Selection and Study of Potent Lactose-Fermenting Yeasts

Whey-fermenting Kluyveromyces cultures were revealed among 105 yeast strains assimilating lactose. Eighteen strains from milk products, showing maximum potency, fermented galactose, sucrose, and raffinose, in addition to lactose. Many yeast strains fermented inulin. Most strains were resistant to cycloheximide and grew in medium containing glucose, NaCl, and ethanol at concentrations of up to 50, 11–12, and 10–12%, respectively (4°C). Three strains had mycocinogenic activity. After fermentation of whey with selected yeast strains at 30°C for 2–3 days, the ethanol concentration was 4–5%.     

Sedimentation characteristics and effect of molasses concentration and medium supplementation on the ethanol productivity of an ethanol tolerant palm wineSaccharomyces

Summary Sedimentation rates ranging from 58.82 to 93.10% were recorded for six palm wineSaccharomyces yeast isolates. Isolates S, J and A₃, gave values of 90.00, 91.95 and 93.10% respectively compared to 81.54% for a standard lager beer yeast. Fermentation of unclarified molasses medium 10–30° Brix with isolated J gave ethanol productivity of 74.35-67.07%. Soyabean, groundnut or castor oil seed meal supplements significantly enhanced ethanol productivity in all the molasses media.     

Factors affecting oviposition and fecundity in the grain miteAcarus siro L. (Acarina:Acaridae), especially temperature and relative humidity

Oviposition and fecundity in the grain miteAcarus siro were studied at 5–30°C and 62.5–90% RH. At and above 20°C, 80% RH, mating and oviposition occurred soon after emergence, but at lower temperatures and humidities egg laying was progressively delayed from one to several days. Females needed to mate repeatedly in order to achieve maximum egg production, optimum conditions for which were 15°C, 90% RH, where total output per female averaged 435 with a maximum of 858. Oviposition rates were highest at higher temperatures, the mean daily rate at 20 and 25°C, 90% RH, rising to maximum levels of 28/29 eggs per female per day on day six.Oviposition followed clearly defined patterns, favourable conditions producing rapid increases in the mean daily oviposition rate to high peak levels reached at an early stage in the oviposition period. Less favourable conditions resulted in reduced outputs and lower, more uniform rates of egg laying. The mean oviposition period, varying with humidity, fell from 72–122 days at 5°C to 9–13 days at 30°C and the mean incubation period from 42–70 days at 5°C to 3–4 days at 30°C. Egg viability increased with increasing humidity but was little affected by temperature and unaffected by age of the female at time of oviposition.Males tended to live longer than females at most conditions; longevity—depending on humidity—averaging 13–15 days at 30°C and 129–175 days at 5°C. Adult life for females averaged 12–19 days at 30°C and 88–169 days at 5°C. An index of suitability, calculated from egg number, viability and duration of the egg stage and oviposition period, indicated that the most favourable conditions for oviposition and hatching were 20–25°C and 80–90% RH.     

Biology of the shore fly Scatella stagnalis in rockwool under greenhouse conditions

The development times and survival of immature stages in rockwool and the fecundity and longevity of adult Scatella stagnalis were determined and stage-specific life-tables constructed for the species at constant 20 and 25 °C and at a fluctuating temperature (23–34 °C, mean 28.5 °C). Development time from egg to adult decreased with temperature, being 15.9±0.1 days at 20 °C, 11.4±0.1 days at 25 °C and 10.1±0.2 days at fluctuating temperature with mean of 28.5 °C. The lower threshold for egg-to-adult development was 6.4±2.7 °C and the total quantity of thermal energy required to complete development was 212.8±.0 °C. The proportion of females in two populations studied was 0.521. High temperature increased the mortality of pupae from 7% (20 °C) and 10% (25 °C) to 29% at 28.5 °C. At 25 °C, female longevity was 15.5±0.7 days and fecundity 315±19 eggs/female (20.4 eggs/female/day). Males lived for 22.0±1.1 days. At constant 25 °C, the net reproductive rate was 126.1 female eggs/female, generation time was 18.4 days, the doubling time of the population 5.3 days, and the intrinsic rate of increase (r _m) 0.263 day^–1.     

Control of diapause in codling moth larvae

Diapause in a New Zealand strain of codling moth (Cydia pomonella Linnaeus [Lepidoptera: Olethreutidae]) was induced in larvae by photoperiods of 15 h or less. Once diapause had been initiated, it could not be terminated by any combination of conditions tested for at least 20 days after cocooning. In diapausing larvae a low rate of pupation occurred at 25 °C under a long day (18 h) photoperiod. A high rate of pupation was achieved under a long day regime when larvae were decocooned, and provided with apple as nourishment. Diapause could be terminated predictably in 94–100% of larvae by 1) conditioning at 15 °C and constant darkness for periods of 40–100 days, then 2) chilling at 2±2 °C and constant darkness for 20–50 days followed by 3) any post-chill condition periods at 25 °C, 18 h photoperiod. Complete diapause termination was achieved when 100 days conditioning was followed by 30 days or 50 days post-chill period. Under these conditions, 76% termination occurred in the post-chill period after 10 days, and 93% after 25 days.To terminate diapause in codling moth larvae, we recommend that a 100 days conditioning followed by 30 days chilling and 50 days post chilling periods be used.     

Production of poly-3-hydroxybutyrate by Bacillus sp. JMa5 cultivated in molasses media

A strain of Bacillus sp. coded JMa5 was isolated from molasses contaminated soil. The strain was able to grow at a temperature as high as 45°C and in 250 g/l molasses although the optimal growth temperature was 35–37°C. Cell density reached 30 g/l 8 h after inoculation in a batch culture with an initial concentration of 210 g/l molasses. Under fed-batch conditions, the cells grew to a dry weight of 70 g/l after 30 h of fermentation. The strain accumulated 25–35%, (w/w) polyhydroxybutyrate (PHB) during fermentation. PHB accumulation was a growth-associated process. Factors that normally promote PHB production include high ratios of carbon to nitrogen, and carbon to phosphorus in growth media. Low dissolved oxygen supply resulted in sporulation, which reduced PHB contents and dry weights of the cells. It seems that sporulation induced by reduced supply of nutrients is the reason that PHB content is generally low in the Bacillus strain.     

Isolation and improvement of a thermotolerant Saccharomyces cerevisiae strain

The ambient temperature is a drawback in industrial ethanol production in Jaffna due to heat killing of yeast during fermentation. Thus a search was initiated for thermotolerant organisms suitable for fermentation in hot climates. The screening of the best wild-type organisms was undertaken as the first step. Thermotolerant strains were selected from environments where there are chances of organisms being exposed to high temperature. The samples were enriched and screened for thermotolerant organisms which survived at 45 °C for 15 h. Among the yeast strains selected from different sources, thermotolerant strains with the capacity to withstand 45 °C for 15 h were found in samples collected from the compost heap and distillery environments. Three colonies from the distillery environment were selected for further studies and named p1, p2 and p3. Exponential phase (18 h) cultures of p1, p2 and p3 were subjected to 15 temperature treatment cycles (at 50 °C each for 3 h) and thermally adapted strains pt1, pt2 and pt3 were obtained, showing 100, 30 and 20% viability at 50 °C for 30 min respectively. The initial round of thermal adaptation cycles increased the duration of 100% viability from 20 h (p1) to 68 h (pt1) when incubated at 40 °C. Very little benefit was obtained when pt1 was treated with u.v. and ethyl methanesulphonate. The selected strain was identified and designated as Saccharomyces cerevisiae S1. The ethanol produced from 100 g glucose l^–1 by S. cerevisiae S1 was 46 g l^–1 (36 h), 38 g l^–1 (48 h) and 26 g l^–1 (48 h) at 40, 43 and 45 °C respectively in rich nutrient medium.     

Fed-batch alcoholic fermentation of sugar cane blackstrap molasses: influence of the feeding rate on yeast yield and productivity

Fed-batch ethanol fermentation tests of sugar cane blackstrap molasses were carried out at 32° C and ph 4.5–5.0, using pressed yeast as inoculum, and with no air supply. Two values of the fermentor filling-up time were adopted: 5 h and 7 h. The feeding rates obeyed equation F=F₀·^–K·t, with K equal to 0.0, 0.2, 0.4, 0.6 and 0.8 h^–1. The average yeast yields and the average yeast productivities increased up to 33% and 45%, respectively, while the ethanol yield (average=76%; standard deviation=4%) was practically unaffected when K increased from 0 to 0.8 h^–1. Correspondence to: E. Aquarone     

Thermotolerant single cell protein production byKluyveromyces marxianus var.marxianus

Summary Amino acid analyses were undertaken on single cell protein (SCP) produced by thermotolerant strains ofKluyveromyces marxianus var.marxianus grown on sugar cane molasses at 40°C. The maximum conversion of available sugars to biomass at 45°C was only 10.8% (g dry wt.·g^–1 total sugars). The amino acid composition of the SCP did not differ markedly from that reported for other yeast species.     

Effects of temperature and salinity on survival of toxigenicVibrio cholerae O1 in seawater

In 1991 and 1992, the Latin American epidemic strain of Vibrio cholerae O1 was isolated from ballast water, bilge water, and sewage taken from cargo ships docked in Mobile Bay, Alabama. The findings raised questions regarding the organism's ability to survive long-term aboard ships and to withstand the exchange of ballast at sea. The effects of temperature (6, 18, and 30°C) and salinity (8, 16, and 32 ppt) on survival of V. cholerae O1 strains C6706 and C6707 and a ballast water isolate in sterile seawater were determined. The ballast water isolate, which had a D-value (number of days required to produce a 1 log₁₀ reduction in colony-forming units per milliliter) of 240 days at 18°C, 32 ppt salinity, had the longest survival time. The range of D-values was 36–240 days at 18°C, 60–120 days at 30°C, and 5–20 days at 6°C. In sterile seawater short-term survival was temperature dependent, whereas long-term survival was salinity dependent. In raw seawater, survival time of the ballast water isolate was reduced to 12–27 days, implying the existence of biological influences. As also shown in our previous work, the organism appeared to be able to survive for several months under relatively stable conditions in ballast water aboard ships; however, viability may be reduced to only a few weeks after the organism is introduced into estuarine or marine environments. Correspondence to: Susan A. McCarthy.     

Effect of torpor on the water economy of an arid-zone marsupial,the stripe-faced dunnart (<Emphasis Type="Italic">Sminthopsis macroura</Emphasis>)

Metabolic rate and evaporative water loss (EWL) were measured for a small, arid-zone marsupial, the stripe-faced dunnart (Sminthopsis macroura), when normothermic and torpid. Metabolic rate increased linearly with decreasing ambient temperature (T_a) for normothermic dunnarts, and calculated metabolic water production (MWP) ranged from 0.85±0.05 (T_a=30°C) to 3.13±0.22 mg H₂O g^–1 h^–1 (T_a=11°C). Torpor at T_a=11 and 16°C reduced MWP to 24–36% of normothermic values. EWL increased with decreasing T_a, and ranged from 1.81±0.37 (T_a=30°C) to 5.26±0.86 mg H₂O g^–1 h^–1 (T_a=11°C). Torpor significantly reduced absolute EWL to 23.5–42.3% of normothermic values, resulting in absolute water savings of 50–55 mg H₂O h^–1. The relative water economy (EWL/MWP) of the dunnarts was unfavourable, remaining >1 at all T_a investigated, and did not improve with torpor. Thus torpor in stripe-faced dunnarts results in absolute, but not relative, water savings.     

Enhancement of biomass and fermentation activity of surplus brewers' yeast in a fed-batch process

The growth of surplus brewers' yeast in a fed-batch process was studied with the aim of increasing the fermentation activity of the yeast cells and of optimizing the growth conditions: 20 h cultivation at 30° C and pH 5.0–5.5 using beet molasses as substrate, with a regulated feeding rate, showed satisfactory results. Under the chosen conditions, the final amount of biomass increased more than fivefold, achieving a specific growth rate of 0.1 h^–1 and substrate yield coefficient of 0.54 g·g^–1. The increase in fermentation activity of yeast cells during cultivation correlated very well with the concentration of reduced glutathione, which increased from 1.2 to 2.7 mg·g^–1 (dry matter). At the same time the fermentation activity increased fivefold, which related to the nitrogen content of the yeast cells. Ethanol formation throughout the cultivation did not exceed 0.5 g·l^–1. Correspondence to: B. Strel     

Microbiological process for the removal of Cr(VI) from chromate-bearing cooling tower effluent

Summary A microbiological process using Pseudomonas mendocina was developed for the removal of Cr(VI) from cooling tower effluent. The process, when carried out in a 20 liter continuous stirred tank reactor removed 25–100 mg chromate/l in 4.5–8 h with >99.9% efficiency in the presence of sugarcane molasses as nutrient. The process could sustain wide variations in pH (6.5–9.5), temperature (25°C–40°C) and was unaffected by commonly used biocides.     

Cheese whey: an alternative growth and protective medium for<Emphasis Type="Italic"> Rhizobium loti</Emphasis> cells

Cheese whey (CW)-based growth medium efficiently protects Rhizobium loti cells during freezing and desiccation and can maintain their growth in a manner similar to that of traditional mannitol-based medium (YEM). The cheese-whey-based medium (CW) improved viability when used to re-suspend cell pellets kept at –20 °C and –80 °C and resulted in the survival of over 90% of the cells. Moreover, bacterial pellets obtained from cells grown in CW withstand desiccation better than cells grown in YEM. Survival was over 60% after 30 days at 4 °C. No differences were observed in nodulation efficiency between YEM-grown and CW-grown cells. Fast protein liquid chromatography (FPLC) protocols are presented for total protein profile analyses of sweet and acid cheese whey.In memoriam of Sylvio Cortina Vicepresident of Fundación COREPRO     

Synchronous Crepuscular Flight of Female Asian Gypsy Moths: Relationships of Light Intensity and Ambient and Body Temperatures

Female gypsy moths (Lymantria dispar) of Asian heritage studied in central Siberia and Germany exhibit a highly synchronous flight at dusk, after light intensity falls to about 2 lux. This critical light intensity sets the timing of flight behaviors independent of ambient temperature. Flight follows several minutes of preflight wing fanning during which females in Germany and those from a laboratory colony (derived from Siberian stock) raised their thoracic temperatures to 32–33°C at ambient temperatures of 19–22°C. Thoracic temperature of females in free flight exceeded the air temperature (19–22°C) by approximately 11–13°C. The duration of wing fanning was strongly dependent on ambient temperature. In Germany, where ambient temperatures at dusk ranged between 21 and 25°C, females wing fanned for only 2.1 ± 0.2 (SE) min; in the much colder temperatures prevalent at dusk in Bellyk, central Siberia (11–13°C), females spent 11.2 ± 0.6 min in preflight wing fanning. The majority (80%) of mated and even virgin females initiated flight during the evening of the day they eclosed. However, in Bellyk, a small proportion (12%) of females wing fanned for an extended time but then stopped, whereas others (8%) never wing fanned and, therefore, did not take flight. Females also were capable of flight when disturbed during the daylight hours in Germany where the maximal temperature was high (27–30°C), but not in Siberia, where temperatures peaked at only 17–19°C. However, Siberian females were able to propel themselves off the tree on which they were perched by executing several vigorous wing flicks when approached by the predaceous tettigoniid, Tettigonia caudata.     

Respiration of individual honeybee larvae in relation to age and ambient temperature

The CO₂ production of individual larvae of Apis mellifera carnica, which were incubated within their cells at a natural air humidity of 60–80%, was determined by an open-flow gas analyzer in relation to larval age and ambient temperature. In larvae incubated at 34 °C the amount of CO₂ produced appeared to fall only moderately from 3.89±1.57 µl mg^–1 h^–1 in 0.5-day-old larvae to 2.98±0.57 µl mg^–1 h^–1 in 3.5-day-old larvae. The decline was steeper up to an age of 5.5 days (0.95±1.15 µl mg^–1 h^–1). Our measurements show that the respiration and energy turnover of larvae younger than about 80 h is considerably lower (up to 35%) than expected from extrapolations of data determined in older larvae. The temperature dependency of CO₂ production was determined in 3.5-day-old larvae, which were incubated at temperatures varying from 18 to 38 °C in steps of 4 °C. The larvae generated 0.48±0.03 µl mg^–1 h^–1 CO₂ at 18 °C, and 3.97±0.50 µl mg^–1 h^–1 CO₂ at 38 °C. The temperature-dependent respiration rate was fitted to a logistic curve. We found that the inflection point of this curve (32.5 °C) is below the normal brood nest temperature (33–36 °C). The average Q₁₀ was 3.13, which is higher than in freshly emerged resting honeybees but similar to adult bees. This strong temperature dependency enables the bees to speed up brood development by achieving high temperatures. On the other hand, the results suggest that the strong temperature dependency forces the bees to maintain thermal homeostasis of the brood nest to avoid delayed brood development during periods of low temperature.Abbreviations m body mass - R rate of development or respiration - T_I inflexion point of a logistic (sigmoid) curve - T_L lethal temperature - T_O temperature of optimum (maximum) developmentCommunicated by G. Heldmaier     

Isolation of thermotolerant,fermentative yeasts growing at 52°C and producing ethanol at 45°C and 50°C

Five thermotolerant, alcohol-producing yeast cultures were isolated from samples obtained from India. Two were identified as ofKluyveromyces marxianus. All five grew on plate-cultures up to 52°C, with maximum growth rates in liquid culture at 40°C. All produced relatively high alcohol concentrations: 5.7 to 7.0% (w/v) at 45°C and 5.0 to 5.5% (w/v) at 50°C when growing on 14.0% (w/v) glucose. All five isolates fermented diluted molasses containing 16.0% (w/v) total sugars, producing 5.6 to 6.0% (w/v) alcohol concentrations. Supplementing the molasses with P, K, Mg and Mn resulted in a 13 to 20% increase in alcohol production at 40°C. The maximum amounts of alcohol produced on supplemented molasses were 7.5 to 8.0 and 6.5 to 7.0% (w/v) at 37°C and 40°C, respectively.     

Studies on the rust,Maravalia cryptostegiae,a potential biological control agent of rubber-vine weed,Cryptostegia grandiflora (Asclepiadaceae: Periplocoideae), in Australia,II: Infection

The parameters which govern infection of rubber-vine weed by the rustMaravalia cryptostegiae were investigated. The infection process, from appressorial formation to sporulation, is described and illustrated. Uredinioid teliospores have an optimum temperature range for germination at 22–27 °C, both in vitro and in vivo. However, germination on the rubber-vine leaf was more than double (81–92%) that in the absence of the host, and appressoria were formed only in vivo. An optimum temperature of 20–22°C and a dew period of 12 hours or more gave the highest level of infection as measured by sporulation density. The latent period from inoculation to pustule formation decreased with increasing temperature; the shortest period (8–11 days) being recorded at 25–27°C. At the lower temperatures (18°C), this was significantly extended (19–21 days). Four successive inoculations significantly reduced plant height and dry weight, although a compensatory growth flush occurred after the third inoculation. The addition of cryoprotectants had a negative affect on spore viability and subsequent infectivity. Cooling dry spores to –196°C at the rate of 10°C min^–1 gave the best results, with high germination (93–65%) up to 8 days after thawing.     

Cold hardiness of the green gemmules of Spongilla lacustris L. (Porifera: Spongillidae)

Most green gemmules of Spongilla lacustris survived enclosure in ice at –20 °C for up to 30 days; however, their rate of germination at 20 °C was less rapid than that of control gemmules. The length of time spent at low temperature had little effect on gemmule survival. In contrast, repeated cooling to –20 °C and warming to 4 °C led to a progressive decline in gemmule viability. These results indicate that cold injury occurs primarily during transitions between high and low temperatures.     

Development of Apheliuus mali an endoparasitoid of woolly apple aphid, Eriosoma lanigerum at different temperatures

Developmental times for both sexes of Aphelinus mali (Haldeman) (Hymenoptera: Aphelinidae) an endoparasitoid of woolly apple aphid, Eriosoma lanigerum (Hausmann) (Hemiptera: Pemphigidae) were studied at 13, 15, 18, 20, 25 and 30°C and compared with those of E. lanigerum. Mean developmental times ranged from 11.7–53.3 days for males, 11.8–55.4 days for females and 11.8–54.4 days for both sexes combined and were significantly inversely related to temperature. A good linear model fit (r²>0.99) between developmental rate and temperature in the range 13–30°C was observed. Results indicate significant differences between developmental times of males and females at 13, 18, 20, and 25°C but no differences at 15 and 30°C. The notional developmental threshold was 8.3°C for both males and females. Compared with its host, A. mali has a higher lower developmental threshold. An average of 252.8, 256.7 and 254.8 degree-days (DD) above the lower threshold were required by A. mali to complete development from time of oviposition to adult emergence for males, females and both sexes combined, respectively. Field experimentation also indicated that the developmental time of A. mali lags significantly behind that of its aphid host throughout the year. These findings are discussed in relation to its status as a biological control agent for E. lanigerum.