Jujube Powder Enhances Cyclophosphamide Efficiency against Murine Colon Cancer by Enriching CD8+ T Cells While Inhibiting Eosinophilia

Cyclophosphamide (CTX) is widely applied in cancer treatment. However, the outcome is often compromised by lymphopenia, myelosuppression, and gut dysbiosis. Here, we used jujube powder to enhance CTX efficiency through nurturing gut microbiota in order to facilitate favorable metabolisms. It was observed that the oral administration of jujube powder enriched CD8+ T cells in mouse MC38 colon tumor microenvironment and increased the diversity of gut microbiota and the abundance of Bifidobacteriales, which is helpful to the production of butyrate in the cecum content. The application of jujube powder also stimulated the production of white blood cells, especially CD8+ T cells in peripheral and bone marrow, while inhibiting the growth of eosinophils in peripheral blood and the production of IL-7 and GM-CSF in serum. All these are conductive to the significant inhibition of the tumor growth, suggesting the high potential of nurturing gut microbiota with natural products for improving the efficiency of chemotherapy.

Common Bean (Phaseolus vulgaris L.) Flour Can Improve the Gut Microbiota Composition and Function in Mice Fed a High-Fat Diet

Objectives To evaluate the effects of whole flour and protein hydrolysate from common bean on gut health in mice fed a High-Fat Diet. Methods BALB/c adults mice (n = 48) where divided in 4 groups (n = 12 each): normal control (NC standard diet AIN-93M); High-Fat Diet (HFD) only; HFD plus bean flour (HFBF) and HFD plus bean protein hydrolysate (HFPH; 700 mg/kg of body/day). After 9 weeks, the animals were euthanized. Cecum weight was measured and cecum content was collected. Cecum content was used to analyze moisture by gravimetric method, lipids by extraction in Soxhlet apparatus, short chain fatty acids (SCFA) by HPLC and DNA extraction and sequencing of the gut microbiota. Total genomic DNA was extracted from fecal samples and loaded using the Illumina MiSeq platform at Argonne National Laboratory. Data were analyzed by ANOVA and post-hoc of Newman-Kews. Nonparametric and independent samples were submitted to Kruskal-Wallis with a Dunn's multiple comparison test (P < 0.05). Results HFBF increased cecum weight (+69%), moisture (+104.6%) and lipids (+11.5%) in the feces compared to HFD group (P < 0.05), and the Beta diversity was different from HFD. Acetic acid concentration decreased (−37.7%) in cecal content of HFBF group compared to HFD group (P < 0.05), and propionic and butyric acids cecal concentration did not differ (P > 0.05) among experimental groups. The abundance of Bacteroidetes increased and the Firmicutes/Bacteroidetes ratio decreased in the HFBF compared to control groups. The operational taxonomic units (OTUs) enriched by HFBF were mainly assigned to Muribaculaceae family, which show high potential to improve gut health. The functional analysis of the microbiota shown beneficial changes in the host's genetic capacity, especially in the metabolic pathways involved with glucose metabolism. KEGG metabolic pathways involved with starch and sucrose metabolism, as well as the galactose metabolism were enriched in the HFBF group compared to the HFD group (P < 0.05). Conclusions The intake of common bean flour modulates the microbiota composition and abundance of SCFA-producing bacteria, and attenuates the effects of HFD, showing potential to improve gut microbiota composition and function of mice. Funding Sources CNPq, CAPES, Fapemig and Fulbright (Brazil).

Mediterraneibacter catenae SW178 sp. nov., an intestinal bacterium of feral chicken

A Gram-positive, coccobacillus, white raised and circular with an entire edge colony, and obligately anaerobic bacterium, strain SW178 was isolated from the cecum content of feral chickens in Brookings, South Dakota, USA. The most closely related strain based on 16S rRNA gene sequence analysis of strain SW178 was Mediterraneibacter torques ATCC 27756T (Ruminococcus torques ATCC 27756T) with 96.94% similarity. The genome of strain SW178 is 3.18 Mbp with G+C content of 46.9 mol%. The optimal temperature and pH for growth in modified brain heart infusion (BHI-M) medium were 45 °C and pH 7.5, respectively. The sole carbon sources of the strain were dextrin, L-fucose, D-galacturonic, α-D-glucose, L-rhamnose and D-sorbitol. The primary cellular fatty acids were C14 : 0, C16 : 0 and C16 : 0 dimethyl acetal (DMA). Based on the genotypic and phenotypic comparison, we proposed that strain SW178 belong to the genus Mediterraneibacter in the family Lachnospiraceae as a novel species, in which the name Mediterraneibacter catenae is proposed. The type strain is SW178 (= DSM 109242T = CCOS 1886T).

Reduction of Salmonella Typhimurium Cecal Colonisation and Improvement of Intestinal Health in Broilers Supplemented with Fermented Defatted ‘Alperujo’, an Olive Oil By-Product

Salmonella spp. contaminates egg and poultry meat leading to foodborne infections in humans. The emergence of antimicrobial-resistant strains has limited the use of antimicrobials. We aimed to determine the effects of the food supplement, fermented defatted ‘alperujo’ (FDA), a modified olive oil by-product, on Salmonella Typhimurium colonisation in broilers. One hundred and twenty 1-day-old broilers were divided into four experimental groups—two control groups and two treated groups, and challenged with S. Typhimurium at day 7 or 21. On days 7, 14, 21, 28, 35, and 42 of life, duodenum and cecum tissue samples were collected for histopathological and histomorphometric studies. Additionally, cecum content was collected for Salmonella spp. detection by culture and qPCR, and for metagenomic analysis. Our results showed a significant reduction of Salmonella spp. in the cecum of 42-day-old broilers, suggesting that fermented defatted ‘alperujo’ limits Salmonella Typhimurium colonization in that cecum and may contribute to diminishing the risk of carcass contamination at the time of slaughter. The improvement of the mucosal integrity, observed histologically and morphometrically, may contribute to enhancing intestinal health and to limiting Salmonella spp. colonisation in the host, mitigating production losses. These results could provide evidence that FDA would contribute to prophylactic and therapeutic measures to reduce salmonellosis prevalence in poultry farms.

Reduction of Salmonella Typhimurium Cecal Colonisation and Improvement of Intestinal Health in Broilers Supplemented with Fermented Defatted ‘Alperujo’, an Olive Oil By-Product

Salmonella spp. contaminates egg and poultry meat leading to foodborne infections in humans. The emergence of antimicrobial-resistant strains limit the use of antimicrobials. We aimed to determine the effects of the food supplement, fermented defatted ‘alperujo’, a modified olive-oil by-product, on Salmonella Typhimurium colonisation in broilers. One hundred and twenty 1-day-old broilers were divided into four experimental groups: 2 control groups and 2 treated groups, and challenged with S. Typhimurium at day 7 or 21. On days 7, 14, 21, 28, 35 and 42 of life, duodenum and caecum tissue samples were collected for histopathological and histomorphometric studies. Additionally, cecum content was collected for Salmonella spp. detection by culture and qPCR, and for metagenomic analysis. Our results showed a significant reduction of Salmonella spp. in the cecum of 42-day-old broilers, suggesting that fermented defatted ‘alperujo’ stimulates Salmonella Typhimurium clearance in that cecum and may contribute to diminishing the risk of carcass contamination at the time of slaughter. Additionally, the improvement of the mucosal integrity suggests that enhancing intestinal health helps to limit Salmonella spp. colonisation in the host, mitigating production losses. These results could provide evidence that FDA would contribute to prophylactic and therapeutic measures to reduce salmonellosis prevalence in poultry farms.

The use of an alternative feed additive, containing benzoic acid, thymol, eugenol, and piperine, improved growth performance, nutrient and energy digestibility, and gut health in weaned piglets

This research evaluated a feed additive (benzoic acid, eugenol, thymol, and piperine), associated or not with colistin, in weaned piglets feeding. The parameters evaluated were growth performance, apparent total tract digestibility (ATTD) of nutrients, diarrhea incidence, intestinal morphology, relative weights of digestive organs, microbial diversity, and the percentages of operational taxonomic units of microorganisms in the cecum content of pigs. One-hundred and eight crossbred piglets (5.3 ± 0.5 kg) were used in a three-phase feeding program (21 to 35, 36 to 50, 51 to 65 d of age) and fed a control diet with no inclusion of growth promoter feed additive, a diet with 40 ppm of colistin, a diet with 0.3% of alternative additive, and a diet with 0.3% of alternative additive and 40 ppm of colistin. The diets were based on corn, soybean meal, dairy products, and spray-dried blood plasma and formulated to provide 3.40, 3.38, and 3.20 Mcal of ME/kg and 14.5, 13.3, and 10.9 g/kg of digestible lysine, in phases 1, 2, and 3, respectively. The piglets were housed three per pen, with nine replicates per diet, in a complete randomized block design based on initial BW. The data were submitted to ANOVA and means were separated by Tukey test (5%), using SAS. Pigs fed diets with the alternative feed additive had greater (P < 0.05) ADG (114.3 vs. 91.8 g) and ADFI (190.1 vs. 163.3 g) in phase 1 than pigs fed diets without the product. The alternative additive improved (P < 0.05) ATTD of crude protein (CP) in phase 1 (71.0% vs. 68.6%), gross energy in phases 1 (77.4% vs. 75.2%) and 3 (79.0% vs. 77.1%), and dry matter in phase 3 (79.1% vs. 77.1%). The antibiotic inclusion in the diets increased (P < 0.05) ATTD of CP in phase 1 (71.5% vs. 68.2%). The alternative feed additive tended (P = 0.06) to increase (46%) normal feces frequency, decreased (P < 0.05) goblet cells count (104.3 vs. 118.1) in the jejunum, and decreased (P < 0.05) small intestine (4.60% vs. 4.93%) and colon (1.41% vs. 1.65%) relative weights, compared with pigs not fed with the alternative additive. There was a tendency (P = 0.09) for a lower concentration of Escherichia–Shigella (1.46% vs. 3.5%) and lower (P < 0.05) percentage of Campylobacter (0.52% vs. 10.21%) in the cecum content of piglets fed diets containing essential oils and benzoic acid compared with pigs fed diets without the alternative feed additive. The alternative feed additive was effective in improving growth performance, diets digestibility, and gut health in piglets soon after weaning.

Bile Acid Profile and its Changes in Response to Cefoperazone Treatment in MR1 Deficient Mice

Mucosal associated invariant T-cells (MAIT cells) are activated following recognition of bacterial antigens (riboflavin intermediates) presented on major histocompatibility complex class I-related molecule (MR1). Our previous study showed that MR1−/− knock-out (KO) mice (lacking MAIT cells) harbor a unique microbiota that is resistant to antibiotic disruption and Clostridioides difficile colonization. While we have characterized the microbiota of this mouse strain, changes in global metabolic activity in these KO mice have not been assessed. Here, LC/MS-based untargeted metabolomics was applied to investigate the differences in the metabolome, specifically in the bile acid (BA) profile of wild-type (WT) and MR1−/− KO mice, as well as how antibiotics change these profiles. BA changes were evaluated in the intestinal content, cecum content, and stool samples from MR1−/− mice and WT mice treated with cefoperazone (Cef). Fecal pellets were collected daily and both intestinal and cecal contents were harvested at predetermined endpoints on day 0 (D0), day 1 (D1), day 3 (D3), and day 5 (D5). KO mice exhibited no changes in 6-hydroxymethyl-8-D-ribityllumazine (rRL-6-CH2OH; an MR1-restricted riboflavin derivative) in the stool samples at either time point vs. D0, while WT mice showed significant decreases in rRL-6-CH2OH in the stool samples on all treatment days vs. D0. Metabolomics analysis from cecal and stool samples showed that KO mice had more total BA intensity (KO/WT = ~1.7 and ~3.3 fold higher) than that from WT mice prior to Cef treatment, while the fold change difference (KO/WT = ~4.5 and ~4.4 fold) increased after five days of Cef treatment. Both KO and WT mice showed decreases in total BA intensity in response to Cef treatment, however, less dramatic decreases were present in KO vs. WT mice. Increases in taurocholic acid (TCA) intensity and decreases in deoxycholic acid (DCA) intensity in the stool samples from WT mice were associated with the depletion of certain gut bacteria, which was consistent with the previously reported microbiome data. Furthermore, the non-detected TCA and relatively higher DCA intensity in the KO mice might be related to Clostridioides difficile infection resistance, although this needs further investigation.

PSVI-10 Essential oils and benzoic acid, associated or not with antibiotic, in diets for weaned piglets on cecum microbiota composition

This study evaluated the effects of an alternative feed additive (benzoic acid and the essential oils of eugenol, thymus and piperine), associated or not with colistin, in weaned piglets feeding on α and β bacterial diversity, and on the percentages of operational taxonomic unit of Lactobacillus, Bifidobacterium, Clostridium, Escherichia, and Campylobacter in the cecum content of pigs. One hundred and eight crossbred piglets (5.3 ± 0.5kg) were housed 3 per pen and fed the following diets: a control diet (CON) with no addition of growth promoter feed additive; CON + 40 ppm of colistin; CON + 0.3% of essential oils and benzoic acid; and CON + 0.3% of essential oils and benzoic acid, and 40 ppm of colistin. The diets were based on corn, soybean meal, dairy products, and spray-dried blood plasma, and were formulated to provide 3.40 Mcal of ME/kg; 220 g of CP/kg; and 14.5 g of digestible lysine/kg. The animals were sanitary challenged, by spraying 2 L of diluted pig manure (60 % of manure to 40 % of water) at each nursery pen two days before the beginning of the trial. At the 14th day of the trial, 9 animals per diet were euthanized, and the cecum content was collected for microbial DNA extraction and sequencing. The data was submitted to ANOVA using the PROC MIXED of SAS. The α and β bacterial diversity, and the percentages of Lactobacillus, Bifidobacterium and Clostridium in the cecum of pigs were not affected (P > 0.05) by the experimental diets. There was a tendency (P = 0.09) for a lower concentration of Escherichia (3.5 vs. 1.46%) and lower percentage of Campylobacter (10.21 vs. 0.52%) in the cecum content of piglets fed diets containing essential oils and benzoic acid compared to pigs fed diets without the alternative feed additive. Inclusion of benzoic acid and essential oils in piglets feeding was effective in controlling intestinal pathogenic bacteria.

Butyrate Enhances Desensitization Induced by Oral Immunotherapy in Cow’s Milk Allergic Mice

Background. In previous studies, we showed that a fructo-oligosaccharide- (FOS-) supplemented diet enhanced oral immunotherapy (OIT) efficacy in a mouse model for cow’s milk allergy. Fermentation of FOS by intestinal bacteria leads to production of short-chain fatty acids (SCFA) including butyrate.Aim. To investigate the contribution of butyrate in the enhanced efficacy of OIT + FOS.Methods. C3H/HeOuJ mice were sensitized and received OIT with or without FOS or butyrate supplementation. After treatment, whole blood was collected to conduct a basophil activation test (BAT) and allergen challenges were performed to measure acute allergic symptoms. CD4 + CD25 + regulatory T cells (Tregs) were isolated from treated mice or differentiatedin vitroand used in a bone marrow-derived mast cell (BMMC) suppression assay. Cecum content was collected to analyze SCFA concentrations.Results. Allergen-induced basophil activation was reduced in OIT + butyrate samples compared to OIT. Accordingly, the acute allergic skin response and mast cell degranulation upon challenge were reduced in OIT + butyrate and OIT + FOS mice compared to sensitized controls. Butyrate was increased in the cecum content of OIT + FOS mice compared to OIT mice and sensitized controls. Treg-mediated BMMC suppression was enhanced afterin vivobutyrate and FOS exposure in combination with OIT but with a more pronounced effect for butyrate.Conclusion. Butyrate supplementation enhanced OIT-induced desensitization of basophils and mast cells and Treg functionality. Only OIT + FOS treatment induced potential microbial alterations, shown by increased butyrate levels in cecum content. Both butyrate and FOS are promising candidates to improve OIT efficacy in human studies to treat food allergies.