An assessment of Salmonella survival in pig manure and its separated solid and liquid fractions during storage.

The objective of this study was to examine Salmonella survival in pig manure and its separated fractions during storage. Salmonella declined, but significant reductions were not observed in the manure and liquid until day 56, whereas counts in the solids were lower by day 7. The Salmonella inoculum initially impacted counts but not after days 28-56. By day 112 Salmonella was undetectable in the manure and liquid but was recovered from the solids. There was no clear dominance of particular serotypes and antibiotic resistance transfer was not found. Storage duration and pH impacted Salmonella counts in all samples, with duration having the greatest effect. Of the nutrients, nitrate had the greatest impact in the manure and, together with phosphate, it also affected counts in the liquid fraction. This study demonstrates that if pig manure or its separated fractions are stored under controlled conditions at 10.5°C for 84-112 days Salmonella is reduced or eliminated, irrespective of the initial load.

In vitro assessment of marine Bacillus for use as livestock probiotics.

Six antimicrobial-producing seaweed-derived Bacillus strains were evaluated in vitro as animal probiotics, in comparison to two Bacillus from an EU-authorized animal probiotic product. Antimicrobial activity was demonstrated on solid media against porcine Salmonella and E. coli. The marine isolates were most active against the latter, had better activity than the commercial probiotics and Bacillus pumilus WIT 588 also reduced E. coli counts in broth. All of the marine Bacillus tolerated physiological concentrations of bile, with some as tolerant as one of the probiotics. Spore counts for all isolates remained almost constant during incubation in simulated gastric and ileum juices. All of the marine Bacillus grew anaerobically and the spores of all except one isolate germinated under anaerobic conditions. All were sensitive to a panel of antibiotics and none harbored Bacillus enterotoxin genes but all, except B. pumilus WIT 588, showed some degree of ß-hemolysis. However, trypan blue dye exclusion and xCELLigence assays demonstrated a lack of toxicity in comparison to two pathogens; in fact, the commercial probiotics appeared more cytotoxic than the majority of the marine Bacillus. Overall, some of the marine-derived Bacillus, in particular B. pumilus WIT 588, demonstrate potential for use as livestock probiotics.

Assessment of the bacteriocinogenic potential of marine bacteria reveals lichenicidin production by seaweed-derived Bacillus spp.

The objectives of this study were (1) to assess the bacteriocinogenic potential of bacteria derived mainly from seaweed, but also sand and seawater, (2) to identify at least some of the bacteriocins produced, if any and (3) to determine if they are unique to the marine environment and/or novel. Fifteen Bacillus licheniformis or pumilus isolates with antimicrobial activity against at least one of the indicator bacteria used were recovered. Some, at least, of the antimicrobials produced were bacteriocins, as they were proteinaceous and the producers displayed immunity. Screening with PCR primers for known Bacillus bacteriocins revealed that three seaweed-derived Bacillus licheniformis harbored the bli04127 gene which encodes one of the peptides of the two-peptide lantibiotic lichenicidin. Production of both lichenicidin peptides was then confirmed by mass spectrometry. This is the first definitive proof of bacteriocin production by seaweed-derived bacteria. The authors acknowledge that the bacteriocin produced has previously been discovered and is not unique to the marine environment. However, the other marine isolates likely produce novel bacteriocins, as none harboured genes for known Bacillus bacteriocins.

A Systematic Review and Individual Patient Data Network Analysis of the Residual Symptom Structure Following Cognitive-Behavioral Therapy and Escitalopram, Mirtazapine and Venlafaxine for Depression.

OBJECTIVE: Consistent evidence suggests residual depressive symptomology are the strongest predictors of depression relapse following cognitive-behavioral therapy (CBT) and antidepressant medications (ADM's). Psychometric network models help detecting and understanding central symptoms that remain post-treatment, along with their complex co-occurrences. However, individual psychometric network studies show inconsistent findings. This systematic review and IPD network analysis aimed to estimate and compare the symptom network structures of residual depressive symptoms following CBT, ADM's, and their combination. METHODS: PsycINFO, PsycArticles, and PubMed were systematically searched through October 2020 for studies that have assessed individuals with major depression at post-treatment receiving either CBT and/or ADM's (venlafaxine, escitalopram, mirtazapine). IPD was requested from eligible samples to estimate and compare residual symptom psychometric network models post-CBT and post-ADM's. RESULTS: In total, 25 from 663 eligible samples, including 1,389 patients qualified for the IPD. Depressed mood and anhedonia were consistently central residual symptoms post-CBT and post-ADM's. For CBT, fatigue-related and anxiety symptoms were also central post-treatment. A significant difference in network structure across treatments (CBT vs. ADM) was observed for samples measuring depression severity using the MADRS. Specifically, stronger symptom occurrences were present amongst lassitude-suicide post-CBT (vs. ADM's) and amongst lassitude-inability to feel post-ADM's (vs. CBT). No significant difference in global strength was observed across treatments. CONCLUSIONS: Core major depression symptoms remain central across treatments, strategies to target these symptoms should be considered. Anxiety and fatigue related complaints also remain central post-CBT. Efforts must be made amongst researchers, institutions, and journals to permit sharing of IPD.Systematic Review Registration: A protocol was prospectively registered on PROSPERO (CRD42020141663; https://www.crd.york.ac.uk/PROSPERO/display_record.php?RecordID=141663).

Prebiotics from marine macroalgae for human and animal health applications.

The marine environment is an untapped source of bioactive compounds. Specifically, marine macroalgae (seaweeds) are rich in polysaccharides that could potentially be exploited as prebiotic functional ingredients for both human and animal health applications. Prebiotics are non-digestible, selectively fermented compounds that stimulate the growth and/or activity of beneficial gut microbiota which, in turn, confer health benefits on the host. This review will introduce the concept and potential applications of prebiotics, followed by an outline of the chemistry of seaweed polysaccharides. Their potential for use as prebiotics for both humans and animals will be highlighted by reviewing data from both in vitro and in vivo studies conducted to date.

Investigation of beta-carotene and lutein transport in Caco-2 cells: carotenoid-carotenoid interactions and transport inhibition by ezetimibe.

Carotenoid bioavailability is influenced by a number of factors including the presence of other carotenoids, which may enhance or inhibit the transport of one another by intestinal cells. Therefore, the objectives of the present study were: first, to determine carotenoid uptake and secretion (i. e. transport) by supplementing differentiated Caco-2 cells with increasing concentrations of either lutein or beta-carotene (0 - 1 microM); second, to assess any interactions between beta-carotene and lutein on their cellular uptake and secretion; and third, to a minor extent, to determine the effects of a carotenoid absorption inhibitor, ezetimibe, on beta-carotene and lutein transport. The carotenoid mixes were used at molar ratios of 1:1 and 4:1. At equimolar concentrations, lutein had a negative impact on beta-carotene transport and vice versa. However, these effects were not seen when the ratios were adjusted to 4:1. Following treatment with ezetimibe (25 - 100 microM) for 16 hours there was a reduction in beta-carotene transport, whereas a non-significant reduction in lutein transport was observed. In conclusion, this study confirmed that beta-carotene and lutein interact during their absorption, depending on the concentration/ratios used, and that carotenoid absorption is partially affected by ezetimibe.

Carotenoid micellarization varies greatly between individual and mixed vegetables with or without the addition of fat or fiber.

Carotenoid bioavailability is influenced by a number of factors, including the type of food matrix and the presence of fat, fiber, and other carotenoids. Therefore, the objectives of the present study were: first, to assess the effects of mixing raw vegetables on the micellarization of beta-carotene, lycopene, beta-cryptoxanthin, and lutein compared with individual vegetables; second, to investigate the effects of adding different oils on carotenoid transfer to the micelles; and third, and to a minor extent, to determine carotenoid micellarization following the addition of fiber. The two mixed vegetable meals were TRS (tomato, red pepper, and spinach) and CRS (courgette/zucchini, red pepper, and spinach). Similar trends in carotenoid micellarization were seen between individual vegetables and the TRS meal but not with the CRS meal. In general, the addition of olive, peanut, or rapeseed oil to the CRS meal significantly enhanced carotenoid micellarization but this effect was not concentration-dependent. In relation to the TRS meal, adding either vegetable oils or fiber (oat bran, wheat bran, and pectin) significantly decreased the micellarization of carotenoids to varying degrees. The results from this study indicate that changes to a combination of raw vegetables, with or without the addition of dietary fat or fiber, can have varying results on carotenoid bioavailability.

Evaluation of the efficacy and safety of a marine-derived Bacillus strain for use as an in-feed probiotic for newly weaned pigs.

Forty eight individual pigs (8.7±0.26 kg) weaned at 28±1 d of age were used in a 22-d study to evaluate the effect of oral administration of a Bacillus pumilus spore suspension on growth performance and health indicators. Treatments (n = 16) were: (1) non-medicated diet; (2) medicated diet with apramycin (200 mg/kg) and pharmacological levels of zinc oxide (2,500 mg zinc/kg) and (3) B. pumilus diet (non-medicated diet + 10(10) spores/day B. pumilus). Final body weight and average daily gain tended to be lower (P = 0.07) and feed conversion ratio was worsened (P<0.05) for the medicated treatment compared to the B. pumilus treatment. Ileal E. coli counts were lower for the B. pumilus and medicated treatments compared to the non-medicated treatment (P<0.05), perhaps as a result of increased ileal propionic acid concentrations (P<0.001). However, the medicated treatment reduced fecal (P<0.001) and cecal (P<0.05) Lactobacillus counts and tended to reduce the total cecal short chain fatty acid (SCFA) concentration (P = 0.10). Liver weights were lighter and concentrations of liver enzymes higher (P<0.05) in pigs on the medicated treatment compared to those on the non-medicated or B. pumilus treatments. Pigs on the B. pumilus treatment had lower overall lymphocyte and higher granulocyte percentages (P<0.001) and higher numbers of jejunal goblet cells (P<0.01) than pigs on either of the other two treatments or the non-medicated treatment, respectively. However, histopathological examination of the small intestine, kidneys and liver revealed no abnormalities. Overall, the B. pumilus treatment decreased ileal E. coli counts in a manner similar to the medicated treatment but without the adverse effects on growth performance, Lactobacillus counts, cecal SCFA concentration and possible liver toxicity experienced with the medicated treatment.

Development of a novel antimicrobial seaweed extract-based hydrogel wound dressing.

The objective of this study was to develop a novel antimicrobial seaweed wound dressing. The seaweed extract was active against nine clinically-relevant wound pathogens. A hydrogel formulation was prepared using polyvinyl alcohol (PVA) and polyvinylpyrrolidone (PVP), followed by addition of 1% seaweed extract. The antimicrobial properties of the novel dressing were tested using agar diffusion assays, with release-profiles examined using gel leaching and gel transfer assays. The dressing was found to be effective against the same microbial strains as the seaweed extract, with similar efficacy to the commonly used silver-based dressing, Acticoat(®). Antimicrobial release-profile assays revealed that the dressing was effective in inhibiting 70-90% of the bacterial population within the first 30 min, followed by a long, sustained released up to 97 h, without leaving a residue following five subsequent transfers of the dressing. Antimicrobial activity was stable for up to 6 months of storage at 4 °C, but activity was reduced slightly after 15 weeks. Following autoclave sterilization, the dressing displayed a slower release profile compared to a non-autoclaved counterpart. Hence, the seaweed dressing may have commercial applications, potentially competing with silver-based dressings at a lower cost per-application. This is the first report of development of a seaweed-based antimicrobial dressing.

Bioaccessibility, uptake, and transport of carotenoids from peppers (Capsicum spp.) using the coupled in vitro digestion and human intestinal Caco-2 cell model.

Spanish bell peppers (Capsicum annuum L.) and chili peppers sourced from Kenya and Turkey were analyzed for their carotenoid content, bioaccessibility, and bioavailability. The order of total carotenoid content in peppers and their respective micelles was red > green > yellow. In terms of cellular carotenoid transport as a percentage of original food and micelle content, the order was yellow peppers > green > red; however, the opposite trend was seen for the actual amount of total carotenoids transported by Caco-2 cells. Although lutein was generally the most abundant carotenoid in the micelles (496.3-1565.7 microg 100 g(-1)), cellular uptake and transport of beta-carotene were the highest, 8.3-31.6 and 16.8-42.7%, respectively. Hence, the actual amount of carotenoids present in the original food and respective micelles seems to reflect the amount transported by Caco-2 cells. Therefore, color influenced the carotenoid profile, bioaccessibility, and bioavailability of carotenoids rather than pepper type.

Cellular transport of lutein is greater from uncooked rather than cooked spinach irrespective of whether it is fresh, frozen, or canned.

Lutein, a carotenoid found in significant levels in spinach, has attracted a great deal of attention owing to its reported function as a shield against the photooxidative effects of blue light. Therefore, the rationale of this study was to examine the effects of various processing and cooking methods on lutein bioavailability from spinach (Spinacia oleracea) using an in vitro digestion procedure coupled with the use of a human intestinal Caco-2 cell model. Fresh, frozen, and canned spinach were analyzed uncooked and after boiling or microwave cooking. Lutein content from the uncooked and cooked digested food (digestate) and appropriate micelles was determined. Micellarized lutein from the spinach samples was adjusted to 0.1 micromol/L and added to Caco-2 cells. Cellular uptake and secretion (cellular transport) of lutein were determined. Our results showed that digestate obtained from uncooked canned spinach had greater lutein content (P < .05) than uncooked fresh or frozen spinach. Microwave cooking, but not boiling, significantly lowered the lutein content of canned spinach digestate and micelles compared with their uncooked counterparts. Interestingly, there were no differences in the micellarization of lutein between the cooking and processing methods. Cellular transport of lutein was greater from uncooked spinach micelles compared with boiled or microwave-cooked spinach. To conclude, although the lutein content of digesta and micelles may have been modified, its micellarization was not significantly affected by any of the cooking or processing methods tested. In general, cellular transport of lutein was greatest in uncooked spinach irrespective of whether the spinach was fresh, frozen, or canned.

Comparison of the uptake and secretion of carotene and xanthophyll carotenoids by Caco-2 intestinal cells.

Carotenoids have been shown to have potential beneficial effects on human health which has led to an increasing interest in the study of their bioavailability. A Caco-2 cell model, as previously described, was employed to examine the percentage transfer of the carotenoids alpha-carotene, beta-carotene, lycopene, astaxanthin, beta-cryptoxanthin, lutein and zeaxanthin through an intact, highly differentiated Caco-2 cell monolayer at a range of different amounts. Our results show that astaxanthin, a carotenoid with powerful antioxidant capacity, had the highest percentage transfer overall. We examined the cellular uptake and secretion of lutein and zeaxanthin to compare two structurally similar carotenoids. Both were efficiently transported through the monolayer with a range between 5.1 (sem 1.2) % to 20.2 (sem 3.3) % and 5.5 (sem 2.5) % to 13.4 (sem 4) % for lutein and zeaxanthin, respectively. These carotenoids were compared to each other at each added amount and no significant difference was observed between the two xanthophylls. The carotene carotenoids alpha-carotene, beta-carotene and lycopene and the xanthophyll beta-cryptoxanthin were also examined and had lower uptake and secretion values when compared to lutein, zeaxanthin and astaxanthin. The xanthophyll beta-cryptoxanthin was also not significantly different when compared to the carotene carotenoids. Data generated from this study compares well with in vivo bioavailability studies. Furthermore, the model provides comparative data on the relative absorption and transfer of seven different carotenoids. Our data indicate that lower amounts of carotenoids were absorbed and transferred more efficiently than higher amounts suggesting a saturation effect at higher exposure.