Use of bacteriophages as biocontrol agents to control Salmonella associated with seed sprouts

Two Salmonella bacteriophages (SSP5 and SSP6) were isolated and characterized based on their morphology and host range, and evaluated for their potential to control Salmonella Oranienburg in vitro and on experimentally contaminated alfalfa seeds. Phages SSP5 and SSP6 were classified as members of the Myoviridae and Siphoviridae families, respectively. Both phages had a broad host range of over 65% of the 41 Salmonella strains tested. During in vitro trials, the phages resulted in incomplete lysis of Salmonella cultures, in spite of high levels of phage remaining in the system. Phage SSP5 was more effective in reducing Salmonella populations. Addition of phage SSP6 to alfalfa seeds previously contaminated with S. Oranienburg caused an approximately 1 log(10) CFU g(-1) reduction of viable Salmonella, which was achieved 3 h after phage application. Thereafter the phage had no inhibitory effect on Salmonella population growth. A second addition of the same (SSP6) or different (SSP5) phage to a Salmonella culture treated with phage SSP6, did not affect Salmonella populations. It was further shown that development of Salmonella permanently resistant to phage was not evident in either seed or in vitro challenge trials, suggesting the existence of a temporary, acquired, non-specific phage resistance phenomenon. These factors may complicate the use of phages for biocontrol.     

Laboratory tests for assessing efficacy of atoxigenic Aspergillus flavus strains as biocontrol agents

Biocontrol by competitive inhibition using atoxigenic Aspergillus flavus strains has been shown to be an effective method for controlling aflatoxin production in peanuts, maize and cottonseed. Selecting biocontrol strains is not straightforward, as it is difficult to assess fitness for the task without expensive field trials. Reconstruction experiments have been generally performed under laboratory conditions to investigate the biological mechanisms underlying the efficacy of atoxigenic strains in preventing aflatoxin production and/or to give a preliminary indication of strain performance when released in the field. The study here described was conducted in order to evaluate the potential of the different atoxigenic A. flavus strains, colonizing the corn fields of the Po Valley, in reducing aflatoxin accumulation when grown in mixed cultures together with atoxigenic strains; additionally, we developed a simple and inexpensive procedure that may be used to scale-up the screening process and to increase knowledge on the mechanisms interfering with mycotoxin production during co-infection.     

Evaluation of atoxigenic isolates of Aspergillus flavus as potential biocontrol agents for aflatoxin in maize

Aflatoxin contamination resulting from maize infection by Aspergillus flavus is both an economic and a public health concern. Therefore, strategies for controlling aflatoxin contamination in maize are being investigated. The abilities of eleven naturally occurring atoxigenic isolates in Nigeria to reduce aflatoxin contamination in maize were evaluated in grain competition experiments and in field studies during the 2005 and 2006 growing seasons. Treatments consisted of inoculation of either grains in vials or ears at mid-silking stage in field plots, with the toxigenic isolate (La3228) or atoxigenic isolate alone and co-inoculation of each atoxigenic isolate and La3328. Aflatoxin B(1) + B(2) concentrations were significantly (p < 0.05) lower in the co-inoculation treatments compared with the treatment in which the aflatoxin-producing isolate La3228 was inoculated alone. Relative levels of aflatoxin B(1) + B(2) reduction ranged from 70.1% to 99.9%. Among the atoxigenics, two isolates from Lafia, La3279 and La3303, were most effective at reducing aflatoxin B(1) + B(2) concentrations in both laboratory and field trials. These two isolates have potential value as agents for the biocontrol of aflatoxin contamination in maize. Because these isolates are endemic to West Africa, they are both more likely than introduced isolates to be well adapted to West African environments and to meet regulatory concerns over their use throughout that region.     

Evaluation of atoxigenic isolates of Aspergillus flavus as potential biocontrol agents for aflatoxin in maize

Aflatoxin contamination resulting from maize infection by Aspergillus flavus is both an economic and a public health concern. Therefore, strategies for controlling aflatoxin contamination in maize are being investigated. The abilities of eleven naturally occurring atoxigenic isolates in Nigeria to reduce aflatoxin contamination in maize were evaluated in grain competition experiments and in field studies during the 2005 and 2006 growing seasons. Treatments consisted of inoculation of either grains in vials or ears at mid-silking stage in field plots, with the toxigenic isolate (La3228) or atoxigenic isolate alone and co-inoculation of each atoxigenic isolate and La3328. Aflatoxin B₁?+?B₂ concentrations were significantly (p?<?0.05) lower in the co-inoculation treatments compared with the treatment in which the aflatoxin-producing isolate La3228 was inoculated alone. Relative levels of aflatoxin B₁?+?B₂ reduction ranged from 70.1% to 99.9%. Among the atoxigenics, two isolates from Lafia, La3279 and La3303, were most effective at reducing aflatoxin B₁?+?B₂ concentrations in both laboratory and field trials. These two isolates have potential value as agents for the biocontrol of aflatoxin contamination in maize. Because these isolates are endemic to West Africa, they are both more likely than introduced isolates to be well adapted to West African environments and to meet regulatory concerns over their use throughout that region.     

Development of biocontrol agents from food microbial isolates for controlling post-harvest peach brown rot caused by Monilinia fructicola

An unconventional strategy of screening food microbes for biocontrol activity was used to develop biocontrol agents for controlling post-harvest peach brown rot caused by Monilinia fructicola. Forty-four microbial isolates were first screened for their biocontrol activity on apple fruit. Compared with the pathogen-only check, seven of the 44 isolates reduced brown rot incidence by >50%, including four bacteria: Bacillus sp. C06, Lactobacillus sp. C03-b and Bacillus sp. T03-c, Lactobacillus sp. P02 and three yeasts: Saccharomyces delbrueckii A50, S. cerevisiae YE-5 and S. cerevisiae A41. Eight microbial isolates were selected for testing on peaches by wound co-inoculation with mixtures of individual microbial cultures and conidial suspension of M. fructicola. Only two of them showed significant biocontrol activity after five days of incubation at 22 degrees C. Bacillus sp. C06 suppressed brown rot incidence by 92% and reduced lesion diameter by 88% compared to the pathogen-only check. Bacillus sp.T03-c reduced incidence and lesion diameter by 40% and 62%, respectively. The two isolates were compared with Pseudomonas syringae MA-4, a biocontrol agent for post-harvest peach diseases, by immersing peaches in an aliquot containing individual microbial isolates and the pathogen conidia. Treatments with isolates MA-4, C06 and T03-c significantly controlled brown rot by 91, 100, and 100% respectively. However, only isolates MA-4 and C06 significantly reduced brown rot by 80% and 15%, respectively when bacterial cells alone were applied. On naturally infected peaches, both the bacterial culture and its cell-free filtrate of the isolate C06 significantly controlled peach decay resulting in 77 and 90% reduction, respectively, whereas the treatment using only the bacterial cells generally had no effect. Isolate C06 is a single colony isolate obtained from a mesophilic cheese starter, and has been identified belonging to Bacillus amyloliquefaciens. The results have clearly demonstrated that isolate C06 has a great potential for being developed into a biocontrol agent.     

Prevalence of bacteriophages infecting Staphylococcus aureus in dairy samples and their potential as biocontrol agents

The prevalence of bacteriophages infecting Staphylococcus aureus in dairy samples was assessed. Fourteen Staph. aureus strains were used in enrichment cultures of 75 dairy samples. All samples grew specific Staph. aureus bacteriophages. According to the host range, 8 different phages were isolated. Three of them, phages ΦH5, ΦG7, and ΦA72, were found in 89% of the samples; all the isolated phages were temperate. Phages ΦH5 and ΦA72 were used in preliminary bacterial challenge tests against Staph. aureus in milk. A phage mixture (1:1) was more effective than each single phage, most likely by preventing the survival of lysogenized cells. Phages inhibited Staph. aureus in UHT and pasteurized whole-fat milk. However, the phages were less active in semi-skimmed raw milk and little inhibition was achieved in whole, raw milk. Killing of Staph. aureus was observed at room temperature and at 37°C, but not at refrigeration temperature.     

Chemistry of sulphiting agents in food.

The main reason for the reactivity of sulphites in food is the nucleophilicity of the sulphite ion. The factors which determine the activity of this nucleophile are summarized and critically evaluated for concentrated systems, e.g. dehydrated foods. The distinction between free and bound sulphite is explained, and reversible binding of the additive in beverages and dehydrated foods is discussed with reference to simple theory of chemical equilibrium. The inhibition of non-enzymic browning reactions accounts for a large proportion of sulphite which undergoes irreversible reaction in concentrated foods. The mechanisms of reactions between sulphite species and intermediates in the model Maillard reaction, glucose+glycine, are considered in depth together with supporting kinetic data. An interesting feature is the fact that sulphites seem to catalyse the reactions they are added to control. Implications of this to the level of use of sulphite are discussed. Reaction products from the inhibition of Maillard browning include 3,4-dideoxy-4-sulphohexosulose which is formed initially and polymeric substances which arise from the reaction of sulphites with melanoidins. A proportion of sulphite added to food becomes converted to sulphate. Mechanisms of autoxidation are critically appraised in view of the presence of considerable concentrations of antioxidants in foods. The autoxidation of sulphite involves reactive free radical intermediates which include effective oxidizing agents. Thus, a pro-oxidant effect by the additive is possible and demonstrable in model system experiments.     

Egyptian honeybee pollen as antimicrobial, antioxidant agents, and dietary food supplements

This paper describes the development of a specific ultra performance LC with electrospray ionization tandem mass spectrometric method for simultaneous determination of 13 quinolones in livestock and fishery products. The procedures involve the sample preparation based on solvent extraction without further clean up procedure. The method was validated according to the FDA guidelines. The limit of detection (LOD) and quantification (LOQ) were lower than 0.1 and 0.4 μg/kg, respectively, and these were below the maximum residue limits (MRLs) established by European Union. Recoveries ranged from 87.5 to 104.7% for livestock products and 83.0 to 100.9% for fishery products. Relative standard deviations (RSD) ranged from 0.4 to 6.0% and 0.9 to 5.7%, respectively. The method was successfully applied to the analysis of quinolones residue. This quantitative method has many advantages including simple preparation, rapid determination, and high sensitivity, which could be applied to the determination of quinolones residues in foodstuff.     

Recent developments in the enhancement of some postharvest biocontrol agents with unconventional chemicals compounds

BackgroundProduce are susceptible to microbial attack from diverse sources and the use of novel BCAs has gained recognition as alternative and sustainable applications to lessen the current emerging problems with synthetic fungicide use. Most researchers recommend the combination of two or more biocontrol agents in postharvest diseases control. To this end, the enhancement of biocontrol agents and the mechanisms of action in biocontrol systems have attracted many research interests. In this regard, there have been remarkable efforts to develop a multifaceted system approach for disease control.Scope and approachIt has been recognized that the various methods of biological control act together additively or synergistically to achieve significant commercial level of 97–99% disease control. The integration of microbial agents with a wide range of unconventional chemicals and their corresponding mechanisms of action to controlling postharvest fungal pathogens of fruits has been proven be successful.Key findings and conclusionsIn this review, the combined strategy of unconventional chemical compounds and other BCAs have contributed to varied degree of postharvest diseases control. The beneficial effects of these methods depend on the appropriate combination of the agents based on adequate knowledge of their mechanisms of action in the biocontrol system. Lastly, efforts to upscale these methods to commercial implementation level must be given the necessary consideration.     

酸奶噬菌体的防治

从上海市光明乳业股份有限公司车间发酵罐中分离出4株酸奶噬菌体,电镜下均呈蝌蚪形。酸奶噬菌体防治实验结果表明:使用来今1207处理液或来今1208处理液,虽然一定浓度的处理液(100mg/kg的来今1207处理液,0.2%的来今1208处理液)可以达到杀灭噬菌体的效果,但是会影响乳酸菌的正常生长,所以处理酸奶发酵生产设备如工厂管道、发酵罐和种子罐时,需要用清水洗去残留液才能投入生产,否则会导致乳酸菌生长和酸奶发酵的不正常。而来今1205处理液的浓度为0.1%时就可以达到杀灭噬菌体的效果,且不影响乳酸菌的生长,适合于作为酸奶噬菌体的杀灭处理液。     

Fungal siderophores function as protective agents of LDL oxidation and are promising anti-atherosclerotic metabolites in functional food

Iron-mediated oxidation of low-density lipoprotein has been implicated in the pathogenesis of vascular disorders such as atherosclerosis. The present investigations were performed to test whether hydrophobic fungal siderophores - hexadentate trihydroxamates desferricoprogen, desferrichrome, desferrirubin, and desferrichrysin - might suppress heme-catalyzed LDL oxidation and the toxic effects of heme-treated LDL on vascular endothelium. Indeed, two of these - desferricoprogen and desferrichrome - markedly increased the resistance of LDL to heme-catalyzed oxidation. In similar dose-response fashion, these siderophores also inhibited the generation of LDL products cytotoxic to human vascular endothelium. When iron-free fungal siderophores were added to LDL/heme oxidation reactions, the product failed to induce heme oxygenase-1, a surrogate marker for the noncytocidal effects of oxidized LDL (not in the case of desferrichrysin). Desferricoprogen also hindered the iron-mediated peroxidation of lipids from human atherosclerotic soft plaques in vitro, and was taken up in the gastrointestinal tract of rat. The absorbed siderophore was accumulated in the liver and was secreted in its iron-complexed form in the feces and urine. The consumption of mold-ripened food products such as aged cheeses and the introduction of functional foods and food additives rich in fungal iron chelators in diets may lower the risk of cardiovascular diseases.     

Quorum-quenching enzymes: Promising bioresources and their opportunities and challenges as alternative bacteriostatic agents in food industry

The problems of spoilage, disease, and biofilm caused by bacterial quorum-sensing (QS) systems have posed a significant challenge to the development of the food industry. Quorum-quenching (QQ) enzymes can block QS by hydrolyzing or modifying the signal molecule, making these enzymes promising new candidates for use as antimicrobials. With many recent studies of QQ enzymes and their potential to target foodborne bacteria, an updated and systematic review is necessary. Thus, the goals of this review were to summarize what is known about the effects of bacterial QS on the food industry; discuss the current understanding of the catalytic mechanisms of QQ enzymes, including lactonase, acylase, and oxidoreductase; and describe strategies for the engineering and evolution of QQ enzymes for practical use. In particular, this review focuses on the latest progress in the application of QQ enzymes in the field of food. Finally, the current challenges limiting the systematic application of QQ enzymes in the food industry are discussed to help guide the future development of these important enzymes.     

南美白对虾产业发展报告

为研究乳酸链球菌素（Nisin）对高水分烤虾常温下贮藏性的影响,在烤虾样品中分别添加20、60和100 mg/kg的Nisin,以感官结合菌落总数、蜡样芽孢杆菌数、挥发性盐基氮、pH等指标对样品品质进行评价。结果表明,烤虾的感官评分在常温贮藏过程中逐渐下降,菌落总数、蜡样芽孢杆菌数、挥发性盐基氮与pH则呈增长趋势。随着Nisin添加量的增加,菌落总数、蜡样芽孢杆菌数、挥发性盐基氮的增长趋慢。第6 d时,未加Nisin的对照组样品菌落总数达到行业标准《调味烤虾》中规定的限值（3×10⁴ CFU/g）,而添加20、60、100 mg/kg Nisin的样品,菌落总数达到规定限值的时间分别被延长至8、12和14 d。本研究表明Nisin对高水分烤虾中蜡样芽孢杆菌等腐败微生物生长有明显的抑制作用,能有效延长产品的货架期。     

串联式生防真菌产孢箱的发酵工艺优化

气生分生孢子是生防真菌应用和剂型化的主要有效成分之一,该研究对自行研制的具有应用潜力的串联式产孢箱发酵生产气生分生孢子的工艺参数进行了优化。通过对球孢白僵菌ChB9菌株在两种常用培养基质(籼米和粳米)以不同重量面积比、不同灭菌方式下发酵得到的孢子粉产量、气生菌丝产量和培养基利用率等指标的统计分析来优化该产孢箱在发酵生产气生分生孢子过程中的相关关键工艺参数。结果表明,粳米、105℃灭菌30 min与重量面积比(g:cm2)为1∶2的组合条件最符合产业应用实际,产量为17.56±2.79 g孢子粉/kg稻米,为最佳选择。该研究获得的优化参数不仅可以指导生防真菌孢子粉的小规模实验室生产,而且可为以麦类等其他发酵基质的发酵生产工艺优化提供参考。     

Synergistic effects of combining biocontrol agents with silicon against postharvest diseases of jujube fruit

The synergistic effects of biocontrol yeasts Cryptococcus laurentii and Rhodotorula glutinis combined with silicon (Si) against Alternaria alternata and Penicillium expansum molds were investigated in jujube fruit (Chinese date, Zizyphus jujuba) stored at 20 and 0 degrees C, respectively. Combinations of C. laurentii and R. glutinis at 5 x 10(7) cells/ml with 2% Si was most effective in controlling the diseases caused by A. alternata and P. expansum on jujube fruit stored at 20 degrees C. When fruits were stored at 0 degrees C, combining C. laurentii and R. glutinis with Si was as effective against P. expansum as was Si or the yeasts applied alone and was more effective in controlling A. alternata. Si may have a fungistatic effect by directly inhibiting pathogen growth, but it did not greatly influence the growth of the antagonists.     

Microbiological screening of food preservatives,cold sterilants and specific antimicrobial agents as potential Silage Additives

Some food preservatives, cold sterilants and specific antimicrobial agents were tested against a range of microorganisms at pH 3, 4, 5 and 6 in a semi-micro assay. The antimicrobial spectra obtained indicated that several of the components might be useful as silage additives. Lactic acid appears to have potential but only when the feed is kept anaerobic as it affords no protection against the activities of yeasts and moulds. Potassium sorbate might have wider application for increasing the aerobic stability of silage, whilst sodium nitrite appears to have application for restricting or suppressing fermentation in silage. The use of sodium benzoate is unlikely to be economic. Formaldehyde and paraformaldehyde would appear to have great potential in the production of non-fermented silage, whereas the relatively high quantities of hexamine and propylene oxide which are required make these chemicals unattractive. The antibiotic pimaricin appears to have potential as an agent for preventing aerobic degradation of silage and the synthetic antimicrobial agent Bronopol for preventing the activity of sporebearing bacteria. Although it appears that tylosin could be used to prevent bacterial growth the quantity required is, for an antibiotic, quite large.