salt stress. In addition, we describe the phosphate-solubilizing ability of Gordonia terrae at high NaCl concentrations.     

Neonatal exposure to bisphenol A advances pubertal development in female rats

Neonatal exposure to bisphenol A (BPA) is hypothesized to advance pubertal development. However, the effects of neonatal BPA exposure on pubertal development has not been described. In this study, female Sprague‐Dawley rats were exposed to 0.05, 0.5, 5, or 10 mg·kg^?1·day^?1 BPA, or corn oil vehicle alone from postnatal day 1 (PND1) to PND10 via subcutaneous injection. We evaluated day of vaginal opening (DVO), ovarian morphology, serum hormone concentrations, and hypothalamic expression of Gnrh1 and Kiss1 in female rats at PND35. DVO was significantly advanced in rats exposed to 5 and 10 mg·kg^?1·day^?1 BPA. Serum hormone concentrations increased as BPA dose increased. Additionally, hypothalamic Gnrh1 and Kiss1 expression were increased with BPA exposure; rats exposed to 10 mg·kg^?1·day^?1 BPA had significantly upregulated hypothalamic Gnrh1 and Kiss1 expressions in terms of both messenger RNA and protein levels. Our results suggest that exposure to a 10 mg·kg^?1·day^?1 dose of BPA might advance pubertal development significantly. In addition, within the range of 0 to 10 mg·kg^?1·day^?1, neonatal exposure to BPA may affect pubertal development in a dose‐dependent manner.     

Arthrospira platensis biomass with high protein content cultivated in continuous process using urea as nitrogen source

Aims: Arthrospira platensis has been studied for single‐cell protein production because of its biomass composition and its ability of growing in alternative media. This work evaluated the effects of different dilution rates (D) and urea concentrations (N₀) on A. platensis continuous culture, in terms of growth, kinetic parameters, biomass composition and nitrogen removal. Methods and results: Arthrospira platensis was continuously cultivated in a glass‐made vertical column photobioreactor agitated with Rushton turbines. There were used different dilution rates (0·04–0·44 day^?1) and urea concentrations (0·5 and 5 mmol l^?1). With N₀ = 5 mmol l^?1, the maximum steady‐state biomass concentration was1415 mg l^?1, achieved with D = 0·04 day^?1, but the highest protein content (71·9%) was obtained by applying D = 0·12 day^?1, attaining a protein productivity of 106·41 mg l^?1 day^?1. Nitrogen removal reached 99% on steady‐state conditions. Conclusions: The best results were achieved by applying N₀ = 5 mmol l^?1; however, urea led to inhibitory conditions at D ≥ 0·16 day^?1, inducing the system wash‐out. The agitation afforded satisfactory mixture and did not harm the trichomes structure. Significance and Impact of the Study: These results can enhance the basis for the continuous removal of nitrogenous wastewater pollutants using cyanobacteria, with an easily assembled photobioreactor.     

Comparison of supplements* to enhance recovery of heat‐injured Salmonella from egg albumen

Aims: The purpose of this study was to determine the proficiency of supplements to enhance the recovery of Salmonella from heat‐treated liquid egg albumen on solid agar media. Methods and Results: Salmonella‐inoculated albumen, heated at 53·3°C for 4 min, was plated on 39 combinations of solid media with or without the addition of 12 supplements. Greater numbers of Salmonella (P < 0·05) recovered with the addition of 1·0 g l^?1 ferrous sulfate (FeSO₄) than with any other supplements, except for 0·5 or 1·0 g l^?1 3′3′‐thiodipropionic acid (TDP), which recovered equivalent populations. Addition of 1·0 g l^?1 sodium pyruvate or 6·0 g l^?1 yeast extract plus 1·0 g l^?1 sodium pyruvate supported greater resuscitation than unsupplemented tryptic soy agar (TSA) or supplementing with 0·01 or 0·1 g l^?1 N‐propyl gallate, 10 g l^?1 activated charcoal, 0·1 g l^?1 KMnO₄ or 50 mg l^?1 ethoxyquin. The remaining supplements supported recovery of equivalent numbers of Salmonella, which were fewer cells than recovered with 1·0 g l^?1 FeSO₄, yet greater populations than recovered with 50 mg l^?1 ethoxyquin. Conclusion: Supplementation of plating media with FeSO₄, TDP or sodium pyruvate enhanced recovery of sublethally injured Salmonella from albumen. Significance and Impact of the Study: Pasteurizing albumen impedes recovery of pathogens. These results suggest that the addition of supplements to plating media may assist resuscitation and colony development of heat‐injured salmonellae.     

In vitro propagation and micromorphological studies of Cleome gynandra: a C4 model plant closely related to Arabidopsis thaliana

We developed a micropropagation protocol for Cleome gynandra, a C₄ model plant with medicinal importance. Surface-sterilized nodal segments obtained from 1 to 2-month-old field grown plant were used as explants for culture establishment and plant regeneration. Multiple shoots differentiated through bud breaking on Murashige and Skoog (MS) medium with different concentrations of benzyladenine (BA) and kinetin (Kin). The optimum shoot differentiation occurred on medium with 1.5 mg l^?1 BA. Out of various concentrations and combinations of cytokinins and auxins, MS medium containing 0.5 mg l^?1 BA and 0.1 mg l^?1 IAA (indole-3-acetic acid) was found best for shoot multiplication. However, the differentiated shoots exhibited hyperhydration, leaf curling and early leaf fall during subculturing. To overcome these problems, regenerated shoots were transferred to the modified MS medium with reduced nitrates (825 mg l^?1 NH₄NO₃ and 950 mg l^?1 KNO₃) and 100 mg l^?1 (NH₄)₂SO₄. The micropropagated shoots were rooted (i) in vitro on one-fourth strength of MS salts with 0.25 mg l^?1 each of IBA (indole-3 butyric acid) and NOA (2-naphthoxyacetic acid) + 100 mg l^?1 activated charcoal, and (ii) ex vitro, by treating the shoot base(s) with 200 mg l^?1 of IBA for 3 min and transferred to soilrite moistened with one-fourth strength of MS macro salts in culture bottles. The plants were hardened in the greenhouse with 85 % survival rate. Micromorphological studies of the plants were conducted during hardening with reference to development and changes in vein spacing, glandular trichome and stomata. In comparison to leaves under in vitro condition, higher density of veins and glandular trichomes was observed in the leaves of hardened plants. In addition, stomata became functional during hardening which were non-functional under in vitro condition.     

The Tolerance and Accumulation of Miscanthus Sacchariflorus (maxim.) Benth., an Energy Plant Species,to Cadmium

Miscanthus sacchariflorus (Maxim.) Benth. is a metallophyte suitable for the phytoremediation of mine wastes. The tolerance and accumulation of M. sacchariflorus to cadmium was studied by pot experiments. The results showed that O₂·^? generation rate, plasma membrane permeability and MDA content of M. sacchariflorus leaves increased with increasing Cd concentrations in soil, but significant effect was only observed when Cd concentrations were ≥ 50 mg·kg^?1. SOD and POD activities increased initially but decreased later on, whereas CAT activity only increased significantly at higher Cd concentrations, 50–100 mg·kg^?1. The content of photosynthetic pigment and growth of M. sacchariflorus were both not significantly affected when Cd concentration was ≤ 25 mg·kg^?1. In contrast, both parameters were significantly affected when Cd concentration was ≥ 50 mg·kg^?1. M. sacchariflorus could accumulate much Cd, but most of the Cd assimilated was retained in the belowground part, suggesting that M. sacchariflorus has poor ability to translocate Cd to the aboveground part. Our results suggested that although M. sacchariflorus was not a hyper-accumulator, it has a strong capacity to tolerate and stabilize the Cd. Therefore, M. sacchariflorus has a certain potential in the phytostabilization of Cd-contaminated soils.     

Characterizing the escape response of juvenile summer flounder Paralichthys dentatus to diel‐cycling hypoxia

Swimming speed, angular correlation and expected displacement were measured in juvenile summer flounder Paralichthys dentatus acclimated to either oxygen saturation (c. 7·8 mg O₂ l^?1; saturation‐acclimated fish) or diel‐cycling hypoxia (cycling between 11·0 and 2·0 mg O₂ l^?1) for 10 days and subsequently exposed to more severe diel‐cycling hypoxia (cycling between 7·0 and 0·4 mg O₂ l^?1). Saturation‐acclimated P. dentatus exhibited an active response to declining dissolved oxygen (DO) by increasing swimming speed, angular correlation and expected displacement to peak levels at 1·4 mg O₂ l^?1 that were 3·5, 5·5 and 4·2 fold, respectively, greater than those at DO saturation. Diel‐cycling hypoxia‐acclimated P. dentatus also exhibited an active response to declining DO, although it was relatively less pronounced. Diel‐cycling hypoxia‐acclimated P. dentatus swimming speed, however, still doubled as DO decreased from 7·0 to 2·8 mg O₂ l^?1. Diel‐cycling hypoxia‐acclimated P. dentatus did not recover as well from low DO exposure as did saturation‐acclimated fish. This was reflected in their relatively more random swimming (low angular correlation between successive moves) and poor maintenance of rank order between individuals during the recovery phase. Even saturation‐acclimated P. dentatus did not resume swimming at speeds observed at saturation until DO was 4·2 mg O₂ l^?1. Paralichthys dentatus were very sensitive to decreasing DO, even at DO levels that were not lethal or growth limiting. This sensitivity and their poor recovery may preclude juvenile P. dentatus from using highly productive nursery habitats affected by diel‐cycling hypoxia.     

Effects of hypoxia on oxygen consumption,swimming velocity and gut evacuation in southern bluefin tuna (<Emphasis Type="Italic">Thunnus maccoyii</Emphasis>)

Of the few measurements of the behavioural and physiological responses of tuna to hypoxia, most are restricted to shallow diving tropical species. Furthermore, when wild tuna experience low dissolved oxygen, they are likely to have an increased oxygen demand associated with the metabolic cost of food digestion and assimilation (specific dynamic action). However the response of postprandial tuna to hypoxia has never been examined. This study focuses on the metabolic and behavioural responses of both fasted and postprandial southern bluefin tuna (Thunnus maccoyii) to low dissolved oxygen. Fasted T. maccoyii were exposed to dissolved oxygen levels of 4.44, 3.23, 2.49 and 1.57 mg·l⁻¹ for 20–21 h. In moderate hypoxia (4.44 and 3.23 mg·l⁻¹), swimming speed was enhanced (1.5 and 1.3 times normoxic speed, respectively) presumably to increase ventilation volume. Routine metabolic rate (R _r ) was similarly elevated (1.3 and 1.2 times normoxic R _r , respectively), most likely due to increased metabolic demand of faster swimming. At 2.49 mg·l⁻¹, swimming speed increased to over double the normoxic speed, possibly as an escape response. At 1.57 mg·l⁻¹, both swimming speed and R _r were reduced (0.8 and 0.9 times normoxic level, respectively), and tuna failed to survive the entire 20 h exposure period. This reveals that the critical oxygen level of T. maccoyii is between 1.57 and 2.49 mg·l⁻¹, demonstrating that they are remarkably well adapted to low dissolved oxygen. Feeding did not greatly influence their hypoxia tolerance with tuna surviving exposure to dissolved oxygen levels of 2.96 and 1.81 mg·l⁻¹ for 21 h, after ingesting a ration of 6.7% body weight of sardines (Sardinops sagax). In a subsequent experiment to determine the effects of hypoxia on digestion rate, T. maccoyii were fed to satiation and exposed to a dissolved oxygen level of 2.84 mg·l⁻¹ for 6.5–8 h. There was no significant difference in swimming speed, R _r and gastric evacuation rates of tuna in hypoxia compared to those in normoxia. This demonstrates that in moderate to severe hypoxia, T. maccoyii are still capable of aerobically supporting maintenance metabolism, routine swimming and specific dynamic action. It is hypothesized that adaptations which support the large metabolic scope of tuna are also likely to be beneficial for oxygen extraction and delivery in conditions of hypoxia.     

Isolation and characterization of actinomycetes from healthy goat faeces

Aims: To isolate and characterize actinomycetes with probiotic activities from healthy goat faeces. Methods and Results: Faecal actinomycetes were isolated by dilution methods and identified by 16S rRNA gene sequence analysis. The hydrolytic enzyme activities were analysed by clear zone formation. The antimicrobial activities and resistance to heavy metals were tested by growth inhibition methods. The isolates belong to a small group of actinobacterial genera, including Streptomyces, Nocardiopsis and Oerskovia. The Oerskovia was the most widely distributed genus among the cultures. The proportion of streptomycete‐like strains producing amylase or protease is significantly higher than those of other actinomycetes (P < 0·05). Compared with streptomycete‐like strains, a higher proportion of (α‐ or β‐) galactase‐producing other actinomycetes was found in goat faeces. More than 50% of streptomycete‐like strains showed activities against test fungi. Streptomycetes could tolerate 0·25 mmol l^?1 Cr₂O₇^2?, 2 mmol l^?1 Ni²⁺; however, other actinomycetes are liable to 40 mmol l^?1 Fe³⁺ and 0·25 mmol l^?1 Cr₂O₇^2? and resistant to 5 mmol l^?1 Ni²⁺ and 2 mmol l^?1 Cu²⁺. Conclusions: The different physiological characteristics of the actinomycetes suggested that the cooperation in the actinomycetes might be involved in their association with goat. Significance and Impact of the Study: Probiotic mixtures based on faecal actinomycetes showed potentials in animal production.     

HOT WATER EXTRACTABLE PHOSPHORUS—AN INDICATOR OF NUTRITIONAL STATUS OF PERIDINIUM CINCTUM (DINOPHYCEAE) FROM LAKE KINNERET (ISRAEL)?1

The intracellular levels of hot water extractable and total phosphorus were determined in the dinoflagellate Peridinium cinctum. f. westii (Lemm.) Lef. for natural samples from the bloom in Lake Kinneret and from laboratory cultures. Amounts of phosphorus (P) in the hot water fraction, relative to total cellular phosphorus, were similar in lake Peridinium and in cells grown in high ambient orthophosphate (P_i) media (3–6 mg P · l^?1). The absolute amounts of hot water extractable P in natural cell and those cultured at lower P_i concentrations (0.02–0.05 mg P · 1^?1) were similar, although average P_i in lake water were 4 μg · l^?1. Under most growth conditions the hot water extract contained approximately equal amounts of molybdate reactive phosphorus (MRP) and non-MRP. Short chain (6–9 units) polyphosphates (mol wt 630–950) probably constituted the bulk of the non-MRP pool, which was hydrolysable by alkaline phosphatase and may serve as a precursor for a more permanent P store. Intracellular P levels and distribution were not directly dependent on external P_i concentrations but may be determined by the N:P atomic ratio or overall external ionic milieu. Peridinium grown in low ambient P_i released significant amounts of non-MRP compounds. In Lake Kinneret, for at least most of the bloom period, Peridinium does not appear to be limited by P supply.     

Inactivation of Cronobacter spp. (Enterobacter sakazakii) in infant formula using lactic acid,copper sulfate and monolaurin

Aims: To investigate the effect of lactic acid (LA), copper (II), and monolaurin as natural antimicrobials against Cronobacter in infant formula. Methods and Results: The effect of LA (0·1, 0·2 and 0·3% v/v), copper (II) (10, 50 and 100 μg ml^?1) and monolaurin (1000, 2000, and 3000 μg ml^?1) suspended into tween‐80? or dissolved in ethanol against Cronobacter in infant formula was investigated. Reconstituted infant formula and powdered infant formula were inoculated with five strains of Cronobacter spp. at the levels of c. 1 × 10⁶ CFU ml^?1 and 1 × 10³ CFU g^?1, respectively. LA at 0·2% v/v had a bacteriostatic effect on Cronobacter growth, whereas 0·3% v/v LA resulted in c. 3 log₁₀ reduction. Copper (II) at the levels of 50 μg ml^?1 and 100 μg ml^?1 elicited c. 1 and 2 log₁₀ reductions, respectively. The combination of 0·2% LA and 50 μg ml^?1 copper (II) resulted in a complete elimination of the organism. Monolaurin exhibited a slight inhibitory activity against Cronobacter (c. 1·5 log₁₀ difference) compared to the control when ethanol was used to deliver monolaurin. Conclusions: A complete elimination of Cronobacter was obtained when a combination of sublethal concentrations of LA (0·2%) and copper (II) (50 μg ml^?1) was used. Significance and Impact of the Study: The use of the synergistic interactive combination of LA and copper (II) could be beneficial to control Cronobacter in the infant formula industry.     

THE EFFECT OF SULFIDE ON THE BLUEGREEN ALGAE OF HOT SPRINGS. I. NEW ZEALAND AND ICELAND1

The concentration of soluble sulfide (H₂S, HS^?, S⁼)² in hot springs of New Zealand and Iceland has a “species-determining” effect which appears to override the effect of all other chemical factors excepting hydrogen ion concentration. The cosmopolitan high-temperature form (HTF) of Mastigocladus laminosus (Ag.) Cohn appears to be sensitive to concentrations of S⁼ over 0.15 mg·l^?1 in New Zealand and 0.25 mg·l^?1 in Iceland. It was absent in thermal streams where S⁼ level was high enough to exceed this concentration at a point below its optimal temperature range (i.e., below ca. 50 C). In low or non-S⁼ springs in these regions this alga formed mats to an upper temperature limit of 63–64 C. In contrast, one bluegreen alga (Oscillatoria amphigranulata van Goor) occurred abundantly only in moderate to high S⁼ springs of New Zealand; this was a species with an upper temperature limit of about 57 C. Field ¹⁴C-HCO₃^? incorporation experiments with various levels of added S⁼ confirmed the sensitivity of HTF Mastigocladus and the great tolerance of O. amphigranulata. In the latter, the photosynthetic incorporation of ¹⁴C-HCO₃^? was sustained or enhanced by the additions of S⁼ in the presence of 5 × 10^?5m DCMU, an inhibitor of photoelectron transport on the reducing side of photosystem II. It is possible, but not proven, that S⁼ may act as a photoreductant in this species.