Control of Listeria monocytogenes in a biofilm by competitive-exclusion microorganisms

Biofilms from drains in food processing facilities with a recent history of no detectable Listeria monocytogenes in floor drains were cultured for microorganisms producing antilisterial metabolites. A total of 413 microbial isolates were obtained from 12 drain biofilm samples and were assayed at 15 and 37 degrees C for activities that were bactericidal or inhibitory to L. monocytogenes, by two agar plate assays. Twenty-one of 257 bacterial isolates and 3 of 156 yeast isolates had antilisterial activity. All 24 isolates which produced metabolites inhibitory to L. monocytogenes were assayed for antilisterial activity in coinoculated broth cultures containing tryptic soy broth with yeast extract (TSB-YE). A five-strain mixture of 10(3) CFU of L. monocytogenes/ml and 10(5) CFU of the candidate competitive-exclusion microorganism/ml was combined in TSB-YE and incubated at 37 degrees C for 24 h, 15 degrees C for 14 days, 8 degrees C for 21 days, and 4 degrees C for 28 days. Substantial inhibition of L. monocytogenes growth (4 to 5 log CFU/ml) was observed for nine bacterial isolates at 37 degrees C, two at 15 and 8 degrees C, and three at 4 degrees C. The inhibitory isolates were identified as Enterococcus durans (six isolates), Lactococcus lactis subsp. lactis (two isolates), and Lactobacillus plantarum (one isolate). The anti-L. monocytogenes activity of these isolates was evaluated in biofilms of L. monocytogenes on stainless steel coupons at 37, 15, 8, and 4 degrees C. Results revealed that two isolates (E. durans strain 152 and L. lactis subsp. lactis strain C-1-92) were highly inhibitory to L. monocytogenes (growth inhibition of >5 log(10) CFU of L. monocytogenes/cm(2)). These two bacterial isolates appear to be excellent competitive-exclusion candidates to control L. monocytogenes in biofilms at environmental temperatures of 4 to 37 degrees C.     

Effects of moisture content on long-term survival and regrowth of bacteria in wastewater sludge.

The effects of moisture content on the survival and regrowth of seeded and indigenous enteric bacteria in raw sludge were determined. Cultures of six strains of fecally associated bacteria grown in sterilized, liquid sludge (5% solids) were all quite stable at this moisture level for over 90 days at 21 degrees C. When the moisture content of the sludge containing these organisms was reduced by evaporation and the samples were stored at 21 degrees C for extended periods, bacterial inactivation rates were generally proportional to the moisture losses of the samples. A dramatic reversal in this effect was observed in samples containing more than 90% solids. In this dried sludge, every bacterial species studied except Proteus mirabilis was found to be extremely stable. Bacteria indigenous to sludge were also found to survive for long periods in dried sludge. Regrowth of bacterial isolates in sterilized raw sludge was found to occur readily at 37 degrees C in samples containing less than or equal to 75% solids, but no growth was observed in samples with greater than or equal to 85% solids. Some growth, but to less than saturation densities, occurred with 80% solids. Growth of seeded Salmonella typhimurium was also found to occur in the presence of indigenous organisms in both liquid and dewatered raw sludges. However, the population density attained was well below that found in sterilized samples of the same sludges. In addition, the number of salmonellae dropped below detectable limits within a few days in sludges containing viable indigenous organisms, whereas little decrease occurred during this time with salmonellae grown in previously sterilized sludges.     

Production of raw cassava starch-degrading enzyme by Penicillium and its use in conversion of raw cassava flour to ethanol

A newly isolated strain Penicillium sp. GXU20 produced a raw starch-degrading enzyme which showed optimum activity towards raw cassava starch at pH 4.5 and 50 °C. Maximum raw cassava starch-degrading enzyme (RCSDE) activity of 20 U/ml was achieved when GXU20 was cultivated under optimized conditions using wheat bran (3.0% w/v) and soybean meal (2.5% w/v) as carbon and nitrogen sources at pH 5.0 and 28 °C. This represented about a sixfold increment as compared with the activity obtained under basal conditions. Starch hydrolysis degree of 95% of raw cassava flour (150 g/l) was achieved after 72 h of digestion by crude RCSDE (30 U/g flour). Ethanol yield reached 53.3 g/l with fermentation efficiency of 92% after 48 h of simultaneous saccharification and fermentation of raw cassava flour at 150 g/l using the RCSDE (30 U/g flour), carried out at pH 4.0 and 40 °C. This strain and its RCSDE have potential applications in processing of raw cassava starch to ethanol.     

Potential microbiological hazards in the production of refined paper products for food applications

This study sought to investigate the significance of raw materials (starch-based glues, raw material papers) at different microbiologically critical stages in the manufacturing process of refined paper products. The study examined the occurrence of microorganisms in the process and in end-product samples. Microbiological surveys verified that the production and use of pasteurized starch-based glue was the most important factor threatening the process hygiene and product safety. Subsequently, the production and use of starch-based glue was changed, and a follow-up programme targeting the microbiological quality of glue was developed as part of a hygiene and safety management system. A total of 33 spore-forming bacterial and 15 enterobacterial isolates were ribotyped, and 22 and 10 different ribogroups (ribotypes), respectively, were generated. These isolates from starch-based glue, raw material paper and end products were atypical and, thus, in many cases physiological, chemotaxonomic (FAME) and molecular (partial 16S rDNA) results did not correspond. The most common spore-forming bacteria (55% of the isolates) were Paenibacillus sp. and within this genus several new species were also proposed. The most common enterobacteria (87%) were Enterobacter cloacae and Citrobacter freundii belonging to bacteria in hazard group 2, or species closely related to them. It was demonstrated that the same spore-forming bacteria (ribotypes) were present in both the glue samples and the end products (45% of isolates). All RiboPrint patterns were saved at the VTT identification library for future use.     

Effect of incubation temperature on isolation of Campylobacter jejuni genotypes from foodstuffs enriched in Preston broth

Preston broth and agar incubated at either 37 or 42 degrees C have been widely used to isolate campylobacters from foodstuffs. The consequences of using either incubation temperature were investigated. Retail packs of raw chicken (n = 24) and raw lamb liver (n = 30) were purchased. Samples were incubated in Preston broth at 37 and 42 degrees C and then streaked onto Preston agar and incubated as before. Two Campylobacter isolates per treatment were characterized. Poultry isolates were genotyped by random amplification of polymorphic DNA (RAPD), pulsed-field gel electrophoresis (PFGE), and flagellin PCR-restriction fragment length polymorphism, and lamb isolates were genotyped by RAPD only. In total, 96% of the poultry and 73% of the lamb samples yielded campylobacters. The lamb isolates were all Campylobacter jejuni, as were 96% of the poultry isolates, with the remainder being Campylobacter lari. The incubation temperature had no significant effect on the number of positive samples or on the species isolated. However, genotyping of the C. jejuni isolates revealed profound differences in the types obtained. Overall (from poultry and lamb), the use of a single incubation temperature, 37 degrees C, gave 56% of the total number of RAPD C. jejuni genotypes, and hence, 44% remained undetected. The effect was especially marked in the poultry samples, where incubation at 37 degrees C gave 47% of the PFGE genotypes but 53% were exclusively recovered after incubation at 42 degrees C. Thus, the incubation temperature of Preston media selects for certain genotypes of C. jejuni, and to detect the widest range, samples should be incubated at both 37 and 42 degrees C. Conversely, genotyping results arising from the use of a single incubation temperature should be interpreted with caution.     

16S rRNA sequences and differences in bacteria isolated from the Muztag Ata glacier at increasing depths

Small subunit 16S rRNA sequences, growth temperatures, and phylogenetic relationships have been established for 129 bacterial isolates recovered under aerobic growth conditions from different regions of a 22-m ice core from the Muztag Ata Mountain glacier on the Pamirs Plateau (China). Only 11% were psychrophiles (grew at 2 degrees C or -2 degrees C up to approximately 20 degrees C), although the majority (82%) were psychrotolerant (grew at 2 degrees C or -2 degrees C up to 37 degrees C). The majority of the isolates had 16S rRNA sequences similar to previously determined sequences, ranging from 85% to 100% identical to database sequences. Based on their 16S rRNA sequences, 42.6% of the isolates were high-G+C (HGC) gram-positive bacteria, 23.3% were gamma-Proteobacteria, 14.7% were alpha-Proteobacteria, 14.7% were Flavobacteria, and 4.7% were low-G+C (LGC) gram-positive bacteria. There were clear differences in the depth distribution, with Proteobacteria, HGC/Cytophaga-Flavobacterium-Bacteroides (CFB), Proteobacteria, LGC/CFB/HGC, Cryobacterium psychrophilum, HGC/CFB, Proteobacteria/HGC/CFB, and HGC/CFB being the predominant isolates from ice that originated from 2.7 to 3.8, 6.2, 7.5, 8.3, 9.0, 9.7, 12.5, and 15.3 m below the surface, respectively. This layered distribution of bacterial isolates presumably reflects both differences in bacteria inhabiting the glacier's surface, differences in bacteria deposited serendipitously on the glacier's surface by wind and snowfall, and nutrient availability within the ice.     

Biological control of apple blue mold with Pseudomonas fluorescens

Pseudomonas fluorescens isolate 1100-6 was evaluated as a potential biological control agent for apple blue mold caused by Penicillium expansum or Penicillium solitum. Both the wild-type isolate 1100-6 and a genetically modified derivative labeled with the gene encoding the green fluorescent protein (GFP) were compared. The P. fluorescens isolates with or without GFP equally reduced the growth of Penicillium spp. and produced large zones of inhibition in dual culture plate assays. Cell-free metabolites produced by the bacterial antagonists reduced the colony area of Penicillium isolates by 17.3% to 78.5%. The effect of iron chelate on the antagonistic potential of P. fluorescens was also studied. The use of iron chelate did not have a major effect on the antagonistic activity of P. fluorescens. With or without GFP, P. fluorescens significantly reduced the severity and incidence of apple decay by 2 P. expansum isolates after 11 d at 20 degrees C and by P. expansum and P. solitum after 25 d at 5 degrees C when the biocontrol agents were applied in wounds 24 or 48 h before challenging with Penicillium spp. Populations of P. fluorescens labeled with the GFP were determined 1, 9, 14, and 20 d after inoculation at 5 degrees C. The log CFU/mL per wound increased from 6.95 at the time of inoculation to 9.12 CFU/mL (P < 0.05) 25 d after inoculation at 5 degrees C. The GFP strain did not appear to penetrate deeply into wounds based on digital photographs taken with an inverted fluorescence microscope. These results indicate that P. fluorescens isolate 1100-6 could be an important new biological control for apple blue mold.     

Improvement of nutritive value of grass pea (Lathyrus sativus) seed meal in the formulated diets for rohu, Labeo rohita (Hamilton) fingerlings after fermentation with a fish gut bacterium

Eight isonitrogenous (35% crude protein approximately) and isocaloric (4.0 kcalg(-1) approximately) diets were formulated incorporating raw and fermented grass pea (Lathyrus sativus) seed meal at 10%, 20%, 30% and 40% levels by weight into a fish meal based diet and fed to rohu, Labeo rohita, fingerlings for 80 days and fish performance was studied. A particular bacterial strain (Bacillus sp.) isolated from the intestine of adult common carp (Cyprinus carpio) reared in the wild having significant amylolytic, cellulolytic, lipolytic and proteolytic activities were used for fermentation of seed meal for 15 days at 37 degrees C. Fermentation of grass pea seed meal was effective in significantly reducing the crude fibre content and anti-nutritional factors, such as tannins, phytic acid and the neurotoxin, beta-ODAP and enhancing the available free amino acids and fatty acids. In terms of growth response, feed conversion ratio and protein efficiency ratio, 30% fermented grass pea seed meal incorporated diet resulted in significantly (P < 0.05) better performance of rohu fingerlings. In general, growth and feed utilization efficiencies of fish fed diets containing fermented seed meal were superior to those fed diets containing raw seed meal. The apparent protein digestibility (APD) values decreased with increasing levels of raw seed meal in the diets. The APD for raw seed meal was lower at all levels of inclusion in comparison to those for the fermented seed meals. The highest deposition of carcass protein was recorded in fish fed the diet containing 40% fermented seed meal. The results indicated that fermented grass pea seed meal can be incorporated in carp diets up to 30% level compared to 10% level of raw seed meal.     

Oat Bran as a Cholesterol-Reducing Dietary Adjunct in a Young,Healthy Population

In a study to examine the effects of dietary oat bran on serum lipids, subjects who ate two oat bran muffins a day for 28 days showed a 5.3% reduction in serum total cholesterol and an 8.7% reduction in low-density-lipoprotein-cholesterol levels, while no changes were noted in subjects consuming wheat or mixed wheat and oat bran muffins. Similarly, those who ingested oat bran muffins showed an 8.3% reduction in serum triglyceride values as contrasted with an overall increase of 6.4% in the other groups combined. These findings suggest that oat bran taken daily can significantly lower serum cholesterol and triglyceride levels in a young, healthy population.     

Inactivation of poliovirus type 1 in mixed human and swine wastes and by bacteria from swine manure.

The persistence of poliovirus type 1 (PO1) in mixed septic tank effluent and swine manure slurry was determined, and the antiviral effects of several bacterial cultures isolated from swine manure slurry were demonstrated. In two field experiments, PO1 was consistently inactivated more rapidly in the mixed waste than in the control Dulbecco's phosphate-buffered saline (D-PBS). D values (time [in days] for a 90% reduction of virus titer) were 18.7 and 29.9 for the mixed waste and 56.5 and 51.8 for the D-PBS control, respectively. The virus inactivation in the mixed waste was temperature dependent. A comparison of PO1 inactivation in raw mixed waste, autoclaved mixed waste, and bacterium-free filtrate of raw mixed waste at the same pH and temperatures provided an initial demonstration that the virus inactivation in the mixed waste is related, at least in part, to microbial activity. At 25 degrees C, the D value was 6.8 for the mixed waste, 11.2 for the autoclaved mixed waste, and 10.5 for the bacterium-free filtrate of raw mixed waste. At 37 degrees C, D values were 1.3, 3.9, and 3.1 for these three suspending media, respectively. Three bacterial isolates which had shown antiviral effects in a screening test each caused virus inactivation in autoclaved mixed waste, in which the effect of other microorganisms was excluded. Inhibition of PO1 inactivation by protease inhibitors suggests that the virus inactivation in the mixed waste was due in part to proteolytic enzymes produced by bacteria in the waste.     

Duckweed (Lemna polyrhiza) leaf meal as a source of feedstuff in formulated diets for rohu (Labeo rohita Ham.) fingerlings after fermentation with a fish intestinal bacterium

Eight isonitrogenous (35% crude protein approximately) and isocaloric (4.2 kcal g(-1) approximately) diets were formulated including raw and fermented duckweed (Lemna polyrhiza) leaf meal at 10%, 20%, 30% and 40% levels. A particular bacterial strain (Bacillus sp.) isolated from carp (Cyprinus carpio) intestine and having extracellular amylolytic, cellulolytic, proteolytic and lipolytic activities was used for leaf meal fermentation for 15 days at 37 degrees C. The fibre content of leaf meal reduced from 11.0% to 7.5% and the antinutritional factors, tannin and phytic acid, were reduced from 1.0% to 0.02% and 1.23% to 0.09%, respectively after fermentation. However, the available reducing sugars, free amino acids and fatty acids increased in the fermented leaf meal. The response of rohu, Labeo rohita, fingerlings fed the experimental diets for 80 days was compared with fish fed a fish meal based reference diet. On the basis of growth response, food conversion ratio and protein efficiency ratio, 30% fermented Lemna leaf meal incorporated in the diet resulted in the best performance of rohu fingerlings. In general, growth and feed utilization efficiencies of fish fed fermented leaf meal containing diets were superior to those fed diets containing raw leaf meal. The apparent protein digestibility (APD) decreased with increasing levels of leaf meal irrespective of treatment. The APD for raw leaf meal was lower at all levels of inclusion in comparison to those for the fermented meals. The highest carcass protein and lipid deposition was recorded in fish fed the diet containing 30% fermented leaf meal. The results showed that fermented Lemna leaf meal can be incorporated into carp diets up to 30% level compared to 10% level of raw meal.     

Inactivation of poliovirus type 1 in mixed human and swine wastes and by bacteria from swine manure.

The persistence of poliovirus type 1 (PO1) in mixed septic tank effluent and swine manure slurry was determined, and the antiviral effects of several bacterial cultures isolated from swine manure slurry were demonstrated. In two field experiments, PO1 was consistently inactivated more rapidly in the mixed waste than in the control Dulbecco's phosphate-buffered saline (D-PBS). D values (time [in days] for a 90% reduction of virus titer) were 18.7 and 29.9 for the mixed waste and 56.5 and 51.8 for the D-PBS control, respectively. The virus inactivation in the mixed waste was temperature dependent. A comparison of PO1 inactivation in raw mixed waste, autoclaved mixed waste, and bacterium-free filtrate of raw mixed waste at the same pH and temperatures provided an initial demonstration that the virus inactivation in the mixed waste is related, at least in part, to microbial activity. At 25 degrees C, the D value was 6.8 for the mixed waste, 11.2 for the autoclaved mixed waste, and 10.5 for the bacterium-free filtrate of raw mixed waste. At 37 degrees C, D values were 1.3, 3.9, and 3.1 for these three suspending media, respectively. Three bacterial isolates which had shown antiviral effects in a screening test each caused virus inactivation in autoclaved mixed waste, in which the effect of other microorganisms was excluded. Inhibition of PO1 inactivation by protease inhibitors suggests that the virus inactivation in the mixed waste was due in part to proteolytic enzymes produced by bacteria in the waste.     

Antimicrobial effects of mustard flour and acetic acid against Escherichia coli O157:H7, Listeria monocytogenes,and Salmonella enterica serovar Typhimurium

This study was designed to investigate the individual and combined effects of mustard flour and acetic acid in the inactivation of food-borne pathogenic bacteria stored at 5 and 22 degrees C. Samples were prepared to achieve various concentrations by the addition of acetic acid (0, 0.5, or 1%) along with mustard flour (0, 10, or 20%) and 2% sodium chloride (fixed amount). Acid-adapted three-strain mixtures of Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella enterica serovar Typhimurium strains (10(6) to 10(7) CFU/ml) were inoculated separately into prepared mustard samples stored at 5 and 22 degrees C, and samples were assayed periodically. The order of bacterial resistance, assessed by the time required for the nominated populations to be reduced to undetectable levels against prepared mustards at 5 degrees C, was S. enterica serovar Typhimurium (1 day) < E. coli O157:H7 (3 days) < L. monocytogenes (9 days). The food-borne pathogens tested were reduced much more rapidly at 22 degrees C than at 5 degrees C. There was no synergistic effect with regard to the killing of the pathogens tested with the addition of 0.5% acetic acid to the mustard flour (10 or 20%). Mustard in combination with 0.5% acetic acid had less bactericidal activity against the pathogens tested than did mustard alone. The reduction of E. coli O157:H7 and L. monocytogenes among the combined treatments on the same storage day was generally differentiated as follows: control < mustard in combination with 0.5% acetic acid < mustard alone < mustard in combination with 1% acetic acid < acetic acid alone. Our study indicates that acidic products may limit microbial growth or survival and that the addition of small amounts of acetic acid (0.5%) to mustard can retard the reduction of E. coli O157:H7 and L. monocytogenes. These antagonistic effects may be changed if mustard is used alone or in combination with >1% acetic acid.     

Effects of temperature on fecundity in vitro, egg hatching and reproductive development of Benedenia seriolae and Zeuxapta seriolae (Monogenea) parasitic on yellowtail kingfish Seriola lalandi

Globally, aquaculture industries involved with commercial culture of kingfish (Seriola spp.) experience outbreaks of monogenean parasites, which can cause heavy stock losses. In Australia and New Zealand, aquaculturists of kingfish Seriola lalandi incur financial losses caused by two monogenean species: Benedenia seriolae and Zeuxapta seriolae which parasitise the skin and gills, respectively. This study provides information on some basic temperature-dependent life-cycle parameters of these problematic monogeneans on S. lalandi. Hatching times and age at maturity were inversely related to water temperature within the range experienced by wild kingfish in New Zealand (13-21 degrees C). Mature B. seriolae in vitro laid on average 37 eggs/day that hatched over approximately 4 days; peak hatching occurred 9, 11 and 22 days post-deposition at temperatures of 21, 17.5 and 13+/-1.0 degrees C, respectively. Z. seriolae in vitro laid on average 246 eggs/day that hatched over 2 days; peak hatching occurred 7, 9 and 15 days post-deposition at these respective temperatures. B. seriolae matured within 20, 25 and 48 days p.i. at 21, 18 and 13 degrees C. Z. seriolae matured within 25, 37 and >52 days p.i. at the same temperatures. This research describes stages in the reproductive development of B. seriolae and Z. seriolae and discusses the inclusion of basic parasitic life-cycle parameters into management strategies designed to maximise treatment efficacy and limit monogenean epizootics in sea-cage kingfish culture.     

Survival of bovine embryos stored at 4 degrees C

These experiments were designed to test the efficacy of storing bovine embryos at 4 degrees C. Of particular interest were the age of embryo at which maximum post-storage survival could be achieved and longevity at 4 degrees C. A greater proportion of day 8 blastocysts developed in vitro at 37 degrees C following refrigeration for 48 hr than did embryos collected 2, 4 or 6 days after estrus (P<0.01). Survival of blastocysts stored at 4 degrees C for 48 hr was similar to that of nonstored blastocysts. In a subsequent experiment, day 8 blastocysts were recovered nonsurgically and assigned to one of the following treatments: (a) immediate transfer; (b) culture at 37 degrees C; or (c) storage at 4 degrees C for 1, 2, 3 or 5 days. Post-storage viability was assessed by either development in culture at 37 degrees C or embryo survival following nonsurgical transfer to synchronized recipients. In vitro survival of nonstored embryos and embryos stored 1 day did not differ. Survival decreased after storage for 2 days (P<0.10) or longer (P<0.05). Similar results were observed for survival after transfer, but embryo viability decreased even more rapidly with increasing duration of storage. In vitro survival was approximately 50% for blastocysts stored for 3 and 5 days, but few pregnancies resulted from transfer of embryos stored for these periods. In another experiment survival after transfer of blastocysts stored at 4 degrees C for up to 2 days was similar to that of nonstored blastocysts.     

新疆胀果甘草内生菌的分离和鉴定

对采自新疆的健康野生胀果甘草不同组织中的内生菌进行分离,确立了分离的条件为5%NaClO4浸5min,分离纯化得到149株细菌和2种真菌。通过形态学观察和革兰氏染色,细菌中芽孢杆菌为93株,杆状菌56株,使用法国梅里埃细菌自动鉴定仪对其进行鉴定,得到鉴定结果的细菌属于13个属,真菌显微形态鉴定属于Penicillium青霉菌属和Fusarium镰刀菌属。     

Campylobacter jejuni survival in chicken meat as a function of temperature.

Recognition of Campylobacter fetus subsp. jejuni (referred to hereafter as C. jejuni) as an important human pathogen and its isolation from meat products indicate the need for knowledge of its survival characteristics in meats. Thermal death times (D-values) for a single strain and a five-strain composite were determined in 1% peptone and autoclaved ground chicken meat at temperatures ranging from 49 to 57 degrees C. Survival was determined for these strains in chicken meat at 4, 23, 37, and 43 degrees C. Survival was also determined on raw chicken drumsticks stored at 4 degrees C in either an ambient or a CO2 atmosphere. D-values were greater in chicken meat than in peptone in all cases. D-values in peptone for strain H-840 at 49, 51, 53, 55, and 57 degrees C were 15.2, 4.90, 1.71, 0,64, and 0.25 min, respectively. The corresponding D-values in ground chicken meat were 20.5, 8.77, 4.85, 2.12, and 0.79 min, respectively. Similar results were obtained with a composite of five strains. When sterile ground chicken meat was inoculated with approximately 10(6) to 10(7) C. jejuni cells per g and stored at 37 degrees C in an ambient atmosphere, a 1-to 2-log count increase occurred during the first 4 days, followed by a gradual decline of about 1 log during the remainder of the 17-day storage period. No growth was observed among similarly inoculated samples that were stored at 4, 23, and 43 degrees C but counts declined by about 1 to 2 logs at 4 degrees C (17 day), by 2.5 to 5 logs at 23 degrees C (17 days), and to undetectable levels at 43 degrees C (between 10 and 16 days). Survival on raw chicken drumsticks stored at 4 degrees C in CO2 and in an ambient atmosphere declined by about 1.5 and 2.0 logs, respectively, during 21 days of storage. The effect of temperature on the survival of C. jejuni in chicken meat was similar to that reported in other natural and laboratory milieus. Ordinary cooking procedures that destroy salmonellae would be expected to destroy C. jejuni.     

PCR-RFLP of ITS rDNA for the rapid identification of Penicillium subgenus Biverticillium species

RFLP of ITS rDNA is proposed as a useful tool for molecular identification of the most common species of biverticillate penicillia. 60 isolates were analysed representing 13 species and 21 unique sequences were produced. The combination of five restriction enzymes was successful in separating 12 species. However, the variety Penicillium purpurogenum var. rubrisclerotium remained indistinguishable from Penicillium funiculosum. P. funiculosum appeared as the most confused species, being mis-identified with Penicillium miniolutum and Penicillium pinophilum, which were originally part of the species, and with P. purpurogenum perhaps because of the common production of red pigment. Penicillium variabile was difficult to investigate as introns were found on half of the isolates. Penicillium piceum, Penicillium rugulosum, Penicillium loliense, Penicillium erythromellis and P. purpurogenum were homogeneous from molecular and morphological positions and corresponded to a well circumscribed taxon. Furthermore, intraspecific variability was evidenced within P. pinophilum and P. funiculosum. The ex-type isolate of P. funiculosum produced a unique pattern. The method is sensitive, rapid and inexpensive and can be used for isolate identification of the biverticillate species. It is recommended particularly when many isolates have to be authentificated prior to analysis for phylogenetic assessment or population genetics.     

The microbiological transformation and degradation of pyridine and alkyl pyridines

The scientific literary data on the microbiological transformation and degradation of pyridine, methylpyridines (picolines), dimethylpyridines (lutidines) and some ethylpyridines are summarized and analysed in this review. These highly toxic compounds are on the one hand the raw materials for medicinal preparation production with microorganisms, on the other hand they are industrial wastes polluting the environment which may be degraded microbiologically.     

Endoglucanase-producing fungi isolated from Cape Evans historic expedition hut on Ross Island, Antarctica

Early explorers of Antarctica's Heroic Era erected wooden buildings and brought large quantities of supplies to survive in Antarctica. The introduction of wood and other organic materials provided nutrient sources for fungi that were indigenous to Antarctica or were brought in with the materials and adapted to the harsh conditions. Seventy-two isolates of filamentous fungi were cultured on selective media from interior structural wood of the Cape Evans historic hut and 27 of these screened positive for the ability to degrade carboxymethyl cellulose (CMC). Four non-CMC-degrading isolates were added to a group of 14 CMC-degrading isolates for further study, and endo-1, 4-beta-glucanase activity was demonstrated in the extracellular supernatant from all of these 18 isolates when grown at 4 degrees C, and also when they were grown at 15 degrees C. Isolates of Penicillium roquefortii and Cadophora malorum showed preference for growth at 15 degrees C rather than 25 degrees C or 4 degrees C indicating psychrotrophic characteristics. These results demonstrate that cellulolytic filamentous fungi found in Antarctica are capable of growth at cold temperatures and possess the ability to produce extracellular endo-1, 4-beta-glucanase when cultured at cold and temperate temperatures.