Impact of Storage Temperature on Microbial Diversity and Probiotic Effect of Liquid Brewers’ Yeast

Using probiotics as animal feed additives instead of antibiotics is gaining momentum to avert adverse negative effects on human health. Liquid brewers yeast (LBY) is an industrial by-product containing probiotic microorganisms and is also used as a protein supplement for dairy animals. Nevertheless, value chain actors lack of appropriate handling practices compromises the by-products quality and safety. This study aimed to determine the effect of variation in temperature on microbial diversity and probiotic effects during the storage time of LBY sampled from distributors and farmers from Githunguri sub-county of Kenya. The samples were stored at 20C, 25C and 30C, then tested on 0, 5, 10, 15 and 20 days. The studys parameters involved determining the pH levels, lactic acid bacteria (LAB), total coliform count (TCC), mould, and yeast in LBY. The rate (k) of the reaction kinetics model was used to extrapolate the expected probiotic shelf life. The LAB and yeast populations were reduced in a first-order reaction at all storage temperatures. The rate of reduction in the numbers of LAB reduced with an increase in temperature (k = 0.019 and 0.023) at 20C and 30C, respectively. Yeasts highest rate of growth reduction was 25C (k = 0.009) and least at 30C (k = 0.043). The minimum effective concentration for probiotics of 106 CFU/mL needed to observe the beneficial physiological impact on farm animals was achieved between 34.9 and 35.5 days at the tested storage temperatures. The study provides insight into the unexploited low-cost probiotic potential of LBY in dairy production. Conversely, handling practices and environmental microbial contamination along the value chain can compromise product quality and safety. There is a need to advocate its use in dairy for improved productivity and sensitize farmers to appropriate hygienic measures along the LBY value chain.

Effect of β-Glucan (Angel Yeast) Compared to a Placebo on Cold and Flu Incidence and Symptoms in an Adult Population—A Double Blind, Randomised Controlled Trial

Background: 1-3, 1-6 β-glucan derived from Baker’s yeast (Saccharomyces cerevisiae) has been widely studied for its immune stimulatory capabilities and safety. Previous studies found β-glucan to have efficacy at reducing incidence of URTIs as well as being a low risk for negative side effects. The current study aimed to examine the effects of yeast β-glucan (Angel Yeast) on cold and flu incidences and symptoms in healthy adults. Methods: Two hundred and thirty-one males and females aged 18 to 65 years old supplemented with either β-glucan or a placebo for 3-months. Participants completed a general health questionnaire every 4 weeks and in addition, if participants experienced any cold or flu symptoms, these were recorded daily (along with severity) until resolved or up to 2 weeks. Results: Supplementation with β-glucan reduced the self-reported severity of sore throats and improved sleep quality compared to the placebo group. Conclusions: Yeast β-glucan supplementation appears to be able to help reduce certain symptoms experienced during a cold or flu episode and is safe and well tolerated.

Dairy Farming Conditions and Utilization Levels of Liquid Brewers’ Yeast in Kenya

Dairy production plays an integral part in supporting smallholder farmers’ livelihoods. The desire to increase the number of dairy cattle is not feasible due to the reduced output of feed resources occasioned by climate change. Consequently, the need to increase productivity per cow is inevitable. Conventional protein supplements are costly;hence, the need to explore affordable nutrientdense alternative feed resources. Liquid brewers’ yeast (LBY), a by-product of the brewing industry, is a rich protein supplement in dairy production. This study aimed to assess the dairy farming conditions and utilization levels of LBY as a feed supplement in Githunguri Sub-county, Kiambu. Semi-structured questionnaires were administered to 457 dairy farmers in a cross-sectional survey. The findings revealed that most farmers (94.2%) fed their cattle on established forage/fodder and crop residues with supplementation. Even though 53.1% of the respondents were aware of the use of LBY, only 30.6% utilized it to supplement dairy cows, most of whom (96.0%) used it fresh without preservation. Membership in farmers’ organizations increased awareness of LBY (r = 0.732). Principal component analysis indicated that the benefits of using LBY outweigh the challenges involved with a loading matrix of 0.891 - 0.954 and 0.681 - 0.807, respectively. The low adoption and use levels of LBY as a source of protein supplements were due to low awareness. There is a need for concerted efforts by stakeholders in the industry to increase farmers’ knowledge base on the utilization and effectiveness of LBY in dairy production.

Recovery of Residual Brewer’s Yeast by Electroactivation

Yeasts resulting from the brewing process (RBY) are a valuable by-product, with an important content of minerals, vitamins and, especially, proteins. The purpose of the research was the electroactivation of RBY and the simultaneous obtaining of two products—protein concentrates and hydrolyzed protein from residual brewer’s yeast. Electroactivation is a non-residual process, without the use of chemical reagents and relatively inexpensive. The variation of the electroactivation conditions allowed the separation of 90% - 94% of the proteins in the form of protein concentrates. During the process, it is attested to increase the pH value and decrease the redox potential, which characterizes the multiple redox processes that take place in the cathode cell, including sedimentation at the isoelectric point. The presence of albumin in the protein fractions of RBY allows the formation of protein complexes with calcium, attributing a higher biological value to the obtained products.

The setereoselective preparation ofβ-keto esters using baker's yeast in the presence of rice wine koji

By the addition of rice wine koji, enhancement of the reactivity was observed for the baker＇s yeast reduction of β-keto esters into （S）-β-hydroxy esters with high enantiomeric purity （73-98%）.

Baker's Yeast Mediated Reduction of Optically Active Diketone

Baker's Yeast mediated reduction of optically active diketone has been described.

Reaction Characteristics of Asymmetric Synthesis of (2S,5S)-2,5-Hexanediol Catalyzed with Baker’s Yeast Number 6

Baker’s yeast number 6 was selected by screening. It showed good catalytic activity and enantioselec-tivity for asymmetric reduction of 2,5-hexanedione to produce (2S,5S)-2,5-hexanediol. Gas chromatography-mass spectrometry (GC-MS) revealed that the intermediate was (S)-5-hydroxyhexane-2-one. Reduction of 2,5-hexanedione proceeded in a two-step reaction. The hydroxyketone was initially formed, and this intermediate was further re-duced to the diol. Factors influencing the product yield and the enantiomeric excess of the reduction of 2,5-hexandione catalyzed by baker’s yeast number 6 were investigated. Higher concentration (≤100 mmol·L-1) of 2,5-hexandione did not influence 5-hydroxyhexane-2-one production, but 2,5-hexanediol production was inhibited by excess accumulation (>30 mmol·L-1) of intermediate. The optimal conditions were glucose as the co-substrate at an initial glucose concentration of 20 g·L-1, 34°C, pH 7.0 and cell concentration 60 g·L-1 (cell dry mass). Under the optimal condition and an initial substrate concentration of 30 mmol·L-1, the yield of 2,5-hexandiol was 78.7% and the enantiomeric excess of (2S,5S)-2,5-hexandiol was 94.4% for 24-h reduction.

A rapid absorbance-based growth assay to screen the toxicity of oligomer Aβ42 and protect against cell death in yeast

Multiple lines of evidence show that soluble oligomer forms of amyloidβprotein(Aβ42)are the most neurotoxic species in the brain and correlates with the degree of neuronal loss and cognitive deficit in Alzheimer’s disease.Although many studies have used mammalian cells to investigate oligomer Aβ42 toxicity,the use of more simple eukaryotic cellular systems offers advantages for large-scale screening studies.We have previously established and validated budding yeast,Saccharomyces cerevisiae to be a simple and a robust model to study the toxicity of Aβ.Using colony counting based methods,oligomeric Aβ42 was shown to induce dose-dependent cell death in yeast.We have adapted this method for high throughput screening by developing an absorbance-based growth assay.We further validated the assay with treatments previously shown to protect oligomer Aβ42 induced cell death in mammalian and yeast cells.This assay offers a platform for studying underlying mechanisms of oligomer Aβ42 induced cell death using gene deletion/overexpression libraries and developing novel agents that alleviate Aβ42 induced cell death.

Helicobacter is preserved in yeast vacuoles! Does Koch's postulates confirm it?

The manuscript titled "Vacuoles of Candida yeast behave as a specialized niche for Helicobacter pylori(H.pylori)" not only has not been prepared in a scientific manner but the methodology used was not adequate, and therefore the conclusion reached was not correct. First of all, "yeast" is a broad terminology covering a great number of genera and species of unicellular micro-organisms. The authors should have defined the organism with its binary scientific name. This measure would allow experiment reproduction by the scientific community. Moreover, the criteria established by Robert Koch to identify a specific microorganism or pathogen was not adopted in the methodology used. Regarding the methodology applied, use of the chicken eggyolk(Ig Y) antibody and PCR of the apparently tainted yeast population to prove H. pylori existence in the yeast vacuoles might be main factors for their wrong conclusions. Bacterial tropism toward yeast extract is a known phenomenon, and yeast extract is one of the main ingredients in culture media. Their internalization through phagocytosis or similar pathways does not seem possible or practical because of the thick and cellulosic yeast wall. While the small size of yeast cells does not support their ability in harboring several H. pylori, other observations such as inefficiency of antifungal therapy as anti-Helicobacter therapy strongly reject the conclusion reached by the above-mentioned article.

Yeast One-hybrid System Used to Identify the Binding Proteins for Rat Glutathione S-transferase P Enhancer I

Objective To detect the trans-factors specifically binding to the strong enhancerelement (GPEI) in the upstream of rat glutathione S-transferase P (GST-P) gene. MethodsYeast one-hybrid system was used to screen rat lung MATCHMAKER cDNA library toidentify potential trans-factors that can interact with core sequence of GPEI(cGPEI).Electrophoresis mobility shift assay (EMSA) was used to analyze the binding of trans-factors to cGPEI. Results cDNA fragments coding for the C-terminal part of thetranscription factor c-Jun and rat adenine nucleotide translocator (ANT) were isolated. Thebinding of c-Jun and ANT to GPEI core sequence were confirmed. Conclusions Rat c-juntranscriptional factor and ANT may interact with cGPEI. They could play an important rolein the induced expression of GST-P gene.

Transcriptional activation function of hepatitis B virus Pre S1 protein in yeast

OBJECTIVE: To explore the feasibility of cloning of the hepatocyte receptor interacting with the Pre Slprotein of HBV by two-hybrid system.METHODS: Yeast expression plasmids encoding fusion proteins of full length or portions of Pre Sl ofHBV and DNA binding domain of yeast protein GAL4 were constructed and used to transform yeastreporter strain SFY526. Reporter gene product β-galactosidase activity was assayed as a measure oftranscriptional activation in yeast, Mammalian expression plasmid encoding fusion proteins of full lengthPre Sl and DNA binding domain of GAL4 was constructed and used to cotransfect hepatoma cell lineHuh-7 together with CAT reporter plasmid. Cell extracts were assayed for CAT activity by thin-layerchromatography.RESULTS: The fusion proteins of full length Pre Sl protein and GAL4 DNA binding domain presentedtranscriptional activation function in yeast. The transcription activating sequence was localized to the 21 to47 amino acids of Pre Sl protein. Fusion proteins of full length Pre Sl and GAL4 DNA binding domaindid not show transcriptional activation function in mammalian cells.CONCLUSIONS: The transcription activating sequence of HBV Pre Sl protein in yeast overlaps thehepatocyte receptor binding site. The transcriptional activation function of HBV Pre Sl protein in yeastmay prevent researchers from using yeast two-hybrid system to clone HBV receptor interacting with Pre Slprotein. However, the Pre Sl protein does not show transcriptional activation function in mammaliancells. Mammalian two-hybrid system may be a practical method to clone the HBV hepatocyte receptorinteracting with Pre Sl protein.

Mec1-Dependent Phosphorylation of the Scc3 Subunit of Cohesin during Mitosis in Budding Yeast

Cohesin is an evolutionary conserved complex that controls chromosome segregation during mitosis. Here we show that, in response to DNA damage, Saccharomyces cerevisiae Scc3, one of the major regulatory subunits of the Smc1-Smc3-Scc1 cohesin ring, is phosphorylated on S/T-Q residues. This event depended on the Mec1 checkpoint kinase as well as on cell cycle arrest triggered by the DNA damage checkpoint network. This phosphorylation event also took place during mitosis of an unperturbed cell cycle. The present finding that S. cerevisiae Scc3 is phosphorylated during mitosis represents a potentially important new regulatory step in cohesin’s mitotic functions.

Effect of Feeding Different Levels of Dietary Protein and Addition of Baker＇s Yeast （Saccharomyces cerevisiae） on Productive Parameters of Awassi Lambs

Forty eight individually fed Awassi male lambs were used in factorial experiment to investigate their responses to feeding concentrate diets containing three levels of dietary crude protein （CP）, each was offered without or with baker＇s yeast （SC） at rate of 0.5% （on dry matter （DM） basis）. Concentrates were offered at rate of 3% of live body weight with free choice of barley straw. Results revealed that higher （P 〈 0.05） digestible dry matter （DDM） and digestible organic matter （DOM） intakes were achieved due to feeding medium level of CP and to the addition of SC. Addition of SC improved （P 〈 0.05） gain, lambs fed medium and high levels gained higher （P 〈 0.05） than those fed the low level of dietary CP. Feed conversion ratio （FCR） based on DM and organic matter （OM） intakes was not significantly affected by level of dietary CP or addition of yeast. Even though, less amount of N required per unit of gain was achieved with low and medium as compared to high levels. Higher DM, nitrogen free extract （NFE） and hemicellulose （P 〈 0.05）, OM, CP, crude fiber （CF） and cellulose （P 〈 0.01） digestibilities were achieved in lambs fed the medium level of CP, whereas, no significant effect was observed on ether extract （EE）, neutral detergent fiber （NDF） and acid detergent fiber （ADF） digestibilities. Results also revealed that digestibility of almost all nutrients was improved with different extent due to addition of SC. Effect of interaction between levels of dietary CP and addition of SC referred to the preferability of addition of SC with medium level of dietary protein.

Isolation and Partial Evaluation of a Potential Indigenous Yeast Strain <i>Pichia kudriavzevii</i>from a Traditional Rice Beer—“Gora” Prepared by the Koloi Tribes of Tripura

As per tradition from hundreds of years the Koloi tribes of Tripura are preparing “Gora”-therice based fermented beer which is very good in taste and aroma applying their traditional indigenous brewing techniques. In this study, an attempt has been made to identify the indigenous yeast which is the main causative agent for fermentation and to investigate its fermentation ability with an industrial Saccharomyces cerevisiaes train. After investigation based on culture dependent phenotypic characteristics like-staining and biochemical characterization, primarily the responsible yeast species was determined as Pichia kudriavzevii and further confirmed followed by 18S rRNA ribotyping and the sequences was deposited at Gene bank and NCBI bearing specific accession number. In the comparative analysis, it has been found a significant similarities in all aspects of nutritional and alcohol percentages with the industrial strain in laboratory condition. The alcohol percentage in the rice beer “Gora” measured 6.40 ± 0.008% v/v. The study may be the first scientific investigation of its kind about this indigenous yeast strain isolated from “Gora” of this Indo-Burma biodiversity region and may provide sufficient background and potentiality for promoting these kinds of indigenous alcoholic beverages for small scale commercialization to strengthen the rural livelihood as well as to maintain immaterial cultural heritage.

Techniques Optimization of Combined Enzymatic Hydrolysis on Brewers＇ Spent Grain from Novozymes

Purpose： To extract protein, decrease the cellulose and facilitate the digestion and absorption of brewers＇ spent grain by animal. Topic： Discuss and optimize the hydrolysis conditions of the combined enzymatic hydrolysis by Novozymes. Method： The fresh brewers＇ spent grain was firstly dried, smashed and sifted. Then as indicators of the protein extraction rate in the enzyme solution and the content of cellulose in the index, the parameters of enzymatic hydrolysis, such as the solid-liquid ratio, reaction temperature, pH, enzyme dosage and reaction time, were investigated in detailed. After hydrolysis, the brewers＇ spent grain was put in the boiling water bath for inactivation for 15 minutes, and centrifuged, the supernatants were volume to 100 mL and the protein content was measured. After the precipitate was dried, the cellulose content was also measured. Achievements： The optimized conditions were with temperature of 50 ℃, pH 6.5, enzyme amount of 30 mg for Novozymes enzyme and 2.5 h for reaction time. Under these conditions, the protein extraction rate in the enzyme reaction reached 41.82%, and the cellulose content reached 13.90%, the degradation rate of cellulose was 18.86%.

Efficacy of Xiaojin Capsules Combined with Selenium Yeast Capsules on 120 Cases of Hashimoto's Thyroiditis with Abnormal Thyroid Function

[Objectives]To explore the efficacy of Xiaojin Capsules combined with Selenium Yeast Capsules in the treatment of Hashimoto's thyroiditis(HT)with abnormal thyroid function.[Methods]A total of 180 HT patients who were treated in the First Affiliated Hospital of Henan University and Henan Provincial People's Hospital from December 2020 to December 2021 were selected as the research subjects and randomly divided into the observation group(n=120)and the control group(n=60)in at a tratio of 2∶1 ratio according to their visiting time.The observation group was treated with Xiaojin Capsules(oral,5 capsules each time,twice a day for 12 weeks)and Selenium Yeast Capsules(oral,1 capsule each time,twice a day for 12 weeks).The control group was treated with only Selenium Yeast Capsules(oral,1 capsule each time,twice a day for 12 weeks).[Results]The difference in the levels of thyroid peroxidase antibody(TPO-Ab),thyroglobulin antibody(TGAb),serum free triiodothyronine(FT_(3)),serum free thyroxine(FT_(4)),and thyroid stimulating hormone(TSH)between the observation group and control group was statistically significant(P<0.05).The diameters of thyroid between the observation group and the control group before,during and after treatment were significantly different(P<0.05).The total effective rate of the observation group was 82.5%(99/120),which was significantly higher than 56.67%(34/60)in the control group,and the difference was statistically significant(P<0.05).The cure rate(22.50%)in the observation group was significantly higher than the control group(3.33%),and the difference was statistically significant(P<0.01).The ineffective rate of the observation group was 17.50%,which was significantly lower than that of the control group(40.00%),and the difference was statistically significant(P<0.01).The markedly effective rate of the observation group was 40.83%,which was significantly higher than that of the control group(31.67%),and the difference was statistically significant(P<0.05).The effective rate in the observation group was 191%,and the effective rate in the control group was 21.67%,and the difference was not statistically significant(P>0.05).[Conclusions]Xiaojin Capsules combined with Selenium Yeast Capsules in the treatment of HT can eliminate or alleviate the clinical symptoms and signs,significantly reduce the levels of TPO-Ab and TGAb in serum,restore thyroid function,improve thyroid shape and structure in the treatment of HT,and the clinical effect is satisfactory.

The Economic Potential of Brewer’s Spent Grain (BSG) as a Biomass Feedstock

This paper analyzes a pro-forma economic market and supply chain system for the reuse of a lignocellulose (brewer’s spent grain) in an industrial biotechnology environment. An extant literature review was conducted, followed by a technical analysis of BSG, and the development of a supply chain system and economic market analysis based upon a participant brewing company and industry experts. In this paper, it was found that, even with the potential for future improvements in the conversion of brewer’s spent grain (BSG) from an efficiency standpoint, this industrial residual is supply chain prohibitive as a biofeedstock in comparison to other lignocellulose materials, therefore, centralized market relationships would not be advantageous for sellers and buyers. Future research should consider the viability of centralized supply chain structures for alternatives that may exist as future bio-feedstocks.

Screening Yeasts from Ruminal Fluid of Dairy Heifer Fed a Different Ratio Roughage to Concentrate Diets

The objective of these studies were to identify ruminal yeast in varying ratios of roughage to concentrate in TMR diets in order to explore yeast diversity by using molecular technique with similarity of rDNA sequence. The experiment was assigned to four 98.6% of cross bred Holstein Friesian heifers with 2 levels and two replicates of roughage to concentrate ratios as： 10：90 （T1） and 50：50 （T2）. The experimental period was 14 days. Rumen fluid sample was collected by stomach tube for total DNA extraction by using silica gel method, and analysis of quantity and quality of DNA by Nanovue and agarose gel electrophoresis. The divergent DI/D2 domain of 26S rDNA was amplified by primers NL-1（5＇-GCA TAT CAA TAA GCG GAG GAA AAG-Y） and NL4 （5＇-GGT CCG TGT TTCAAG ACG G-3＇） by polymerase chain reaction （PCR）. The nucleotide sequences of D l/D2 domain of 26S rDNA were determined using PCR products. Generated sequences were aligned with related species by using the CLUSTAL W. The result showed that an average dry matter intake of TI was 7.00 kg/d and T2 was 6.99 kg/d. DNA concentrate from TIRI, TIR2, T2RI and T2R2 were 106, 131.5, 84 and 182.5 ng//aL, respectively. The purity of DNA was 1.57, 1.76, 1.78 and 1.86, respectively. The divergent D1/D2 domain of 26S rDNA of treatment could be amplified for T1R1 and T2R1 but could not for T1R2 and T2R2. The sequences of D1/D2 domain of 26S rDNA were compared with nucleotide database by BLAST programs （http：//www.ncbi.nlrn.nih.gov/BLAST）, the T2RI yeast-strain was closest to Yarrowia lipolytica. However, yeast strain in T1R1 could not be specifically identified because D 1/132 domain of 26S rDNA seem to represent variable region with number of nucleotide sequence showing 2-3 substitution from known species. The phylogenetic tree based on the sequences of the DI/D2 domain of 26S rDNA showed that TIR! was related to Pichia and Candida （96%） and T2R1 was related to Yarrowia lypolytica （100%）. This study indicated that ruminal yeast strains could be found varying in different ratio of roughage to concentrate.

富铬酵母中14种元素的ICP-AES测定

本文用ＩＣＰ－ＡＥＳ法同时测定富铬酵母中Ｋ、Ｆｅ、Ｐ、Ｚｎ、Ｃｒ、Ｃｕ、Ｍｎ、Ｃｏ、Ｂａ、Ｓｒ、Ｖ、Ｎａ、Ｍｇ、Ｃａ１４种元素的含量。结果表明，富铬酵母中的铬对其他元素的浓度有一定影响，并且，Ｋ、Ｍｎ、Ｐ、Ｖ、Ｍｇ、Ｃａ的浓度明显低于普通酵母。

荧光微孔板法检测啤酒酵母胞内脯氨酸含量

高浓酿造是啤酒生产中一种常用的高效经济技术,而在高浓酿造过程中啤酒酵母在各个时期存在不同的环境压力。脯氨酸被认为是啤酒酵母中的一种应激保护剂,赋予了酵母细胞更高的胁迫耐受性。酵母胞内脯氨酸含量可以作为筛选环境耐受能力强菌株的参考,因此有必要建立一种能够快速检测啤酒酵母胞内脯氨酸含量的方法。该研究基于次氯酸钠氧化和邻苯二甲醛/N-乙酰半胱胺酸荧光检测脯氨酸的通用方法,结合细胞破碎的优化,成功建立了一种可以准确测定啤酒酵母胞内脯氨酸含量的荧光微孔板法,其线性范围和回收率分别为5~40μmol/L(0.58~4.6 mg/L)和73.33%~105.5%。通过比较该检测方法与氨基酸专用高效液相色谱法,发现两者测定胞内脯氨酸含量绝对值相比无显著性差异,但是荧光微孔板法操作简便快捷。因此,该研究建立的荧光微孔板法为测定酵母胞内脯氨酸提供了一种新思路,同时为适应高浓酿造胁迫的啤酒酵母筛选提供了一种新策略。