A natural carbohydrate fraction Actigen™ from Saccharomyces cerevisiae cell wall: effects on goblet cells, gut morphology and performance of broiler chickens

A study was conducted to evaluate a natural carbohydrate fraction Actigen™ (NCF), derived from mannanoligosaccharide, in feed on growth performance, intestinal morphology and goblet cell number and area of male broilers'. Dietary treatments included: 1) control diet (antibiotic and NCF free), 2) NCF at 200g/t, 3) NCF at 400g/t, and 4) NCF 800g/t. Two hundred and forty birds were placed into 12 replicate pens per treatment (5 birds/pen), sixty birds per treatment. Body weight and feed intake were recorded weekly up to day 42. At this time a 2.5cm section of jejunum and duodenum were excised post mortem for morphological analysis. Birds fed 200g/t and 800g/t NCF were significantly (P<0.01) heavier from day 14 onwards than the control birds. Feed intake was significantly higher in birds fed 200g/t NCF compared to those fed the control at 21 and 35 days (P<0.05). Diets containing 200g/t and 800g/t of NCF significantly decreased broiler feed conversion ratio (FCR) compared to the control in the first phase (1-14 days) (P<0.01) and levels of NCF decreased FCR (P<0.05) in the second phase (15-28 days). NCF had no significant effect on villus height, villus width, crypt depth or villus to crypt ratio in either duodenum or jejunum. NCF did not significantly affect goblet cell area or goblet cell number in the duodenum, however, in the jejunum, 800g/t NCF significantly (P<0.05) increased goblet cell area over the control. In conclusion, NCF showed a positive effect on broiler performance in the starter and grower phases, and increased goblet cell area in the jejunum, suggesting higher levels of mucin production. This indicated that the performance benefit of NCF could be age-dependent, with younger birds responding more than the older ones. There were no additional benefits to performance when feeding NCF for a longer period (after 28 d of age), however it is postulated that birds fed NCF would have greater defence to pathogenic challenge through increased storage capacity of muci

Influence of rate of inclusion of microalgae on the sensory characteristics and fatty acid composition of cheese and performance of dairy cows

Modification of milk and cheese fat to contain long-chain n-3 fatty acids (FA) by feeding microalgae (ALG) to dairy cows has the potential to improve human health, but the subsequent effect on the sensory attributes of dairy products is unclear. The objective was to determine the effect of feeding dairy cows different amounts of ALG that was rich in docosahexaenoic acid (DHA) on milk and cheese FA profile, cheese sensory attributes, and cow performance. Twenty Holstein dairy cows were randomly allocated to 1 of 4 dietary treatments in a 4 × 4 row and column design, with 4 periods of 28 d, with cheddar cheese production and animal performance measurements undertaken during the final 7 d of each period. Cows were fed a basal diet that was supplemented with ALG (Schizochytrium limancinum) at 4 rates: 0 (control, C), 50 (LA), 100 (MA), or 150 g (HA) of ALG per cow per day. We found that both milk and cheese fat content of DHA increased linearly with ALG feed rate and was 0.29 g/100 g FA higher in milk and cheese from cows fed HA compared with C. Supplementation with ALG linearly reduced the content of saturated FA and the ratio of n-6:n-3 FA in milk and cheese. Supplementation with ALG altered 20 out of the 32 sensory attributes, with a linear increase in cheese air holes, nutty flavor, and dry mouth aftertaste with ALG inclusion. Creaminess of cheese decreased with ALG inclusion rate and was positively correlated with saturated FA content. We also observed a quadratic effect on fruity odor, which was highest in cheese from cows fed HA and lowest in LA, and firmness and crumbliness texture, being highest in MA and lowest in HA. Supplementation with ALG had no effect on the dry matter intake, milk yield, or live weight change of the cows, with mean values of 23.1, 38.5, and 0.34 kg/d respectively, but milk fat content decreased linearly, and energy-corrected milk yield tended to decrease linearly with rate of ALG inclusion (mean values of 39.6, 38.4, 37.1, and 35.9 g/kg, and 41.3, 41.3, 40.5, and 39.4 kg/d for C, LA, MA, and HA, respectively). We conclude that feeding ALG to high-yielding dairy cows improved milk and cheese content of DHA and altered cheese taste but not cow performance, although milk fat content reduced as inclusion rate increased

Composites Curriculum Development: tackling the skills gap in UK advanced composites

The aim of this project is to generate an industrially relevant and academically rigorous curriculum which could be deployed to tackle the significant skills gap in composites professionals, vital for delivering on the UK’s National Composite Strategy and allowing the industry to grow to its full potential, forecast by the Composites Leadership Forum to grow by a factor of 5 by 2030. A Masters’ level curriculum of short, industrially focused units has been specified and a small number of trial units developed. Engagement of academics in this novel collaborative curriculum development, utilising each institution’s expertise, has been very good and feedback from industry and participants in pilot units has been positive. Consortium participants are investigating numerous options for developing this further and have begun to put plans in place for the next stage

Characterisation of Bacteriophage-Encoded Depolymerases Selective for Key <i>Klebsiella pneumoniae</i> Capsular Exopolysaccharides.

Capsular polysaccharides enable clinically important clones of Klebsiella pneumoniae to cause severe systemic infections in susceptible hosts. Phage-encoded capsule depolymerases have the potential to provide an alternative treatment paradigm in patients when multiple drug resistance has eroded the efficacy of conventional antibiotic chemotherapy. An investigation of 164 K. pneumoniae from intensive care patients in Thailand revealed a large number of distinct K types in low abundance but four (K2, K51, K1, K10) with a frequency of at least 5%. To identify depolymerases with the capacity to degrade capsules associated with these common K-types, 62 lytic phage were isolated from Thai hospital sewage water using K1, K2 and K51 isolates as hosts; phage plaques, without exception, displayed halos indicative of the presence of capsule-degrading enzymes. Phage genomes ranged in size from 41-348 kb with between 50 and 535 predicted coding sequences (CDSs). Using a custom phage protein database we were successful in applying annotation to 30 - 70% (mean = 58%) of these CDSs. The largest genomes, of so-called jumbo phage, carried multiple tRNAs as well as CRISPR repeat and spacer sequences. One of the smaller phage genomes was found to contain a putative Cas type 1E gene, indicating a history of host DNA acquisition in these obligate lytic phage. Whole-genome sequencing (WGS) indicated that some phage displayed an extended host range due to the presence of multiple depolymerase genes; in total, 42 candidate depolymerase genes were identified with up to eight in a single genome. Seven distinct virions were selected for further investigation on the basis of host range, phage morphology and WGS. Candidate genes for K1, K2 and K51 depolymerases were expressed and purified as his6-tagged soluble protein and enzymatic activity demonstrated against K. pneumoniae capsular polysaccharides by gel electrophoresis and Anton-Paar rolling ball viscometry. Depolymerases completely removed the capsule in K-type-specific fashion from K. pneumoniae cells. We conclude that broad-host range phage carry multiple enzymes, each with the capacity to degrade a single K-type, and any future use of these enzymes as therapeutic agents will require enzyme cocktails for utility against a range of K. pneumoniae infections

Compaction, aeration and addition of mycotoxin contaminated silage alters the fermentation profile, mycotoxin content and aerobic stability of ryegrass (Lolium perenne) silage

This study investigated the effect of compaction, aeration and addition of a naturally mycotoxin contaminated ryegrass silage (MCS) containing 1803 µg/kg DM penicillic acid, on the nutritional value and mycotoxin content after ensiling and subsequent aerobic stability of ryegrass Lolium perenne silage (second-cut, June 2020, UK). Mini silos (30 L) were filled with differential compaction (500 kg FW/m3 and 333 kg FW/m3), aeration by injection of air (1L per 48h for 30d then 1L per 7d) and addition of MCS (1.5 g/kg FW ensiled forage) in a 2×2×2 factorial design. During ensilage, mean CO2% (kg FW) in the aerated silos increased with low compaction. Crude protein (CP) increased and ash decreased with aeration. Mean silage fermentation end products acetic (AA), lactic (LA) and propionic acid (PA) concentrations increased with MCS. PA concentration increased with aeration/low compaction. LA decreased and ethanol increased with low compaction. Mycotoxin profiles differed between the silages on opening and after 14-days incubation in aerobic conditions with disappearance of fusarenon X and penicillic acid and appearance of mycophenolic acid and roquefortine C (318 µg/kg DM and 890 µg/kg DM). Addition of MCS, increased the concentration of penicillic acid on opening with an interaction with aeration (80.6µg/kg DM MCS × aerated, 40.0 µg/kg DM in the MCS × sealed). Aerobic stability was affected by aeration and low compaction with reduced time taken to heat to +5°C and +10°C above ambient temperature, higher rate of increase in pH and higher cumulative temperatures to the first peak temperature. Higher mycotoxin concentration at opening had a similar effect increasing time to heat +5°C and +10°C above ambient temperatures in aerobic conditions. Regression analysis showed predominantly direct relationships between silage fermentation end-product concentrations and aerobic stability. This study revealed interactions between silage bacteria and fungi activity from the concentrations of fermentation end-products and mycotoxins during ensilage and subsequent aerobic spoilage. The results supported current best practice for silage making, promoting conditions for improved preservation and aerobic stability. The addition of MCS had unexpected positive effects. However, factors associated with the MCS benefiting aerobic stability were not determined

Characterisation of Bacteriophage-Encoded Depolymerases Selective for Key Klebsiella pneumoniae Capsular Exopolysaccharides.

Capsular polysaccharides enable clinically important clones of Klebsiella pneumoniae to cause severe systemic infections in susceptible hosts. Phage-encoded capsule depolymerases have the potential to provide an alternative treatment paradigm in patients when multiple drug resistance has eroded the efficacy of conventional antibiotic chemotherapy. An investigation of 164 K. pneumoniae from intensive care patients in Thailand revealed a large number of distinct K types in low abundance but four (K2, K51, K1, K10) with a frequency of at least 5%. To identify depolymerases with the capacity to degrade capsules associated with these common K-types, 62 lytic phage were isolated from Thai hospital sewage water using K1, K2 and K51 isolates as hosts; phage plaques, without exception, displayed halos indicative of the presence of capsule-degrading enzymes. Phage genomes ranged in size from 41-348 kb with between 50 and 535 predicted coding sequences (CDSs). Using a custom phage protein database we were successful in applying annotation to 30 - 70% (mean = 58%) of these CDSs. The largest genomes, of so-called jumbo phage, carried multiple tRNAs as well as CRISPR repeat and spacer sequences. One of the smaller phage genomes was found to contain a putative Cas type 1E gene, indicating a history of host DNA acquisition in these obligate lytic phage. Whole-genome sequencing (WGS) indicated that some phage displayed an extended host range due to the presence of multiple depolymerase genes; in total, 42 candidate depolymerase genes were identified with up to eight in a single genome. Seven distinct virions were selected for further investigation on the basis of host range, phage morphology and WGS. Candidate genes for K1, K2 and K51 depolymerases were expressed and purified as his6-tagged soluble protein and enzymatic activity demonstrated against K. pneumoniae capsular polysaccharides by gel electrophoresis and Anton-Paar rolling ball viscometry. Depolymerases completely removed the capsule in K-type-specific fashion from K. pneumoniae cells. We conclude that broad-host range phage carry multiple enzymes, each with the capacity to degrade a single K-type, and any future use of these enzymes as therapeutic agents will require enzyme cocktails for utility against a range of K. pneumoniae infections

The Effect of a Mycotoxin Deactivation Product on Growth of Juvenile Rainbow Trout Fed Distillers Dried Grains

Distillers dried grains (DDG) with solubles (DDGS) is a product that has shown potential as a protein source for some fish species, but high inclusion rates of DDGS have not always been successfully achieved for Rainbow Trout Oncorhynchus mykiss. Our objective was to determine whether inclusion of a mycotoxin deactivation product (Biofix Plus) could improve the ability of high-protein DDG (HPDDG) to replace a portion of the fish meal in diets for Rainbow Trout. The 2 × 2 factorial feeding trial examined protein source (menhaden fish meal [MFM] or HPDDG) with or without Biofix Plus. A control diet (42% digestible protein, 20% crude lipid, 25% MFM) was compared to a test diet in which HPDDG replaced 12% of the total MFM on a digestible-protein basis (24% HPDDG inclusion). Diets were fed to juvenile Rainbow Trout (initial weight: mean ± SE = 30.5 ± 1.6 g) in four replicate tanks per treatment for 9 weeks in a 15°C recirculating system. At the conclusion of the feeding trial, we observed no negative effects of fish meal replacement on growth or feed conversion ratio; no benefit of Biofix Plus supplementation was observed. These data indicate that when Rainbow Trout diets containing a high-quality DDGS product are balanced for digestible protein, lysine, methionine, and threonine, dietary fish meal levels can be successfully reduced to 13% without compromising growth and without the need for mycotoxin deactivator inclusion

Action potentials in abscisic acid-deficient tomato mutant generated spontaneously and evoked by electrical stimulation

Action potentials generated spontaneously (SAPs) and evoked by electrical stimulation (APs) in tomato plants (Solanum lycopersicum L.) cv. Micro-Tom ABA-deficient mutants (sitiens—MTsit) and its wild type (MTwt) were characterized by continuous monitoring of electrical activity for 66 h and by application of an electrical stimulation supplied extracellularly. MTsit generated SAPs which spread along the stem, including petioles and roots with an amplitude of 44.6 ± 4.4 mV, half-time (t½) of 33.1 ± 2.9 s and velocity of 5.4 ± 1.0 cm min−1. Amplitude and velocity were 43 and 108 % higher in MTsit than in MTwt, respectively. The largest number of SAPs was registered in the early morning in both genotypes. MTsit was less responsive to electrical stimuli. The excitation threshold and the refractory period were greater in MTsit than in MTwt. After current application, APs were generated in the MTwt with 21.2 ± 2.4 mV amplitude and propagated with 5.6 ± 0.5 cm min−1 velocity. Lower intensity stimuli did not trigger APs in these plants. In MTsit APs were measured with amplitude of 26.8 ± 4.8 mV and propagated with velocity of 8.5 ± 0.1 cm min−1

Herpesviruses carrying a Brainbow cassette reveal replication and expression of limited numbers of incoming genomes

Whether all the infectious herpesvirus particles entering a cell are able to replicate and/or express their genomes is not known. Here, we developed a general method to determine the number of viral genomes expressed in an infected cell. We constructed and analysed fluorophore expression from a recombinant pseudorabies virus (PRV263) carrying a Brainbow cassette (Cre-conditional expression of different fluorophores). Using three isogenic strains derived from PRV263, each expressing a single fluorophore, we analysed the colour composition of cells infected with these three viruses at different multiplicities. We estimate that fewer than seven incoming genomes are expressed per cell. In addition, those templates that are expressed are the genomes selected for replication and packaging into virions. This finite limit on the number of viral genomes that can be expressed is an intrinsic property of the infected cell and may be influenced by viral and cellular factors