Control of postharvest decay on cherry tomatoes by marine yeast Rhodosporidium paludigenum and calcium chloride

Aims: In this study, the potential of calcium chloride (CaCl₂) application to improve the efficacy of the marine antagonist Rhodosporidium paludigenum in controlling postharvest diseases of cherry tomatoes was assessed. Methods and Results: CaCl₂ alone was found not to have any direct influence on the population growth of R. paludigenum in NYDB cultures or in cherry tomato wounds. However, the combined treatments with 1 × 10⁸ cells ml^?1R. paludigenum and CaCl₂ at the concentration from 0·5 to 2% showed high activities to reduce black rot caused by Alternaria alternata in cherry tomato wounds, significantly higher than those of R. paludigenum or CaCl₂ alone. Meanwhile, 0·5% CaCl₂ in combination with 1 × 10⁸ cells ml^?1R. paludigenum greatly inhibited the natural decay of cherry tomatoes in 21 days’ storage at 25°C. Conclusions: The combination of R. paludigenum and CaCl₂ enhances the inhibition of black rot and natural decay of postharvest cherry tomatoes. The results from this study provide a new way to improve the efficiency of R. paludigenum in maintaining the quality of postharvest fruits and vegetables. Significance and Impact of the Study: The marine yeast R. paludigenum combined with CaCl₂ has greatly potential use as an alternative to chemical fungicides in inhibiting postharvest decay on cherry tomatoes.     

The combined effect of 1‐methylcyclopropene and citral suppressed postharvest grey mould of tomato fruit by inhibiting the growth of Botrytis cinerea

1‐Methylcyclopropene (1‐MCP, 1 μl/L) and 1 × minimum fungicidal concentration (MFC) citral alone and in combination were used to treat on postharvest tomato fruits to investigate their influence on disease incidence and postharvest quality during fruit storage, which were stored at 90%–95% relative humidity and 25 ± 2°C. Weight loss, pH, hue angle (Hue^°), total soluble solid (TSS), ascorbic acid content, firmness and antioxidant enzyme activities were evaluated after each storage period. 1 μl/L 1‐MCP or 1 × MFC citral reduced weight loss, retarded peel colour changes and retained postharvest fruit quality. 1 μl/L 1‐MCP + 1 × MFC citral could better maintain firmness and ascorbic acid content and increase antioxidant enzyme activities, compared to other treatments. Disease incidence of tomato fruit was significantly decreased, and spore germination and mycelia growth of Botrytis cinerea were suppressed by the combined treatment with 1 μl/L 1‐MCP and 1 × MFC citral. These results indicate that the combined treatment could effectively delay postharvest tomato fruits senescence and inhibit postharvest pathogens in vitro.     

Biological Control of Green Mould Decay in Postharvest Chinese Bayberries by Pichia membranaefaciens

The effect of the yeast antagonist Pichia membranaefaciens for control of green mould decay caused by Penicillium citrinum or Verticicladiella abietina and natural decay in postharvest Chinese bayberries (Myrica rubra Seib & Zucc.), and the possible mechanisms were investigated. The results showed that 1 × 10⁹ colony‐forming units (CFU)/ml of washed cell suspensions of the yeast provided better control of green mould decay than yeast in culture broth at the same concentration. Treatment with cell‐free culture filtrates or autoclaved cell cultures had little effect on disease incidence. The concentration of a washed cell suspension of P. membranaefaciens had a significant effect on efficacy in controlling disease incidence. At a concentration range from 1 × 10⁶ to 1 × 10⁹ CFU/ml, the higher the concentration of the antagonist, the lower was the incidence of the disease. In the inoculated wounds of Chinese bayberries, populations of P. membranaefaciens increased by approximately 145‐ and 41‐fold, respectively, after incubation at 20°C for 2 day or at 1°C for 8 day. P. membranaefaciens significantly induced activities of two defence‐related enzymes chitinase and β‐1, 3‐glucanase in Chinese bayberries. The in vitro experiment showed that spore germination and germ tube elongation of the two pathogens were markedly inhibited by washed cell suspensions of P. membranaefaciens. In addition, P. membranaefaciens significantly reduced natural decay in Chinese bayberries. These results indicate that P. membranaefaciens can effectively reduce fruit decay possibly by directly inhibiting pathogen growth and indirectly by inducing disease resistance. Thus, we suggest that P. membranaefaciens has potential as a biocontrol agent to control fruit decay in Chinese bayberries during postharvest storage.     

Molecular characterization,virulence and horizontal transmission of Beauveria pseudobassiana from Dendroctonus micans (Kug.) (Coleoptera: Curculionidae)

The great spruce bark beetle, Dendroctonus micans (Kugelann) (Coleoptera: Curculionidae), has been a potential threat for Turkey and the entire Eurasian spruce forests for many years. Control strategies which have been applied so far are still insufficient to prevent its damage. A previous study has shown that a Beauveria isolate (ARSEF 9271) proved to be an efficient microbial control agent against the great spruce bark beetle. In this study, this isolate was identified as B. pseudobassiana based on the partial sequence of EF1‐α and ITS sequence. A conidial suspension (1 × 10⁸/ml) of this fungus caused 100% mortality on both larvae and adults of D. micans within 5 and 6 days, respectively. Also, it caused 100% mycosis value on both larvae and adults. Mortality values of horizontal transmission experiments between larvae and adults which were contaminated with 1 × 10⁶/ml spore suspension at 25%, 50%, 75% and 100% rates were determined as 100% after 15 days at 20°C under the laboratory conditions. We also determined the decrease of the damage in spruce wood block (15 × 25 cm) when the contamination rate of the larvae was increased. Our results indicate that B. pseudobassiana ARSEF 9271 seems to be a very promising biocontrol agent against D. micans.     

Dietary Mannan-oligosaccharides potentiate the beneficial effects of Bifidobacterium bifidum in broiler chicken

This study investigated the effects of dietary Bifidobacterium bifidum (BFD) and mannan-oligosaccharide (MOS), as a synbiotic, on the production performance, gut microbiology, serum biochemistry, antioxidant profile and health indices of broiler chicken. Six dietary treatments were T1 (negative control), T2 (positive control-20 mg antibiotic BMD kg⁻¹ diet; BMD: bacitracin methylene disalicylate), T3 (0·1% MOS + 10⁶ CFU BFD per g feed), T4 (0·1% MOS + 10⁷ CFU BFD per g feed), T5 (0·2% MOS + 10⁶ CFU BFD per g feed) and T6 (0·2% MOS + 10⁷ CFU BFD per g feed). Significantly (P < 0·01) better growth performance and efficiency was observed in birds supplemented with 0·2% MOS along with 10⁶ CFU BFD per g of feed compared to BMD and control birds. Supplementation with 0·2% MOS along with either 10⁶ or 10⁷ CFU BFD per g feed reduced (P < 0·01) the gut coliform, Escherichia coli, total plate count, and Clostridium perfringens count and increased the Lactobacillus and Bifidobacterium count. Significantly (P < 0·01) higher serum and liver antioxidant enzyme pool, serum HDL cholesterol and lower serum glucose, triglyceride, total cholesterol, cardiac risk ratio, atherogenic coefficient and atherogenic index of plasma were observed in birds supplemented with 0·2% MOS along with 10⁶ CFU BFD per g of feed compared to control or BMD supplemented birds. Better production performance, gut microbial composition, serum biochemistry, antioxidant profile and health indices were depicted by broiler chicken supplemented with 0·2% MOS and 10⁶ CFU BFD per g of feed.     

First description of the environmental niche of the epibenthic dinoflagellate species Coolia palmyrensis,C. malayensis,and C. tropicalis (Dinophyceae) from Eastern Australia

Environmental variables such as temperature, salinity, and irradiance are significant drivers of microalgal growth and distribution. Therefore, understanding how these variables influence fitness of potentially toxic microalgal species is particularly important. In this study, strains of the potentially harmful epibenthic dinoflagellate species Coolia palmyrensis, C. malayensis, and C. tropicalis were isolated from coastal shallow water habitats on the east coast of Australia and identified using the D1‐D3 region of the large subunit (LSU) ribosomal DNA (rDNA). To determine the environmental niche of each taxon, growth was measured across a gradient of temperature (15–30°C), salinity (20–38), and irradiance (10–200 μmol photons · m^?2 · s^?1). Specific growth rates of Coolia tropicalis were highest under warm temperatures (27°C), low salinities (ca. 23), and intermediate irradiance levels (150 μmol photons · m^?2 · s^?1), while C. malayensis showed the highest growth at moderate temperatures (24°C) and irradiance levels (150 μmol photons · m^?2 · s^?1) and growth rates were consistent across the range of salinity levels tested (20–38). Coolia palmyrensis had the highest growth rate of all species tested and favored moderate temperatures (24°C), oceanic salinity (35), and high irradiance (>200 μmol photons · m^?2 · s^?1). This is the first study to characterize the environmental niche of species from the benthic harmful algal bloom genus Coolia and provides important information to help define species distributions and inform risk management.     

Antagonistic Yeasts for Biocontrol of the Banana Postharvest Anthracnose Pathogen Colletotrichum musae

The biocontrol potentials of Candida tropicalis YZ1, C. tropicalis YZ27 and Saccharomyces cerevisiae YZ7 against the postharvest anthracnose pathogen Colletotrichum musae were investigated. Treatments with all the three biocontrol agents (1 × 10⁸ CFU/ml) significantly reduced the natural anthracnose disease severity of harvested banana fruits stored at ambient condition. Germination and survival of C. musae spores were markedly inhibited by all the three yeast strains in in vitro tests. The niche overlap index (NOI) was used to determine the interaction between the antagonists and C. musae, and the results (high NOI values) suggest competitive exclusion of C. musae by the yeast strains. C. tropicalis YZ27 inoculated on banana wounds exhibited rapid colonization and maintenance of its population on the inoculated site. The biocontrol efficacy was also observed as a function of concentration of the antagonist applied. The fruits treated with C. tropicalis YZ27, 36 h before pathogen inoculation, showed the best results with 96.0% disease inhibition followed by those treated 24 h before with 84.0% inhibition. The above results point to competition for nutrients and space as the main mechanism of antagonistic action of C. tropicalis YZ27 against C. musae.     

Analysis of broth-cultured Bacillus atrophaeus and Bacillus cereus spores

Aims: To compare physical properties of spores that were produced in broth sporulation media at greater than 10⁸ spores ml⁻¹. Methods and Results: Bacillus atrophaeus reproducibly sporulated in nutrient broth (NB) and sporulation salts. Microscopy measurements showed that the spores were 0·68 ± 0·11 μm wide and 1·21 ± 0·18 μm long. Coulter Multisizer (CM3) measurements revealed the spore volumes and volume-equivalent spherical diameters, which were 0·48 ± 0·38 μm³ and 0·97 ± 0·07 μm, respectively. Bacillus cereus reproducibly sporulated in NB, sporulation salts, 200 mmol l⁻¹ glutamate and antifoam. Spores were 0·95 ± 0·11 μm wide and 1·31 ± 0·17 μm long. Spore volumes were 0·78 ± 0·61 μm³ and volume-equivalent spherical diameters were 1·14 ± 0·11 μm. Bacillus atrophaeus spores were hydrophilic and B. cereus spores were hydrophobic. However, spore hydrophobicity was significantly altered after treatment with pH-adjusted bleach. Conclusions: The utility of a CM3 for both quantifying Bacillus spores and measuring spore sizes was demonstrated, although the volume between spore exosporium and spore coat was not measured. This study showed fundamental differences between spores from a Bacillus subtilis- and B. cereus-group species. Significance and Impact of the Study: This is useful for developing standard methods for broth spore production and physical characterization of both living and decontaminated spores.     

Characterization and flocculation properties of biopolymeric flocculant (glycosaminoglycan) produced by Cellulomonas sp. Okoh

Aims

Bioflocculant production potential of an actinobacteria isolated from a freshwater environment was evaluated and the bioflocculant characterized.

Methods and Results

16S rDNA nucleotide sequence and BLAST analysis was used to identify the actinobacteria and fermentation conditions, and nutritional requirements were evaluated for optimal bioflocculant production. Chemical analyses, FTIR, 1H NMR spectrometry and SEM imaging of the purified bioflocculant were carried out. The 16S rDNA nucleotide sequences showed 93% similarities to three Cellulomonas species (strain 794, Cellulomonas flavigena DSM 20109 and Cellulomonas flavigena NCIMB 8073), and the sequences was deposited in GenBank as Cellulomonas sp. Okoh (accession number HQ537132 ). Bioflocculant was optimally produced at an initial pH 7, incubation temperature 30°C, agitation speed of 160 rpm and an inoculum size of 2% (vol/vol) of cell density 1·5 × 10⁸ cfu ml^?1. Glucose (88·09% flocculating activity; yield: 4·04 ± 0·33 g l^?1), (NH₄)₂NO₃ (82·74% flocculating activity; yield: 4·47 ± 0·55 g l^?1) and MgCl₂ (90·40% flocculating activity; yield: 4·41 g l^?1) were the preferred nutritional source. Bioflocculant chemical analyses showed carbohydrate, protein and uronic acids in the proportion of 28·9, 19·3 and 18·7% in CPB and 31·4, 18·7 and 32·1% in PPB, respectively. FTIR and 1H NMR indicated the presence of carboxyl, hydroxyl and amino groups amongst others typical of glycosaminoglycan. SEM imaging revealed horizontal pleats of membranous sheets closely packed.

Conclusion

Cellulomonas sp. produces bioflocculant predominantly composed of glycosaminoglycan polysaccharides with high flocculation activity.

Significance and Impact of the Study

High flocculation activity suggests suitability for industrial applications; hence, it may serve to replace the hazardous flocculant used in water treatment.     

In vitro assays on the susceptibility of four species of nematophagous fungi to anthelmintics and chemical fungicides/antifungal drug

Using nematophagous fungi for the biological control of animal parasitic nematodes will become one of the most promising strategies in the search for alternative chemical drugs. The purpose of this study was to check the in vitro activity of four anthelmintics, four chemical fungicides and two antifungal drugs on the spore germination of nematophagous fungi: Duddingtonia flagrans (SF170), Arthrobotrys oligospora (447), Arthrobotrys superba (435) and Arthrobotrys sp. (PS011). A modified 24-well cell culture plate assay was conducted to evaluate the susceptibility of nematophagous fungi against drugs tested by calculating the effective middle concentrations (EC₅₀) of each tested drug to inhibit the germination of fungal spores. EC₅₀ ranged between 0·7 and 47·2 μg ml⁻¹ for fenbendazole, thiabendazole and ivermectin, except levamisole (546·5–4057·8 μg ml⁻¹). EC₅₀ of tested fungicides was 0·6–2·3 μg ml⁻¹ for carbendazim, 55·9–247·4 μg ml⁻¹ for metalaxyl, 24·4–45·2 μg ml⁻¹ for difenoconazole, and 555·9–1438·3 μg ml⁻¹ for pentachloronitrobenzene (PCNB). EC₅₀ of two antifungal drugs was 0·03–3·4 μg ml⁻¹ for amphotericin B and 0·3–10·9 μg ml⁻¹ for ketoconazole. The results showed that 10 tested drugs, except for levamisole and PCNB, had in vitro inhibitory effects on nematophagous fungi. The chlamydospores of D. flagrans had the highest sensitivity to nine tested drugs, except for ketoconazole.     

Ethyl formate fumigation and ethyl formate plus cold treatment combination as potential phytosanitary quarantine treatments of Drosophila suzukii in blueberries

Spotted wing drosophila, Drosophila suzukii (Diptera: Drosophilidae), is a serious invasive pest of berries and cherries in the U.S. and Europe and has become a major phytosanitary trade barrier. In this pilot study, we explored the potential of using stand-alone ethyl formate (EF) treatment and a combinatory treatment of EF and cold temperature as postharvest control options for D. suzukii in imported blueberries. Stand-alone EF fumigations were effective against D. suzukii with LCt_99% of 207.7 and 168.5 g·h·m^?3 for eggs, the most tolerant life stage, at 5 and 21 °C, respectively. In a scale-up (10 m³) trial conducted at 5 °C, complete control of D. suzukii eggs placed inside and outside of blueberry boxes was achieved using 70 g·m^?3 EF for 4 h with 5% blueberry loading ratio without deleterious impact on blueberry appearance such as soft spot or berry shrivel. In small scale pilot studies, 9-d stand-alone cold treatment at 5 °C was sufficient for complete control of D. suzukii eggs and larvae tested, but not pupae. The efficacy of this cold treatment appeared to be improved when D. suzukii eggs were first treated with low-dose EF (LCt_50% level) prior to the cold treatment. The combination treatment resulted in complete mortality of D. suzukii eggs, larvae, and pupae tested after 7, 5, and 9 d of cold treatment, respectively. Together, these results suggest that stand-alone EF treatment, or the combination treatment of low-dose EF and cold as a systems approach may have a potential as postharvest treatments for D. suzukii in blueberries.     

Enhancing the adhesion of Epicoccum nigrum conidia to peach surfaces and its relationship to the biocontrol of brown rot caused by Monilinia laxa

Aims: To find a formulation of Epicoccum nigrum conidia that enhances its adhesion to peach surfaces and improves its biocontrol efficacy against brown rot caused by Monilinia laxa. Methods and Results: The stickers, glycerol, sodium alginate and methylcellulose; the desiccants, silica powder and talc; and a commercial adhesive (NU FILM 17^®) were added at two different points during the production of an E. nigrum conidial formulation to improve conidial adhesion to peach surfaces. Conidial adhesion levels were determined from the number of E. nigrum conidia that adhered to glass slides or peach surfaces and conidial viability of adherent E. nigrum conidia was determined from the number of colony‐forming units of glass or peach‐adherent E. nigrum that grew on Petri dishes that contained potato dextrose agar. Compared to dried E. nigrum conidia without additives, the adhesion and viability of adherent E. nigrum conidia to peach surfaces were enhanced when either 1·25% sodium alginate or 2·5% methylcellulose was added to the conidial mass after fluid‐bed drying, and when 2·5% methylcellulose was added to the conidial mass after its production and before fluid‐bed drying. Epicoccum nigrum conidial formulations with 2·5% methylcellulose were more effective than dried E. nigrum conidia without additives in reducing the incidence of brown rot in peaches caused by M. laxa. Conclusions: When 2·5% methylcellulose is incorporated into an E. nigrum conidial formulation, the adhesion of E. nigrum conidia to peach surfaces improves and results in efficacious biocontrol of brown rot. Significance and Impact of the Study: A new improved formulation of a biocontrol agent has been developed to improve the control of M. laxa on peaches.     

Optimization of selected cultivation parameters for Cordyceps guangdongensis

Aims: To increase the fruit body production of Cordyceps guangdongensis, selected cultivation conditions, especially nutritional parameters were optimized. Methods and Results: Cordyceps guangdongensis was inoculated on potato dextrose agar slants with pH values from 4·5 to 9·0 and cultivated in artificial media with different carbon and nitrogen supplements. Primordium formation in C. guangdongensis was favoured by slightly acidic conditions. Fruit body yields and biological efficiency (BE) recorded were all highest in cultures of C. guangdongensis supplemented with sucrose and KNO₃ as carbon and nitrogen supplements, respectively. Highest fruit body yields and BE values were recorded with C : N ratio of 12 : 1. The optimal medium consisted of (g l^?1) 20·0 sucrose, 4·0 soya bean powder, 5·0 beef extract and 10·0 KNO₃. Cultivation experiments using this medium confirmed its reliability; 18·35% of BE was obtained, compared with a calculated maximum BE of 18·65% based on orthogonal test data. Conclusions: Cordyceps guangdongensis preferred sucrose and potassium nitrate as best carbon and nitrogen supplements. It produced satisfying yield of fruit body with optimized medium. Significance and Impact of the Study: Optimized artificial cultivation conditions could promote the yield of C. guangdongensis and decreased the cost of production.     

Competitive exclusion as a mode of action of a novel Bacillus cereus aquaculture biological agent

Aims: To determine the contribution of potential modes of action of a Bacillus cereus aquaculture biological control agent in inhibition of the fish pathogen, Aeromonas hydrophila. Methods and Results: When B. cereus was tested in plate well inhibition studies, no production of antimicrobial compounds was detected. Bacillus cereus had a high growth rate (0·96 h^?1), whereas Aer. hydrophila concentration decreased by c. 70% in co‐culture experiments. In nutrient limitation studies, B. cereus had a significantly higher growth rate when cultured under glucose (P < 0·05) and iron (P < 0·01) limitation in comparison with Aer. hydrophila. Bacillus cereus glucose (0·30 g l^?1 h^?1) and iron (0·60 mg l^?1 h^?1) uptake rates were also significantly higher (P < 0·01) than the Aer. hydrophila glucose (0·14 g l^?1 h^?1) and iron (0·43 mg l^?1 h^?1) uptake rates. Iron uptake was facilitated by siderophore production shown in time profile studies where relative siderophore production was c. 60% through the late exponential and sporulation phases. Conclusions: Competitive exclusion by higher growth rate, competition for organic carbon and iron, facilitated by siderophore production, could be identified as mechanisms of pathogen growth inhibition by B. cereus. Significance and Impact of the Study: This study is the first elucidation of the mechanism of action of our novel B. cereus biological agent in growth attenuation of pathogenic Aer. hydrophila. This study enhances the application knowledge and attractiveness for adoption of B. cereus NRRL 100132 for exploitation in aquaculture.     

Improving probiotic spore yield using rice straw hydrolysate

Spore-forming Bacillus sp. has been extensively studied for their probiotic properties. In this study, an acid-treated rice straw hydrolysate was used as carbon source to produce the spores of Bacillus coagulans. The results showed that this hydrolysate significantly improved the spore yield compared with other carbon sources such as glucose. Three significant medium components including rice straw hydrolysate, MnSO₄ and yeast extract were screened by Plackett–Burman design. These significant variables were further optimized by response surface methodology (RSM). The optimal values of the medium components were rice straw hydolysate of 27% (v/v), MnSO₄ of 0·78 g l⁻¹ and yeast extract of 1·2 g l⁻¹. The optimized medium and RSM model for spore production were validated in a 5 l bioreactor. Overall, this sporulation medium containing acid-treated rice straw hydrolysate has a potential to be used in the production of B. coagulans spores.     

Antilisterial activity of lactose monolaurate in milk,drinkable yogurt and cottage cheese

Lactose monolaurate (LML) was previously found to be an antimicrobial against Listeria monocytogenes in culture medium at concentrations between 3 and 5 mg ml^?1. In this study, the microbial inhibitory activity of LML in dairy products inoculated with a 5‐strain cocktail of clinical isolates of L. monocytogenes was investigated. Addition of LML at a concentration of 5 mg ml^?1 resulted in 4·4, 4·0 and 4·2 log reductions in 0·5% fat, 1% fat and 3·25% fat milks, respectively; 4·1, 4·4, and 3·5 log reductions in nonfat, 1% fat, and 1·5% fat yogurts, respectively; and 4·0 log reductions in both nonfat and 2% fat cottage cheese. The inhibitory effect of LML was only observed at 37°C and not 5°C. Experiments suggest that both the lauric acid and the esterified lactose moiety of LML play roles in the growth inhibition.

Significance and Impact of the Study

A novel sugar ester, lactose monolaurate, inhibited the growth of a five‐strain cocktail of Listeria monocytogenes in milk, yogurt and cottage cheese. This is the first report of the use of a sugar ester to inhibit the growth of Listeria in food systems.     

Effects of ozone treatment on the quality of kiwifruit during postharvest storage affected by Botrytis cinerea and Penicillium expansum

The objective was to reveal the effects of ozone treatment on quality maintenance and resistance to Botrytis cinerea and Penicillium expansum in kiwifruit during postharvest storage. Kiwifruits were treated with 79.44 ppm gaseous ozone for 1 hr once a day for 7 day at 0°C to determine the effects of ozone treatment on the quality and disease incidence caused by B. cinerea and P. expansum in vivo and the growth of B. cinerea and P. expansum in vitro. Ozone treatment significantly reduced the disease incidence of kiwifruit and inhibited the mycelial development and spore germination of B. cinerea and P. expansum. High levels of fruit firmness and titratable acidity were maintained in the ozone‐treated kiwifruit, and the activities of the defence‐related enzymes were remarkably enhanced. Therefore, ozone treatment may be an effective method to maintain the quality of kiwifruit and control its decay during postharvest storage.     

Deciphering the origin of Aspergillus flavus NRRL21882, the active biocontrol agent of Afla-Guard®

Single nucleotide polymorphisms (SNPs) of genome sequences of eight Aspergillus flavus and seven Aspergillus oryzae strains were extracted with Mauve, a multiple-genome alignment programme. A phylogenetic analysis with sequences comprised of concatenated total SNPs by the unweighted pair group method with arithmetic mean (UPGMA) of MAFFT adequately separated them into three groups, A. flavus S-morphotype, A. flavus L-morphotype and A. oryzae. Divergence time inferred for A. flavus NRRL21882, the active agent of the biocontrol product Afla-Guard^®, and S-morphotype was about 5·1 mya. Another biocontrol strain, A. flavus AF36, diverged from aflatoxigenic L-morphotype about 2·6–3·0 mya. Despite the close relatedness of A. oryzae to A. flavus, A. oryzae strains likely evolved from aflatoxigenic Aspergillus aflatoxiformans (=A. parvisclerotigenus). A survey of A. flavus populations implies that prior Afla-Guard^® applications are associated with prevalence of NRRL21882-type isolates in Mississippi fields. In addition, a few NRRL21882 relatives were identified. A. flavus Og0222, a biocontrol ingredient of Aflasafe™, was verified as a NRRL21882-type strain, having identical sequence breakpoints that led to deletion of aflatoxin and cyclopiazonic acid gene clusters. A similar UPGMA analysis suggests that the occurrence of NRRL21882-type strains is a more recent event.     

Temperature × light interaction and tolerance of high water temperature in the planktonic freshwater flagellates Cryptomonas (Cryptophyceae) and Dinobryon (Chrysophyceae)

Using microcosm experiments, we investigated the interactive effects of temperature and light on specific growth rates of three species each of the phytoplanktonic genera Cryptomonas and Dinobryon. Several species of these genera play important roles in the food web of lakes and seem to be sensitive to high water temperature. We measured growth rates at three to four photon flux densities ranging from 10 to 240 μmol photon · m^?2 · s^?1 and at 4–5 temperatures ranging from 10°C to 28°C. The temperature × light interaction was generally strong, species specific, and also genus specific. Five of the six species studied tolerated 25°C when light availability was high; however, low light reduced tolerance of high temperatures. Growth rates of all six species were unaffected by temperature in the 10°C–15°C range at light levels ≤50 μmol photon · m^?2 · s^?1. At high light, growth rates of Cryptomonas spp. increased with temperature until the temperature optimum was reached and then declined. The Dinobryon species were less sensitive than Cryptomonas spp. to photon flux densities of 40 μmol photon · m^?2 · s^?1 and 200 μmol photon · m^?2 · s^?1 over the entire temperature range but did not grow under a combination of very low light (10 μmol photon · m^?2 · s^?1) and high temperature (≥20°C). Among the three Cryptomonas species, cell volume declined with temperature and the maximum temperature tolerated was negatively related to cell size. Since Cryptomonas is important food for microzooplankton, these trends may affect the pelagic carbon flow if lake warming continues.     

Chitosan and hot water treatments reduce postharvest green mould in ‘Murcott’ tangor

This study aims to evaluate the efficacy of hot water and chitosan treatments to control green mould caused by Penicillium digitatum in 'Murcott' tangor. P. digitatum conidial germination and mycelial growth were evaluated in assays in vitro to verify whether chitosan (0.5, 1 and 2%) or hot water (45, 50, and 55°C, for 30 s, 1, 2, and 5 min) acts directly on fungus development. In vivo assays consisted of inoculating the fruit with P. digitatum (10⁵ conidia ml⁻¹) 4 hr before the chitosan and hot water treatments. Subsequently, green mould incidence and severity were evaluated in fruits stored at 25°C/80% RH for 4 days. Also, treatments combining chitosan and hot water were investigated for controlling green mould and the effect on postharvest quality of fruit stored at 5°C/90% RH. The results showed that P. digitatum conidia germination and mycelial growth were significantly reduced by the hot water treatments especially at 50°C/5 min and 55°C/2 or 5 min in the first case and 50 and 55°C/5 min in the second. These two treatments, when applied alone, 1 min dipping in 2% chitosan or hot water at 50°C/5 min, significantly reduced green mould development in fruit kept at 25°C/80% RH or refrigerated. However, the hot water dip combined with chitosan did not improve green mould control on ‘Murcott’ tangor at room temperature or under refrigeration. Besides, chitosan and hot water did not impair fruit quality. Thus, chitosan and hot water could be an alternative to synthetic fungicides to control green mould in citrus while also contributing to a decrease in the postharvest losses of ‘Murcott’ tangor.