Inhibition of hyphal development and kill of Candida albicans blastospores by noxythiolin in vitro

The cidal activity of the antimicrobial agent, noxythiolin, was investigated against a laboratory strain and a fresh isolate of Candida albicans. The order of resistance to noxythiolin was hyphal form (isolate) ≥ 25°C-grown blastospores (isolate) ≥ 37°C-grown blastospores (isolate) > laboratory strain blastospores. Noxythiolin activity was superior to that of 'equivalent' formaldehyde concentrations. Mycelial transformation in C. albicans was examined by light and scanning electron microscopy and measured in terms of percentage germination and hyphal extension. Noxythiolin, 2·5%, in contact for 30 min prevented germination of the blastospore population whereas the decomposition products, formaldehyde and N-methylthiourea, showed no appreciable effect in the expected concentrations. The implications of these results are discussed in relation to the observed clinical efficacy of noxythiolin.     

Inhibition of hyphal development and kill of Candida albicans blastospores by noxythiolin in vitro

The cidal activity of the antimicrobial agent, noxythiolin, was investigated against a laboratory strain and a fresh isolate of Candida albicans. The order of resistance to noxythiolin was hyphal form (isolate) greater than or equal to 25 degrees C-grown blastospores (isolate) greater than 37 degrees C-grown blastospores (isolate) greater than laboratory strain blastospores. Noxythiolin activity was superior to that of 'equivalent' formaldehyde concentrations. Mycelial transformation in C. albicans was examined by light and scanning electron microscopy and measured in terms of percentage germination and hyphal extension. Noxythiolin, 2.5%, in contact for 30 min prevented germination of the blastospore population whereas the decomposition products, formaldehyde and N-methylthiourea, showed no appreciable effect in the expected concentrations. The implications of these results are discussed in relation to the observed clinical efficacy of noxythiolin.     

Effect of Temperature on the Formation of Microsclerotia of Verticillium dahliae

Microsclerotium formation by six isolates of Verticillium dahliae was studied at different temperatures both in vitro and in Arabidopsis thaliana . In vitro mycelial growth was optimal at 25°C, but microsclerotium formation was greatest at 20°C (two isolates) or 15–20°C (one isolate). Seedlings of A. thaliana were root-dipped in a conidial suspension, planted, and either placed at 5, 10, 15, or 25°C, or left at 20°C until the onset of senescence, after which some of the plants were placed at 5, 10, 15, or 25°C. The amount of microsclerotia per unit of shoot weight was assessed in relation to isolate and temperature. The optimal temperature for production of microsclerotia was 15–25°C. Two isolates each produced about 10 times more microsclerotia than each of the other four isolates. For these isolates, high R ²_adj.-values of 0.77 and 0.66 were obtained, with temperature and its square as highly significant (P   < 0.001) independent variables. R ²_adj.-values for the other isolates varied between 0.28 and 0.39. Moving plants to different temperatures at the onset of senescence led to microsclerotial densities that were intermediate between densities on plants that had grown at constantly 20°C and plants grown at other temperatures. This suggests that vascular colonization rate and rate of microsclerotium formation are similarly affected by temperature. The senescence rate of plants appeared unimportant except for plants grown at 25°C, which showed the highest amounts of microsclerotia per unit of plant weight in the most rapidly senescing plants.     

Factors affecting the adhesion and uptake of an oculo-genital strain of Chlamydia trachomatis to McCoy cells

Abstract We have studied adhesion and uptake of C. trachomatis serovar E in McCoy cells under various infection conditions. Adhesion and uptake of chlamydiae was completed about 3 h after the initiation of stationary infection at 37°C, but ingestion of cell membrane-attached organisms was finished within 0.5 h at 37°C. Reincubated chlamydiae, not attached after 3 h at 37°C, attached readily to fresh McCoy cell monolayers, but to a lesser extent than the original inoculum. Our results indicate that the lack of further attachment after 3 h incubation at 37°C under stationary infection conditions has complex causes, involving both host cell and parasite. Centrifugation did not affect the uptake of chlamydiae already bound to the cell membrane, suggesting that the uptake phase of C. trachomatis serovar E by McCoy cells is unaffected by centrifugation.     

Isolation and characterization of nisin-producing Lactococcus lactis subsp. lactis from bean-sprouts

Bacterial isolates from bean-sprouts were screened for anti- Listeria monocytogenes bacteriocins using a well diffusion method. Thirty-four of 72 isolates inhibited the growth of L.monocytogenes Scott A. One, HPB 1688, which had the biggest inhibition zone against L.monocytogenes Scott A, was selected for subsequent analysis. Both ribotyping and DNAsequencing of 16S ribosomal RNA gene demonstrated that the isolate was Lactococcus lactis subsp. lactis . Polymerase chain reaction and nucleotide sequencing revealed that thegenomic DNA of the bean-sprout isolates contained a nisin Z structural gene. In MRS broth,bean-sprout isolate HPB 1688 survived at 3–4·5°C for at least 20 d, grew at 4°Cand produced anti-listerial compoundsat 5°C. When co-cultured with L. monocytogenes in MRS broth, the isolate inhibited thegrowth of L. monocytogenes at 4°C after 14d and at 10°C after 2 d. When co-inoculatedwith 10²cells g⁻¹ of L.monocytogenes on fresh-cut ready-to-eat Caesar salad, L. lactis subsp. lactis (10⁸cells g⁻¹) was able to reduce the number of L. monocytogenes by 1–1·4 logs after storage for 10 d at 7° and 10°C. A bacteriocin-producing Enterococcusfaecium was also able to reduce the numbers of L. monocytogenes onCaesar salad, butdid not act synergistically when co-inoculated with L. lactis subsp. lactis .     

Effects of Temperature on Germination of Peronospora parasitica Conidia and Infection of Brassica oleracea

The effect of temperature on germination of a South African isolate of Peronospora parasitica , and infection of Brassica oleracea was studied. The optimum condition for germination was 20° C at 100% relative humidity. The percentage germination obtained was 80–98% and 70–80% between 15 and 25° C at 100% relative humidity, after a 12 and 6h incubation period, respectively. Optimum temperature for germ tube growth was also 20° C. The temperature range for maximum infection of seedlings of a highly susceptible cabbage cultivar and subsequent disease development in vitro was 15–25° C and 90–100% infection was achieved after 48 h of incubation. At<15°C and 26–30° C infection percentage was decreased to 40–50% and 35–40%, respectively. No disease incidence was recorded at temperatures above 35° C. A scanning electron microscope study of the infection process showed that penetration of cotyledons by germ tubes was mostly via stomata and occasionally directly through the cuticle. Results are discussed in relation to the need for future studies of P. parasitica in South Africa.     

Effects of development, temperature and salinity on metabolism in eggs and yolk-sac larvae of milkfish, Chanos chanos (Forsskål)

Oxygen uptake rates and yolk-inclusive dry weiGhts were measured during the egg and yolk-sac larval stages of milkfish, Chanos chanos (Forsskal). Oxygen uptake by eggs and yolk-sac larvae was measured to assess the effects of four salinities (20,25,30,35 ppt) at 28°C. The effects of three temperatures (23,28,33°C) on oxygen uptake by yolk-sac larvae were determined at a salinity of 35 ppt. Dry weights were measured throughout embryonic development at 28°C and the yolk-sac stage at 23.28 and 33°C.
Oxygen uptake rates of eggs increased more than fivefold during embryogenesis (0.07±0.03 to 0.40 ± 03 μl O₂ egg ⁻¹ h ⁻¹;blastula to prehatch stage). Larval oxygen uptake did not change with age but was affected by rearing temperature (0.33 ± 0.08, 0.44 ± 0.07 and 0.63 ± 0.13 μl O₂ larva ⁻¹ h⁻¹ at 23, 28 and 33°C, respectively; Q₁₀= 1.93). Acute temperature changes from 28 to 33°C caused significant increases in oxygen uptake by embryos (Q ₁₀= 1.69–3.58) and yolk-sac larvae (Q ₁₀=2.55). Salinity did not affect metabolic rates.
Dry weight of eggs incubated at 28°C decreased 13% from fertilization to hatching. Incubation temperatures from 23–33°C did not affect dry weights at hatching. Rearing temperatures significantly affected the rate of larval yolk absorption (Q ₁₀= 2.25).     

Oxygen uptake by rainbow trout blood, Salmo gairdneri

Oxygen uptake rate by rainbow trout red cells was measured in the temperature range 5° to 40° C. Between 5° and 20° C there was a steady increase in rate and at 20° C a maximum rate of 2.5 μl O₂/min/ml red cells occurred, followed by a decline in the rate as the temperature increased. A morphological investigation of the red cell was conducted and possible functions of the nucleus in fish red blood cells are discussed.     

Thermolabile ribonucleases from antarctic psychrotrophic bacteria: detection of the enzyme in various bacteria and purification from Pseudomonas fluorescens

Abstract Thirteen terrestrial psychrotrophic bacteria from Antarctica were screened for the presence of a thermolabile ribonuclease (RNAase-HL). The enzyme was detected in three isolates of Pseudomonas fluorescens and one isolate of Pseudomonas syringae . It was purified from one P. fluorescens isolate and the molecular mass of the enzyme as determined by SDS-PAGE was 16 kDa. RNAase-HL exhibited optimum activity around 40°C at pH 7.4. It could hydrolyse Escherichia coli RNA and the synthetic substrates poly(A), poly(C), poly(U) and poly(A-U). Unlike the crude RNAase from mesophilic P. fluorescens and pure bovine pancreatic RNAase A which were active even at 65°C, RNAase-HL was totally and irreversibly inactivated at 65°C.     

Giraudia sphacelarioides (Phaeophyceae) at the Canary Islands and in Danish waters: a study in ecotypic differentiation and its biogeographical implications

Comparative culture studies on isolates from Lanzarote (Canary Islands) and from Danish waters of Giraudia sphacelarioides show that temperature plays the key role to determine its geographical distribution. Experiments show that the upper lethal temperature is the same for both isolates 26.5—-31.1°C for microthalli, and formation of macrothalli is inhibited between 23.2 and 26.5°C. The lower lethal temperature for both micro- and macrothalli in the Lanzarote isolate is between 10.8° C and 7.6°C, while the Danish isolate survives 54°C. The microthallus is thus the more persistent stage in relation to temperature, while the macrothalli occur only when more suitable conditions exist in nature. In the North Atlantic Ocean G. sphacelarioides is distributed over approximately 30° latitudes with severe winter temperatures especially in Danish waters. This distribution can only be explained by ecotypic differentiation. In Danish waters salinity decreases from c. 30 ‰ in the northern Kattegat to below 8 ‰ in the inner parts and therefore becomes an important factor for the immigration of species into the Baltic Sea. Combined salinity (30–20–10–5 ‰) and temperature (4 to 30°C) experiments show that the Lanzarote isolate only grows at 20 ‰ in an attenuated temperature interval and at 10 ‰ only fertile microthalli are observed, however, with limited liberation of swarmers. The Danish isolate grows and reproduces also at salinities from 20–5 ‰ both as microand macrothalli, but in a still more attenuated temperature interval, and the morphology of the macrothalli becomes increasingly aberrant vs. decreasing salinity. This is in agreement with the fact that G. sphacelarioides is reported from the northern part of Kattegat with salinity from 28—-25 ‰, and it is only recorded a few times in the southern part of Kattegat and limited by the 10 ‰ isohaline.     

Shifts in the optimal temperature for nocturnal CO2 uptake caused by changes in growth temperature for cacti and agaves

When the day/night air temperatures were raised from 10°C/10°C to 30°C/30°C, the optimal tempearture for nocturnal CO² uptake by six species of cacti and three species of agave shifted from an average of 12°C to an average of 20°C. The maximum rate of CO² uptake was higher for Agave americana at the higher ambient temperature, lower for A. deserti , and much lower for A. utahensis , consistent with the relative mean temperatures of their native habitats. For the cactus Coryphantha vivipara , which had the greatest temperature shift observed (13°C), the halftime was 8 days for the upward shift and 4 days for the downward shift. The halftimes for the comparable shifts averaged 1.6 days for three other species of cacti and less than 1 day for two agave species. The shifts in the optimal temperature for nocturnal CO² uptake were in response to changes in nighttime temperature, at least for C. vivipara , and reflected temperature responses of both the stomates and the chlorenchyma.     

The influence of some ecological factors on the metabolism—temperature curve of the larvae of Limnephilus rhombicus (Trichoptera, Limnephilidae)

SUMMARY. The oxygen uptake of Limnephilus rhombicus , a typical inhabitant of lentic waters, was measured in three different experimental situations: in still water with a plastic mesh as a substrate, in stirred water with a plastic mesh and in still water without any substrate. Water flow caused a small increase in oxygen uptake at all temperatures but principally above 15°C. The presence or absence of the plastic substrate had little influence on oxygen uptake; L. rhombicus can be active over the whole range of temperatures studied but its activity is always small. Its metabolism—temperature curve stands out in having a plateau between 5 and 10°C, with a steeper part between 10 and 20°C, the slope of which varies according to the conditions. Maximum oxygen uptake at high temperatures was relatively low (1235 ± 153 μl O₂g dry wt-1h-1 at 25°C).
The results are compared with those obtained previously for the larvae of Microptema testacea which inhabit flowing zones in brooks. L. rhombicus larvae appear both as cold-water animals and as animals from highly variable biotopes. They are in fact very ubiquitous and occupy all lentic biotopes from cold and mountainous districts to temperate lowlands.     

Isolation and characterization of refuse methanogens

Four mesophilic, irregular, rod-shaped methanogenic bacteria were isolated from decomposing refuse recovered from laboratory-scale reactors and a municipal solid waste landfill. H₂/CO₂ was the only substrate on which the isolates could grow in a complex medium. Isolates grew between either 25° or 30° and 45°C and between pH 6 and 8. One isolate exhibited growth at pH 5. Growth of each isolate was enhanced by yeast extract and inhibited by anaerobic sewage sludge supernatant fluid. No isolate showed greater than 25% lysis on exposure to 1% sodium dodecyl sulphate (SDS) for 24 h, as is typical of methanogens with a proteinaceous cell wall. The physiological traits of the methanogens isolated here are similar to many previously characterized isolates.     

Aerobic growth and survival of Campylobacter jejuni in food and stream water

When 40 Campylobacter jejuni isolates from human clinical cases, raw chicken and water were tested, 29 (72·5%) could be adapted to grow on nutrient agar under aerobic conditions. Once adapted, these isolates could grow on repeated aerobic subculture. An aerobically-grown Camp. jejuni isolate survived almost as well as the same isolate grown microaerophilically in sterile chicken mince at 5 °C, and survival of a cocktail of Camp. jejuni isolates under both atmospheres was comparable at 25 °C. However, at 37 °C, the decline in numbers of the aerobically-grown cells was greater. Survival of cells on chicken nuggets was poorer than in chicken mince. In filter-sterilized stream water incubated aerobically at 5 °C, survival of inocula grown under different atmospheres was again similar, but slightly better with the microaerophilically-grown cells. Adaptation to aerobic growth was not found to enhance survival under aerobic conditions.     

Nisin induces changes in membrane fatty acid composition of Listeria monocytogenes nisin-resistant strains at 10°C and 30°C

Listeria monocytogenes isolates resistant to 10⁵ IU ml^-1 nisin were obtained at 30°C (NR30) and at 10°C (NR10). Nisin prolonged the lag phase of isolate NR30 at 10°C. Isolates NR30 and NR10 did not produce a nisinase. Protoplasts of isolate NR30 were unaffected by exposure to nisin. The fatty acid composition from the wild-type strain and NR isolates was determined. As expected, temperature-induced differences in the C15/C17 fatty acid ratios were found. Growth of the NR strains in the presence of nisin resulted in significantly different C15/C17 ratios and a significant increase in the percentage of C16:0, C16: 1, C18:0 and C18: 1 fatty acids at 10°C and 30°C. Both the NR10 and NR30 isolates had similar growth rates at low temperatures, but these were slower than the wild-type strain. These results indicate that 'nisin resistance'is an environmentally defined phenotype and that nisin induces changes in the fatty acid composition of the membrane in L. monocytogenes nisin-resistant isolates regardless of the growth temperature.     

A thermotolerant and high acetic acid-producing bacterium Acetobacter sp. I14–2

A thermotolerant bacterium with high production of acetic acid was isolated from spoiled banana in Taiwan. The isolate, I14–2 ,was considered to be an Acetobacter sp. according to phenotypic and chemotaxonomic characteristics. Optimal cultural conditions for Acetobacter sp. I14–2 to produce acetic acid were studied under cultivation in a medium containing 2 mg l⁻¹ acetic acid and 5% ethanol at 30 °C. Acetic acid productivity by Acetobacter sp. I14–2 was almost two and three times the amount produced by Acet. aceti IFO3283 and Acetobacter sp. CCRC 12326, respectively. The isolate retained 22% residual acetic acid-producing activity after 3 d incubation in a medium containing 8% ethanol, and produced acetic acid in a medium containing 10 g l⁻¹ acetic acid. This bacterium is thermotolerant and retained 97% and 68% of acetic acid-producing activity after 3 d incubation at 35 °C and 37 °C, respectively, compared with that when incubated at 30 °C.     

Decrease in adherence of bacteria and yeasts to human mucosal epithelial cells by noxythiolin in vitro

Adherence of buccal and vaginal isolates of Candida albicans to buccal epithelial cells and the adherence of urine isolates of Escherichia coli and Staphylococcus saprophyticus to uroepithelial cells was quantified by light microscopy. The antimicrobial agent noxythiolin reduced the adherence of these micro-organisms in both exponential and stationary growth phases. Adherence of both the blastospore and pseudohyphal forms of C. albicans was reduced. Treatment of epithelial cells and/or micro-organisms with noxythiolin resulted in decreased adherence. No anti-adherence effect was observed with formaldehyde and N-methylthiourea, the degradative products of noxythiolin.     

Proteolytic activity of rumen fungi belonging to the genera Neocallimastix and Piromyces

Proteolytic activity of two rumen fungal isolates Neocallimastix sp. strain N1 and Piromyces sp. strain P1 was examined. Proteases are active between pH 6.5 and 9.0 with maximum at 7.9 for isolate N1 and between 6.5 and 10.5 with maximum at 8.8 for isolate P1. Proteolytic activity increased as temperature increased until 50°C and a sudden decrease at 60°C was observed in both isolates. EDTA, 1,10-phenanthroline, p -chloromercuribenzoate (PCMB), merthiolate and phenylmethylsulphonyl fluoride (PMSF) were effective proteolytic inhibitors against both isolates.     

Effect of temperature, salinity, and dissolved oxygen concentration upon respiratory activity of the larva of Philanisus plebeius (Trichoptera)

Records of oxygen uptake of marine larvae of Philanisus in different salinities and at different temperatures show that a maximum is reached in normal sea water at 25°C. A close relation exists between oxygen uptake and the rhythm of ventilatory movements, leading to the conclusion that changes in oxygen uptake are affected by alteration in the frequency rather than the amplitude of the ventilatory movements. Ventilation of the case is stimulated by a fall in oxygen tension but inhibited at low oxygen levels. The ventilatory efficiency of Philanisus in saturated sea water at 25°C is about 26 per cent.     

Klebsiella pneumoniae haemolysin adsorption to red blood cells

It was necessary to incubate the Klebsiella pneumoniae haemolysin with erythrocytes at 37°C to produce the whole lytic action. The amount of attached klebolysin at 4°C increased as its concentration in the medium increased, until the erythrocyte surface was saturated. Treatment with 2-mercaptoethanol was necessary to permit the adsorption; it was inhibited by low concentrations of cholesterol. Klebsolysin was immunogenic and its antiserum neutralized its own haemolytic effect and streptolysin O. Anti-streptolysin serum also neutralized klebsolysin. Streptolysin O attachment to erythrocytes impeded the posterior klebolysin adsorption in the same way that klebsolysin adsorption interfered with streptolysin O attachment.