Ecology ofAeromonas hydrophila in a South Carolina cooling reservoir

Densities ofAeromonas hydrophila were determined monthly from December 1975 to December 1977 in a South Carolina cooling reservoir which receives heated effluent from a single nuclear production reactor. Selected water quality parameters and prevalence of red-sore disease among largemouth bass were monitored simultaneously.Higher densities ofA. hydrophila were observed in areas of the reservoir receiving effluent from the reactor. Densities ofA. hydrophila generally were heterogeneous in the water column. The sediments had lower densities ofA. hydrophila than water immediately above.A. hydrophila could not be isolated from sediments greater than 1 cm from the water interface. Temperature, redox potential, pH, and conductivity were all significantly correlated with densities ofA. hydrophila in the water column. The temporal and spatial distribution and abundance ofA. hydrophila in water were not related to total organic carbon, dissolved organic carbon, particulate organic carbon, inorganic carbon, or dissolved oxygen. High densities ofA. hydrophila were observed in mats of decomposingMyriophyllum spicatum and, enterically, in largemouth bass, several other species of fish, turtles, alligators, and snails. The greatest densities ofA. hydrophila in water occurred during March and June with a second peak in October. The mean monthly densities ofA. hydrophila were positively correlated with the incidence of infection in largemouth bass. Largemouth bass from thermally altered parts of the reservoir had a significantly higher incidence of infection. It is concluded that thermal effluent significantly affects the ecology ofA. hydrophila and the epizootiology of red-sore disease within Par Pond.     

Chemotaxis ofAeromonas hydrophila to the surface mucus of fish

Isolates ofAeromonas hydrophila from various sources show different chemotactic responses to mucus from the surface of freshwater fish. Some isolates were nonchemotactic to fish surface mucus. Isolates ofA. hydrophila from fish lesions had a significantly higher chemotactic index than isolates ofA. hydrophila from water. Maximum chemotactic responses occurred more often to diluted fish mucus than to undiluted samples. Fish which were experimentally stressed did not produce mucus that was more or less chemotactic than that of unstressed fish. Fish with red-sore lesions produced surface mucus which was not chemotactic toA. hydrophila. Differences between fish, for any isolate, were also not significant. The chemotactic substance(s) in fish mucus has a molecular weight of approximately 100,000 and did not appear to be labile when heated to 56°C.     

Effects of cultural conditions on protease production by Aeromonas hydrophila

Production of extracellular proteolytic activity by Aeromonas hydrophila was influenced by temperature, pH, and aeration. Conditions which produced maximal growth also resulted in maximal protease production. Enzyme production appeared to be modulated by an inducer catabolite repression system whereby NH4+ and glucose repressed enzyme production and complex nitrogen and nonglucose, carbon energy sources promoted it. Under nutritional stress, protease production was high, despite poor growth.     

Fed-batch culture ofAeromonas hydrophila for the production of poly(3-hydroxubutyrate-co-3-hydroxyhexanoate) using two carbon sources

To produce polyhydroxyalkanoate (PHA) copolymer which consists of 3-hydroxy-butyrate (3HB) and 3-hydroxyhexanoate (3HHx) by cultivation ofAeromonas hydrophila, fed-batch cultures were done under several nutrient limiting conditions. With the results from flask cultures, fed-batch cultures were carried out to produce large amounts of PHA. In the fed-batch culture, firstly glucose was fed to grow cell, and then, oleic acid fed to stimulate PHA in the cell. The final cell concentration, PHA content, PHA concentration, and 3-hydroxy-hexanoate fraction in 38 hr were 48.9 g/L, 15.05 wt%, 7.36 g/L and 12.2 wt%, respectively, resulting in the productivity of 0.19 g/L-h under phosphate-limiting condition.     

Effects of cultural conditions on protease production by Aeromonas hydrophila.

Production of extracellular proteolytic activity by Aeromonas hydrophila was influenced by temperature, pH, and aeration. Conditions which produced maximal growth also resulted in maximal protease production. Enzyme production appeared to be modulated by an inducer catabolite repression system whereby NH4+ and glucose repressed enzyme production and complex nitrogen and nonglucose, carbon energy sources promoted it. Under nutritional stress, protease production was high, despite poor growth.     

Occurrence ofAeromonas hydrophila in limnetic environments: Relationship of the organism to trophic state

The densities ofAeromonas hydrophila in various natural waters were found to be strongly correlated with a relative index proposed for use in trophic state assessments of fresh waters. No such correlation was found with the recoverable heterotrophic population of whichA. hydrophila is a part.A. hydrophila was found to be seasonally distributed with maximal densities occurring during summer through early fall. It was also found to be spatially distributed within a pond with the most consistent densities occurring from 1 m depth down to that depth in the water column where the temperature reaches 16°C. The densities of the organism correlated most strongly with total phosphorus, chlorophylla, and Secchi depth. Moderate correlations were found with dissolved phosphorus, Kjeldahl nitrogen, and dissolved organic carbon. Little or no correlation was obtained with ammonia, orthophosphate, pH, alkalinity, or dissolved oxygen. The discriminating ability that theA. hydrophila density measurements provide in the oligotrophic through mesotrophic range appears to exceed those of presently available methods. The facility and sensitivity of the enumeration method forA. hydrophila should make it a useful tool for trophic state assessments.     

Effect of dithiocarbamates on cellulase activity in culture filtrates of Trichoderma reesei

Different concentrations of tetramethylthiuram disulfide (TMTD), sodium dimethyldithiocarbamate (NaDDC), and zinc dimethyldithiocarbamate (ZnDDC) affected the amount of cellulase(s) activity in the culture of Trichoderma reesei. After eight days incubation at 28 degrees C the greatest increase in Avicelase, CMCase, and beta-glucosidase over the control were observed at 0.1 ppm (TMTD) and 0.4 ppm (NaDDC and ZnDCC). There was decrease in the growth in the ZnDDC, but beta-glucosidase activity was reduced considerably. Total protein in the culture filtrate increased with the increase in cellulase(s) activity. No change in pH was observed at eight days incubation but pH increased (not exceeding 5.9) at 12 days incubation.     

A model for the density ofAeromonas hydrophila in Albemarle Sound,North Carolina

The abundance ofAeromonas hydrophila was measured monthly at 29 sites in Albemarle Sound, North Carolina and its tributaries from April 1977 through July 1979. Simultaneous measurements included heterotrophic plate count bacteria, fecal coliform bacteria, and 18 physical and chemical parameters. Using only 6 water quality parameters, multiple correlation and regression analysis of the data produced a best-fit regression which explained 38% of the variation observed inA. hydrophila density. The 6 water quality parameters included dissolved oxygen, temperature, orthophosphate, chlorophyll A trichromatic, total Kjeldahl nitrogen, and ammonia. Heterotrophic plate count bacteria and fecal coliform densities were highly correlated withA. hydrophila density, but made the model very unstable. The model was successfully tested against similar data collected for 2 other North Carolina reservoirs, Lake Norman and Badin Lake. Data from 10 sites in Badin Lake over 18 months and from 7 sites on Lake Norman over 5 months were not significantly different from the Albemarle Sound model. Conditions of water quality that may give rise to “blooms” ofA. hydrophila will simultaneously contribute to the probability of increased epizootics in fish in the southeastern United States.     

Effect of the culture filtrates ofStreptomyces on growth and productivity of wheat plants

Streptomyces olivaceoviridis, S. rimosus andS. rochei proved to possess a high capacity for the production of auxins, gibberellins and cytokinin-like substances, together with substantial levels of α-amylase and proteinase. Grain priming with culture filtrates ofS. olivaceoviridis, S. rimosus orS. rochei appeared to enhance growth vigor and crop yield of wheat plants. In the majority of cases, the culture filtrate ofS. olivaceoviridis appeared to be the most effective in this respect. The present results are discussed in relation to the indirect role played by these bacteria in producing plant growth-regulating substances and their effects on growth and yield of wheat.     

Occurrence of a procellulase in the culture filtrates of Penicillium janthinellum

The endo-1,4-β-d-glucanase [cellulase, 1,4-(1,3:1,4)-β-d-glucan 4-glucanohydrolase, EC 3.2.1.4] activity of two-day old culture filtrates of Penicillium janthinellum has been enhanced four-fold by incubating with a 10-day old culture filtrate of Penicillium funiculosum grown on the same medium. An inactive protein isolated by fractionation of two-day old culture filtrate of P. janthinellum using preparative isoelectric focusing, showed 30- to 50-fold enhancement of endo-1,4-β-d-glucanase activity. This fraction has been designated the ‘procellulase’ in the present paper. The purity of the procellulase was confirmed by analytical isoelectric focusing and sodium dodecyl sulphate-polyacrylamide gel electrophoresis. It had a molecular weight of 68 000 and an isoelectric point of pH 3.7.     

Effect of a lactic starter culture on the growth and protease activity of Aeromonas hydrophila

Lactococcus lactis subsp. lactis strain Lac 388 (isolated from goats' cheese made without starter) had inhibitory activity against three strains of Aeromonas hydrophila either by the plating method or by the associative culture approach (broth, skim milk and ewes' milk). Low pH was only partially responsible for the antagonism. Inhibition due to hydrogen peroxide and nutrient depletion was excluded. This inhibitory compound(s) was not destroyed by proteolytic enzymes. In mixed cultures the strains of Aer. hydrophila did not produce detectable amounts of protease.     

Comparison of lethalities in mouse versus goldfish for clinical and food isolates ofAeromonas hydrophila

Summary The lethality of 16 clinical or food isolates ofAeromonas hydrophila was assessed by determination of LD₅₀ (i.p.) in mice and goldfish. In mice LD₅₀ values for the variousA. hydrophila strains were similar, ranging from 1.2–21.0×10⁸ cells/animal. A wider range of LD₅₀ values, 0.03–11.8×10⁸ cells/animal, was observed with goldfish. Lethality was not correlated between the two test animals. Further, cytotoxic response in Y-1 adrenal cells did not correlate with lethality in either test animal. It appears that lethality is not a good measure of potential enterotoxigenicity, but may be useful in assessing the invasive character of isolates causing systemic infections in immunocompromised hosts.     

Enzymatic hydrolysis of cellulose: Evaluation of cellulase culture filtrates under use conditions

Culture filtrates from three mutant strains of Trichoderma reesei grown on lactose and on cellulose were compared under use conditions on four cellulose substrates. Cellulose culture filtrates contained five to six times as much cellulase as lactose culture filtrates. Unconcentrated cellulose culture filtrates produced up to 10% sugar solutions from 15% cellulose in 24 h. Specific activity in enzyme assays and efficiency in saccharification tests were low for enzymes from all the mutants. Over a wide range the percent saccharification of a substrate in a given times was directly proportional to the logarithm of the ratio of initial concentrations of enzyme and substrate. As a result of this, dilute enzyme is more efficient than concentrated enzyme, but if high sugar concentrations are desired, very large quantities of enzyme are required. Since the slopes of these plots varied, the relative activity of cellulase on different substrates may be affected by enzyme concentration.