Innovative technology of the process of disinfection and inactivation of antinutrients in compound feeds using liquid components

Научный и практический интересы представляют процессы обеззараживания и инактивации антипитательных веществ в комбикормах. Известны разные технологии обеззараживания (например, нагрев, давление, растворы антимикробных препаратов и другие новые технологии) для повышения эффективности снижения риска заболеваний не только животных и птиц, но и людей. Процессы обеззараживания и инактивации антипитательных веществ может применяться к широкому набору пищевых продуктов и к сырью комбикормовой промышленности. Однако необходимо провести дальнейшие исследования, чтобы понять кинетику инактивации антипитательных веществ в комбикормах для достижения приемлемой технологии обеззараживания. Исследовано влияние гидротермической обработки на поверхностную и глубинную микрофлору зернового сырья. С увеличением времени обработки пшеница и ячмень, искусственно зараженные, а также зараженные естественным способом, были обеззаражены предлагаемым способом на 100 %. Получены химико-технологические характеристики кондиционированного зерна, результаты которых представлены для процессов пропаривания и охлаждения. Установлено, что с увеличением температуры влажность пропаренных и увлажненных зерен пшеницы возрастает с 15,2 до 21,7 %, для зерен ячменя - с 15,3 до 12,8 %. При этом исходная влажность пшеницы составила 10,8 %, а ячменя - 11,2 %. Определено, что при увеличении продолжительности пропаривания и увлажнения зерен пшеницы давление уменьшается с 0,15 до 0,19 МПа и уменьшается расход пара с 85 до 60 кг/т. Анализ физико-механических свойств для пшеницы и ячменя показал, что объемная масса исследуемых образцов возросла. Угол естественного откоса составил для пшеницы 45-48, для ячменя 43-50. Трехстадийную технологию гидротермической обработки комбикормов рекомендуется использовать в трехсекционном реакторе-кондиционере, в котором продукт равномерно кондиционируется с инактивацией антипитательных веществ, обеззараживается и частично желатинизируется. The processes of anti-nutrients disinfection and inactivation in compound feeds are of scientific and practical interest. Various disinfection technologies are known (for example, heating, pressure, solutions of antimicrobial preparations and other new technologies) to increase the effectiveness of diseases risk reducing not only for animals and birds, but for humans as well. The processes of anti-nutrients desinfection and inactivation can be applied to a wide variety of food products and feedstock raw materials. However, further research is necessary to understand the kinetics of anti-nutrients inactivation in compound feeds to achieve an acceptable disinfection technology. The influence of hydrothermal treatment on the surface and deep microflora of grain raw materials was investigated in the work. With an increase in the processing time, both artificially infected and naturally infected wheat and barley were 100 % disinfected with the method proposed. Chemical- and technological characteristics of conditioned grain were received. Their results are presented for the steaming and cooling processes. It was found out that with an increase in temperature, the humidity of steamed and moistened wheat grains increases from 15.2 to 21.7 %, and of barley grains from 15.3 to 12.8 %. At the same time, the initial moisture content of wheat was 10.8 %, and of barley - 11.2 %. It was determined that with an increase in the duration of steaming and moistening of wheat grains, the pressure decreases from 0.15 to 0.19 MPa, and the steam consumption decreases from 85 to 60 kg/t. Analysis of wheat and barley physical- and mechanical properties revealed that the bulk density of the samples studied increased. The repose angle was 45-48 for wheat and 43-50 for barley. It is recommended to use a three-stage technology of compound feed hydrothermal treatment in a three-section reactor-conditioner. The product is uniformly conditioned with anti-nutrients inactivation, disinfected and partially gelatinized in it.

Isolation, Characterization and Acid/Alkaline Tolerance of Lactic Acid Bacteria Present in Fermented Rye, Wheat, Oat and Barley

Objective: This study determined the lactic acid bacteria (LAB) present in fermented rye, wheat, oat and barley grains, and evaluated their survival in simulated gastric juice and pancreatic juice. Methods: Samples of rye, wheat, oat and barley grains were fermented for 72 hours at room temperature. Lactic acid bacteria (LAB) strains were isolated using MRS agar and were enumerated. Isolated LAB strains were cultured with MRS broth and the fermentation patterns of the isolated strains were characterized using API 50 CH kit (Biomerieux, France). Each isolated LAB strain was exposed to simulated gastric juice at pH of 2.0 for 80 minutes at 370C, followed by exposure to simulated pancreatic juice at pH of 8.0 for 120 minutes at 370C. Aliquots were taken at 0 minute and 80 minutes at pH of 2.0 and 0 minutes and 120 minutes at pH of 8.0 for enumeration of LAB strains. Results: The total LAB cell count ranged from 6.6 * 108 ± 11 cfu/ml in the rye sample to 9.5*109 ± 7 cfu/ml in the oat sample. 13 LAB strains were isolated from the four selected cereal grains and were characterized as six strains of Lactobacillus plantarum1, five strains of L. brevis 1 and one strain each of L. collinoides and Leuconostoc citreum. All the isolated LAB strains from the four selected cereals survived in the simulated gastric juice at pH of 2.0 (before and after incubation at 0min and 80a min) and after addition of simulated pancreatic juice at pH of 8.0 (before and after incubation at 80b min and 200 min respectively). The mean viable counts of all the strains ranged from 2.0 *108 in R3 at 80b min to 1.54 * 1010 in B4 at 80b minutes. Conclusion: LAB associated with fermentation of rye, wheat, oat and barley grains are likely to survive transport through the harsh acidic and alkaline conditions of the GIT.

Haze in Beer: Its Formation and Alleviating Strategies, from a Protein–Polyphenol Complex Angle

Beer is one of the oldest and most widely consumed alcoholic beverages. Haze formation in beer is a serious quality problem, as it largely shortens the shelf life and flavor of beer. This paper reviews the factors affecting haze formation and strategies for reducing haze. Haze formation is mainly associated with specific chemical components in malt barley grains, such as proteins. The main factor causing haze formation is a cross-linking of haze active (HA) proteins and HA polyphenols. Many HA proteins and their editing genes or loci have been identified by proteomics and quantitative trait locus (QTL) analysis, respectively. Although some technical approaches have been available for reducing haze formation in beer, including silica and polyvinylpolypyrrolidone (PVPP) adsorbent treatments, the cost of beer production will increase and some flavor will be lost due to reduced relevant polyphenols and proteins. Therefore, breeding the malt barley cultivar with lower HA protein and/or HA polyphenols is the most efficient approach for controlling haze formation. Owing to the completion of barley whole genome sequencing and the rapid development of modern molecular breeding technology, several candidate genes controlling haze formation have been identified, providing a new solution for reducing beer haze.

The Influence of Germinated Hull-less Barley Sordough Fermentation Conditions on the Microbiota Development

Sourdough plays an important role in quality assurance, especially during developing wholegrain wheat bread products. The value of barley and hull-less barley grains is determined by the low lipid and high carbohydrate (starch and resistant starch) content, balanced amino acid profile, dietary fibre and phenolic compounds. During germination (24 h) the amount of sugar, amino acids and vitamins increase in the grains. Therefore, it could be optimal environment for microbiota growth in sourdough. The goal of the research was to find optimal technological parameters and to study microorganism growth dynamic during the fermentation of sourdough with germinated hull-less barley. The study was performed using three steps of fermentation, with different temperature and time parameters. At each step pH, dynamics of lactic acid bacteria (LAB), yeast-like fungi and total plate count (TPC) growth were determined using standard methods. The results of the study showed that the optimal fermentation conditions for the first stage are 48 h and 26 ± 1 °C, for the second stage 8 h and 26 ± 1 °C and for the third stage 20 h and 28 ± 1 °C. During fermentation of the sourdough, the pH 3.91 is reached, LAB 8.8 log10 CFU g−1, and the yeast-like fungi 8.7 log10 CFU g−1 was determined. Obtained data show that more than 80 bacteria and 50 microscopic fungal species were detected in hull-less barley flour, but after germination their diversity decreased and in the germinated hull-less barley sourdoughs dominated species were Pediococcus pentosaceus.

First Report of Fusarium verticillioides Causing Fusariosis on Triticale Grain in Serbia

Triticale (Triticosecale Wittmack) is a cereal crop species developed to combine positive traits of wheat and rye into a single plant (Arendt and Zannini, 2013). Due to its high protein content, favourable amino acid composition and nutritional values higher than maize, triticale is an excellent component for preparing feed for domestic animals (Đekić et al. 2019). In May 2017, discoloured spikes of triticale were observed in a field in Zemun Polje, Serbia, with a disease incidence between 15 and 20%. A total of 400 kernels obtained from discoloured spikes were surface sterilised with 1% sodium hypochlorite solution for 3 minutes and then rinsed 3 times with sterilized water and plated on potato dextrose agar (PDA) at 25°C incubated in the dark for 7 days. A total of 15 isolates of genus Fusarium on PDA formed cottony white to greyish purple colony with dark yellow to purple grey reverse and average colony. Obtained isolates were transferred to carnation leaf agar (CLA) and synthetic nutrition agar (SNA), for 7-day incubation in the dark at 25°C. Ten isolates formed “rabbit ear” monophialides which is the main characteristic of Fusarium verticillioides (Sacc.) Nirenberg. Microconidia are formed from monophialids in long chains. Their shape is oval with a flattened base, without septa (4.0-18.0 x 1.5-4.5 µm). Shape of macroconidia is slightly falcate to almost straight, (3.5-4.5 x 31-58 µm). Chlamydospores are not produced (Leslie and Summerell, 2006). The genomic DNA from one F. verticillioides isolate was extracted using the DNeasy Plant Mini kit (Qiagen, Hilden) according to the manufacturer’s protocol. Molecular identification was confirmed by sequencing the internal transcribed spacer (ITS), translation elongation factor-1 alpha (EF-1α) and RPB2 (O′Donell, 2000). Sequence was deposited in GenBank as MZ664391, MZ666958 and MZ666957. This sequence was compared with the sequences of F. verticillioides strains registered in the GenBank database based on nucleotide similarity. It showed 100% identity to the sequences MT180471.1 (ITS), MN861767.1 (EF-1 α) and MT264836.1 (RPB2). Pathogenicity of all isolates was tested on 20 randomly selected triticale spikes in four replicates (Mesterházy et al. 1999). Inoculation was performed when half of the plants reached the flowering stage, by spraying with 20 ml of spore suspension (1 × 106 spores/ml from 7-day-old cultures on PDA). F. verticillioides RBG 1603 Q27 was used as a positive control. Inoculated spikes were covered with PVC bags for 48h. Discoloured spikes were observed after 3 week incubation in a greenhouse at 20°C with a 16h light/8h dark photoperiod. All inoculated spikes showed symptoms, similar to those from field infections. Control spikes were symptomless. The fungus was reisolated and was morphologically identical to the original isolates, thus completing Koch’s postulates. Based on morphological, molecular and pathogenic features the isolated fungus was identified as F. verticillioides. Fusarium species infect crop plants worldwide resulting in yield loss and reduced grain quality due to mycotoxin contamination (Asam et al, 2017). In Serbia F. verticillioides was isolated from wheat and barley grains (Stanković et al., 2012) and to the best of our knowledge, this is the first report of F. verticillioides on triticale grain in Serbia.

Plant Growth Regulators INCYDE and TD-K Underperform in Cereal Field Trials

Using plant growth regulators to alter cytokinin homeostasis with the aim of enhancing endogenous cytokinin levels has been proposed as a strategy to increase yields in wheat and barley. The plant growth regulators INCYDE and CPPU inhibit the cytokinin degrading enzyme cytokinin oxidase/dehydrogenase (CKX), while TD-K inhibits the process of senescence. We report that the application of these plant growth regulators in wheat and barley field trials failed to enhance yields, or change the components of yields. Analyses of the endogenous cytokinin content showed a high concentration of trans-zeatin (tZ) in both wheat and barley grains at four days after anthesis, and statistically significant, but probably biologically insignificant, increases in cisZ-O-glucoside, along with small decreases in cZ riboside (cZR), dihydro Z (DHZ), and DHZR and DHZOG cytokinins, following INCYDE application to barley at anthesis. We discuss possible reasons for the lack of efficacy of the three plant growth regulators under field conditions and comment on future approaches to manipulating yield in the light of the strong homeostatic mechanisms controlling endogenous cytokinin levels.

Effects of background food on alternative grain uptake and zinc phosphide efficacy in wild house mice

BACKGROUND: House mice (Mus musculus) cause significant, ongoing losses to grain crops in Australia, particularly during mouse plagues. Zinc phosphide (ZnP) coated grain is used for control, but with variable success. In a laboratory setting, we tested if mice would (1) switch from consumption of one grain type to another when presented with an alternative, and (2) consume ZnP-treated grains when presented as a choice with a different grain. RESULTS: Mice readily switched from their background grain to an alternative grain, preferring cereals (wheat or barley) over lentils. Mice readily consumed ZnP-coated barley grains. Their mortality rate was significantly higher (86%, n=30) in the presence of a less-favoured grain (lentils) compared to their mortality rate (47%, n=29; and 53%, n=30) in the presence of a more-favoured grain (wheat and barley, respectively). Mice died between 4-112 h (median = 18 h) after consuming one or more toxic grains. Independent analysis of ZnP-coated grains showed variable toxin loading indicating that consumption of a single grain would not guarantee intake of a lethal dose. There was also a strong and rapid behavioural aversion if mice did not consume a lethal dose on the first night. CONCLUSIONS: The registered dose rate of 25 g ZnP/kg wheat; ~ 1 mg ZnP/grain in Australia needs to be re-evaluated to determine what factors may be contributing to variation in efficacy. Further field research is also required to understand the complex association between ZnP dose, and quantity and quality of background food on efficacy of ZnP baits.

Genome-wide association studies uncover candidate genes for Zinc, Iron, and Selenium concentration in barley grains

Background Barley (Hordeum vulgare L.) is one of the most important staple food crops worldwide. Mineral concentrations in cereals are important for human health; hence improving Zn, Fe and Se accumulation in grains is an imperative need. This study was designed to understand the genetic architecture of Zn, Fe and Se grain concentrations in barley grains. Results We performed a genome-wide association study (GWAS) for grain Zn, Fe and Se concentrations in 216 spring barley accessions, using field data from 2 years. All the accessions were genotyped with a high-density 9K SNPs array from IlluminaTM. The mean values of estimated BLUEs for Zn, Fe and Se were 38.37, 35.56 and 39.45 µg g− 1 dry weight, respectively. High heritability was equaled 75.65% for Fe across the two environments, while moderate heritability values were detected for Zn and Se. Notably, wide genetic variation was found among genotypes for Zn, Fe and Se concentrations. A total of 222 SNPs associated with Zn, Fe and Se were detected on all chromosomes, where the highest significant associations is linked to Fe accumulation. Three genomic regions include newly identified putative candidate genes, which are related to Zn uptake and transport or represent Homeobox leucine zipper protein. Additionally, several significant associations were physically located inside or near genes which are potentially involved in Zn and Fe homoeostasis of which two candidate genes at 5H (502,454,312–502,455,148 bp) and 7H (205,216,091–205,221,133 bp) were found to be involved in Basic helix loop helix (BHLH) family transcription factor and Squamosa promoter binding-like protein, respectively. Conclusions These findings provide new insights into the genetic basis of Zn, Fe and Se concentration in barley grains that in turn may help plant breeders to select high Zn, Fe and Se-containing genotypes to improve human consumption and grain quality.