Curative activity of postharvest GRAS salt treatments to control citrus sour rot caused by Geotrichum citri-aurantii.

The effectiveness of the "generally recognized as safe" (GRAS) salts potassium sorbate (PS), sodium benzoate (SB), sodium ethylparaben (SEP) and sodium methylparaben (SMP) to control sour rot, caused by Geotrichum citri-aurantii, was assessed by dipping economically important citrus species and cultivars in aqueous solutions for 30, 60 or 150 s at 20 °C, followed by examination after 8 d of storage at 28 °C. Curative activity was determined because the fruit were inoculated 24 h prior to treatment. Dipping fruit for 60 s in SMP (200 mM), SEP (200 mM) or SB (3% w/v) were very effective and reduced sour rot incidence and severity by up to 90%. Their effectiveness was similar or superior to that of the conventional fungicide propiconazole (PCZ). In contrast, PS (200 mM) did not control sour rot on 'Oronules' or 'Ortanique' mandarins, but it reduced sour rot incidence on 'Barnfield' oranges by 50% compared to inoculated, water-treated control fruit. Sour rot was better controlled on oranges than on mandarins. Furthermore, heating the solutions to 50 °C enhanced their effectiveness, while post-treatment rinsing of the fruit with tap water reduced their effectiveness. Dipping 'Valencia Late' oranges in SB (3% w/v) or SMP (200 mM) for 60 s followed by long storage for up to 8 weeks at 5 °C and 90% RH, reduced sour rot incidence from 55% among water-treated control fruit to 2 to 6%, and matched the effectiveness of PCZ. No fruit in any test were visibly harmed. Both SB and SMP salts could be potential alternatives to conventional fungicides, such as PCZ or guazatine, for the integrated postharvest management of citrus sour rot.

Production of volatile compounds by yeasts using hydrolysed grape seed oil obtained by immobilized lipases in continuous packed-bed reactors.

Lipases CAL-B, TLL, and RML were used in the synthesis of free fatty acids of grape seed oil as heterogeneous substrate. The best enzyme was used to optimize the reaction variables temperature, enzyme content, and molar ratio of water:oil in batch reactions using experimental planning. The ideal conditions to produce free fatty acids using pure RML were 45 °C, 12:1 substrate molar ratio, and 15% enzyme, resulting in 66% of oil hydrolysis and a productivity of 0.54 mol L-1 min-1 in 4 h of reaction at 180 rpm. Repeated batches of reaction were performed testing the operational stability of RML, results showing that this enzyme could be used for at least 20 cycles keeping more than 80% of its initial activity, suggesting its potential use in industrial processes. The synthesis of free fatty acids was then evaluated in continuous reactions using packed-bed reactor (PBR). The highest productivity in the continuous process was 6.85 mol L-1 min-1, using only RML, showing an operational stability higher than 80% of its initial conversion capacity after 11 days of operation, at a flow rate of 0.13 mL min-1 at 45 °C. We evaluated the use of this hydrolyzed oil as substrate for lactone bioproduction using Galactomyces geotrichum UFMG-CM-Y3276, G. geotrichum UFMG-CM-Y3558, and Geotrichum klebahnii UFMG-CM-Y3014 screened for their oil-hydrolysis ability. Volatile compounds were qualitatively identified in GC-MS as γ-octalactone and γ-nonalactone.

Biofilm production by Aureobasidium pullulans improves biocontrol against sour rot in citrus.

The biocontrol efficiency of Aureobasidium pullulans strain ACBL-77 against Geotrichum citri-aurantii, causal agent of sour rot in citrus, and their interactions were evaluated. For this, were evaluated the incorporation of nutrients in optimizing the antagonistic activity of the yeast, the competition for nutrients between microorganisms, the effect of nutrients on yeast cell and biofilm production and their correlation in the biocontrol of disease, the survival of yeast in citrus fruits and the interaction between microorganisms using scanning electron microscopy. Micronutrients (boric acid, cobalt chloride and ammonium molybdate) favoured the antagonistic action of A. pullulans. Ammonium sulfate 1% and sucrose 0.5% favoured the yeast during the competition between the microorganisms. The addition of ammonium sulfate (1%) in the yeast culture stimulated biofilm production and increased the antagonistic activity against the disease, as also allowed the better survival of yeast in wounded sites of citrus fruit. The yeast was found to be able to form biofilms on citrus, deforming the pathogen hyphae. These results showed the importance of the addition of nutrients in A. pullulans based-formulations when aiming for their use on a commercial scale. This is the first report of a positive correlation between the increase in the quantity of biofilm produced by A. pullulans, with increased antagonistic activity.

Geotrichum candidum as a possible cause of bovine abortion.

Geotrichum spp. are ubiquitous, saprotrophic fungi found in soil, organic matter, and silage, as a contaminant in food products and in the digestive tracts of mammals. The current study reports a case of Geotrichum candidum infection with dermatitis in an aborted bovine fetus with skin and lung lesions. A 6-month-old aborted male Holstein Friesian fetus displayed unusual lesions on the skin of the abdomen, thorax, and head, which was excessively thickened and wrinkled. These changes corresponded to orthokeratotic hyperkeratosis, neutrophil accumulation in the stratum corneum, a pyogranulomatous inflammatory infiltrate, and superficial dermal necrosis. Moderate suppurative multifocal pneumonia was observed. Large numbers of mononuclear cells and occasional fibrin thrombi within blood vessels were found in the lungs, brain, and cerebellum. Gridley staining revealed fungal structures within the skin lesions. The mycological exam demonstrated the growth of G. candidum, and phase contrast microscopy conducted on the abomasal fluid revealed hyphae compatible with this agent. The skin lesions observed, in association with the fungus isolated, indicated that the abortion was due to G. candidum infection of the bovine fetus.

Influência do uso de cultura adjunta nas características físico-químicas e sensoriais do queijo tipo Camembert / Effect of the addition of a supplementary culture on the physical-chemical and sensory characteristics of Camembert-type cheese

O presente estudo avaliou o efeito da adição do Geotrichum candidum, como cultura adjunta, na aceitação e composição do queijo tipo Camembert. O leite utilizado na fabricação dos queijos foi submetido à pasteurização e adicionado de Lactococcus lactis subsp. Lactis e Lactococcus lactis subsp. cremoris. Foram realizados quatro tratamentos: controle (T1) sem adição de Geotrichum candidum; e T2, T3 e T4, com concentrações, respectivamente, de 0,5 U, 1 U e 1,5 U do microorganismo. Após 1, 9, 17, 25, 33 e 41 dias de fabricação, os queijos foram avaliados quanto aos parâmetros: acidez, pH, proteína, NS a pH 4,6, NS em TCA 12%, umidade, e para gordura e sal no queijo com um dia de preparação. A análise de aceitação foi realizada aos 30 dias de maturação. Os queijos com e sem adição de Geotrichum candidum não diferiram (P > 0,05) nas análises físico-químicas. Os quatros tratamentos diferiram (P < 0,05) em todas as análises no decorrer do tempo de maturação. Com o aumento na concentração do Geotrichum candidum, houve aumento na média de aceitação dos provadores. A utilização do Geotrichum candidum é uma boa alternativa para as indústrias que desejam um produto com boa aceitabilidade e características semelhantes ao tradicional queijo Camembert.(AU)

This study evaluated the effect of Geotrichum candidum addition as an adjunct culture on the acceptance and composition of Camembert-type cheese. The milk used for cheese manufacturing was pasteurized and supplemented with Lactococcus lactis subsp. Lactis and Lactococcus lactis subsp. cremoris. Four treatments were performed: control (T1) without adding Geotrichum candidum; T2-T3-T4: containing microorganisms at concentrations of 0.5 U, 1 U and 1.5 U, respectively. After 1, 9, 17, 25, 33 and 41 days of manufacture, the cheese samples were analyzed on the following parameters: acidity, pH, protein, soluble nitrogen at pH 4.6, TCA 12 % soluble nitrogen, and moisture, and fat and salt contents in cheese after 1-day production. Sensory analysis was performed after 30 days of maturing. No significant difference was found in the physical-chemical analysis in cheeses produced with and without adding Geotrichum candidum (P > 0.05). However, significant differences (P < 0.05) were detected in all of the analyses throughout the maturing period. The acceptance of cheese by tasters increased with the rise of Geotrichum candidum concentration added into the cheese. The use of Geotrichum candidum is a valuable alternative for industries seeking for a product with favorable acceptability and with characteristics similar to the traditional Camembert-type cheese.(AU)

Proteinase and phospholipase activities and development at different temperatures of yeasts isolated from bovine milk.

The presence of yeasts in milk may cause physical and chemical changes limiting the durability and compromising the quality of the product. Moreover, milk and dairy products contaminated by yeasts may be a potential means of transmission of these microorganisms to man and animals causing several kinds of infections. This study aimed to determine whether different species of yeasts isolated from bovine raw milk had the ability to develop at 37°C and/or under refrigeration temperature. Proteinase and phospholipase activities resulting from these yeasts were also monitored at different temperatures. Five genera of yeasts (Aureobasidium sp., Candida spp., Geotrichum spp., Trichosporon spp. and Rhodotorula spp.) isolated from bovine raw milk samples were evaluated. All strains showed one or a combination of characteristics: growth at 37°C (99·09% of the strains), psychrotrophic behaviour (50·9%), proteinase production (16·81% of the strains at 37°C and 4·09% under refrigeration) and phospholipase production (36·36% of the isolates at 37°C and 10·9% under refrigeration), and all these factors may compromise the quality of the product. Proteinase production was similar for strains incubated at 37°C (16·81% of the isolates) and room temperature (17·27%) but there was less amount of phospholipase-producing strains at room temperature (15·45% of the isolates were positive) when compared with incubation at 37°C (36·36%). Enzymes production at 37°C by yeasts isolated from milk confirmed their pathogenic potential. The refrigeration temperature was found to be most efficient to inhibit enzymes production and consequently ensure better quality of milk. The viability of yeasts and the activity of their enzymes at different temperatures are worrying because this can compromise the quality of dairy products at all stages of production and/or storage, and represent a risk to the consumer.

Inhibition of citrus fungal pathogens by using lactic acid bacteria.

The effect of lactic acid bacteria (LAB) on pathogenic fungi was evaluated and the metabolites involved in the antifungal effect were characterized. Penicillium digitatum (INTA 1 to INTA 7) and Geotrichum citri-aurantii (INTA 8) isolated from decayed lemon from commercial packinghouses were treated with imazalil and guazatine to obtain strains resistant to these fungicides. The most resistant strains (4 fungal strains) were selected for evaluating the antifungal activity of 33 LAB strains, among which only 8 strains gave positive results. The antifungal activity of these LAB strains was related to the production of lactic acid, acetic acid, and phenyllactic acid (PLA). A central composite design and the response surface methodology were used to evaluate the inhibitory effect of the organic acids produced by the LAB cultures. The antifungal activity of lactic acid was directly related to its concentration; however, acetic acid and PLA showed a peak of activity at 52.5 and 0.8 mM, respectively, with inhibition rates similar to those obtained with Serenade((R)) (3.0 ppm) imazalil (50 ppm) and guazatine (50 ppm). Beyond the peak of activity, a reduction in effectiveness of both acetic acid and PLA was observed. Comparing the inhibition rate of the organic acids, PLA was about 66- and 600-fold more effective than acetic acid and lactic acid, respectively. This study presents evidences on the antifungal effect of selected LAB strains and their end products. Studies are currently being undertaken to evaluate the effectiveness in preventing postharvest diseases on citrus fruits.

Sequential use of ammonium and leucine as nitrogen sources during growth of Geotrichum candidum on a glucose based medium / Uso secuencial del amonio y leucina como fuentes de nitrógeno durante el crecimiento de Geotrichum candidum en un medio basado en glucosa

Geotrichum candidum growth on ammonium and leucine as nitrogen sources and glucose as a carbon source was examined. A clear preference of G. candidum for ammonium over leucine as a nitrogen source was shown. Indeed, ammonium was completely exhausted at the end of exponential growth after less than 35 hrs of culture; in contrast only 5 percent of leucine was concomitantly assimilated. Growth continued at slower rates on glucose and leucine as carbon and nitrogen sources respectively, and at the end of culture (185 hrs), leucine was completely exhausted.

Growth of lignocellulosic-fermenting fungi on different substrates under low oxygenation conditions.

Four soil fungi able to grow under low oxygenation conditions were selected and used in studies to determine the production of enzymes that promote the degradation of lignocellulosic materials. The capacity of these fungi to ferment such materials was also investigated. The fungi were grown in sugarcane bagasse and sawdust at final concentrations of 4 and 10%, as the carbon sources. The strains were cultivated under microaerophilic and combined conditions of oxygenation (aerobic followed by microaerophilic conditions). The results obtained with the basidiomycete specie, Trichocladium canadense, Geotrichum sp., and Fusarium sp. suggest that they prefer lower oxygen concentration for growth and enzyme production. Lignocellulolytic activities were detected in all strains but varied with the carbon source used. The highest levels of these activities were produced by the Basidiomycete specie and Fusarium sp. Ethanol and other nongaseous fermentation products were detected following high-performance liquid chromatography analysis using a supelcogel C-610H column, demonstrating the fermentative capability of these strains. In view of their ability to produce enzymes necessary for the breakdown of lignocellulosic materials and to utilize most of the degradation products for growth, these strains have a great potential for biotechnological application.

Fungal bioturbation paths in a compact disk.

We report here on bioturbation traces, with micro-dendrite textures, composed of a mixture of altered aluminum and polycarbonate, which have been developed in a common compact disk (CD), destroying information pits. Fungal hyphae proliferated in these deteriorated zones, and Geotrichum-type fungus was isolated from surface-sterilized CD fragments. The severe biodeterioration described is attributed to the slow growth of this arthroconidial fungus on the CD material in the tropical indoor environment of Belize, Central America (approximately 30 degrees C, approximately 90% humidity).