Improvement of nisin production in pH feed-back controlled,fed-batch culture by <Emphasis Type="Italic">Lactococcus lactis</Emphasis>subsp<Emphasis Type="Italic">. lactis</Emphasis>

Nisin production by Lactococcus lactis subsp. lactisin fed-batch culture was doubled by using a pH feed-back controlled method. Sucrose concentration was controlled at 10 g l^–1 giving 5010 IU nisin ml^–1 compared to 2660 IU nisin ml^–1 in batch culture.     

Study of physicochemical factors limiting the growth ofKluyveromyces marxianus

Summary The effects of various physicochemical parameters on the growth of twoKluyveromyces marxianus strains were investigated, including: pH values, sodium chloride, water activity in the medium and temperature. Both yeast strains were unaffected by pH changes. Optimal pH for growth was found to be 4 with both strains, but they were able to develop within the pH 3–8 range. Suitable growth was obtained at temperatures of 4–44°C and the optimal temperature for growth was 36°C for both strains. Modelling of this latter parameter is described. Growth of both microorganisms was considerably modified by increased NaCl or decreased water activity in the medium.     

Does indoleacetic acid promote growth via cell wall acidification?

Growth of Triticum aestivum L. cv. Cappelle Desprez coleoptiles is promoted by 5.7×10^–5 M indole acetic acid (IAA) as effectively in pH 3.4 buffer as in water, but IAA is not effective in the presence of buffer at pH 3.0 or 3.2 A combination of 5.7×10^–5 M IAA and pH 3.4 buffer promotes growth to a greater extent than pH 3.2 buffer alone, which is optimal for acid-induced growth. IAA employed at 10^–7 M is still effective at promoting growth in the presence of pH 3.4 buffer, moreover, IAA at 10^–7 M interacts synergistically with the acidic buffer to promote growth. It is concluded that IAA and acid promote growth via separate mechanisms, and that IAA does not promote cell wall loosening by rendering the cell wall more acid.Abbreviation IAA Indoleacetic acid     

Mass cultivation of the nitrogen-fixing cyanobacteriumGloeotrichia natans,indigenous to rice-fields

Gloeotrichia natans, a nitrogen fixing cyanobacterium common in rice fields in the Philippines, was used for studies to establish key features of its physiology and potential production in outdoor cultures. Under optimal growth conditions (38 °C, pH 8.0, no carbon enrichment) the specific growth rate of rice-field isolate was 0.076 h^–1. The pH of the medium (between 6.5 and 9.0) did not influence the growth rate, but it did affect phycobiliprotein content, as reflected by a change in colour. At pH 7.0 the culture was green-brown, with phycobiliproteins constituting up to 10% of the total protein, while at pH 9.0 the culture was brownish-black and the pigment content was as high as 28% of the total protein. In outdoor cultures the specific growth rate was related directly to cell density in the range of 0.7–1.5 g dry weight 1^–1 at a rate of stirring of 30 rpm, and inversely related to cell density at half this rate. At a stirring of 30 rpm, daily production of outdoor cultures harvested to maintain cell densities of 0.7, 1.15 andw 1.5 g 1^–1 were 14.7, 17.1 and 18.1 g m^–2 d^t, respectively. This rate of production was maintained for more than 45 days. Phycobiliprotein content in the culture kept at a density of 1.5 g 1^–1 reached 14% of the total biomass.     

Isolation and study of two strains ofLeptospirillum-like bacteria from a natural mixed population cultured on a cobaltiferous pyrite substrate

Two strains ofLeptospirillum-like bacteria, L6 and L8, have been isolated from a mixed inoculum, also containingThiobacillus ferrooxidans andT. thiooxidans, cultured for one year with a colbaltiferous pyrite as energy substrate in a 100 I continuous bioleaching laboratory unit. Several physiological properties of the strains are described. The vibrio-shaped microorganisms grew at pH values lower than 1.3. Their growth rate was maximum between 2.5 and 8.0 g l¹ ferrous iron. The optimal growth temperature was 37.5° C. Ferric iron had a stimulative effect on bacterial development up to 8 g l^–1, and growth was as rapid at 14 g l^–1 ferric iron as at 8 g l^–1. The negative influence of cobalt on the final cell concentration was observed at 0.5 g l^–1, but the growth rate was not affected up to 2 g l^–1. The G + C content of strains L8 is 55.6 mol%.     

n-3 Polyunsaturated fatty acid productivity of the marine microalgaIsochrysis galbana. Growth conditions and phenotypic selection

Two set of isolates were obtained in an isolation/selection programme to select eicosapentaenoic acid (EPA) and/or docosahexaenoic acid (DHA) rich strains ofIsochrysis galbana. EPA content was improved up to an average of 5.3% (d.wt) for the second set of isolates. On the other hand, with the aeration rate, pH and irradiance kept at low levels, the growth rate was slow and EPA synthesis was enhanced, but productivity increased when growth rates were maximum. A model relating steady-state dilution rates in chemostat cultures ofI. galbana to internal average irradiance is proposed. The greatest productivities were obtained between 0.0295 h^–1 and 0.0355 h^–1 with 300 mg m^–3 h^–1 for EPA and 130 mg m^–3 h^–1 for DHA.     

The optimum and tolerance pH range is correlated to colonial morphology in isolates of the entomopathogenic fungus Beauveria bassiana – a potential biopesticide

Beauveria bassiana is an insect pathogenic fungus used as a biopesticide in crop pest management. It exists saprophytically in the soil. The pH of the soil and the insect cuticle and its haemocoel may influence fungal survival and infection potential. Therefore the tolerance and optimum pH ranges were studied in 29 isolates of B. bassiana. Germination and growth bioassays in liquid culture medium adjusted to pH values in the range 3–14 at unit intervals were studied. A pH of 3 was found to be toxic to all isolates – conidia germinated at this pH but growth was totally inhibited. All isolates tolerated a pH of 5–13. Some isolates showed tolerance to a pH of 4 and/or 14 as well. Sixteen isolates showed a wide range of pH optimum of 5–13 while others had a narrower optimal range. The relation of the pH tolerance and optimum range of an isolate to its phenotypic characters (colonial morphology and growth-rate) was studied. All isolates with a chalky type colonial morphology had an intermediate growth rate and showed a wide optimum range of pH 5–13 or 5–14. All isolates with dusty type colonial morphology were found to have a high growth rate and a wide pH tolerance range of 4–14. Both chalky and dusty colonial morphology are due to a growth pattern characterized by iterated conidiation with a very brief hyphal stage. The correlation observed between these traits based on external phenotype would facilitate prediction of the pH tolerance characters of an isolate of B. bassiana.     

Effects of carbon and nitrogen sources,carbon-to-nitrogen ratio,and initial pH on the growth of nematophagous fungus <Emphasis Type="Italic">Pochonia chlamydosporia</Emphasis> in liquid culture

The effects of carbon and nitrogen sources, carbon-to-nitrogen ratio (C:N) and initial pH value on the growth and sporulation of the nematophagous fungus Pochonia chlamydosporia in liquid culture were examined. Among the 21 carbon sources and 15 nitrogen compounds tested, the optimal carbon and nitrogen sources for mycelial growth were sweet potato and L-tyrosine, and for sporulation were sweet potato and casein peptone. A C:N ratio of 10:1 at pH 3.7 gave the maximum yield of conidia and a C:N ratio of 40:1 at pH 6.8 gave the maximum biomass. The initial pH value had a significant effect on mycelial growth and conidial production, with the optimal ranges being 3.5–4.5 for sporulation and 5–6 for growth. Maximum conidial production was obtained at an initial pH of 4.0 and the maximum biomass at pH 6.0. The results also showed that the final pH after 7 days cultivation was always higher than the initial value. The variability in growth and sporulation of seven strains of P. chlamydosporia in liquid culture was also compared and discussed.     

Pullulanases of alkaline and broad pH range from a newly isolated alkalophilicBacillus sp. S-1 and aMicrococcus sp. Y-1

Summary Two highly alkalophilic bacteria, and potent producers of alkaline pullulanase, were isolated from Korean soils. The two isolates, identified asBacillus sp. S-1 andMicrococcus sp. Y-1, grow on starch under alkaline conditions and effectively secrete extracellular pullulanases. The two isolates were extremely alkalophilic since bacterial growth and enzyme production occurred at pH values ranging from pH 6.0 to 12.0 forMicrococcus sp. Y-1 and pH 6.0 to 10.0 forBacillus sp. S-1. Both strains secrete enzymes that possess amylolytic and pullulanolytic acitivities. Extracellular crude enzymes of both isolates gave maltotriose as the major product formed from soluble starch and pullulan hydrolysis. Compared to other alkalophilic microbes such asMicrococcus sp. (0.57 units ml^–1),Bacillus sp. KSM-1876 (0.56 units ml^–1) andBacillus No. 202-1 (1.89 units ml^–1) these isolates secreted extremely high concentrations (7.0 units ml^–1 forBacillus sp. S-1 and 7.6 units ml^–1 forMicrococcus sp. Y-1) of pullulanases in batch culture. The pullulanase activities from both strains were mostly found in the culture medium (85–90%). The extracellular enzymes of both bacteria were alkalophilic and moderately thermoactive; optimal activity was detected at pH 8.0–10.0 and between 50 and 60°C. Even at pH 12.0, 65% of original Y-1 pullulanase activity and 10% of S-1 pullulanase activity remained. The two newly isolated strains had broad pH ranges and moderate thermostability for their enzyme activities. These result strongly indicate that these new bacterial isolates have potential as producers of pullulanases for use in the starch industry.     

Chondrus crispus (Gigartinaceae,Rhodophyta) tank cultivation: optimizing carbon input by a fixed pH and use of a salt water well

The injection of exogenous carbon into intensively cultivated algal tanks is necessary to insure a maximum growth rate by stabilizing the dissolved inorganic carbon (DIC) pool, but represents the major part of the cultivation cost (ca. 73%). This study was conducted in paddle-wheel tanks ranging in size from 260 m² to 1000 m². Additional carbon was provided by carbon dioxide mixed into the incoming sea water through a tubular reactor. Production vs pH was analysed on 120 growth measurements covering two years of continuous cultivation. Whereas production peaked at pH 8.0–8.2, the economic optimum for pH regulation was in the range 8.4–8.5, where CO₂ injection was greatly reduced (–29%) for only a slight decrease in production (–4%). Expressed as a function of pH level, the specific carbon injection (g c gdw^–1 of Chondrus produced) showed an inverse exponential relationship, whereas gross photoconversion ratio (gdw mol photons^–1) varied according to a second degree equation with a low amplitude. The photoconversion ratio was not improved when the culture was maintained at a DIC concentration higher than the natural equilibrium (0.64 ± 0.11 gdw mol photons^–1 at 2.35 mM and 0.65 ± 0.15 gdw mol photons^–1 at 3.19 MM).A complementary source of carbon was found in underground salt water with a high and stable DIC concentration (10.15 ± 0.25 mmole Cl^–1). The mixing of the well water with natural sea water allowed another economy of CO₂ (–20% at pH 8.5) and nutrients (–12%), the total unitary cost of production being cut by about 17%.     

Effect of temperature,hydrogen ion concentration and osmotic potential on oospore germination of fivePythium spp. isolated from pond water

Oospore germination occurred over a temperature ranging of 15–35°C forPythium coloratum, 10–35°C forP. diclinum, 15–30°C forP. dissotocum, 7–30°C forP. monospermum, and 10–30°C forP. pleroticum. Optimum temperature was 25°C for all species tested. In case of pH, oospore germination occurred over a range of 4.76–8.55 with an optimum of 6.40–7.40. The least germination occurred at pH 4.76 forP. coloratum, P. diclinum, P. monospermum andP. pleroticum, whileP. dissotocum germinated from pH 5.02. Oospores of the all tested pythia were able to germinate at –0.13 to –1.65 MPa and could not germinate at –3.40 MPa, with the highest germination rate at –0.27 to –0.47 MPa. The effect of temperature, pH and osmotic potential on oospore germination was discussed in relation to pollution of pond water.     

Optimization of culture conditions for the production of an extracellular ribonuclease by <Emphasis Type="Italic">Aspergillus niger</Emphasis> in a benchtop bioreactor

SummarySelf-directing optimization was successfully employed to determine the optimal combination of engineering parameters, viz., pH, aeration rate and agitation rate, for extracellular ribonuclease production by Aspergillus niger SA-13-20 in a batch bioreactor. Maximal RNase production of 5.38 IU ml^–1 was obtained at controlled pH of 2.33, aeration rate of 1.67 v/v/m and agitation rate of 850 rev/min. The effect of oxygen on the fermentation was also investigated. With increase in volumetric oxygen transfer coefficients (K_La), cell growth and RNase production first increased and then decreased. RNase production was further increased to 7.10 IU ml^–1 and the fermentation time was shortened from 96 to 72 h by controlling dissolved oxygen concentration at 10% saturation by aerating oxygen after about 28 h of fermentation under the above optimal condition. The kinetic model showed that RNase production by A. niger SA-13-20 was growth-associated.     

Biodegradation of p-nitrophenol by methyl parathion-degrading Ochrobactrum sp. B2

Ochrobactrum sp. B2, a methyl parathion-degrading bacterium, was proved to be capable of using p-nitrophenol (PNP) as carbon and energy source. The effect of factors, such as temperature, pH value, and nutrition, on the growth of Ochrobactrum sp. B2 and its ability to degrade p-nitrophenol (PNP) at a higher concentration (100 mg l⁻¹) was investigated in this study.The greatest growth of B2 was observed at a temperature of 30 °C and alkaline pH (pH 9–10). pH condition was proved to be a crucial factor affecting PNP degradation. Enhanced growth of B2 or PNP degradation was consistent with the increase of pH in the minimal medium, and acidic pH (6.0) did not support PNP degradation. Addition of glucose (0.05%, 0.1%) decreased the rate of PNP degradation even if increased cell growth occurred. Addition of supplemental inorganic nitrogen (ammonium chloride or ammonium sulphate) inhibited PNP degradation, whereas organic nitrogen (peptone, yeast extract, urea) accelerated degradation.     

Comparative effects of Glomus mosseae and P fertilizer on foliar mineral composition of Acacia senegal seedlings inoculated with Rhizobium

Summary A factorial experiment with two controlled factors was conducted in the greenhouse with Acacia Senegal seedlings. The substrate was a degraded sandy soil (Dior soil) poor in available P (11 ppm — Olsen). The first controlled factor was soil sterilization, with two levels: (A) sterilized soil; (B) non-sterilized soil. The second factor was fertilization, with six levels: (1) uninoculated control; (2) inoculation with Rhizobium (ORS 1007); (3) inoculation with Glomus mosseae; (4) double inoculation with ORS 1007 and G. mosseae; (5) inoculation with ORS 1007 and 30 ppm phosphorus per plant; (6) inoculation with ORS 1007 and 60 ppm phosphours. The combination of the two factors and their levels led to 12 different plant treatments (A1–A6 and B1–B6). Compared to the control B1, the B5 and B6 treatments containing phosphorus increased: nodule dry weight about 7 times ; leaf dry weight about 4 times ; total N, P and Mg 4–5 times; total K and Ca 3–4 times. The mycorrhizal inoculation had the same positive effect on plant growth and mineral composition but with lower values. Plants inoculated with Rhizobium alone gave the lowest results. The A1 treatment gave lower values than B1. Foliar mineral contents varied within a narrow range (20–30%).     

The effect of temperature on growth and cellulase (β-1,4-endoglucanase) production in the compost fungusChaetomium thermophile var.dissitum

Chaetomium thermophile var.dissitum, isolated from an experimental urban refuse compost, had the following growth characteristics: Minimum temperature, 27±1°C; optimum, 45–50°C; maximum, 57±1°C; pH optimum 5.5–6.0.A number of carbohydrates could be used for growth, but cellulase formation measured with carboxymethylcellulose as substrate was initiated only on cellulose or xylan. With cellulose as the carbon source, cellulase accumulation in the culture filtrate followed closely that of growth, when the temperature was varied. pH optimum for the cellulase system was 5.0.The optimum temperature for cellulase activity with carboxymethylcellulose as substrate varied between 77°C with 1/2 h incubation time and 58°C with 10 h incubation time.With cotton as substrate, the optimum temperature was 58°C regardless of incubation time. Carboxymethylcellulose had a higher stabilizing effect on the enzyme than cotton. The temperature stability of the cellulase was highest at pH 6.0.     

Effects of dilution rate and pH on the ruminal cellulolytic bacterium Fibrobacter succinogenes S85 in cellulose-fed continuous culture

The ruminal cellulolytic bacterium Fibrobacter succinogenes S85 was grown in cellulose-fed continuous culture at 22 different combinations of dilution rate (D, 0.014–0.076 h^-1) and extracellular pH (6.11–6.84). Effects of pH and D on the fermentation were determined by subjecting data on cellulose consumption, cell yield, product yield (succinate, acetate, formate), and soluble sugar concentrationto response surface analysis. The extent of cellulose conversion decreased with increasing D. First-order rate constants at rapid growth rates were estimated as 0.07–0.11 h^-1, and decreased with decreasing pH. Apparent decreases in the rate constant with increasing D was not due to inadequate mixing or preferential utilization of the more amorphous regions of the cellulose. Significant quantities of soluble sugars (0.04–0.18 g/l, primarily glucose) were detected in all cultures, suggesting that glucose uptake was rather inefficient. Cell yields (0.11–0.24 g cells/g cellulose consumed) increased with increasing D. Pirt plots of the predicted yield data were used to determined that maintenance coefficient (0.04–0.06 g cellulose/g cells · h) and true growth yield (0.23–0.25 g cells/g cellulose consumed) varied slightly with pH. Yields of succinate, the major fermentation endproduct, were as high as 1.15 mol/mol anhydroglucose fermented, and were slightly affected by dilution rate but were not affected by pH. Comparison of the fermentation data with that of other ruminal cellulolytic bacteria indicates that F. succinogenes S85 is capable of rapid hydrolysis of crystalline cellulose and efficient growth, despite a lower _max on microcrystalline cellulose.     

Effects of acidity on the growth of two Euglena species

Our study separates the effects of elevated protons (at pH <3) and elevated metals (Al, Cd, Cu, Fe, Ni, Zn) on the growth of E. mutabilis Schmitz, a pioneering phototroph in acid mine drainage (AMD) and E. gracilis Klebs, a closely-related species rarely found in severely AMD-impacted sites. Both species were acid tolerant, growing optimally at pH 2.5–7. At pH values typical of AMD (pH 2.5–4) in the absence of elevated metals, E. gracilis outcompeted E. mutabilis (growth rates of 1.0 and 0.8 div d^–1, respectively). Relative metal toxicities were evaluated based on the Effective Exposure causing 50% growth reduction (= EE50). With total metal additions similar to AMD levels, E. mutabilis demonstrated significantly greater tolerance to all metals, except Cu. E. gracilis showed two-fold higher tolerance to Cu²⁺ than E. mutabilis (EE50 of 91.6 vs. 45.7 pmol cell^–1). The EE50 for Zn²⁺ was similar for both species (368 pmol cell^–1 for E. gracilis and 423 pmol cell^–1 for E. mutabilis). With Cd and Ni, E. mutabilis tolerated an order of magnitude higher exposure than E. gracilis(EE50 of 1.6 vs. 0.2 pmol Cd²⁺ cell^–1; EE50 of 942 vs. 87 pmol Ni²⁺ cell^–1). Al and Fe were tolerated at high total metal concentrations (up to 100 mM) by E. mutabilis, but toxicity was evident with E. gracilisat much lower levels. E. mutabilis grew at double the Al³⁺ exposure tolerated by E. gracilis (EE50 of 398 vs. 188 pmol Al³⁺ cell^–1). There was an 18-fold difference in Fe tolerance levels between E. mutabilis and E. gracilis with EE50s of 8773 and 502 pmol Fe²⁺ cell^–1, respectively. We conclude that differential metal tolerance, particularly to Fe²⁺, accounts for the mutually exclusive distribution of E. gracilis and E. mutabilis in AMD-impacted habitats.     

Filamentous algae in fish ponds of the Třeboň Biosphere Reserve-ecophysiological study

Filamentous algae in eutrophic carp ponds in South Bohemia (Central Europe) were studied from 1988 to 1990. High biomass (490 g DW m^-2) was attained by Cladophora fracta (O. F. Müll. ex Vahl) Kütz. after two months of growth. This marked growth depleted inorganic carbon in the water, but did not decrease the concentration of tissue nutrients. Laboratory measurements of final pH indicate that all the filamentous algae studied, except for Tribonema, are very efficient HCO₃ ^- users. An extremely high pH of 11.6 and oxygen concentration of 32 mg l^-1 were measured in the algal mats. High pH resulted in CaCO₃ precipitation, visible as white incrustations on algal filaments. The amount of precipitated CaCO₃ reached 134 kg ha^-1. After reaching peak biomass, 90% of the Cladophora decomposed over the next 95 days.The highest net photosynthetic rate in C. fracta was measured between pH range 8.5–10.0 and oxygen concentrations of 7–12 mg l^-1. Optimum temperature for photosynthesis was between 17–22°C.     

Effects of soil pH and nitrogen fertility on the population dynamics of Thielaviopsis basicola

Black root rot of tobacco, caused by Thielaviopsis basicola, is generally severe at soil pH values >5.6 and suppressed under more acidic conditions (pH < 5.2). Soil acidifying fertilizers containing NH₄–N are generally recommended for burley tobacco production in North Carolina, but the effects of N form and application rate on development of black root rot and on the population dynamics of T. basicola have not been determined. Greenhouse and laboratory studies were conducted to evaluate the effects of N form (NH₄ ⁺ or NO₃ ^–) and rate on pathogen and disease parameters at several initial soil pH levels. A moderately-conducive field soil, initial pH 4.7, was adjusted to a pH of 5.5 or 6.5 by the addition of CaOH₂, then amended with the desired nitrogen form and rate. Pathogen populations were determined over time. In addition, spore production in extracts of roots from plants grown in the various nitrogen and pH treatments was determined. Finally, because tobacco responds to acidic soil conditions and exposure to NH₄–N by accumulating high concentrations of the polyamine putrescine, the toxicity of putrescine on vegetative growth and reproduction of T. basicola was investigated. Low soil pH and high levels of NH₄–N suppressed reproduction of T. basicola in soil and in root extract, while use of NO₃–N and depletion of NH₄–N resulted in rapid increases in populations of T. basicola. At 20 mM, putrescine inhibited hyphal growth by 60% and aleuriospore production by 98%. Fertilizers that reduced soil pH also reduced reproduction by T. basicola, and thus have potential for management of black root rot by suppressing populations of T. basicola over multiple years of crop production. The suppression of T. basicola and black root rot observed with NH₄–N amendments may partially be due to development of an inhibitory environment in the root and not solely to changes in rhizosphere pH.     

Phenotypic diversity amongst strains of Pleurotus sajor-caju: implications for cultivation in arid environments

In arid regions, biodiversity and biomass are limited by water availability, and this problem has been compounded by desertification associated with global climate change. The saprotrophic macrofungi that are indigenous to hot subtropical and tropical regions, such as Pleurotus spp., can play key roles in water sequestration, nutrient cycling, human nutrition, and bioremediation of waste materials. We studied 15 strains of Pleurotus sajor-caju, a widespread and phenotypically-diverse species, to establish variability in growth response and primordium development over a range of stress parameters: osmotic potential (−0.5 to −5 MPa), temperature (5–40 °C) and pH (2–12). The initiation of primordia precedes basidiome production and therefore represents a key stage in bioremediation strategies and fungi-driven nutrient cycles. Primordia were produced at low pH (4–6), at suboptimal growth temperatures (≤25 °C), and under moderate water stress (−0.5 to −3.5 MPa). Although the growth windows for different strains were similar, their maximum growth rates and the optimum conditions for growth varied. We discuss the phenotypic diversity of Pleurotus strains and discuss their potential for cultivation, bioremediation and ecological regeneration.