Insights from modelling sixteen years of climatic and fumonisin patterns in maize in South Africa.

Mycotoxin contamination of agricultural commodities is a global public health problem that has remained elusive to various mitigation approaches, particularly in developing countries. Climate change and its impact exacerbates South Africa's vulnerability to mycotoxin contamination, and significantly threatens its's food systems, public health, and agro-economic development. Herein we analyse sixteen years (2005/2006-2020/2021) of annual national meteorological data on South Africa which reveals both systematic and erratic variability in critical climatic factors known to influence mycotoxin contamination in crops. Within the same study period, data on fumonisin (FB) monitoring show clear climate-dependent trends. The strongest positive warming trend is observed between 2018/2019 and 2019/2020 (0.51 °C/year), and a strong positive correlation is likewise established between FB contamination and temperature (r ranging from 0.6 to 0.9). Four machine learning models, viz support vector machines, eXtreme gradient boosting, random forest, and orthogonal partial least squares, are generalized on the historical data with suitable performance (RMSE as low as 0.00). All the adopted models are able to predict future FB contamination patterns with reasonable precision (R2 ranging from 0.34 to 1.00). The most important model feature for predicting average FB contamination (YA) is the historical pattern of average FB contamination in maize within the region (ΣFBs_avg). The two most significant features in modelling maximum FB contamination (YM) are minimum temperature from the CMIP6 data (Pro_tempMIN) and observed precipitation from the CRU data (O_prep). Our study provides strong evidence of the impact of climate change on FB in South Africa and reiterates the significance of machine learning modelling in predicting mycotoxin contamination in light of changing climatic conditions, which could facilitate early warnings and the adoption of relevant mitigation measures that could help in mycotoxin risk management and control.

Unraveling root and rhizosphere traits in temperate maize landraces and modern cultivars: Implications for soil resource acquisition and drought adaptation.

A holistic understanding of plant strategies to acquire soil resources is pivotal in achieving sustainable food security. However, we lack knowledge about variety-specific root and rhizosphere traits for resource acquisition, their plasticity and adaptation to drought. We conducted a greenhouse experiment to phenotype root and rhizosphere traits (mean root diameter [Root D], specific root length [SRL], root tissue density, root nitrogen content, specific rhizosheath mass [SRM], arbuscular mycorrhizal fungi [AMF] colonization) of 16 landraces and 22 modern cultivars of temperate maize (Zea mays L.). Our results demonstrate that landraces and modern cultivars diverge in their root and rhizosphere traits. Although landraces follow a 'do-it-yourself' strategy with high SRLs, modern cultivars exhibit an 'outsourcing' strategy with increased mean Root Ds and a tendency towards increased root colonization by AMF. We further identified that SRM indicates an 'outsourcing' strategy. Additionally, landraces were more drought-responsive compared to modern cultivars based on multitrait response indices. We suggest that breeding leads to distinct resource acquisition strategies between temperate maize varieties. Future breeding efforts should increasingly target root and rhizosphere economics, with SRM serving as a valuable proxy for identifying varieties employing an outsourcing resource acquisition strategy.

Assessment of Agricultural Practices for Controlling Fusarium and Mycotoxins Contamination on Maize Grains: Exploratory Study in Maize Farms.

Maize is a significant crop to the global economy and a key component of food and feed, although grains and whole plants can often be contaminated with mycotoxins resulting in a general exposure of the population and animals. To investigate strategies for mycotoxins control at the grain production level, a pilot study and exploratory research were conducted in 2019 and 2020 to compare levels of mycotoxins in grains of plants treated with two fertilizers, F-BAC and Nefusoil, under real agricultural environment. The 1650 grains selected from the 33 samples were assessed for the presence of both Fusarium species and mycotoxins. Only fumonisins and deoxynivalenol were detected. Fumonisin B1 ranged from 0 to 2808.4 µg/Kg, and fumonisin B2 from 0 to 1041.9 µg/Kg, while deoxynivalenol variated from 0 to 465.8 µg/Kg. Nefusoil showed to be promising in regard to fumonisin control. Concerning the control of fungal contamination rate and the diversity of Fusarium species, no significant differences were found between the two treatments in any of the years. However, a tendency for was observed Nefusoil of lower values, probably due to the guaranteed less stressful conditions to the Fusarium spp. present in the soil, which do not stimulate their fumonisins production.

Assessment of Maize Hybrids Resistance to Aspergillus Ear Rot and Aflatoxin Production in Environmental Conditions in Serbia.

Aflatoxin, a naturally occurring toxin produced by the fungus Aspergillus flavus, is the most economically important mycotoxin in the world, with harmful effects on human and animal health. Preventive measures such as irrigation and planting dates can minimize aflatoxin contamination most years. However, no control strategy is completely effective when environmental conditions are extremely favorable for growth of the fungus. The most effective control method is growing maize hybrids with genetic resistance to aflatoxin contamination. The aim of this research was to evaluate the sensitivity of different maize hybrids to A. flavus infection and aflatoxin accumulation. Twenty commercial maize hybrids were evaluated in field trials with artificial inoculations using the colonized toothpicks method. The mycotoxin production potential of A. flavus isolates was confirmed by cluster amplification patterns (CAPs) analysis. The results of this research indicated the existence of significant differences in maize hybrids susceptibility to Aspergillus ear rot and aflatoxin B1 accumulation. No hybrid included in this research showed complete resistance in all conditions, but some hybrids showed partial resistance. Different hybrids also responded differently depending on the sowing date. This research showed that infection intensity is not always consistent with aflatoxin levels, and therefore visual evaluation is not enough to assess maize safety.

Aflatoxin Contamination of Maize, Groundnut, and Sorghum Grown in Burkina Faso, Mali, and Niger and Aflatoxin Exposure Assessment.

Aflatoxin contamination of staple crops by Aspergillus flavus and closely related fungi is common across the Sahel region of Africa. Aflatoxins in maize, groundnut, and sorghum collected at harvest or from farmers' stores within two weeks of harvest from Burkina Faso, Mali, and Niger were quantified. Thereafter, aflatoxin exposure values were assessed using per capita consumption rates of those crops. Mean aflatoxin concentrations in maize were high, 128, 517, and 659 µg/kg in Mali, Burkina Faso, and Niger, respectively. The estimated probable daily intake (PDI) of aflatoxins from maize ranged from 6 to 69, 29 to 432, and 310 to 2100 ng/kg bw/day in Mali, Burkina Faso, and Niger, respectively. Similarly, mean aflatoxin concentrations in sorghum were high, 76 and 259 µg/kg in Mali and Niger, respectively, with an estimated PDI of 2-133 and 706-2221. For groundnut, mean aflatoxin concentrations were 115, 277, and 628 µg/kg in Mali, Burkina Faso, and Niger, respectively. Aflatoxin exposure values were high with an estimated 9, 28, and 126 liver cancer cases/100,000 persons/year in Mali, Burkina Faso, and Niger, respectively. Several samples were extremely unsafe, exceeding manyfold regulatory levels of diverse countries (up to 2000 times more). Urgent attention is needed across the Sahel for integrated aflatoxin management for public health protection, food and nutrition security, and access to trade opportunities.

Biosensor approach for electrochemical quantitative assessment and qualitative characterization of the effect of fusaric acid on a culture-receptor.

Fusaric acid (FA) is a secondary fungal metabolite, which is widespread on corn and corn-based feed and food; FA has non-specific toxicity. Biosensor method is an express and easy-to-use method for quantitative and qualitative assessment of FA effect. Search for cultures has been performed for the formation of laboratory models of FA biosensor with the Clark-type oxygen electrode as transducer: respiration intensity of chosen cultures changed in the presence of FA. Resting cells of Fusarium oxysporum f. sp. vasinfectum and Bacillus subtilis were used as receptors of the amperometric biosensor for FA determination in aqueous solution. To enhance the sensitivity of detection, induction by substrate was performed for Bacillus subtilis. Response-concentration linear dependencies were obtained in a range of 0.5-500 FA mg/L. Biosensor models were applied to characterize influence of FA on microbial cells and investigate some features of FA transport. The dependences of the cells' response to FA on FA concentration were obtained; the kinetic parameters S0.5 and Vmax were determined for each culture. Inhibition-threshold FA (Sit) concentrations were similar for both studied cultures. At concentrations lower than Sit, the process of simple diffusion governed FA transport into cells and caused the cells' response to FA for non-induced culture.

Assessment of the Potential of a Native Non-Aflatoxigenic Aspergillus flavus Isolate to Reduce Aflatoxin Contamination in Dairy Feed.

Aspergillus species can produce aflatoxins (AFs), which can severely affect human and animal health. The objective was to evaluate the efficacy of reducing AF contamination of a non-aflatoxigenic isolate of A. flavus experimentally coinoculated with different aflatoxigenic strains in whole plant (WP), corn silage (CS), immature grains (IG) and in culture media (CM). An L-morphotype of A. flavus (CS1) was obtained from CS in a dairy farm located in the Mexican Highland Plateau; The CS1 failed to amplify the AFs biosynthetic pathway regulatory gene (aflR). Monosporic CS1 isolates were coinoculated in WP, CS, IG and CM, together with A. flavus strains with known aflatoxigenic capacity (originating from Cuautitlán and Tamaulipas, Mexico), and native isolates from concentrate feed (CF1, CF2 and CF3) and CS (CS2, CS3). AF production was evaluated by HPLC and fungal growth rate was measured on culture media. The positive control strains and those isolated from CF produced a large average amount of AFs (15,622 ± 3952 and 12,189 ± 3311 µg/kg), whereas A. flavus strains obtained from CS produced a lower AF concentration (126 ± 25.9 µg/kg). CS1 was efficient (p < 0.01) in decreasing AF concentrations when coinoculated together with CF, CS and aflatoxigenic positive control strains (71.6−88.7, 51.0−51.1 and 63.1−71.5%) on WP, CS, IG and CM substrates (73.9−78.2, 65.1−73.7, 63.8−68.4 and 57.4−67.6%). The results suggest that the non-aflatoxigenic isolate can be an effective tool to reduce AF contamination in feed and to minimize the presence of its metabolites in raw milk and dairy products intended for human nutrition.

Can it be all more simple? Manufacturing aflatoxin biocontrol products using dry spores of atoxigenic isolates of Aspergillus flavus as active ingredients.

Aflatoxin contamination of staple crops, commonly occurring in warm areas, negatively impacts human and animal health, and hampers trade and economic development. The fungus Aspergillus flavus is the major aflatoxin producer. However, not all A. flavus genotypes produce aflatoxins. Effective aflatoxin control is achieved using biocontrol products containing spores of atoxigenic A. flavus. In Africa, various biocontrol products under the tradename Aflasafe are available. Private and public sector licensees manufacture Aflasafe using spores freshly produced in laboratories adjacent to their factories. BAMTAARE, the licensee in Senegal, had difficulties to obtain laboratory equipment during its first year of production. To overcome this, a process was developed in Ibadan, Nigeria, for producing high-quality dry spores. Viability and stability of the dry spores were tested and conformed to set standards. In 2019, BAMTAARE manufactured Aflasafe SN01 using dry spores produced in Ibadan and sent via courier and 19 000 ha of groundnut and maize in Senegal and The Gambia were treated. Biocontrol manufactured with dry spores was as effective as biocontrol manufactured with freshly produced spores. Treated crops contained safe and significantly (P < 0.05) less aflatoxin than untreated crops. The dry spore innovation will make biocontrol manufacturing cost-efficient in several African countries.

Distribution, Genetic Diversity and Biocontrol of Aflatoxigenic Aspergillus flavus in Serbian Maize Fields.

Maize is one of the leading export products in the Republic of Serbia. As a country where economic development depends on agriculture, maize production plays a critical role as a crop of strategic importance. Potential aflatoxin contamination of maize poses a risk to food and feed safety and tremendous economic losses. No aflatoxin contamination of maize samples harvested in 2019 and 2020 in different localities in the Republic of Serbia was detected by the Enzyme-Linked Immunosorbent Assay (ELISA) test and High-Performance Liquid Chromatography (HPLC) method. On the other hand, the Cluster Amplification Patterns (CAP) analyses of the isolated Aspergillus flavus strains from 2019 maize samples confirmed the presence of key biosynthesis genes responsible for aflatoxin production. Artificial inoculation and subsequent HPLC analysis of the inoculated maize samples confirmed the high capacity of the A. flavus strains for aflatoxin production, pointing to a high risk of contamination under favorable conditions. Prevention of aflatoxin contamination is primarily based on A. flavus control, where biocontrol agents play a significant role as sustainable disease management tools. In this study, antagonistic activity screening of the novel strains belonging to the Bacillus genus indicated superior suppression of A. flavus strains by two Bacillus strains isolated from the rhizosphere of Phaseolus vulgaris.

Validation and Application of a Low-Cost Sorting Device for Fumonisin Reduction in Maize.

Fumonisin mycotoxins are a persistent challenge to human and livestock health in tropical and sub-tropical maize cropping systems, and more efficient methods are needed to reduce their presence in food systems. We constructed a novel, low-cost device for sorting grain, the "DropSort", and tested its effectiveness on both plastic kernel models and fumonisin-contaminated maize. Sorting plastic kernels of known size and shape enabled us to optimize the sorting performance of the DropSort. The device sorted maize into three distinct fractions as measured by bulk density and 100-kernel weight. The level of fumonisin was lower in the heaviest fractions of maize compared to the unsorted samples. Based on correlations among fumonisin and bulk characteristics of each fraction, we found that light fraction 100-kernel weight could be an inexpensive proxy for unsorted fumonisin concentration. Single kernel analysis revealed significant relationships among kernel fumonisin content and physical characteristics that could prove useful for future sorting efforts. The availability of a low-cost device (materials~USD 300) that can be used to reduce fumonisin in maize could improve food safety in resource-limited contexts in which fumonisin contamination remains a pressing challenge.

Metagenomics Assessment of Soil Fertilization on the Chemotaxis and Disease Suppressive Genes Abundance in the Maize Rhizosphere.

Soil fertility is a function of the level of organic and inorganic substances present in the soil, and it influences the activities of soil-borne microbes, plant growth performance and a host of other beneficial ecological functions. In this metagenomics study, we evaluated the response of maize microbial functional gene diversity involved in chemotaxis, antibiotics, siderophores, and antifungals producing genes within the rhizosphere of maize plants under compost, inorganic fertilizer, and unfertilized conditions. The results show that fertilization treatments at higher compost manure and lower inorganic fertilizer doses as well as maize plants itself in the unfertilized soil through rhizosphere effects share similar influences on the abundance of chemotaxis, siderophores, antifungal, and antibiotics synthesizing genes present in the samples, while higher doses of inorganic fertilizer and lower compost manure treatments significantly repress these genes. The implication is for a disease suppressive soil to be achieved, soil fertilization with high doses of compost manure fertilizer treatments as well as lower inorganic fertilizer should be used to enrich soil fertility and boost the abundance of chemotaxis and disease suppressive genes. Maize crops also should be planted sole or intercropped with other crops to enhance the rhizosphere effect of these plants in promoting the expression and abundance of these beneficial genes in the soil.

Cyanobacterial inoculation as resource conserving options for improving the soil nutrient availability and growth of maize genotypes.

Harnessing the benefits of plant-microbe interactions towards better nutrient mobilization and plant growth is an important challenge for agriculturists globally. In our investigation, the focus was towards analyzing the soil-plant-environment interactions of cyanobacteria-based formulations (Anabaena-Nostoc consortium, BF1-4 and Anabaena-Trichoderma biofilm, An-Tr) as inoculants for ten maize genotypes (V1-V10). Field experimentation using seeds treated with the formulations illustrated a significant increase of 1.3- to 3.8-fold in C-N mobilizing enzyme activities in plants, along with more than five- to six-fold higher values of nitrogen fixation in rhizosphere soil samples. An increase of 22-30% in soil available nitrogen was also observed at flag leaf stage, and 13-16% higher values were also recorded in terms of cob yield of V6 with An-Tr biofilm inoculation. Savings of 30 kg N ha-1 season-1 was indicative of the reduced environmental pollution, due to the use of microbial options. The use of cyanobacterial formulations also enhanced the economic, environmental and energy use efficiency. This was reflected as 37-41% reduced costs lowered GHG emission by 58-68 CO2 equivalents and input energy requirement by 3651-4296 MJ, over the uninoculated control, on hectare basis. This investigation highlights the superior performance of these formulations, not only in terms of efficient C-N mobilization in maize, but also making maize cultivation a more profitable enterprise. Such interactions can be explored as resource-conserving options, for future evaluation across ecologies and locations, particularly in the global climate change scenario.

Potential use of machine learning methods in assessment of Fusarium culmorum and Fusariumproliferatum growth and mycotoxin production in treatments with antifungal agents.

Fusarium-controlling fungicides are necessary to limit crop loss. Little is known about the effect of antifungal formulations at sub-lethal doses, and their interaction with abiotic factors, on Fusarium culmorum and F. proliferatum development and on zearalenone and fumonisin biosynthesis, respectively. In the present study different treatments based on sulfur, trifloxystrobin and demethylation inhibitor fungicides (cyproconazole, tebuconazole and prothioconazole) under different environmental conditions, in Maize Extract Medium, are assayed in vitro. Several machine learning methods (neural networks, random forest and extreme gradient boosted trees) have been applied for the first time for modeling growth of F. culmorum and F. proliferatum and zearalenone and fumonisin production, respectively. The most effective treatment was prothioconazole, 250 g/L + tebuconazole, 150 g/L. Effective doses of this formulation for reduction or total growth inhibition ranged as follows ED50 0.49-1.70, ED90 2.57-6.02 and ED100 4.0-8.0 µg/mL, depending on the species, water activity and temperature. Overall, the growth rate and mycotoxin levels in cultures decreased when doses increased. Some treatments in combination with certain aw and temperature values significantly induced toxin production. The extreme gradient boosted tree was the model able to predict growth rate and mycotoxin production with minimum error and maximum R2 value.

Prevalence of mycotoxigenic fungi and assessment of aflatoxin contamination: a multiple case study along the integrated corn-based poultry feed supply chain in Malaysia.

BACKGROUND: Corn, a main feed ingredient in the livestock industry, is one of the most susceptible crops to fungal infection and aflatoxin contamination. Livestock feeding on aflatoxin (AF)-contaminated feed have been shown to experience feed refusal, and decreased growth rate, milk production, and feed efficiency. In poultry, AF poisoning causes weight loss, poor feed efficiency, and reduced egg production and egg weight. The present work therefore aimed to determine the prevalence of mycotoxigenic fungi and the occurrence of AF contamination along the integrated corn-based poultry feed supply chain in Malaysia. A total of 51 samples were collected from different points along the feed supply chain from integrated poultry feed companies. The samples were subjected to mycological analyses (fungal isolation, enumeration, identification), and AFs were quantified by high-performance liquid chromatography equipped with a fluorescence detector (HPLC-FLD). RESULTS: Samples collected from sampling point 1 (company A) and sampling point 9 (company B) yielded the highest total fungal load (>log 4 CFU g-1 ). The prevalent fungal genera isolated were Aspergillus, Fusarium, and Penicillium spp. Aflatoxin B1 was detected in 8.3% of corn samples, and 7.4% of corn-based poultry feed samples along the feed supply chain, whereas AFs B2 , G1 , and G2 were not detected. CONCLUSION: The incidence of mycotoxigenic fungi along the integrated poultry feed supply chain warrant continuous monitoring of mycotoxin contamination to reduce the exposure risk of mycotoxin intake in poultry. © 2020 Society of Chemical Industry.

Analysis of the Genome of the Heavy Metal Resistant and Hydrocarbon-Degrading Rhizospheric Pseudomonas qingdaonensis ZCR6 Strain and Assessment of Its Plant-Growth-Promoting Traits.

The Pseudomonas qingdaonensis ZCR6 strain, isolated from the rhizosphere of Zea mays growing in soil co-contaminated with hydrocarbons and heavy metals, was investigated for its plant growth promotion, hydrocarbon degradation, and heavy metal resistance. In vitro bioassays confirmed all of the abovementioned properties. ZCR6 was able to produce indole acetic acid (IAA), siderophores, and ammonia, solubilized Ca3(PO4)2, and showed surface active properties and activity of cellulase and very high activity of 1-aminocyclopropane-1-carboxylic acid deaminase (297 nmol α-ketobutyrate mg-1 h-1). The strain degraded petroleum hydrocarbons (76.52% of the initial hydrocarbon content was degraded) and was resistant to Cd, Zn, and Cu (minimal inhibitory concentrations reached 5, 15, and 10 mM metal, respectively). The genome of the ZCR6 strain consisted of 5,507,067 bp, and a total of 5055 genes were annotated, of which 4943 were protein-coding sequences. Annotation revealed the presence of genes associated with nitrogen fixation, phosphate solubilization, sulfur metabolism, siderophore biosynthesis and uptake, synthesis of IAA, ethylene modulation, heavy metal resistance, exopolysaccharide biosynthesis, and organic compound degradation. Complete characteristics of the ZCR6 strain showed its potential multiway properties for enhancing the phytoremediation of co-contaminated soils. To our knowledge, this is the first analysis of the biotechnological potential of the species P. qingdaonensis.

Occurrence and Probabilistic Risk Assessment of Fumonisin B1, Fumonisin B2 and Deoxynivalenol in Nixtamalized Maize in Mexico City.

Fumonisins (FB1+FB2) and deoxynivalenol (DON) are mycotoxins produced by Fusarium species that might be present in maize and maize products. Knowledge on their occurrence in nixtamalized maize from Mexico together with an accompanying risk assessment are scarce, while nixtamalized maize is an important food in Mexico. This study presents the occurrence of FB1 + FB2 and DON in nixtamalized maize samples collected in Mexico City and analyses their distribution and resulting estimated daily intake for Mexican consumers by a probabilistic approach using a two-dimensional Monte-Carlo simulation. The results obtained reveal that for FB1 + FB2, 47% of the Mexican men and 30% of the Mexican women might exceed the provisional tolerable daily intake (PMTDI) of 2 µg/kg bw/day for fumonisins and for DON, 9% of men and 5% of women would be exceeding the PMTDI of 1 µg/kg bw/day, corresponding to the high consumers. The results raise a flag for risk managers in Mexico, to consider regulations and interventions that lower mycotoxin levels in nixtamalized maize for human consumption.

When to use one-dimensional, two-dimensional, and Shifted Transversal Design pooling in mycotoxin screening.

While complex sample pooling strategies have been developed for large-scale experiments with robotic liquid handling, many medium-scale experiments like mycotoxin screening by Enzyme-Linked Immunosorbent Assay (ELISA) are still conducted manually in 48- and 96-well plates. At this scale, the opportunity to save on reagent costs is offset by the increased costs of labor, materials, and risk-of-error caused by increasingly complex pooling strategies. This paper compares one-dimensional (1D), two-dimensional (2D), and Shifted Transversal Design (STD) pooling to study whether pooling affects assay accuracy and experimental cost and to provide guidance for when a human experimentalist might benefit from pooling. We approximated mycotoxin contamination in single corn kernels by fitting statistical distributions to experimental data (432 kernels for aflatoxin and 528 kernels for fumonisin) and used experimentally-validated Monte-Carlo simulation (10,000 iterations) to evaluate assay sensitivity, specificity, reagent cost, and pipetting cost. Based on the validated simulation results, assay sensitivity remains 100% for all four pooling strategies while specificity decreases as prevalence level rises. Reagent cost could be reduced by 70% and 80% in 48- and 96-well plates, with 1D and STD pooling being most reagent-saving respectively. Such a reagent-saving effect is only valid when prevalence level is < 21% for 48-well plates and < 13%-21% for 96-well plates. Pipetting cost will rise by 1.3-3.3 fold for 48-well plates and 1.2-4.3 fold for 96-well plates, with 1D pooling by row requiring the least pipetting. Thus, it is advisable to employ pooling when the expected prevalence level is below 21% and when the likely savings of up to 80% on reagent cost outweighs the increased materials and labor costs of up to 4 fold increases in pipetting.

Draft Genome Sequence Resource for Phyllachora maydis-An Obligate Pathogen That Causes Tar Spot of Corn with Recent Economic Impacts in the United States.

Phyllachora maydis is an important fungal pathogen that causes tar spot of corn and has led to significant yield loss in the United States and other countries. P. maydis is an obligate biotroph belonging to the Sordariomycetes class of Ascomycota. Due to the challenges posed by their obligate nature, there is no genome sequence available in the Phyllachora genus. P. maydis isolate PM01 was collected from a corn field in Indiana and the genome was determined by next-generation sequencing. The assembly size is 45.7 Mb, with 56.46% repetitive sequences. There are 5,992 protein-coding genes and 59 are predicted as effector proteins. This genome resource will increase our understanding of genomic features of P. maydis and will assist in studying the corn-P. maydis interaction and identifying potential resistant candidates for corn breeding programs.

Assessment of inoculating various epiphytic microbiota on fermentative profile and microbial community dynamics in sterile Italian ryegrass.

AIMS: To evaluate the effects of various epiphytic microbiota from Italian ryegrass (IR), maize (MZ) and sorghum (SG) on fermentative profile and microbial community dynamics in sterile IR. METHODS AND RESULTS: Using microbiota transplantation, the irradiated IR was treated with the following: (i) sterile water; (ii) epiphytic microbiota on IR (IRIR); (iii) epiphytic microbiota on MZ (IRMZ); (iv) epiphytic microbiota on SG (IRSG). After 60 days of ensiling, MZ and SG microbiota significantly (P < 0·05) decreased lactic acid (LA) and acetic acid (AA) concentrations compared to IR microbiota, while SG microbiota notably (P < 0·05) reduced the ratio of LA to AA than MZ and IR microbiota. Apparently (P < 0·01) higher amounts of Lactobacillus genus were observed in IRIR and IRMZ groups on 60 day compared to IRSG group, and the dominant Lactococcus genus on 3 day was eventually replaced by Enterobacteriaceae and Lactobacillus in IRSG group. CONCLUSIONS: Exogenous microbiota could evidently affect the fermentative profile and microbial community dynamics of IR silage. The numbers of Enterobacteriaceae and Lactobacillus were mainly responsible for this. SIGNIFICANCE AND IMPACT OF THE STUDY: Identifying the role of microbe during ensiling is of great significance to manipulate the fermentation products and improve the preservation of silage.