Ida Skaar

Diversity and Significance of Mold Species in Norwegian Drinking Water

ABSTRACT In order to determine the occurrence, distribution, and significance of mold species in groundwater- and surface water-derived drinking water in Norway, molds isolated from 273 water samples were identified. Samples of raw water, treated water, and water from private homes and hospital installations were analyzed by incubation of 100-ml membrane-filtered samples on dichloran-18% glycerol agar. The total count (number of CFU per 100 ml) of fungal species and the species diversity within each sample were determined. The identification of mold species was based on morphological and molecular methods. In total, 94 mold species belonging to 30 genera were identified. The mycobiota was dominated by species of Penicillium, Trichoderma, and Aspergillus, with some of them occurring throughout the drinking water system. Several of the same species as isolated from water may have the potential to cause allergic reactions or disease in humans. Other species are common contaminants of food and beverages, and some may cause unwanted changes in the taste or smell of water. The present results indicate that the mycobiota of water should be considered when the microbiological safety and quality of drinking water are assessed. In fact, molds in drinking water should possibly be included in the Norwegian water supply and drinking water regulations.

The study of fungi in drinking water

The occurrence of fungi in drinking water has received increased attention in the last decades, and fungi are now generally accepted as drinking water contaminants. The knowledge about the occurrence and diversity of fungi in water has increased considerably from a low knowledge base. However, the relevance of waterborne fungi for water quality and human health is poorly understood and still conflicting. Scientific reports on effective treatment against fungi in water are also few. This article presents a review of the literature on fungal water studies, including some general results, and considerations of significance, limits, contradictions, precautions, and practical consequences.

The presence of Penicillium and Penicillium mycotoxins in food wastes

A total of 97 samples (48 summer and 49 winter) of food waste from private households were investigated for Penicillium and for mycotoxins. Twenty-five Penicillium species were isolated and Penicillium crustosum, Penicillium brevicompactum, Penicillium chrysogenum, Penicillium expansum, Penicillium roqueforti, Penicillium spinulosum, Penicillium viridicatum, Penicillium commune, Penicillium citrinum and Penicillium solitum were, in decreasing order, the most frequently identified species. Mycotoxins produced by several of these species, including mycophenolic acid, roquefortine C, penitrems A-F and thomitrems A and E, were detected. Of the 48 summer samples, 36 were severely infected and contained more than 10(5) colony forming units (CFU) Penicillium/g sample. The levels of mycotoxins in these samples were in the range 75-19000 microg/kg mycophenolic acid, 40-920 microg/kg roquefortine C, 35-7500 microg/kg penitrem A, 20-2100 microg/kg thomitrem A and 20-3300 microg/kg thomitrem E. Of the 49 winter samples, only one was found to contain mycophenolic acid (4800 microg/kg) and roquefortine C (190 microg/kg), and this sample was severely infected with P. roqueforti. Thirty samples of food waste collected from the food manufacturing industry were also investigated. The number of Penicillium in these samples was between 10(5) and 10(6) colony forming units (CFU)/g sample. Seven of these samples contained mycophenolic acid ranging from 50 to 600 microg/kg and three of these samples also contained roquefortine C in the range 100-250 microg/kg.

Occurrence of moulds in drinking water

Aims:  In order to determine the occurrence of filamentous fungi in public drinking water systems in Norway, water from 14 water supply networks from all over the country was sampled and analysed. Networks with both ground and surface water sources were included in this study.

Moulds contaminants on Norwegian dry-cured meat products.

Dry-cured meat production has a long tradition in Norway. However, uncontrolled mould growth on the surface of the dry-cured meat products is causing significant quality problems. As some moulds are mycotoxigenic, their growth on the dry-cured meat products could also pose a serious health risk. Such quality problems and potential health risks can be better handled if the types of moulds growing on the products are known. In total, 161 samples were collected from the ripening and packaging stages of production with the aim of identifying moulds contaminating smoked and unsmoked Norwegian dry-cured meat products. Moulds were isolated either by transferring aerial mycelium of each visible mould colonies on the products or by directly plating pieces of meat on suitable agar media. The isolates were identified at a species level by a polyphasic approach. In total, 264 isolates belonging to 20 species and four fungal genera were identified. The genus Penicillium covered 88.3% of the total isolates. This genus contributed to the isolates of smoked and unsmoked products by 91% and 84% respectively. Penicillium nalgiovense was the dominant species isolated from both smoked and unsmoked products and covered 38% of the total isolates. Penicillium solitum and P. commune were the next most frequently isolated species with a contribution of 13% and 10% respectively. Species of Cladosporium and Eurotium contributed to the total isolates by 6% and 4.9% respectively. Smoking seems to affect the growth of these dominating species differently. An increase in the isolation frequency of P. nalgiovense accompanied by the reduction in the occurrence of P. solitum, P. commune and species of Cladosporium was observed on smoked products. The survey showed that the species of Penicillium are associated with Norwegian dry-cured meat products in general. Penicillium nalgiovense, the dominating mould species, is a potential producer of penicillin and its presence could represent a threat to allergic consumers.

Mould growth on the Norwegian semi-hard cheeses Norvegia and Jarlsberg

Visible moulds were isolated and identified from 102 samples of each of the Norwegian types of semi-hard cheeses called Norvegia and Jarlsberg. Penicillium species made up 98.1 and 89.2% of the isolates from the Jarlsberg and Norvegia cheeses, respectively. The most frequently occurring species on both was P. roqueforti subspecies roqueforti. The four species Penicillium roqueforti subspecies roqueforti, P. commune, P. palitans and P. solitum made up 69.8% of the total number of isolates from the Norvegia cheese and 81.0% of the total number of isolates from the Jarlsberg cheese.

Mould contaminants on Jarlsberg and Norvegia cheese blocks from four factories.

Visible mould from 225 blocks of the Norwegian semi-hard cheeses Jarlsberg and Norvegia from four factories were subcultured and identified. Altogether 23 different fungal species were detected. The two most important contaminating species were Penicillium commune and P. palitans, constituting 21.4% and 17.9% of the total isolates, respectively. The other dominating contaminants were P. roqueforit spp. roqueforti, Geotrichum candidum, P. solitum and P. crustosum. These species, together with P. commune and P. palitans, represented 80.9% of the total isolates. P. commune, P. palitans, P. roqueforti spp. roqueforti and P. solitum were most common contaminants on cheese produced in all four factories, while G. candidum was found to be important on Jarlsberg cheese from only one factory. P. crustosum was one of the dominating species on Norvegia cheese.

Streptococcus phocae sp. nov., a New Species Isolated from Clinical Specimens from Seals

A new beta-hemolytic streptococcal species, Streptococcus phocae, was isolated from organ specimens obtained from seals. This taxon is described on the basis of the results of a study of 22 strains. S. phocae was serologically somewhat heterogeneous (group antigen -/F/C). Strains belonging to this species exhibited high levels of DNA-DNA homology to each other, as determined by DNA-DNA hybridization, but low levels of DNA-DNA homology to the type strains of other streptococcal species. A simple scheme for the differentiation of S. phocae from other beta-hemolytic streptococci is presented. Strain 8399 H1 (= NCTC 12719) is the type strain of S. phocae.

Pathogenicity of Saprolegnia spp. to Atlantic salmon, Salmo salar L., eggs

Live and dead Atlantic salmon eyed eggs were challenged with eight different Saprolegnia isolates, selected because of their varied origins, known morphological characteristics and growth/germination pattern. Some isolates were also tested for pathogenicity to Atlantic salmon parr. Challenge of eggs was performed by exposure to spores in suspension or by co-incubation of live eggs with infected dead eggs. The phenotypic characteristics of the isolates were evaluated in relation to their observed pathogenicity from the challenge experiment, to identify possible virulence factors leading to egg-infection by Saprolegnia. The results from the experiments confirm that live eggs are refractory to infection with Saprolegnia spores in suspension and that an infection of live eggs can only occur from an infection nucleus represented by dead eggs or debris. It was observed that strains pathogenic to salmon parr were not particularly infective towards eggs, and the isolates that gave the highest infection rates to eggs were species considered to be saprotrophs.

Fungal growth pattern, sources and factors of mould contamination in a dry-cured meat production facility

The aims of this study were to investigate the patterns of fungal growth on dry-cured meat products, identify the important sources and factors of contamination and recommend intervention measures. The production processes of two smoked dry-cured hams and one unsmoked dry-cured leg of lamb were studied. A longitudinal observational study was performed to collect 642 samples from the meat, production materials, room installations and indoor and outdoor air of the production facility. Standard mycological isolation and identification procedures were followed. Totally, 901 fungal isolates were obtained; of which 57% were moulds while 43% were yeast. Yeasts were dominant on meat surfaces by covering 64% of the isolates. Mould growth was not observed until late in the dry-ripening stage. Yeasts and moulds were isolated from half of the environmental samples, of which moulds contributed by 80%. More than 39 mould species were isolated from the entire production process with a 77% contribution by the species of Penicillium. Penicillium nalgiovense dominated the species composition of moulds isolated from the products and the production environment. A preliminary bioassay analysis on bacterial colonies indicated that most of the P. nalgiovense isolates have the ability to produce penicillin. Such isolates might produce penicillin on the products and can become potential food safety hazards. Improper pressing at the salting process, the air quality in salting, brining and smoking rooms and activities in the sorting room were identified as important factors and sources of fungal contamination. Technical solutions and organized production activities that reduce crack formation, airborne spore concentration and improve air circulation in the facility are recommended as intervention measures.