Spoilage fungi in a bread factory in Brazil: Diversity and incidence through the bread-making process.

This study aimed to verify the main fungal species involved in the deterioration of different types of bread and to identify the possible sources of contamination of these products. Samples of raw materials (n = 127), environmental air (n = 50) and moldy bread (n = 90) were analyzed. Aspergillus candidus, Wallemia sebi, and Penicillium roqueforti were the predominant species in the raw materials and were isolated in samples of wheat flour, in two-thirds of the samples of rye and 62.5% of the wheat flour. Penicillium roqueforti was isolated from all types of moldy bread analyzed and Hyphopichia burtoni was also present in samples of moldy wheat and rye bread. These two species were also recovered during air sampling from baking industry facilities (cooling and slice and package areas), which may be crucial for product contamination after baking. Hygienic measures to reduce airborne contamination during the cooling and packaging of food should be taken to prevent the early deterioration of bread.

Ochratoxin A production by Aspergillus westerdijkiae in Italian-type salami.

The occurrence of ochratoxin A (OTA) in matured meat products can be attributed to mycotoxin coming from raw materials and/or molds that develop on the product surface during ripening. This work aimed to evaluate OTA production by Aspergillus westerdijkiae inoculated on the surface of sausages and its diffusion into the product throughout ripening, study the effect of relative humidity (RH) on the production of this mycotoxin, and investigate the presence of OTA in dry fermented sausages that naturally present intense contamination by A. westerdijkiae. Italian-type sausages were surface inoculated with A. westerdijkiae and two distinct experiments were performed. In the first, the sausages were matured in a chamber with decreasing RH (from 95 to 75%) for 35 days. In the other, the sausages were incubated under different RH (79, 85 or 95%) for 21 days. Samples were taken at the beginning of the experiments and every 7 days, subdividing into casing, outer border, and core for analyses. Sausage samples naturally spoiled by ochratoxigenic fungi were collected during sanitary inspection. Even in the presence of A. westerdijkiae mycelia, no OTA was detected for up to 7 days of sausage maturation. On the other hand, this study demonstrated that the growth of A. westerdijkiae on salami surface produces high amounts of OTA on the casing and allows its diffusion through the casing with contamination to the outer border of sausages. In the same way, it shows that under similar water activity values of substrate, RH influences the amount of OTA produced. Conversely, OTA was restricted to the casing in the naturally contaminated sausages.

Antifungal activity of commercial sanitizers against strains of Penicillium roqueforti, Penicillium paneum, Hyphopichia burtonii, and Aspergillus pseudoglaucus: Bakery spoilage fungi.

Information on the sensitivity of spoilage fungi of bakery products to sanitizing agents is scarce in the literature. Thus, the aim of this study was to evaluate the antifungal activity of different classes of commercial sanitizers, which have permitted use in the food industry, on the main fungi involved in spoiling bakery products. The tests were carried out according to the protocol for testing the antifungal effect of chemical sanitizers of the European Committee for Standardization (CEN), with adaptations. Different strains of six isolated fungal species responsible for spoiling bakery products (Penicillium roqueforti, Penicillium paneum, Hyphopichia burtonii, and Aspergillus pseudoglaucus) were tested against five sanitizers at three concentrations: benzalkonium chloride (0.3%, 2.5%, 5%), biguanide (2%, 3.5%, 5%), peracetic acid (0.15%, 1.5%, 3%), quaternary ammonium (0.3%, 2.5%, 5%), and sodium hypochlorite (0.01%, 0.1%, 0.2%). Peracetic acid was the most effective sanitizes considering the genera, species, and concentrations evaluated, generally being capable of reductions between 2 and 4 logs of initial control tested. Biguanide should not be the compound of choice when the main goal of the bakery industry is fungal control.

Survival and stability of Lactobacillus fermentum and Wickerhamomyces anomalus strains upon lyophilisation with different cryoprotectant agents.

The stability of microorganisms along the time is important for allowing their industrial use as starter agents, improving fermentation processes. This study aimed to evaluate the survival and maintenance of the cell viability of the lactic acid bacteria Lactobacillus fermentum IAL 4541 and the yeast Wickerhamomyces anomalus IAL 4533, both isolated from wheat sourdough, after lyophilisation with different cryoprotectant and storage at room temperature along a year. Treatments involved adding control solution (S1 = 0.1% peptone water), and four cryoprotectant solutions S2 (10% sucrose), S3 (5% trehalose), S4 (10% skim milk powder) and S5 (10% skim milk powder plus 5% sodium glutamate) to the microbial cells previously of freeze drying processing. To verify the effect of lyophilisation on the number of microbial cells recovered, microbiological analyses were performed and cell viability was calculated before and after lyophilisation and regularly during a storage period of 365 days at room temperature. Viability after freeze-drying was influenced by the cryoprotectant agent employed, as well the microbial stability conferred along the storage. Differences on the microorganism response to some protectors were observed between the lactic acid bacteria and the yeast evaluated. W. anomalus was more affected by absence of cryoprotectant (S1) during freeze drying processing, but this microorganism was more stable than L. fermentum along the storage without the presence of protectant agents. For L. fermentum, S5 was the best protectant, allowing the recovering of 100% of the bacterial cells after lyophilisation and 87% of cell viability was observed after one year storage, followed by S4 (96 and 74%, respectively). S4 and S5 were the best protectant to W. anomalus (viability >80% after 1 year), but no increase in the yeast cell viability was conferred by addition of glutamate (S5) to skim milk. After 1 year of storage, trehalose was much more effective on protection of the yeast than bacteria (72% and 7% of viability, respectively). S2 was the less protectant agent among the tested, and their effectiveness was higher in L. fermentum (allowing 14% of cell recovering up to 120 days of storage) if compared to W. anomalus (25% of viability until 90 days of storage). Our results demonstrate that freeze-drying is a realistic technology for the stability and maintenance of the potential sourdough starter L. fermentum and W. anomalus for long time; however, the choice of cryoprotectant will influence the process effectiveness.

Mycological quality of pecan nuts from Brazil: absence of aflatoxigenic fungi and aflatoxins / Qualidade micológica de nozes pecan do Brasil: ausência de fungos aflatoxigênicos e aflatoxinas

ABSTRACT: In recent years, Brazil has encouraged the cultivation of pecans to meet both demands of domestic and international market of nuts. New genetic varieties of pecans have been selected in recent years, but available scientific information on the occurrence of fungi and aflatoxins in the international literature is out of date. Therefore, the present study aimed to quantify and identify fungal microbiota and the presence of aflatoxins in pecan nuts cultivated in southern Brazil. Fifty-two pecan nut lots (Barton variety) were obtained from producers from 19 cities of Rio Grande do Sul State and analyzed by direct plating in Agar Dichloran Glycerol 18% (DG18) and Aspergillus Flavus and Parasiticus Agar (AFPA), following incubation at 25 °C for 7 days. Aflatoxins analyses were carried out using HPLC coupled with a mass spectrometer. Results revealed at least 10 different genera of fungi. Aspergillus, Penicillium, Fusarium, and Cladosporium were predominant. Xerophilic species of Aspergillus (A. wentii, A. ruber, A. pseudoglaucus, and A. chevalieri) were commonly reported in the samples. No potential aflatoxin-producing species was isolated and no aflatoxins were detected (LOQ=1 μg/kg and LOD=0.1 μg/kg for AFB1 and AFB2, and 0.3 μg/kg for AFG1 and AFG2) in the evaluated samples. The absence of this carcinogenic group of mycotoxins is highly positive and could boost the investments in the sector, as well as stimulate the commercialization and consumption of this variety of nut.

RESUMO: Nos últimos anos, o Brasil tem incentivado o cultivo de pecãs para atender a demanda do mercado nacional e internacional de nozes. Novas variedades genéticas de pecãs foram selecionadas nos últimos anos, mas informações científicas disponíveis sobre a ocorrência de fungos e aflatoxinas na literatura internacional estão desatualizadas. Portanto, o presente estudo objetivou quantificar e identificar a microbiota fúngica e a presença de aflatoxinas em nozes cultivadas no sul do Brasil. Cinquenta e dois lotes de nozes (variedade Barton) foram obtidos de produtores de 19 municípios do Estado do Rio Grande do Sul e analisados ​​por meio de Ágar Dicloran Glicerol 18% (DG18) e Aspergillus Flavus e Parasiticus Agar (AFPA), após incubação em 25 °C durante 7 dias. Análises de aflatoxinas foram realizadas usando HPLC acoplado a um espectrômetro de massa. Os resultados revelaram pelo menos 10 gêneros diferentes de fungos. Aspergillus, Penicillium, Fusarium e Cladosporium foram predominantes. Espécies xerofílicas de Aspergillus (A. wentii, A. ruber, A. pseudoglaucus e A. chevalieri) foram comumente encontradas nas amostras. Nenhuma espécie potencial produtora de aflatoxinas foi isolada e nenhuma aflatoxina foi detectada (LOQ=1 μg/kg e LOD=0,1 μg/kg para AFB1 e AFB2; e 0,3 μg/kg para AFG1 e AFG2) nas amostras avaliadas. A ausência desse grupo carcinogênico de micotoxinas é altamente positiva e pode impulsionar os investimentos no setor, além de estimular a comercialização e o consumo dessa variedade de nozes.

Fungi in spices and mycotoxigenic potential of some Aspergilli isolated.

The aim of this study was to identify fungal species present in 200 samples of rosemary, fennel, cinnamon, clove, pepperoni, black and white pepper and oregano and evaluate the mycotoxigenic potential of the some Aspergilli isolated. Clove, black and white peppers were analyzed by direct plating. For rosemary, cinnamon, fennel, pepperoni pepper and oregano samples were used spread plate. Mycotoxigenic capacity was verified by the agar plug method. With the exception of clove, all the spices showed high fungal contamination, especially by Aspergillus sp., Penicillium sp. and Cladosporium sp. Frequency of toxigenic Aspergillus spp. was intense in white and black peppers, with presence of Aspergillus flavus (up to 32%), Aspergillus nomius (up to 12%), Aspergillus parasiticus (up to 4%), Aspergillus niger complex (up to 52%), Aspergillus ochraceus (up 12%) and Aspergillus carbonarius (up to 4%). 14,2% of A. flavus isolated from black pepper were aflatoxins producers. In the white pepper, 66.7% of A. flavus isolates and 100% of A. nomius were aflatoxigenic. Oregano showed the highest number of A. niger complex isolates (49), however, only 2.04% produced ochratoxin A. This study showed a huge fungal presence in spices, which could compromise the sensorial quality of these products and represent a hazard for consumers.

Incidence, populations and diversity of fungi from raw materials, final products and air of processing environment of multigrain whole meal bread.

This study aimed to assess the incidence, to quantify and to assess the diversity of fungi in a multigrain whole meal bread processing plant. Two hundred and eight one (n=281) samples were analyzed, including raw materials (n=120), air samples (n=136) and multigrain breads (n=25). Among the raw materials, the whole corn flour showed the highest counts of fungi (4.8logCFU/g), followed by whole-wheat flour (3.1logCFU/g). The counts of fungi in the air of processing environment were higher in post-baking steps (oven output, cooling, slicing, packaging) than in pre-baking steps (weighing and mixer) (p<0.05). Species of fungi isolated from spoiled bread samples stored at 5, 20, 25 and 30, and 40°C corresponded mostly to Penicillium paneum and Penicillium polonicum isolated from 20 and 24% of samples, respectively. These species were also isolated from raw materials (P. paneum and P. polonicum) and air collected at different processing sampling points (P. polonicum). The high counts of filamentous fungi in raw materials and air samples in processing steps such as cooling, slicing, and packaging, suggest that contamination that may occur in these steps can be critical for the shelf life of breads. The results of this study highlight that the prevention of contamination of breads by fungal spores is still a challenge for bakery industries and that other strategies such as control of germination and growth of spoilage fungi through the development of more stable formulations have to be developed.

Influence of storage temperature on growth of Penicillium polonicum and Penicillium glabrum and potential for deterioration of frozen chicken nuggets.

The practice of freezing food is one of the main processes used by the industry to prolong the shelf life of foods. Its use has expanded in recent years due to the increased consumption of convenience products, many of which are sold in frozen form. The temperature at which these foods are maintained during marketing in supermarkets or stored in the consumer's home is critical to ensure microbiological stability of products. Temperature abuse can allow microbial growth, especially growth of filamentous psychrophilic fungi. Besides economic losses in the industrial sector due to the return of products and loss of confidence by consumers, the development of fungi in foods is a public health problem due to the possibility of mycotoxin production. The aim of this study was to assess the growth at temperatures of 5, 0, -5 and -18°C for two species of fungi involved in the deterioration of frozen chicken nuggets, Penicillium polonicum (33/12 NGT) and Penicillium glabrum (29/12 NGT), inoculated both in culture medium and in the food. The results demonstrated that P. polonicum was able to form microcolonies on potato dextrose agar plates at 0°C and form visible colonies on the surface of the frozen chicken nuggets kept at -5°C for 120 days, regardless of brand. For P. glabrum the limiting growth temperature was 5°C in the culture medium and 0°C on frozen chicken nuggets, regardless of the brand analyzed. Thus, it is essential to adhere to the storage temperatures recommended to ensure the stability and safety of this food product.