Temperature effect on Fusarium oxysporum f.sp. melonis survival during horticultural waste composting

AIMS: The aim of this work was to study the effect of high temperatures generated during composting process, on the phytopathogen fungus Fusarium oxysporum f.sp. melonis. This investigation was achieved by both in vivo (semipilot-scale composting of horticultural wastes) and in vitro (lab-scale thermal treatments) assays. METHODS AND RESULTS: Vegetable residues infected with F. oxysporum f.sp. melonis were included in compost piles. Studies were conducted in several compost windrows subjected to different treatments. Results showed an effective suppression of persistence and infective capacity, as this process caused complete fungal elimination after 2-3 days of composting. In order to confirm the effect of high temperature during this process, in vitro experiments were carried out. Temperature values of 45, 55 and 65 degrees C were tested. All three treatments caused the elimination of fungal persistence. Treatment at 65 degrees C was especially effective, whereas 45 degrees C eliminated fungal persistence only after 10 days. CONCLUSIONS: The composting process is an excellent alternative for the management of plant wastes after harvesting, as this procedure is able to suppress infective capacity of several harmful phytopathogens such as F. oxysporum f.sp. melonis. SIGNIFICANCE AND IMPACT OF THE STUDY: Fusarium oxysporum f.sp. melonis is a plant pathogen fungus specially important in the province of Almería (south-east Spain), where intensive greenhouse horticulture is very extended. High temperatures reached during composting of horticultural plant wastes ensure the elimination of phytopathogen microorganisms such as F. oxysporum f.sp. melonis from vegetable material, providing an adequate hygienic quality in composts obtained.     

A Sinapic Derivative as an Induced Defence Compound of Date Palm Against Fusarium oxysporum f.sp. albedinis, the Agent Causing Bayoud Disease

A non-constitutive phenolic compound was isolated from date palm ( Phoenix dactylifera L.) roots and calli derived from offshoots. It inhibits greatly conidial germination as well as infectious germ tube growth and must be considered as a phytoalexin. Its accumulation occurred when calli turned brown or when roots were in contact with Fusarium oxysporum f.sp. albedinis , the agent causing the date palm vascular fusariosis'bayoud'. Tentatively, this compound has been identified as sinapoyl-amide by acid and alkaline hydrolysis, dansylation of amines and polyamines and by ninhydrin treatment. The relationship between this methoxy-substituted hydroxycinnamic acidamide and biotic and abiotic stress leading to tissue browning or vascular disease resistance is discussed.     

Identification of phenylacetic acid produced by Fusarium oxysporum f. sp. albedinis, the causal agent of bayoud, using GC-MS

These studies are concerned with the isolation and identification of secondary metabolites produced by Fusarium oxysporum f. sp. albedinis (F. o. a.), the causal agent of bayoud, the wilt disease of the date palm (Phoenix dactylifera L.). Fungal secondary metabolites are chemical compounds identified in a limited number of species. They consist of toxins, antibiotics and antifungal agents. Among the metabolites we could isolate from the pathogen grown in a liquid medium, and then identify by gas chromatography coupled with mass spectrometry (GC-MS), phenylacetic acid has been distinguished. This compound is widely described in the literature as having antimicrobial, antifungal, phytotoxic properties and also endowed with hormonal activity similar to that of indole acetic acid (IAA). To date, this metabolite has never been reported in F. o. a.     

Molecular characterization of Syrian date palm cultivars using plasmid-like DNA markers

Date palm (Phoenix dactylifera L.) is one of the most important domesticated fruit trees in the Near East and North African countries. This tree has been, for several decades, in serious threat of being completely destroyed by the "Bayoud" disease caused by Fusarium oxysporum f. sp. albedinis. In this study, 18 Syrian date palm cultivars and four male trees were analyzed according to the identity of mitochondrial plasmid-like DNAs. A PCR strategy that employs plasmid-like DNAs-specific primer pair was used. These primers amplify a product of either 373-bp or 265-bp that corresponds to the S-(Bayoud-susceptible) or the R-plasmid (Bayoud-resistant), respectively. Generated data revealed that only six cultivars ('Medjool', 'Ashrasi', 'Gish Rabi', 'Khineze', and yellow- and red-'Kabkab') have the S-plasmid, suggesting their susceptibility to the fusariosis, while the remaining 12 cultivars and the four male trees contain the R-plasmid, suggesting their resistance to the fusariosis. The PCR process applied here has been proved efficient for the rapid screening for the presence of the S and R DNAs in Syrian date palm. PCR markers developed in this study could be useful for the screening of date palm lines growing in the field. The availability of such diagnostic tool for plasmid characterization in date palm would also be of great importance in establishing propagation and breeding programs of date palm in Syria.     

Bio-degradation of olive oil husks in composting aerated piles

In this study, the composting performance of two olive oil husk piles was compared using two different aeration processes: aeration by mechanical turning or by forced air-injection. The results showed that after the maturation phase both piles had a similar chemical composition and the same level of organic matter degradation (around 55%). However, the time necessary to reach the thermophilic phase was shorter for the turned pile (2 months in comparison to 3 months for the air-injected pile) and the humification degree achieved was slightly higher (94% versus 83%). Both piles showed the presence of an active microbial community, with an increase by 2-3 orders of magnitude in the number of cultured microbial colonies during the composting process. No significant difference in quantitative or qualitative terms was found in the microbial populations of both piles, nor was a clear succession between a mesophilic and a thermophilic population observed. In terms of industrial application, the mechanical turning process seems to be better since it is a lower energy and time consuming process.     

Antifungal activity of rhizospheric bacteria

Fluorescent Pseudomonad spp. were isolated from the rhizosphere of potato plants (Algeria) by serial dilutions of rhizosphere soils on Kings B medium and were tested for their antifungal activity. The antifungal activity of the Pseudomonas isolated from Potatoes rhizosphere was tested against Pythium ultimum, Rhizoctonia solani and Fusarium oxysporum in dual culture with bacteria on PDA. The Petri dish was divided into tow, on one the bacteria was spread and on the opposite side fungal plugs were inoculated and incubated for one week. Fourteen bacteria were isolated; only one isolate inhibited the growth of Pythium ultimum, Rhizoctonia solani, Fusarium solani; Fusarium oxysporum f.sp. albedinis and Fusarium oxysporum f. sp. Lycopersici with inhibition zones of 39.9, 33.7, 30.8, 19.9 and 22.5 mm respectively.     

Identification des acides caféoylshikimiques des racines du palmier dattier, principaux composés fongitoxiques vis-à-vis de Fusarium oxysporum f. sp. albedinis

5-caffeoylshikimic acid and its isomeres of position (dactylifric acid) found in the roots of date-palms ( Phoenix daetylijera L.) inhibit the germination of conidia and the growth of the mycelium of Fusarium oxysporum f. sp. albedinis . The acid content accumulated in resistant cultivars are toxic to fungi, whereas that accumulated in sensitive cultivars does not seem to affect growth and development of the parasite. This well demonstrates the implication of caffeoylshikimic acid in the resistance of the date-palm to this fusarium disease. Moreover, these results will be of great use, especially for screening resistant genotypes obtained from directed crossings or from in vitro cultures.     

应用PCR-RFLP和巢式PCR检测黄瓜尖镰孢菌

以3株黄瓜尖镰孢菌(Fusarium oxysporum f.sp.cucumarinum)、23株镰孢菌属(Fusariumspp.)真菌和分离自土壤的20株真菌、6株细菌和7株放线菌为材料,采用化学裂解法提取总DNA,进行PCR-RFLP和巢式PCR检测,试验证明PCR-RFLP程序不能完全区分Fusarium属内不同种,而巢式PCR对黄瓜尖镰孢菌具有特异性.运用优化的PCR-RFLP和巢式PCR检测程序对染病黄瓜组织进行了检测,结果表明,两种方法均可在接种发病早期(未显症时)检测出黄瓜枯萎病菌,PCR-RFLP在感病品种接种后3d即可检测到病原菌,而巢式PCR在接种后5d才能检测到病原菌.     

Composting of de-inking paper sludge with poultry manure at three nitrogen levels using mechanical turning: behavior of physico-chemical parameters.

De-inking paper sludge (DPS) is rich in carbon (C) but poor in nitrogen (N). Thus, it has a high C:N ratio which limits the composting process. Accordingly, the goal of this study was to investigate the effect of three N treatments on DPS composting. Compost piles of 100 m3 were formed by mixing raw DPS with poultry manure and chicken broiler floor litter, giving on average 0.6%, 0.7% and 0.9% total N. The changes in physico-chemical parameters, total weight and fiber losses, and maturity of composting piles of DPS were monitored during 24 weeks. The compost piles had a neutral to alkaline pH throughout the study. Inorganic N decreased whereas organic N increased over time for all treatments. These changes in magnitude were different among N treatments resulting in a final total N content of 0.9% for the 0.6% N treatment whereas final total N contents of 0.7% and 0.9% N were measured for the 0.7% and 0.9% N treatments. The total weight, cellulose and hemicellulose losses were higher in 0.6% N treatment giving the lowest C:N ratio after 24 weeks of composting. However, none of the 24 week-old composts of DPS were mature based on their final C:N ratio and colorimetric test of maturity. Except for copper, their final total trace element contents meet most known standards or guidelines for organic soil conditioners. Overall, 0.6% N treatment was the best to enhance DPS composting using mechanical turning, but a period of more than 24 weeks was required to reach compost maturity.     

Chitinous components of the cell wall of Fusarium oxysporum.

The cell wall of Fusarium oxysporum f. sp. lycopersici was digested with chitinase to analyze the structure of its chitinous components. In spite of a similar acetylation degree of the cell wall components to that of 25-35% acetylated chitosan, only N-acetylglucosamine disaccharide [(GlcNAc)2] was obtained from chitinase hydrolyzate of the fungal cell wall by CM-Sephadex C-25 column chromatography, while (GlcNAc)2 and several types of deacetylated chitooligosaccharides were separated from that of 25-35% acetylated chitosan. The results indicate that N-acetylglucosamine residues in the polysaccharide chains of the fungal cell wall are most likely condensed into some region, while acetylated residues are more scattered in 25-35% acetylated chitosan.     

Co-composting of soybean residues and leaves in Hong Kong

The goal of this project was to evaluate the feasibility of co-composting of soybean residues and leaves and the effects of turning frequency on compost quality. Soybean residues were mixed with leaves and sawdust in 1:1:3 (w/w wet weight) for achieving a C/N ratio of about 30. Three heaps of about 4 m3 of compost mixtures were prepared receiving a turning frequency of daily (pile A), 3-day (pile B) and weekly (pile C) turning. Different turning frequencies did not significantly affect the changes in pH and volatile solids throughout the composting period. High turning frequency caused a lower electrical conductivity and NH4-N contents as well as a shorter duration of thermophilic phase, because of a high heat loss by evaporation and volatilization of ammonia in the pile. The highest C decomposition of 4% occurred in the pile with a 3-day turning period, which coincided with the higher-nitrogen content in this treatment. All treatments with different turning frequencies reached maturation at 63 days as indicated by the soluble organic carbon, soluble NH4-N, C/N ratio and cress seed germination index. However, increasing the aeration during composting period was beneficial in accelerating the maturation process. Taking into consideration less labour and lower operation costs as compared to daily turning, it can be suggested that a 3-day turning frequency would be more appropriate for reaching acceptable quality of compost and ease in operation.     

Community structure and population dynamics of ammonia oxidizers in composting processes of ammonia-rich livestock waste

This study investigated the relationship between the population dynamics of ammonia-oxidizing bacteria (AOB) and archaea (AOA), and changes in the concentrations of nitrogenous compounds during ammonia-rich livestock waste-composting processes. The data showed that ammonia in beef and dairy cow livestock waste-composting piles was slowly oxidized to nitrite and nitrate after approximately 21–35 days under thermophilic or moderately thermophilic and mesophilic conditions. Real-time quantitative PCR (qPCR) assays showed a relative abundance of betaproteobacterial AOB during ammonia oxidation but did not detect AOA in any composting stage. Furthermore, real-time qPCR and terminal-restriction fragment length polymorphism (T-RFLP) analyses for the AOB in two composting processes (beef and dairy cow livestock waste) out of the three studied found that thermophilic or moderately thermophilic uncultured betaproteobacterial AOB from the “compost AOB cluster” contributed to ammonia oxidation during hot composting stages. Non-metric multidimensional scaling analyses of the data from T-RFLP showed that only a few analogous species predominated during composting of beef, dairy cow and pig livestock wastes, and thus, the AOB community structures in the three composting piles operating under different conditions were similar. AOB-targeted clone library analyses revealed that uncultured members of the “compost AOB cluster”, which could be clearly distinguished from the authentic species of the genus Nitrosomonas, were the major constituents of the AOB populations. These results suggested that a limited and unique species of AOB played a role in ammonia oxidation during the composting of ammonia-rich livestock waste.     

Respiration profiles in monitoring the composting of by-products from the olive oil agro-industry

The composting of olive press cake (OPC) repeatedly mixed either with olive mill wastewater (OPC+OMW) or with tap water (OPC+W) was studied using the thermogradient respirometer, an apparatus that determines the respiration rates from a substrate over a wide range of different temperatures (respiratory profile). The composting processes took place over a period of five months during which nine moistenings of the OPC were performed with the respective liquids. The composting resulted in detoxification of the materials used in both treatments, as indicated by seed germination tests. However, the repeated applications of OMW resulted in recurring thermophilic phases (following each application) and in greater pH and conductivity increases in the final product, as compared to water applications. Respiration measurements performed at 35 degrees C were good indicators of the mean metabolic potential in the compost piles (the mean respiration derived from the whole respiration profile over a wide range of environmental temperatures). However, respiration measurements at higher temperatures (48.5 degrees C) were better indicators of the respiration activity occurring in situ. Following the initial thermophilic phase, the respiration potential of the composts at high temperatures (42-63 degrees C) increased drastically compared to their respiration potential at lower temperatures (17-42 degrees C) indicating the establishment of a thermophilic microflora. Subsequently, only the periodic new substrate-C applications in the form of OMW resulted in increased ratios of low temperature-to-high temperature respiration potential. These ratios decreased again following the respective thermophilic phase that each new OMW application had induced.     

Improving the quality of municipal organic waste compost

The effects of different municipal organic waste (MOW) management practices (shredding, addition of carbon-rich materials and inoculation with earthworms) on organic matter stabilization and compost quality were studied. Four static piles were prepared with: (i) shredded MOW; (ii) shredded MOW+woodshavings; (iii) non-shredded MOW; and (iv) non-shredded MOW+woodshavings. After 50 days, a part of each pile was separated for vermistabilization, while the rest continued as traditional thermophilic composting piles. At different sampling dates, and in the finished products, the following parameters were measured: pH, electrical conductivity, carbon dioxide evolution, and concentrations of organic matter, total nitrogen, water-soluble carbon, nitrate nitrogen, ammonium nitrogen, and extractable phosphorus. Shredded treatments exhibited faster organic matter stabilization than non-shredded treatments, evidenced specially by earlier stabilization of carbon dioxide production and shorter thermophilic phases. Woodshavings addition greatly increased quality of final products in terms of organic matter concentration, and pH and electrical conductivity values, but decreased total nitrogen and available nutrient concentrations. Vermicomposting of previously composted material led to products richer in organic matter, total nitrogen, and available nutrient concentrations than composting only, probably due to the coupled effect of earthworm activity and a shorter thermophilic phase.     

sti35, a stress-responsive gene in Fusarium spp.

A stress-induced mRNA was identified in the phytopathogenic fungus Fusarium oxysporum f. sp. cucumerinum. Treatment of the fungus with ethanol resulted in the induction of a major mRNA species encoding a protein of approximate Mr 37,000. A full-length cDNA clone of the induced message was obtained. RNA blot analysis indicated that the mRNA was induced by various other stresses, including treatment with copper(II) chloride and heat (37 degrees C). However, it was not greatly induced by treatment with phaseollinisoflavan, an antifungal isoflavonoid produced by Phaseolus vulgaris (French bean). In contrast, phaseollinisoflavan induced the homologous mRNA in the related bean pathogen Fusarium solani f. sp. phaseoli. A genomic clone of the F. solani f. sp. phaseoli gene was obtained, and both this and the cDNA clone from F. oxysporum f. sp. cucumerinum were sequenced. The latter indicated an open reading frame of 320 codons encoding a 34,556-dalton polypeptide. The corresponding reading frame in F. solani f. sp. phaseoli was 324 codons, 89% identical to the F. oxysporum f. sp. cucumerium sequence, and was interrupted by a short intron. The gene was designated sti35 (stress-inducible mRNA). Although computer homology searches were negative, the cloned gene was observed to cross-hybridize to DNAs of other filamentous fungi, Saccharomyces cerevisiae, and soybean. Thus, sti35 appears to be a common gene among a variety of eucaryotes.     

Risk Levels of Invasive Fusarium oxysporum f. sp. in Areas Suitable for Date Palm (Phoenix dactylifera) Cultivation under Various Climate Change Projections

Global climate model outputs involve uncertainties in prediction, which could be reduced by identifying agreements between the output results of different models, covering all assumptions included in each. Fusarium oxysporum f.sp. is an invasive pathogen that poses risk to date palm cultivation, among other crops. Therefore, in this study, the future distribution of invasive Fusarium oxysporum f.sp., confirmed by CSIRO-Mk3.0 (CS) and MIROC-H (MR) GCMs, was modeled and combined with the future distribution of date palm predicted by the same GCMs, to identify areas suitable for date palm cultivation with different risk levels of invasive Fusarium oxysporum f.sp., for 2030, 2050, 2070 and 2100. Results showed that 40%, 37%, 33% and 28% areas projected to become highly conducive to date palm are under high risk of its lethal fungus, compared with 37%, 39%, 43% and 42% under low risk, for the chosen years respectively. Our study also indicates that areas with marginal risk will be limited to 231, 212, 186 and 172 million hectares by 2030, 2050, 2070 and 2100. The study further demonstrates that CLIMEX outputs refined by a combination of different GCMs results of different species that have symbiosis or parasite relationship, ensure that the predictions become robust, rather than producing hypothetical findings, limited purely to publication.     

The Palmier date palm and Fusarium infection. II. Report on the stereochemistry of monosaccharides and their various derivatives and the growth of Fusarium oxysporum f.sp. albedinis (Killian et Maire) Gordon]

Fusarium oxysporum albedinis was grown in the presence of 20 different sugars or their derivatives. The results obtained were correlated with the stereochemical structure of the main oses. Osmotic phenomena as well as active and selective transport were used to explain differences in utilization of oses of very near structure. Based on the affinity of the fungus for mannose, a hypothesis is formulated concerning the fungus action in the host.     

Variation in microbial population during composting of agro-industrial waste

Two compost piles were prepared, using two ventilation systems: forced ventilation and ventilation through mechanical turning. The material to compost was a mixture of orange waste, olive pomace, and grass clippings (2:1:1 v/v). During the composting period (375 days), samples were periodically taken from both piles, and the enumeration of fungi, actinomycetes, and heterotrophic bacteria was carried out. All studied microorganisms were incubated at 25 and 55 °C after inoculation in appropriate growth media. Fungi were dominant in the early stages of both composting processes; heterotrophic bacteria proliferated mainly during the thermophilic stage, and actinomycetes were more abundant in the final stage of the composting process. Our results showed that the physical and chemical parameters: temperature, pH, moisture, and aeration influenced the variation of the microbial population along the composting process. This study demonstrated that composting of these types of wastes, despite the prolonged mesophilic stage, provided an expected microbial variation.     

Toxin-based in-vitro selection and its potential application to date palm for resistance to the bayoud Fusarium wilt

Date palm (Phoenix dactylifera L.) is qualified as a 'tree' of great ecological and socio-economical importance in desert oases. Unfortunately, it is being decimated, especially in Morocco and Algeria, by a fusariosis wilt called bayoud and caused by Fusarium oxysporum f. sp. albedinis (Fao). Controlling this disease requires the implementation of an integrated management program. Breeding for resistance is one of the most promising component strategies of this program. Few naturally resistant cultivars with a mediocre fruit quality (dates) are known. Conventional and non-conventional methods are under development and have to use the simplest and easiest methods to screen for resistant individuals. The use of pathogen toxins as selective agents at the tissue culture step might be a source of variability that can lead to the selection of individuals with suitable levels of resistance to the toxin and/or to the pathogen among the genetic material available. Foa produces toxins such as fusaric, succinic, 3-phenyl lactic acids and their derivatives, marasmins and peptidic toxins. These toxins can be used bulked or separately as selective agents. The aim of this contribution was to give a brief overview on toxins and their use as a mean to select resistant lines and to initiate a discussion about the potential use of this approach for the date palm-Foa pathosystem. This review does not pretend to be comprehensive or exhaustive and was prepared mainly to highlight the potential use of Foa toxins for selecting date palm individuals with a suitable resistance level to bayoud using toxin-based selective media.     

Simple emission-reducing measures in an open biological waste treatment plant

In the course of composting biological waste, concentrations of various thermophilic and thermotolerant microorganisms increase. Moving piles of compost results in increased emissions of Actinomycetes and fungi. The present investigation deals with the reduction of airborne microorganism emission and immission in large-scale composting plants with open piles. Simple measures were introduced in order to reduce the release of bioaerosols when turning piles and the release of dust and bioaerols at large. These measures included the sealing of turning machinery with rubber mats, the wetting of piles before and after turning and regular cleaning and wetting driveways during the dry season.Concentrations of airborne microorganisms during the summer season were determined on 5 days before and after the introduction of emission-reducing measures using the six-stage Andersen cascade sampler. The investigation showed that following the introduction of emission-reducing measures there was, at all locations, a highly significant reduction not only of all culturable indicator organisms (thermophilic actinomycetes and Aspergillus fumigatus) but also of total microorganism concentrations (p < 0,001). The introduction of the simple emission-reducing measures mentioned above, however, reduced the immission in the vicinity of the plant to such a degree that the natural background levels were reached at a distance of 150 m.