Factors regulating excystment of Alexandrium in Puget Sound,WA, USA

Factors regulating excystment of a toxic dinoflagellate in the genus Alexandrium were investigated in cysts from Puget Sound, Washington State, USA. Experiments were carried out in the laboratory using cysts collected from benthic seedbeds to determine if excystment is controlled by internal or environmental factors. The results suggest that the timing of germination is not tightly controlled by an endogenous clock, though there is a suggestion of a cyclical pattern. This was explored using cysts that had been stored under cold (4 °C), anoxic conditions in the dark and then incubated for 6 weeks at constant favorable environmental conditions. Excystment occurred during all months of the year, with variable excystment success ranging from 31–90%. When cysts were isolated directly from freshly collected sediments every month and incubated at the in situ bottom water temperature, a seasonal pattern in excystment was observed that was independent of temperature. This pattern may be consistent with secondary dormancy, an externally modulated pattern that prevents excystment during periods that are not favorable for sustained vegetative growth. However, observation over more annual cycles is required and the duration of the mandatory dormancy period of these cysts must be determined before the seasonality of germination can be fully characterized in Alexandrium from Puget Sound. Both temperature and light were found to be important environmental factors regulating excystment, with the highest rates of excystment observed for the warmest temperature treatment (20 °C) and in the light.     

Inhibition of Bacillus licheniformis spore growth in milk by nisin, monolaurin, and pH combinations

The effects of nisin and monolaurin, alone and in combination, were investigated on Bacillus licheniformis spores in milk at 37 degrees C. In the absence of inhibitors, germinated spores developed into growing vegetative cells and started sporulation at the end of the exponential phase. In the presence of nisin (25 IU ml-1), spore outgrowth was inhibited (4 log10 reduction at 10 h). Regrowth appeared between 10 and 24 h and reached a high population level (1.25 x 10(8) cfu ml-1) after 7 d. Monolaurin (250 micrograms ml-1) had a bacteriostatic effect during the first 10 h but thereafter, regrowth occurred slowly with a population level after 7 d (4 x 10(5) cfu ml-1) lower than that of nisin. Different combined effects of nisin (between 0 and 42 IU ml-1), monolaurin (ranging from 0 to 300 micrograms ml-1), pH values (between 5.0 and 7.0) and spore loads (10(3), 10(4), 10(5) spores ml-1) were investigated using a Doehlert matrix in order to study the main effects of these factors and the different interactions. Results were analysed using the Response Surface Methodology (RSM) and indicated that nisin and monolaurin had no action on spores before germination; only pH values had a significant effect (P < or = 0.001), i.e. spore count decreased as the pH value increased in relation to germination. Sublethal concentrations of nisin (30 IU ml-1) and monolaurin (100 micrograms ml-1) in combination acted synergistically on outgrown spores and vegetative cells, showing total inhibition at pH 6.0, without regrowth, within 7 d at 37 degrees C.     

Relationship of sporulation, enterotoxin formation, and spoilage during growth of Clostridium perfringens type A in cooked chicken

Sporulation and enterotoxin formation were determined for 17 strains of Clostridium perfringens type A in autoclaved chicken dark meat and in Duncan-Strong sporulation medium. The mean numbers of heat-resistant spores detected after 24 h at 37 degrees C were log10 1.13 to log10 7.64/ml in Duncan-Strong medium and log10 4.93 to log10 6.59/g in chicken. Of 17 strains, 7 formed enterotoxin in Duncan-Strong culture supernatant (1.0 to 60 microgram/ml) and 8 produced enterotoxin in chicken (0.21 to 24 microgram/g). Additional studies with chicken were conducted with C. perfringens NCTC 8239. With an inoculum of 10(6) cells per g, greater than log10 7.99 vegetative cells per g were detected by 4 h in chicken at 37 degrees C. Heat-resistant spores occurred by 4 and 6 h and enterotoxin occurred by 8 and 6 h in autoclaved chicken dark meat and barbecued chicken drumsticks, respectively. Enterotoxin was detected in autoclaved dark meat after incubation at 45 degrees C for 1.5 h followed by 37 degrees C for 4.5 h, but not after incubation at 45 degrees C for 1.5 to 8 h. With an inoculum of 10(2) cells per g in oven-cooked or autoclaved chicken, greater than log10 8.00 vegetative cells per g were detected by 6 to 8 h at 37 degrees C, heat-resistant spores were detected by 8 h, and enterotoxin was detected by 12 h. A statistical analysis of odor determinants of chicken after growth of C. perfringens indicated that, at the 95% confidence level, the product was considered spoiled (off or unwholesome odor) by the time spores or enterotoxin were formed.     

Relationship of sporulation, enterotoxin formation, and spoilage during growth of Clostridium perfringens type A in cooked chicken.

Sporulation and enterotoxin formation were determined for 17 strains of Clostridium perfringens type A in autoclaved chicken dark meat and in Duncan-Strong sporulation medium. The mean numbers of heat-resistant spores detected after 24 h at 37 degrees C were log10 1.13 to log10 7.64/ml in Duncan-Strong medium and log10 4.93 to log10 6.59/g in chicken. Of 17 strains, 7 formed enterotoxin in Duncan-Strong culture supernatant (1.0 to 60 microgram/ml) and 8 produced enterotoxin in chicken (0.21 to 24 microgram/g). Additional studies with chicken were conducted with C. perfringens NCTC 8239. With an inoculum of 10(6) cells per g, greater than log10 7.99 vegetative cells per g were detected by 4 h in chicken at 37 degrees C. Heat-resistant spores occurred by 4 and 6 h and enterotoxin occurred by 8 and 6 h in autoclaved chicken dark meat and barbecued chicken drumsticks, respectively. Enterotoxin was detected in autoclaved dark meat after incubation at 45 degrees C for 1.5 h followed by 37 degrees C for 4.5 h, but not after incubation at 45 degrees C for 1.5 to 8 h. With an inoculum of 10(2) cells per g in oven-cooked or autoclaved chicken, greater than log10 8.00 vegetative cells per g were detected by 6 to 8 h at 37 degrees C, heat-resistant spores were detected by 8 h, and enterotoxin was detected by 12 h. A statistical analysis of odor determinants of chicken after growth of C. perfringens indicated that, at the 95% confidence level, the product was considered spoiled (off or unwholesome odor) by the time spores or enterotoxin were formed.     

Life-history stages of natural bloom populations and the bloom dynamics of a tropical Asian ribotype of Alexandrium minutum

In 2015, a remarkably high density bloom of Alexandrium minutum occurred in Sungai Geting, a semi-enclosed lagoon situated in the northeast of Peninsular Malaysia, causing severe discoloration and contaminated the benthic clams (Polymesoda). Plankton and water samples were collected to investigate the mechanisms of bloom development of this toxic species. Analysis of bloom samples using flow cytometry indicated that the bloom was initiated by the process of active excystment, as planomycetes (>4C cells) were observed in the early stage of the bloom. Increase in planozygotes (2C cells) was evident during the middle stage of the bloom, coinciding with an abrupt decrease in salinity and increase of temperature. The bloom was sustained through the combination of binary division of vegetative cells, division of planozygotes, and cyst germination through continuous excystment. Nutrient depletion followed by precipitation subsequently caused the bloom to terminate. This study provides the first continuous record of in situ life-cycle stages of a natural bloom population of A. minutum through a complete bloom cycle. The event has provided a fundamental understanding of the pelagic life-cycle stages of this tropical dinoflagellate, and demonstrated a unique bloom development characteristic shared among toxic Alexandrium species in coastal embayments.     

Role of temporary cysts in the population dynamics of Alexandrium taylori (Dinophyceae)

Although temporary cyst stages are common in dinoflagellates,their role remains unclear. Every year Alexandrium taylori (Dinophyceae)forms dense patches (10⁶ cells l^-1) along La Fosca beach (Spain,northwest Mediterranean), which last for 2 months (July, August).One of the characteristics of the life history of A. tayloriis the shift from a vegetative motile stage to non-motile temporarycysts. Here we present the temporal changes in the abundanceof temporary cysts in sediments and their in situ encystmentand excystment rates. The in situ encystment rate of temporarycysts from the water column to the sediment ranged from 1.8x 10⁶ to 4.4 x 10⁶ cysts m^-2 day^-1, whereas the excystment ratewas between 0.9 x 10⁶ to 2.7 x 10⁶ cysts m^-2 day^-1 during thebloom period. Some of the temporary cysts in the sediment tookmore than 1 day to produce vegetative cells and remained viablefor at least 4 days. We propose that temporary cyst formationin this species is a tool for reducing population losses. Theproduction of temporary cysts can be an advantage since partof the population is stored in the sediments.     

In vivo development of Echinostoma malayanum Leiper, 1911 with notes on effects of population density, chemical composition and pathogenicity and in vitro excystment of the Metacercaria (Trematoda: Echinostomatidae)

In vivo development of Echinostoma malayanum Leiper, 1911 was studied in white rats and the developmental process was arbitrarily divided into four stages: organogeny, vitellogenesis, formation of Mehlis' gland complex and cirrus sac, and oviposition. The percentage of development was 86-94. Population density affected the prepatent period of flukes and the normal prepatent period of 13-16 days was altered to 18-23 days in infection with 500-800 flukes. The majority of flukes in heavy infection were undersized and in the immature stage of development at patency. Data from chemical analysis of flukes revealed that protein, lipids, calcium and ash decreased quantitatively in flukes from higher population densities but no such change was observed as regards glycogen. Pathological changes in the rat intestine included lysis and destruction of mucosa, increased activity of goblet cells, oedematous and reticulated appearance of lamina propria and slight to moderate hyperplasia of epithelial cells. The metacercariae excysted in the medium containing trypsin plus sodium cholate an pepsin, though not essential for a high percentage of excystment, affected the rate. The reductant sodium dithionite substantially enhanced the rate and percentage of excystment. Excystation was optimal at pH 8, and 42 degrees C was more effective than 39 degrees C.     

Seawater temperature measured at the surface and at two depths (7 and 12 m) in one coral reef at Culebra Bay, Gulf of Papagayo, Costa Rica

Superficial seawater temperature (SST) and at two depths (7 and 12 m) were measured non-continuously during the study of the corals and coral reefs of Culebra Bay (1993-1996). SST showed seasonal variations of approximately 4 degrees C. The highest average temperatures (27.0 +/- 0.1, range 23-29 degrees C) were during the rainy season from May to November and the lowest (22.9 +/- 0.3 degrees C, 15.5-29 degrees C) during the dry season from December to April. Cold fronts with 2-3 degrees C differences in SST frequently pass into the bay and remain there for several hours according to the tidal cycles. Differences of approximately 3 degrees C between SST and the bottom (5-10 m depth) were usually found, particularly at locations where bottom topography and tidal circulation produced tidal bores. The average temperatures recorded by data loggers placed at 7 and 12 m depth on a coral reef at the outer shores of Culebra Bay, were 27.1 +/- 0.02 degrees C (20.5-31.6 degrees C) and 25.8 +/- 0.04 (9.9-31.5 degrees C) respectively. The seasonal pattern of higher and lower temperatures corresponds respectively to the rainy and dry season of the northern Pacific coast of Costa Rica. Water temperature at 12 m was < 14 degrees C for some hours during an upwelling event whilst minimum temperatures at 7 m were > or = 22 degrees C. Negative temperature anomalies coincided with an increase of the NE-E winds intensity and there is a lunar and tidal component which influence diumal variations of temperature. These results suggest that coral reefs built by branching species (e.g. Pocillopora spp.) in Culebra Bay could be limited by both the influence of cold fronts and by seasonal upwellings which affect negatively those coral species, as reported for other locations in the tropical eastern Pacific.     

Algal cyst dormancy: a temporal escape from herbivory

Many phytoplankton species form resting cysts and remain dormant for part of the year. The subsequent excystment is regulated by the external environment and internal maturation processes. Here we assessed the excystment of the dinoflagellates Ceratium hirundinella and Peridinium aciculiferum in relation to herbivores and temperature in laboratory and field studies. C. hirundinella, which has a grazer-resistant morphology, forms summer blooms, whereas P. aciculiferum, which is vulnerable to grazers, grows underneath the ice during winter. In our study, herbivore abundance, and thereby grazing pressure, was low during periods when water temperatures were low, and the abundance of P. aciculiferum was high. In the laboratory experiment, excystment of C. hirundinella occurred at high temperatures irrespective of whether zooplankton exudate was added or not, whereas at intermediate temperatures, excystment was lower if zooplankton exudate was added. Germination of P. aciculiferum cysts was lower in the presence of exudate from a zooplankton culture than in controls at all temperatures. Our studies suggest that dinoflagellates use the presence of zooplankton in addition to temperature as a cue to determine when to excyst. Consequently, not only abiotic factors, but also the composition of the food web, may determine succession and composition of phytoplankton communities.     

Cysts of Danish Gymnodinium nolleri Ellegaard et Moestrup sp. ined. (Dinophyceae): studies on encystment, excystment and toxicity

Life cycle dynamics of Gymnodinium nolleri Ellegaard et Moestrupsp. ined. were studied under different temperature and nutrientconditions. Five culture strains originating from cysts foundin Danish marine sediments were used for the experiments. Bothencystment and excystment were found to vary with temperature.Maximal encystment occurred at 22–28°C, with no cystsformed below 13°C or above 33°C. Cyst production wasslightly higher under phosphorus limitation than under combinednitrogen and phosphorus limitation. Maximal excystment occurredat 26.5°C with negligible excystment under 11°C andover 35°C. The resting period for cyst maturation was typically3–4 weeks. Cysts produced under both phosphorus and nitrogenlimitation were tested for paralytic shellfish poisoning (PSP)toxins by HPLC, and none were detected. It remains unclear whyvegetative cells of this species have not yet been recordedin plankton samples from Scandinavia, despite the widespreaddistribution of the cysts.     

Differential effects of temperature and starvation on induction of the viable-but-nonculturable state in the coral pathogens Vibrio shiloi and Vibrio tasmaniensis

We compared induction of the viable-but-nonculturable (VBNC) state in two Vibrio spp. isolated from diseased corals by starving the cells and maintaining them in artificial seawater at 4 and 20 degrees C. In Vibrio tasmaniensis, isolated from a gorgonian octocoral growing in cool temperate water (7 to 17 degrees C), the VBNC state was not induced by incubation at 4 degrees C after 157 days. By contrast, Vibrio shiloi, isolated from a coral in warmer water (16 to 30 degrees C), was induced into the VBNC state by incubation at 4 degrees C after 126 days. This result is consistent with reports of low-temperature induction in several Vibrio spp. A large proportion of the V. tasmaniensis population became VBNC after incubation for 157 days at 20 degrees C, and V. shiloi became VBNC after incubation for 126 days at 20 degrees C. Resuscitation of V. shiloi cells from cultures at both temperatures was achieved by nutrient addition, suggesting that starvation plays a major role in inducing the VBNC state. Our results suggest that viable V. shiloi could successfully persist in the VBNC state in seawater for significant periods at the lower temperatures that may be experienced in winter conditions, which may have an effect on the seasonal incidence of coral bleaching. For both species, electron microscopy revealed that prolonged starvation resulted in transformation of the cells from rods to cocci, together with profuse blebbing, production of a polymer-like substance, and increased membrane roughness. V. shiloi cells developed an increased periplasmic space and membrane curling; these features were absent in V. tasmaniensis.     

Induction of Synchronized Fissions in Telotrochidium henneguyi

SYNOPSIS. Cultures of Telotrochidium henneguyi , begun with logarithmic phase cells, were employed in an effort to produce synchronized fission by heat treatment. The cells tolerated a temperature range of 20–50 C; temperatures above 50 were lethal. When cells were exposed to a single shock for 30 min, 30–40 produced 0–50% encystment with total excystment after 10 min exposure to room temperature (heat shock range). No encystment occurred between 20–30 (intershock range). Encystment and excystment time varied directly with temperature between 40–50.
The most effective procedure for inducing synchronized fission consisted of 6 cycle program of 38/28 C (shock temperature/intershock temperature) administered for 15/15 (shock/intershock duration in min). Division indices (DI = cells dividing/total population X 100 =%) ranged from 12–66% with a mean of 37.25%. In control cells, division indices ranged from 2–20% with an average of 12%. Inferences from these independently derived findings are discussed.     

Effects of nisin and temperature on survival, growth, and enterotoxin production characteristics of psychrotrophic Bacillus cereus in beef gravy.

The presence of psychrotrophic enterotoxigenic Bacillus cereus in ready-to-serve meats and meat products that have not been subjected to sterilization treatment is a public health concern. A study was undertaken to determine the survival, growth, and diarrheal enterotoxin production characteristics of four strains of psychrotrophic B. cereus in brain heart infusion (BHI) broth and beef gravy as affected by temperature and supplementation with nisin. A portion of unheated vegetative cells from 24-h BHI broth cultures was sensitive to nisin as evidenced by an inability to form colonies on BHI agar containing 10 micrograms of nisin/ml. Heat-stressed cells exhibited increased sensitivity to nisin. At concentrations as low as 1 microgram/ml, nisin was lethal to B. cereus, the effect being more pronounced in BHI broth than in beef gravy. The inhibitory effect of nisin (1 microgram/ml) was greater on vegetative cells than on spores inoculated into beef gravy and was more pronounced at 8 degrees C than at 15 degrees C. Nisin, at a concentration of 5 or 50 micrograms/ml, inhibited growth in gravy inoculated with vegetative cells and stored at 8 or 15 degrees C, respectively, for 14 days. Growth of vegetative cells and spores of B. cereus after an initial period of inhibition is attributed to loss of activity of nisin. One of two test strains produced diarrheal enterotoxin in gravy stored at 8 or 15 degrees C within 9 or 3 days, respectively. Enterotoxin production was inhibited in gravy supplemented with 1 microgram of nisin/ml and stored at 8 degrees C for 14 days; 5 micrograms of nisin/ml was required for inhibition at 15 degrees C. Enterotoxin was not detected in gravy in which less than 5.85 log10 CFU of B. cereus/ml had grown. Results indicate that as little as 1 microgram of nisin/ml may be effective in inhibiting or retarding growth of and diarrheal enterotoxin production by vegetative cells and spores of psychrotrophic B. cereus in beef gravy at 8 degrees C, a temperature exceeding that recommended for storage or for most unpasteurized, ready-to-serve meat products.     

DNA, RNA, and protein biosynthesis in cells of Yersinia pseudotuberculosis at various cultivation temperatures

Y. pseudotuberculosis cells cultivated at temperatures of 37 degrees C and 8 degrees C were found to be capable of incorporating exogenic precursors into DNA, RNA and protein. The linear growth of thymidine incorporation occurred during 8 hours of cultivation at 37 degrees C, then the amount of the incorporated label decreased. At 8 degrees C the level of thymidine incorporation into DNA gradually increased for 80 hours and longer, but not reaching the level of incorporation observed at 37 degrees C. The incorporation of uridine into RNA of Y. pseudotuberculosis cells reached its maximum after 4 hours of cultivation at 37 degrees C, at a lower temperature of cultivation the incorporation of uridine into bacterial cells was almost linear, though slower, and lasted for 20 hours. The content of radioactive alanine in Y. pseudotuberculosis protein increased during 16 hours of cultivation at a high temperature, while at 8 degrees C the growth of the incorporation level lasted for at least 40 hours. For all precursors under study the incorporation rate into the cell biopolymers at the initial stages of cultivation was higher at 37 degrees C, than at a lower temperature.     

Changes in regulation of ribosomal protein synthesis during vegetative growth and sporulation of Saccharomyces cerevisiae.

When diploid Saccharomyces cerevisiae cells logarithmically growing in acetate medium were placed in sporulation medium, the relative rates of synthesis of 40 or more individual ribosomal proteins (r-proteins) were coordinately depressed to approximately 20% of those of growing cells. These new depressed rates remained constant for at least 10 h into sporulation. If yeast nitrogen base was added 4 yh after the beginning of sporulation to shift the cells back to vegetative growth, the original relative rates of r-protein synthesis were rapidly reestablished. this upshift in the rates occurred even in diploids homozygous for the regulatory mutation rna2 at the restrictive temperature for this mutation (34 degrees C). However, once these mutant cells began to bud and grow at 34 degrees C, the phenotype of rna2 was expressed and the syntheses of r-proteins were again coordinately depressed. At least one protein whose rate of synthesis was not depressed by rna2 in vegetative cells did have a decreased rate of synthesis during sporulation. Another r-protein whose synthesis was depressed by rna2 maintained a high rate of synthesis at the beginning of sporulation. These data suggest that the mechanism responsible for coordinate control of r-protein synthesis during sporulation does not require the gene product of RNA2 and thus defines a separate mechanism by which r-proteins are coordinately controlled in S. cerevisiae.     

Properties of glucose uptake in vegetative and sporulating cells of Saccharomyces cerevisiae

The effect of digitonin, acetic acid, urea and ethanol treatment on the glucose uptake of vegetative cells and of sporulating cells (3 h after transfer to sporulation medium) was examined in Saccharomyces cerevisiae. Both glucose uptake activities decreased at a similar rate, and a slightly different rate, in treatment with various concentrations of digitonin and of acetic acid, respectively, at 25 degrees C for 10 min. The glucose uptake activity of the sporulating cells was much more stable to urea treatment than that of the vegetative cells; the activity decreased about 36% and 76% in the sporulating cells and the vegetative cells, respectively, under conditions of 2.5 M urea at 25 degrees C for 10 min. The glucose uptake activity of the vegetative cells was more stable to ethanol treatment than that of the sporulating cells; the activity decreased about 56% and 88% in the vegetative cells and the sporulating cells, respectively, in 25% ethanol at 25 degrees C for 10 min.     

科尔沁沙地15种禾本科植物种子萌发特性比较

在实验室条件下,观测了科尔沁沙地乌兰敖都地区禾本科植物当年新采种子的萌发特点．15种植物中,披碱草、糙隐子草、冠芒草、大油芒、芦苇、虎尾草、野古草、狼尾草的发芽率超过80％,水稗草、牛鞭草、虱子草、狗尾草的发芽率不足10％．1～3d开始发芽的植物有大油芒、画眉、芦苇、虎尾草、披碱草、冠芒草、毛马唐、糙隐子草．超过10d基本不发芽的植物包括狗尾草、虱子草、牛鞭草．发芽持续期小于10d的植物包括毛马唐、水稗草、芦苇、画眉草、大油芒;发芽持续期21～30d的植物有披碱草、冠芒草．发芽种子超过总发芽种子的50％需要天数为虎尾草2d,芦苇3d,大油芒4d,披碱草5d,糙隐子草5d,野古草7d,冠芒草7d,狼尾草10d．与英国Sheffield地区相比,乌兰敖都地区一年生禾草发芽率低的所占比重更大,发芽更为缓慢;乌兰敖都地区多年生禾草的发芽率差别很小,但发芽更为缓慢．杂草植物萌发的风险分摊能力相对明显,因此抗干扰能力相对较强．     

Difference in sporogenous bacterial populations in thermophilic (55 degrees C) and mesophilic (35 degrees C) anaerobic sewage digestion

Spores, sporeforming vegetative cells, and asporogenous populations were enumerated in two semicontinuous anaerobic fermentors digesting municipal primary sludge at 35 and 55 degrees C for more than 87 days. In the 35 degrees C fermentor, the anaerobic total population was 312.5 X 10(6)/ml, with 25.0 X 10(6)/ml being sporogenous. The populations that digest casein, starch, pectin, and cellulose were 23.1 X 10(6), 59.2 X 10(6), 26.2 X 10(6), and 7.3 X 10(6)/ml, respectively, with 2.8 X 10(6), 6.7 X 10(6), 3.4 X 10(6), and 1.5 X 10(6)/ml being sporogenous, respectively. The sporeformers accounted for 8.0 to 20.0% of each of the respective populations. In the 55 degrees C fermentor, the anaerobic total population was 512.5 X 10(6)/ml, with 336.6 X 10(6)/ml being sporogenous. The populations that digest casein, starch, pectin, and cellulose were 97.7 X 10(6), 190.7 X 10(6), 75.8 X 10(6), and 11.2 X 10(6)/ml, respectively, with 47.8 X 10(6), 110.6 X 10(6), 43.3 X 10(6), and 5.1 X 10(6)/ml, respectively, being sporogenous. The sporeformers represented 45.5 to 65.7% of each of the respective populations. The numbers of thermophilic sporeforming vegetative cells in the 55 degrees C fermentor were 9.0 to 19.8 times higher than their counterparts in the 35 degrees C fermentor. Most sporeformers were in the vegetative state in the 35 and 55 degrees C fermentors. After 18 days of fermentation at 55 degrees C, sporeformers carried out most of the digestion; however, the digestion was shared by both sporeformers and asporogenous bacteria after 87 days of fermentation. In the 35 degrees C fermentor, asporogenous bacteria digested most of the sludge. During the 18- and 87-day experimental periods, sporeformers were never predominant.     

Difference in sporogenous bacterial populations in thermophilic (55 degrees C) and mesophilic (35 degrees C) anaerobic sewage digestion.

Spores, sporeforming vegetative cells, and asporogenous populations were enumerated in two semicontinuous anaerobic fermentors digesting municipal primary sludge at 35 and 55 degrees C for more than 87 days. In the 35 degrees C fermentor, the anaerobic total population was 312.5 X 10(6)/ml, with 25.0 X 10(6)/ml being sporogenous. The populations that digest casein, starch, pectin, and cellulose were 23.1 X 10(6), 59.2 X 10(6), 26.2 X 10(6), and 7.3 X 10(6)/ml, respectively, with 2.8 X 10(6), 6.7 X 10(6), 3.4 X 10(6), and 1.5 X 10(6)/ml being sporogenous, respectively. The sporeformers accounted for 8.0 to 20.0% of each of the respective populations. In the 55 degrees C fermentor, the anaerobic total population was 512.5 X 10(6)/ml, with 336.6 X 10(6)/ml being sporogenous. The populations that digest casein, starch, pectin, and cellulose were 97.7 X 10(6), 190.7 X 10(6), 75.8 X 10(6), and 11.2 X 10(6)/ml, respectively, with 47.8 X 10(6), 110.6 X 10(6), 43.3 X 10(6), and 5.1 X 10(6)/ml, respectively, being sporogenous. The sporeformers represented 45.5 to 65.7% of each of the respective populations. The numbers of thermophilic sporeforming vegetative cells in the 55 degrees C fermentor were 9.0 to 19.8 times higher than their counterparts in the 35 degrees C fermentor. Most sporeformers were in the vegetative state in the 35 and 55 degrees C fermentors. After 18 days of fermentation at 55 degrees C, sporeformers carried out most of the digestion; however, the digestion was shared by both sporeformers and asporogenous bacteria after 87 days of fermentation. In the 35 degrees C fermentor, asporogenous bacteria digested most of the sludge. During the 18- and 87-day experimental periods, sporeformers were never predominant.     

Culturability, injury and morphological dynamics of thermophilic Campylobacter spp. within a laboratory-based aquatic model system

AIMS: To study the survival processes of thermophilic Campylobacter spp. within a modelled aquatic system and particularly the involvement and survival potential of viable but non-culturable forms. METHODS AND RESULTS: The survival and morphological characteristics of populations of thermophilic Campylobacter species exposed to simulated aquatic conditions were examined using a combination of cultural and microscopic techniques. Populations underwent progressive decay when exposed to simulated aquatic conditions. The rates of population decay were observed to be significantly greater at the higher temperature (20 degrees C) with a rapid transition of the dominant sub-populations from non-stressed to dead cells occurred within 3 days. At 10 degrees C the rate of culturability loss was much reduced with substantial development (approx. 80% of total population) of viable but non-culturable (VBNC) populations by all species within 3 days, declining to represent approximately 5-25% of the total population at day 60. Significant differences (P < 0.001) were identified between decay rates as a consequence of different species, sub-populations and temperature but not between sub-populations of different species. Morphological variants including spiral, elongated spirals and rods, short rods and coccoid forms were identified. The endpoints of morphological transition were temperature-independent and isolate-specific yet the rate of morphological transition was directly related to temperature and approximately equivalent between species. CONCLUSION: The VBNC state is a transitory stage in the degeneration of Campylobacter population within the aquatic environments simulated during this study. SIGNIFICANCE AND IMPACT OF THE STUDY: VBNC cells form the most persistent, viable, potentially pathogenic sub-population of Campylobacter populations exposed to aquatic stress conditions.