Vplyv aditív na mykotoxickú kontamináciu kukuričných siláží

Addition of silage additives is commonly concentrated on the improvement of nutritive and fermentative features of silages, but in recent years, mycotoxic contamination is also a feature monitored after the treatment. This study aimed to evaluate the effect of silage additives, in the concrete inoculant consisting of a mixture of Lactobacillus buchneri LN40177, Lactobacillus casei LC329090 and urea, on the concentration of major mycotoxins in maize silages. The ensilage mass was made in three variants, which consisted of maize silage (control variant C), maize silage with the addition of a commercial inoculant (variant A) and maize silage with the addition of urea (variant U). The commercial inoculant was added to the cut mass at a dose of 1 g/t of mass and urea at a dose of 5 kg/t of mass. After application of silage additives, the mass was stored and sealed in silage units. Mycotoxin analysis was performed using Veratox tests with ELISA reader, whereas average samples were prepared according to established protocols. The immuno-enzymatic method revealed that all samples of maize silage showed 100% contamination. The maize silages were characterized by the highest concentration of deoxynivalenol regardless of the treatment. The results confirmed the effect of silage additives on mycotoxic contamination of maize silages. Compared to the control variant, the commercial inoculant had negative increasing effect (P<0.05) on deoxynivalenol, ochratoxins and zearalenone, and a positive decreasing effect (P<0.05) on T-2 toxin and fumonisins. The urea addition resulted in significant reduction of T-2 toxin and increase of zearalenone.Prídavkom silážnych aditív sa bežne sleduje zlepšenie nutričných a fermentačných vlastností siláží, ale v posledných rokoch je mykotoxická kontaminácia tiež sledovaným znakom po ošetrení. Táto štúdia bola zameraná na vyhodnotenie účinku silážnych aditív, konkrétne inokulantu pozostávajúceho zo zmesi Lactobacillus buchneri LN40177, Lactobacillus casei LC329090 a močoviny, na koncentráciu hlavných mykotoxínov v kukuričnej siláži. Silážovaná hmota kukurice bola vyhotovená v troch variantoch, ktoré pozostávali z kukuričnej siláže (kontrolný variant C), kukuričnej siláže s prídavkom komerčného inokulantu (variant A) a kukuričnej siláže s prídavkom močoviny (variant U). Komerčný inokulant bol do narezanej hmoty pridaný v dávke 1g/t silážnej hmoty a močovina v dávke 5 kg/t silážnej hmoty. Po aplikácií silážnych aditív sa hmota uskladnila a zasilážovala do silážnych jednotiek. Analýza mykotoxínov bola vykonaná pomocou Veratox testov na ELISA reader, pričom priemerné vzorky boli upravené podľa stanovených protokolov. Výsledky potvrdili efekt silážnych aditív na mykotoxickú kontamináciu kukuričných siláží. V porovnaní s kontrolným variantom mal komerčný inokulant negatívny stúpajúci efekt (P<0,05) na deoxynivalenol, ochratoxíny a zearalenon, a pozitívny klesajúci efekt (P<0,05) na T-2 toxín a fumonizíny. Výsledkom pridania močoviny bolo preukazné zníženie T-2 toxínu a zvýšenie zearalenonu

Vplyv prídavku baktérií mliečneho kvasenia na mikrobiotu a výskyt mykotoxínov v ražných silážach

In the rye silages (control and with additive) microbiota isolation and identification (total count of bacteria, coliform bacteria, enterococci, lactic acid bacteria and microscopic filamentous fungi) and occurrence of mycotoxins were evaluated. Total count of bacteria on Plate count agar at 30 °C for 48-72 hours, coliform bacteria on McConkey agar at 37 °C for 24-48 hours, enterococci on Enterococcus selective agar at 37 °C for 48-72 hours, lactic acid bacteria on De Man, Rogosa and Sharpe agar, Mayeux, Sandine and Elliker and All Purpose TWEEN® agar 37 °C for 48-72 hours in microaerophilic condition and microscopic filamentous fungi on Malt extract agar at 25 °C for 7 days were evaluated and identified with mass spectrometry. The most isolated family from both types of silage was Lactobacillaceae. The most isolated species from the control group was Lentilactobacillus buchneri (22%) and lowest number of isolated species was Mucor spp. The most isolated species from silage with additive was Lactobacillus gasseri (16%). The ELISA method (enzyme-linked immunosorbent assay) with using reader at wavelength 650 nm for detection and quantification of mycotoxins (total aflatoxins, total ochratoxins, total fumonisins, deoxynivalenol, zearalenone and T-2/HT-2 toxin) was used. The rye silages of control and with additive were characterized by occurrence of all analyzed mycotoxins, whereas deoxynivalenol was mycotoxin with the highest concentration. The rye silages with additive had significantly lower content of total aflatoxins compared to the control, while significantly higher concentration of total ochratoxins, deoxynivalenol, zearalenone and T-2/HT-2 toxin was determined.V ražných silážach (kontrolných aj s aditívom) bola hodnotená izolácia a identifikácia mikrobioty (celkový počet baktérií, koliformných baktérií, enterokokov, baktérií mliečneho kvasenia a mikroskopických vláknitých húb) a výskyt mykotoxínov. Pomocou hmotnostnej spektrometrie bol hodnotený a identifikovaný celkový počet baktérií na Plate count agare pri teplote 30 °C počas 48-72 hodín, koliformných baktérií na McConkeyho agare pri teplote 37 °C počas 24- 48 hodín, enterokokov na Enterococcus selektívnom agare pri teplote 37 °C počas 48-72 hodín, baktérií mliečneho kvasenia na De Man, Rogosa a Sharpe agare, Mayeux, Sandine a Elliker a All Purpose TWEEN® agare pri 37 °C počas 48-72 hodín v mikroaerofilných podmienkach a mikroskopické vláknité huby na Sladinovom agare pri 25 °C počas 7 dní. Najviac izolovanou čeľaďou z oboch typov siláže bola Lactobacillaceae. Najviac izolovaným druhom z kontrolnej skupiny bol Lentilactobacillus buchneri (22%) a najnižší počet z izolovaných druhov mal Mucor spp. Najviac izolovaným druhom zo siláže s aditívami bol Lactobacillus gasseri (16%). Na detekciu a kvantifikáciu mykotoxínov (celkové aflatoxíny, celkové ochratoxíny, celkové fumonizíny, deoxynivalenol, zearalenon a T-2/HT-2 toxín) sa použila ELISA metóda (enzyme-linked immunosorbent assay) s využitím čítačky pri vlnovej dĺžke 650 nm. Ražné siláže kontroly a s prídavkom aditíva sa vyznačovali výskytom všetkých analyzovaných mykotoxínov, pričom deoxynivalenol bol mykotoxín s najvyššou koncentráciou. Ražné siláže s aditívom mali výrazne nižší obsah celkových aflatoxínov v porovnaní s kontrolou, pričom bola stanovená výrazne vyššia koncentrácia celkových ochratoxínov, deoxynivalenolu, zearalenonu a T-2/HT-2 toxínu

The Change in Microbial Diversity and Mycotoxins Concentration in Corn Silage after Addition of Silage Additives

In our study the presence of bacteria, yeast, and microscopic fungi was evaluated. Three forms of corn silage were made including silage without additive, silage with microbial additive (lactic acid bacteria), and silage with nutritional additive (urea). Silage additives were applied to the matter within the recommended dosage, then the matter was ensiled into plastic bags and stored at a constant temperature. After 5.5 months of storage, average samples for microbial and mycotoxins analysis were taken. From microbiological points, the plate count agar method for enumeration of total count of bacteria, lactic acid bacteria, enterococci, yeasts, and microscopic fungi and mass spectrometry for microbiota identification were used. In total, 43 species of bacteria and yeasts and 6 genera of microscopic fungi were identified from all samples of corn silages. The most isolated species were Lentilactobacillus buchneri and Kazachstania exigua from bacteria resp. yeasts and Aspergillus and Penicillium from microscopic fungi. Mycotoxins were determined by HPLC-MS/MS and divided into two groups as regulated and emerging. In the corn silages only Fusarium mycotoxins were observed. All corn silages, regardless of the addition of the additive, were the highest in nivalenol content. Deoxynivalenol and beauvericin with the highest concentrations were present in silage with urea. Although the mycotoxins content of the variants changed, these changes were not statistically significant. In general, addition of lactic acid bacteria Lentilactobacillus buchneri and Lacticaseibacillus casei and urea as silage additives affect the microbial diversity; however, the hygienic quality of whole crop corn silage was not negatively changed

Significance of phosphate nano-fertilizers foliar application: A brief real-field study of quantitative, physiological parameters, and agro-ecological diversity in sunflower

One of the challenges in agriculture practices is guaranteeing an adequate and bioavailable phosphorus supply for plants on phosphorus-deficient soils. A promising alternative lies in the utilization of phosphate nano-fertilizers (NFs) through spray applications. Therefore, this short-term study aimed to investigate the yet undetermined widespread impact of P-NFs on crops characterized by broad leaves, an intensive rate of photosynthesis, and belonging to the oilseed plant, sunflower (Helianthus annuus L.). To achieve this, NFs were applied at lower concentrations of various phosphate based NFs, including (i) nano-hydroxylapatite (nano-Hap) and (ii) a mixture of nano-calcium zinc phosphate and macro-sized parascholzite (nano/macro-ZnPhos), in comparison to the NF-free control. The study was carried out under authentic field conditions during the 2022 vegetation season at the Dolná Malanta site within the Central European Region. The empirical evidence presented herein indicates that the utilization of biocompatible and bioactive nano-Hap, initially engineered for biomedical applications, and nano/macro-ZnPhos, now foliarly applied at reduced concentrations, elicited a statistically significant elevation in quantitative parameters and seasonal physiological responses. The parameters analyzed included head diameter, dry head weight, seed yield per hectare, nutritional seed oiliness, etc. as well as the physiological normalized difference vegetation index (NDVI), stomatal conductance index (Ig), and crop water stress index (CWSI). In terms of agro-ecological terrestrial bio/diversity, it was evident that the nano/macro-ZnPhos was the most hospitable variant for the terrestric insect community, but surprisingly, the agronomically more popular nano-Hap showed only statistically insignificant changes in the diversity of the detected communities. However, the relevance of outcomes highlighted using nano-fertilizers, supporting the concept of precision and sustainable agriculture under field conditions.Web of Science1310art. no. 260