Synergistic Antifungal Activity of Chitinolytic Enzymes and Prodigiosin Produced by Biocontrol Bacterium, Serratia marcescens Strain B2 against Gray Mold Pathogen, Botrytis cinerea

Serratia marcescens strain B2 is an effective biocontrol agent against gray mold of cyclamen, caused by Botrytis cinerea Persoon. Strain B2 has strong antifungal activity against B. cinerea in vitro. The culture filtrate was found to contain different types of chitinolytic enzyme activity, including endochitinase and chitobiase activity. Four chitinolytic enzymes were detected among the extracellular proteins of strain B2. Two major enzymes, a 58-kDa endochitinase and a 98-kDa chitobiase, were purified by chromatography and electrofocusing, respectively. Both enzymes inhibited spore germination of B. cinerea. And a red pigment, prodigiosin, extracted and purified from the bacterial cells, also inhibited spore germination. When prodigiosin and the chitinolytic enzymes were applied in concert, a synergistic inhibitory effect was observed. S. marcescens strain B2 has multiple modes of action against the pathogen. Received 14 February 2001/ Accepted in revised form 31 May 2001     

Long-term observations of the vertical distributions of mineral elements and phosphorus dynamics in sediments in a shallow eutrophic lake in Japan

Purpose

To explore the mechanisms in the deposition and release of phosphorus (P) in the sediment of a shallow eutrophic lake using preserved samples, we investigated the vertical and temporal changes in P, manganese (Mn), sulfur (S), iron (Fe), aluminum (Al), calcium (Ca), and magnesium (Mg) in the sediment samples and the phosphate in the sediment pore water samples over a period of 6 years.

Materials and methods

The upper 15 cm of sediment from Lake Kasumigaura in Japan was collected monthly from 2003 to 2008 from the center of the lake. Sediment cores were divided into seven depth segments and were acid-digested for an elemental analysis via inductively coupled plasma atomic emission spectroscopy. Phosphate concentrations of the sediment pore water were determined using the molybdenum blue method. A multiple regression analysis was conducted by setting the P content as the response variable and Mn, S, Fe, Al, Ca, and Mg as explanatory variables.

Results and discussion

The results of the multiple regression analysis demonstrated that P co-precipitates with Fe and Al oxides and accumulates on the sediment surface. The vertical distributions of Mn and S suggest that Mn reduction occurs within the 0–1-cm-depth layer of the sediment and that iron sulfide is actively formed in the 6–10-cm-depth layer of the sediment. These findings imply that the layer in which ferric oxides are reduced to ferrous ions is present near the 1–6-cm-depth layer of the sediment. This layer corresponds to the layer in which the maximum phosphate concentration of the sediment pore water often occurred (the 2–6-cm-depth layer). These results indicate that vertical distributions of mineral elements are useful for assessing P dynamics in sediments.

Conclusions

The lake sediments record the dynamics of P in the sediment. Our analytical approach using long-term observation data demonstrated that the accumulation and release of P associated with a change in the redox state can be assessed based on the vertical distributions of mineral elements in the lake sediments.

     

Chemical characterization of conducive and suppressive soils for potato scab in Hokkaido,Japan

Potato common scab induced by Streptomyces scabies is a serious constraint for potato-producing farmers and the incidence of potato scab depends on the soil chemical properties. We examined the chemical characteristics of conducive and suppressive soils to potato common scab with reference to the chemical properties of nonallophanic Andosols, recently incorporated into the classification system of cultivated soils in Japan. Allophanic Andosols with a ratio of pyrophosphate-extractable aluminum (Al_p) to oxalate-extractable aluminum (Al_o) of less than 0.3–0.4 were “conducive” soils with a high allophane content of more than 3%. On the other hand, nonallophanic Andosols with a Al_p/Al_a ratio higher than this critical value were “suppressive” soils, and their allophane content was less than 2%. The concentration of water-soluble aluminum (AI) was also a useful index for separating conducive from suppressive soils as well as the Al_p/Al_a value and allophane content. The suppressive soils showed a much higher concentration of water-soluble Al at pH 4.5 to 5.5 than the conducive soils. The high concentration of water-soluble Al may be responsible for the control of the incidence of potato common scab in Andosols.     

The response of adipose tissues to Mycoplasma pulmonis and Sendai virus infection in C57BL/6 and DBA/2 mice

Adipose tissues in mammals are categorized into white and brown adipose tissues in which cellular morphology, cell functions, and tissue distribution are different. White adipose tissue (WAT) plays a major role in energy reservation, while brown adipose tissue (BAT) mainly relates to the thermoregulation of the body. One interesting function of adipose tissue is the response to the infection, especially the pathogens that cause pneumonia. We have previously reported that DBA/2 (D2) mice are susceptible to pathogens causing pneumonia, Mycoplasma (M.) pulmonis and Sendai virus (SeV), whereas C57BL/6 (B6) mice are resistant to them. Furthermore, morphological alteration of mediastinal fat tissue (MFT) was seen after infection of M. pulmonis in D2 mice but not in B6 mice. In this study, we aimed to exhibit the difference in adipose tissue response in other areas, including interscapular brown adipose tissue (iBAT), inguinal white adipose tissue (ingWAT), and perigonadal WAT (perigoWAT) between resistant strain, B6 and susceptible strain, D2 after challenging them with M. pulmonis and SeV. Compared with B6 mice, D2 mice showed an increase in fat-associated lymphoid cluster in MFT, an increase in BAT in both iBAT and ingWAT after M. pulmonis and SeV infection. The results of this study indicate that pneumonia caused by M. pulmonis and SeV infection induces browning of adipocyte, suggesting that BAT plays a role in pathogen infection and inflammation.     

Differentiation of feline coronavirus type I and II infections by virus neutralization test

Feline coronavirus (FCoV) is divided into two types I and II, based on their growth in vitro and antigenicity. In this study, virus neutralization (VN) test was applied for type differentiation of FCoV infections. Sera of cats which were clinically and serologically diagnosed as feline infectious peritonitis (FIP) possessed significantly higher VN titers to type I FCoV, and sera from cats experimentally infected with FIPV type II had high VN titers to type II but not type I viruses. A total of 79 cat sera collected in the years between 2004 and 2005 were examined to evaluate seroprevalence by the VN test, showing the following results: (1) 50 cats (63.3%) were sero-positive to FCoV; (2) of the 50 FCoV positive cat serum samples, 49 (98%) showed significantly higher titers to type I virus and only one (2%) for type II virus. These results indicate that the VN test described here can be used for serological differentiation of FCoV infections of cats, and that FCoV type I is a dominant type in recent years of Japan.     

Estimating glomerular filtration rate in healthy dogs using inulin without urine collection

The goals of this study were to determine if the glomerular filtration rate (GFR) in dogs could be estimated by plasma inulin clearance and/or infusion inulin clearance analyses without urine collection, and to compare these results with GFR values obtained by urinary inulin clearance analysis. The dogs included in this study were healthy 20 beagles. Inulin clearance values were obtained by urinary inulin clearance, infusion inulin clearance, and plasma inulin clearance techniques. Urinary inulin clearance was 4.09±0.52 ml min(-1) kg(-1) (body weight); infusion inulin clearance, 4.01±0.49 ml min(-1) kg(-1); and plasma inulin clearance, 4.14±0.66 ml min(-1) kg(-1). The urinary inulin clearance was strongly correlated with infusion inulin clearance and weakly correlated with plasma inulin clearance. The GFR for dogs can be estimated by infusion and plasma inulin clearance analyses by blood sampling alone, without urine collection.