Protective Effects of Novel Lactobacillaceae Strains Isolated from Chicken Caeca against Necrotic Enteritis Infection: In Vitro and In Vivo Evidences.

The present study aimed to show the benefits of novel lactic acid bacteria (LAB) strains isolated from the caeca of healthy chickens. These novel strains, identified as Limosilactobacillus reuteri and Ligilactobacillus salivarius, displayed high levels of lactic acid production, capability of biofilm formation, high aggregation and adhesion scores, and significant survival rates under conditions mimicking the chicken gastrointestinal tract (GIT). In addition, these novel Lactobacillaceae isolates were neither hemolytic nor cytotoxic. In vivo trials were able to establish their ability to reduce necrotic enteritis. Notably, a significant weight gain was registered, on day 10 of treatment, in the group of chickens fed with a mixture of L. reuteri ICVB416 and L. salivarius ICVB430 strains, as compared with the control group. This group has also shown a reduced number of lesions in the gut compared with other infected chicken groups. This study provides in vitro and in vivo evidence supporting the benefits of these novel Lactobacillaceae isolates for their use in poultry livestock as protective cultures to control the bacterial necrotic enteritis (NE) Clostridium perfringens.

Secondary Composite Breast Augmentation: Concept and Outcomes, Introduction to a Layered Approach.

BACKGROUND: Secondary procedures following breast augmentation are often more difficult than primary cases because the soft-tissue envelope changes over time. OBJECTIVES: This study was conducted to confirm the utility of a composite technique in breast revisional surgery. METHODS: This was a 9-year retrospective chart and photographic data study of one surgeon's experience with the combined use of fat and implants in revisional cases. The 148 patients had a follow-up at least 1 year after surgery. Our approach consists of a detailed analysis of the different layers covering the implant and yields a treatment plan addressing all issues involving the secondary breast. RESULTS: On average, revisional surgery was performed 8.66 years after the first augmentation. The mean age of the patients at revision surgery was 42 years (range, 22.2-70.7 years). The mean fat harvest was 600 mL (range, 100-3000 mL) and the mean volume of fat reinjected was 153 mL (range, 60-400 mL). The mean volume before and after revision was the same (288 mL vs 289 mL). At the original surgery, the breast implants were located in a subpectoral pocket in 78.7% of the patients and, at the revision surgery, in a subglandular pocket in 74.8% of the patients. Within the first 2 years, 13 patients (8.7%) underwent reoperation for additional fat grafting. Among 45 preoperative breast capsular contractures, there were 8 recurrences in the first 3 years resulting in 4 reoperations. CONCLUSIONS: Secondary breast augmentation cannot rely solely on implant exchange. Because the soft-tissue envelope also ages over time, fat grafting is mandatory in the vast majority of secondary cases. A rigorous preoperative analysis enables breast defects to be treated appropriately.

Effects of selenium feed supplements on functional properties of eggs.

The aim of the present study was to assess the effects of selenium feed supplements on the functional properties of eggs. The hens in experimental groups were fed diets supplemented with 0.2 mg/kg selenium from sodium selenite (Group 1), selenium-enriched yeast (Group 2), synthetic L-selenomethionine (Group 3), or hydroxy analog of selenomethionine (Group 4). The Control Group (Group C) was fed with basal feed without supplementation. The highest values of albumen gel firmness were shown in Group C eggs; differences with all experimental groups were significant (p < 0.001 to p = 0.009). It was ascertained that albumen gel firmness correlated with albumen pH (rs = 0.490; p < 0.001), which was highest in eggs from non-supplemented hens. Group 1 eggs and Group C eggs showed lower albumen foaming capacity (p < 0.001) compared to eggs from other groups. Both albumen foaming capacity and albumen foam stability were higher in Group 2 eggs than in Group C eggs (p < 0.001). The highest yolk foaming capacity was found in Group 2 eggs (p < 0.001). Sponge cakes baked with Group C eggs had a smaller volume than those baked with eggs from Group 2 (p = 0.005), Group 3 (p = 0.004) and Group 4 (p = 0.024). The results of the study confirmed that selenium added to the laying hen feed significantly affected the monitored functional properties of both albumen and yolk. The most distinctive effect of selenium was shown in eggs from the group supplemented with selenium-enriched yeast, for which the results of albumen foam capacity and stability and yolk foaming capacity were the best.

Effect of live yeast Saccharomyces cerevisiae supplementation on the performance and cecum microbial profile of suckling piglets.

One mechanism through which S. cerevisiae may improve the performance of pigs is by altering the composition of the gut microbiota, a response that may be enhanced by early postnatal supplementation of probiotics. To test this hypothesis, newborn piglets (16 piglets/group) were treated with either S. cerevisiae yeast (5 x 109 cfu/pig: Low) or (2.5 x 1010 cfu/piglet: High) or equivalent volume of sterile water (Control) by oral gavage every other day starting from day 1 of age until weaning (28±1 days of age). Piglet body weight was recorded on days 1, 3, 7, 10, 17, 24 and 28 and average daily gain (ADG) calculated for the total period. At weaning, piglets were euthanized to collect cecum content for microbial profiling by sequencing of the 16S rRNA gene. ADG was higher in both Low and High yeast groups than in Control group (P<0.05). Alpha diversity analyses indicated a more diverse microbiota in the Control group compared with Low yeast group; the High yeast being intermediate (P < 0.01). Similarly, Beta diversity analyses indicated differences among treatments (P = 0.03), mainly between Low yeast and Control groups (P = 0.02). The sparse Partial Least Squares Discriminant Analysis (sPLS-DA) indicated that Control group was discriminated by a higher abundance of Veillonella, Dorea, Oscillospira and Clostridium; Low yeast treated pigs by higher Blautia, Collinsella and Eubacterium; and High yeast treated pigs by higher Eubacterium, Anaerostipes, Parabacteroides, Mogibacterium and Phascolarctobacterium. Partial Least Squares (PLS) analysis showed that piglet ADG was positively correlated with genus Prevotella in High yeast group. Yeast supplementation significantly affected microbial diversity in cecal contents of suckling piglets associated with an improvement of short chain fatty acid producing bacteria in a dose-dependent manner. In conclusion, yeast treatment improved piglet performance and shaped the piglet cecum microbiota composition in a dose dependent way.

Cryptosporidium parvum-Infected Neonatal Mice Show Gut Microbiota Remodelling Using High-Throughput Sequencing Analysis: Preliminary Results.

BACKGROUND: During the last decade, the scientific community has begun to investigate the composition and role of gut microbiota in normal health and disease. These studies have provided crucial information on the relationship between gut microflora composition and intestinal parasitic infection, and have demonstrated that many enteric pathogen infections are associated with altered gut microflora composition. In this study, we investigated the effects of Cryptosporidium parvum infection (zoonotic protozoan affecting a large range of vertebrates) on both qualitative and quantitative composition of gut microbiota in a CD-1 neonatal mouse model. METHODS: 5-day-old neonate mice were experimentally infected with 105Cryptosporidium parvum Iowa oocysts by oesophageal gavage. The intestinal microbiota of both infected (Cp+) and uninfected (Cp-) mice groups was examined by high-throughput sequencing of the bacterial 16S rDNA gene V3-V4 hypervariable region. RESULTS: The most consistent change in the microbiota composition of Cp+ mice was the increased proportion of bacterial communities belonging to the Phylum Bacteroidetes. In contrast, the microbiota of Cp- mice was associated with increased proportions of several Firmicutes and Actinobacteria phyla members. CONCLUSION: For the first time, our study provides evidence of an association between cryptosporidial infection and gut dysbiosis, thus contributing valuable knowledge to the as-yet little-explored field of Cryptosporidium-microbiota interactions in a neonatal mouse model.

Benefits and Inputs From Lactic Acid Bacteria and Their Bacteriocins as Alternatives to Antibiotic Growth Promoters During Food-Animal Production.

Resistance to antibiotics is escalating and threatening humans and animals worldwide. Different countries have legislated or promoted the ban of antibiotics as growth promoters in livestock and aquaculture to reduce this phenomenon. Therefore, to improve animal growth and reproduction performance and to control multiple bacterial infections, there is a potential to use probiotics as non-antibiotic growth promoters. Lactic acid bacteria (LAB) offer various advantages as potential probiotics and can be considered as alternatives to antibiotics during food-animal production. LAB are safe microorganisms with abilities to produce different inhibitory compounds such as bacteriocins, organic acids as lactic acid, hydrogen peroxide, diacetyl, and carbon dioxide. LAB can inhibit harmful microorganisms with their arsenal, or through competitive exclusion mechanism based on competition for binding sites and nutrients. LAB endowed with specific enzymatic functions (amylase, protease…) can improve nutrients acquisition as well as animal immune system stimulation. This review aimed at underlining the benefits and inputs from LAB as potential alternatives to antibiotics in poultry, pigs, ruminants, and aquaculture production.

Supplementation with live yeast increases rate and extent of in vitro fermentation of nondigested feed ingredients by fecal microbiota.

Two studies were conducted to investigate the effect of live yeast (LY) on the in vitro fermentation characteristics of wheat, barley, corn, soybean meal (SBM), canola meal, and distillers dried grains with solubles (DDGS). In Study 1, LY yeast was added directly to in vitro fermentations inoculated with feces from lactating sows, whereas as in study 2, feces collected from lactating sows fed LY as a daily supplement was used. Selected feedstuffs were digested and the residue added to separate replicated (n = 3) fermentation reactions. Study 1 was conducted in two blocks, whereas study 2 was conducted using feces collected after a period of 3 (Exp. 1) or 4 wk (Exp. 2) of LY supplementation. Accumulated gas produced over 72 h was modeled for each substrate and the kinetics parameters compared between LY and control groups. The molar ratio of the volatile fatty acids (VFAs) produced in vitro were also compared at 12 and 72 h of incubation. In study 1, in vitro addition of yeast increased (P < 0.001) the rate of gas production (Rmax). However, a yeast × substrate effect (P < 0.05) observed for total gas accumulated (A), time to half asymptote (B), and time required to reach maximum rate of fermentation (Tmax) suggested that yeast-mediated increases in extent and rate of fermentation varied by substrate. Greater total gas production was observed only for corn and SBM, associated with greater B and Tmax. Supplementation with LY appeared to increase A and Rmax although with variation between experiments and substrates. In Exp. 1, LY decreased (P < 0.05) B and Tmax. However, a yeast × substrate effect (P < 0.05) was observed for only A (for wheat, barley, corn, and corn DDGS) and Rmax (wheat, barley, corn, and wheat DDGS). In Exp. 2, LY increased (P < 0.0001) A and decreased B. However, an interaction (P < 0.05) with substrates was observed for Rmax (except SBM) and Tmax. With exception of the DDGS samples, LY supplementation increased (P < 0.05) VFA production at 12 and 72 h of incubation. Yeast increased (P < 0.05) the molar ratios of acetic acid and branch-chain fatty acids at 12 h of incubation; however, this response was more variable by substrate at 72 h. In conclusion, LY supplementation increased the rate and extent of in vitro fermentation of a variety of substrates prepared from common feedstuffs. Greater effects were observed when LY was fed to sows than added directly in vitro, suggesting effects on fermentation were not mediated directly.

Assembly of the Entire Carbon Backbone of a Stereoisomer of the Antitumor Marine Natural Product Hemicalide.

A strategy for the assembly of the entire carbon backbone of a stereoisomer of the antitumor marine natural product hemicalide has been investigated. The devised convergent approach relies on Horner-Wadsworth-Emmons and Julia-Kocienski olefination reactions for the construction of the C6=C7 and C34=C35 double bonds, respectively, an aldol reaction to create the C27-C28 bond, and a Suzuki-Miyaura cross-coupling as the endgame to form the C15-C16 bond.

Effect of live yeast (Saccharomyces cerevisiae) supplementation on rumen fermentation and metabolic profile of dairy cows in early lactation.

The study evaluated dietary supplementation with live yeast (LY) Saccharomyces cerevisiae (CNCM I-4407, 1010  CFU/g, Actisaf; Phileo Lesaffre Animal Care, France) on rumen fermentation and serum metabolic profile in lactating dairy cows. Fifty Holstein cows received a total mixed ration with (Live Yeast Diet, LYD, n = 25) or without (Control Diet, CD, n = 25) 5 × 1010  CFU/cow/day of LY from 3 to 19 weeks of lactation. Rumen fermentation and serum metabolic profile were measured in eight cows per treatment at 3, 7, 11, 15, 19 weeks post-partum. LYD showed an increased daily milk yield (+4%) over CD (p < 0.05). Mean rumen pH at 4 hr after morning meal was higher in LYD (6.59) than CD (6.32) (p < 0.01). Total volatile fatty acids (VFA) and acetate molar proportion were higher in LYD (114.24 mM; 25.04%) than CD (106.47 mM; 24.73%) (p < 0.05). Propionate and butyrate molar proportions, acetate to propionate ratio, ammonia levels did not differ between LYD and CD. Ruminal lactate was lower in LYD than CD (9.3 vs. 16.4 mM) (p < 0.001), with a 53% decrease in LYD. During peak lactation, LYD had lower serum NEFA (p < 0.05, 0.40 vs. 0.48 mM) and BHBA (p < 0.01, 0.47 vs. 0.58 mM) than CD, lower liver enzyme activities (AST 1.39 vs. 1.54 ukat/L) (p < 0.05). Serum glucose was higher in LYD at peak lactation (3.22 vs. 3.12 mM, and 3.32 vs. 3.16 mM respectively) (p < 0.05). The results confirmed a reducing effect of LY on lactate accumulation in rumen fluid, associated with an increase in rumen pH. Lower serum levels of lipomobilization markers, liver enzyme activities and higher glucose levels may suggest that live yeast slightly mitigated negative energy balance and had a certain liver protective effect.

Efficacy of chitosan, a natural polysaccharide, against Cryptosporidium parvum in vitro and in vivo in neonatal mice.

Cryptosporidiosis is a zoonotic disease caused by species in the genus Cryptosporidium. In young ruminants, Cryptosporidium parvum causes economically significant disease with mild to severe clinical signs and occasional death. The typical clinical course in animals aged 1-3 weeks old is acute diarrhoea. Currently there are no available treatments that are fully effective against cryptosporidiosis in either humans or animals. Therefore there is a critical need for the development of new therapeutic agents. We adapted two in vitro culture systems (HCT-8 and Caco-2 cell lines) for C. parvum infection to investigate the "anticryptosporidial" activity of two chitosans; Chitosan NAG and Chitosan Mix. Chitosan-a naturally-occurring polysaccharide compound-has been found to be active against a variety of diseases, possessing both antimicrobial and anticancer properties. We investigated both chitosan's toxicity and effects on C. parvum in the two in vitro models. To evaluate chitosan's effects on oocyst shedding in vivo, CD-1 neonate mice were orally inoculated with C. parvum oocysts (Iowa strain), treated with chitosan, and compared to infected non-treated animals. Paromomycin, a classical drug used in veterinary medicine, was used as a reference compound. Immunofluorescence techniques were used to analyse the parasites. Our results showed significant reductions in Cryptosporidium oocyst viability (>95%) after oocyst pre-incubation with either paromomycin (P < 0.001), Chitosan Mix or Chitosan NAG (P < 0.001), for 24 h at 37 °C. Additionally, paromomycin, Chitosan Mix, and Chitosan NAG significantly inhibited C. parvum multiplication in HCT-8 and Caco-2 cell lines (P < 0.005). These effects were dose-dependent. In in vivo studies, treatment with both chitosans (Chitosan NAG, Chitosan Mix) or paromomycin sulfate significantly reduced parasite shedding in infected treated newborn mice (-56%, -34.5% and -58%, respectively). In conclusion, these findings provide the first in vitro and in vivo evidence of the anticryptosporidial activities of this natural polysaccharide.

Effects of dietary live yeast supplementation on growth performance, diarrhoea severity, intestinal permeability and immunological parameters of weaned piglets challenged with enterotoxigenic Escherichia coli K88.

This study aimed to investigate the effects of dietary live yeast (LY) supplementation on growth, intestinal permeability and immunological parameters of piglets challenged with enterotoxigenic Escherichia coli K88 (ETEC). Piglets weaned at 21 d were allocated into three treatments with six pens and six piglets per pen, receiving the control diet (CON), diets supplemented with antibiotics plus zinc oxide (ANT-ZnO) and LY (Saccharomyces cerevisiae strain CNCM I-4407), respectively, for a period of 2 weeks. On day 8, thirty-six piglets were selected as control without ETEC (CON), CON-ETEC, ANT-ZnO-ETEC and LY-ETEC groups challenged with ETEC until day 10 for sample collections. Piglets fed ANT-ZnO diet had the highest average daily gain and average daily feed intake (P<0·05) during the 1st week, but ADG of piglets fed the ANT-ZnO diet was similar as piglets fed LY diet during the second week. Piglets with LY-ETEC or ANT-ZnO-ETEC had markedly lower diarrhoea score (P<0·05) than piglets with CON-ETEC during the 24 h after ETEC challenge. Relative to piglets with CON, the counts of E. coli, urinary ratio of lactulose to mannitol, plasma IL-6 concentration, mRNA abundances of innate immunity-related genes in ileum and mesenteric lymph node tissues were increased (P<0·05), whereas the villous height of jejunum and relative protein expression of ileum claudin-1 were decreased (P<0·05) in piglets with CON-ETEC; however, these parameters did not markedly change in piglets with LY-ETEC or ANT-ZnO-ETEC. In summary, dietary LY supplementation could alleviate the severity of diarrhoea in piglets with ETEC, which may be associated with the improved permeability, innate immunity and bacterial profile.

Draft Genome Sequence of Enterococcus faecalis DD14, a Bacteriocinogenic Lactic Acid Bacterium with Anti-Clostridium Activity.

We report the draft genome sequence of Enterococcus faecalis DD14, a strain isolated from meconium of a healthy newborn at Roubaix Hospital (France). The strain displayed antagonism against a set of Gram-positive bacteria through concomitant production of lactic acid and bacteriocin. The genome has a size of 2,893,365 bp and a 37.3% G+C ratio and is predicted to contain at least 2,755 coding sequences and 62 RNAs.

Subfascial Primary Breast Augmentation with Fat Grafting: A Review of 156 Cases.

BACKGROUND: Composite breast augmentation with fat grafting and an implant has become very popular in the past 5 years. This achieves the core volume projection of an implant complemented by the natural appearance and feel of fat. However, no study has looked at the complications and reoperation rates of this technique. METHODS: A retrospective chart review examined all patients who underwent the combined use of an implant and fat grafting for primary breast augmentation. RESULTS: The study identified 156 patients between 2007 and 2013. The mean patient age was 31.7 years and the average body mass index was 18.85 kg/m. The average implant size was 252 cc. Patients received a mean of 126 cc of fat (range, 30 to 250 cc) in subcutaneous soft tissue. Follow-up averaged 22.25 months (range, 1 to 86 months). The total complication rate was 7.7 percent and the reoperation rate was 9.94 percent. Baker grade II/III contracture was the most common complication [Baker grade II, n = 4 (2.56 percent); Baker grade III, n = 2 (2 percent)], followed by infections [n = 2 (1.28 percent)], hematoma [n = 2 (1.28 percent)], and malrotation [n = 1 (0.64 percent)]. Delayed reoperation was performed in nine patients (9.94 percent) after a mean interval of 31.7 months. Two patients who developed Baker grade III contractures needed surgery to correct the problem. Three cases (1.92 percent) required additional fat grafting for insufficient soft-tissue coverage. The mean volume of fat reinjection was 170 cc. CONCLUSIONS: Composite breast augmentation is a valuable, stable, reliable technique in breast aesthetic surgery with good, natural-appearing results. It provides long-term aesthetic benefits and avoids the submuscular plane. CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, IV.

The safe enterocin DD14 is a leaderless two-peptide bacteriocin with anti-Clostridium perfringens activity.

Enterococcus faecalis 14, a strain previously isolated from meconium, displayed activity against four Clostridium perfringens isolates when co-cultured on agar plates. The anti-Clostridium activity was ascribed to the production of enterocin DD14, which was subsequently purified. The minimum inhibitory concentration (MIC) of enterocin DD14 against one collection strain and one clinical C. perfringens strain was determined at 50 µg/mL. Furthermore, using the intestinal epithelial cell line IPEC-1, it was shown that E. faecalis 14 was not cytotoxic after 24 h of contact, and no cytotoxicity was observed when IPEC-1 cells were incubated with pure enterocin DD14 for 4 h. Enterocin DD14 was characterised using mass spectrometry and was shown to consist of two small proteins of 5200.74 Da and 5206.41 Da, respectively. The two peptides (DD14A and DD14B) have highly similar amino acid sequences and no signal peptide, which classifies enterocin DD14 as a class IIb leaderless two-peptide bacteriocin. The genes encoding DD14A and DD14B were sequenced and were shown to be 100% identical to other previously described enterocins MR10A and MR10B, in contrast to the producing strains, which are different. Consequently, the present in vitro study supports the potential of this E. faecalis 14 strain and/or its purified enterocin DD14 as putative anti-C. perfringens compounds in chickens.

Effects of Colistin and Bacteriocins Combinations on the In Vitro Growth of Escherichia coli Strains from Swine Origin.

Escherichia coli strains from swine origin, either susceptible or resistant to colistin, were grown under planktonic and biofilm cultures. After which, they were treated with antibacterial agents including nisin and enterocin DD14 bacteriocins, colistin and their combinations. Importantly, the combination of colistin, enterocin DD14 and nisin eradicated the planktonic and biofilm cultures of E. coli CIP54127 and the E. coli strains with colistin-resistance phenotype such as E. coli 184 (mcr-1 +) and E. coli 289 (mcr-1 -), suggesting therefore that bacteriocins from lactic acid bacteria could be used as agents with antibiotic augmentation capability.

Alternatives to Antibiotics to Prevent Necrotic Enteritis in Broiler Chickens: A Microbiologist's Perspective.

Since the 2006 European ban on the use of antibiotics as growth promoters in animal feed, numerous studies have been published describing alternative strategies to prevent diseases in animals. A particular focus has been on prevention of necrotic enteritis in poultry caused by Clostridium perfringens by the use of microbes or microbe-derived products. Microbes produce a plethora of molecules with antimicrobial properties and they can also have beneficial effects through interactions with their host. Here we review recent developments in novel preventive treatments against C. perfringens-induced necrotic enteritis in broiler chickens that employ yeasts, bacteria and bacteriophages or secondary metabolites and other microbial products in disease control.

A Comparison of the Beneficial Effects of Live and Heat-Inactivated Baker's Yeast on Nile Tilapia: Suggestions on the Role and Function of the Secretory Metabolites Released from the Yeast.

Yeast is frequently used as a probiotic in aquaculture with the potential to substitute for antibiotics. In this study, the involvement and extent to which the viability of yeast cells and thus the secretory metabolites released from the yeast contribute to effects of baker's yeast was investigated in Nile tilapia. No yeast, live yeast or heat-inactivated baker's yeast were added to basal diets high in fishmeal and low in soybean (diet A) or low in fishmeal and high in soybean (diet B), which were fed to fish for 8 weeks. Growth, feed utilization, gut microvilli morphology, and expressions of hsp70 and inflammation-related cytokines in the intestine and head kidney were assessed. Intestinal microbiota was investigated using 16S rRNA gene pyrosequencing. Gut alkaline phosphatase (AKP) activity was measured after challenging the fish with Aeromonas hydrophila. Results showed that live yeast significantly improved FBW and WG (P < 0.05), and tended to improve FCR (P = 0.06) of fish compared to the control (no yeast). No significant differences were observed between inactivated yeast and control. Live yeast improved gut microvilli length (P < 0.001) and density (P < 0.05) while inactivated yeast did not. The hsp70 expression level in both the intestine and head kidney of fish was significantly reduced by live yeast (P < 0.05) but not inactivated yeast. Live yeast but not inactivated yeast reduced intestinal expression of tnfα (P < 0.05), tgfß (P < 0.05 under diet A) and il1ß (P = 0.08). Intestinal Lactococcus spp. numbers were enriched by both live and inactivated yeast. Lastly, both live and inactivated yeast reduced the gut AKP activity compared to the control (P < 0.001), indicating protection of the host against infection by A. hydrophila. In conclusion, secretory metabolites did not play major roles in the growth promotion and disease protection effects of yeast. Nevertheless, secretory metabolites were the major contributing factor towards improved gut microvilli morphology, relieved stress status, and reduced intestinal inflammation of Nile tilapia fed diets supplemented with baker's yeast.