Deoxygenation of ClSO2CF2COOMe with Triphenylphosphine for the Metal-Free Direct Electrophilic Difluoroalkylthiolation of Various Heterocycles.

A direct electrophilic difluoroalkylthiolation reaction of indole derivatives and other electron-rich heterocycles using methyl 2,2-difluoro-2-(chlorsulfonyl)acetate (ClSO2CF2COOMe) derived from Chen's reagent (FSO2CF2COOMe) is described. The ester group in the product can be further utilized in subsequent versatile transformations. The reactions provide good yields of the corresponding difluoroalkylthiolation products and exhibit high functional group compatibility. It is expected to serve as an alternative and practical protocol for difluoroalkylthiolation of various heterocycles.

Comparison of regression and fat-free diet methods for estimating ileal and total tract endogenous losses and digestibility of fat and fatty acids in growing pigs.

An experiment was conducted to 1) compare the regression and fat-free diet methods for estimating total or basal endogenous losses of fat (ELF) and fatty acids (ELFA) and true digestibility (TD) or standardized digestibility (SD) of fat and fatty acids in growing pigs and 2) compare these estimated values at the end of the ileum and over the entire intestinal tract. Ten barrows (initial body weight: 45.1 ± 2.8 kg) were surgically fitted with a T-cannula in the distal ileum and allotted to one of five experimental diets in a three-period Youden Square design. A fat-free diet was formulated using cornstarch, soy protein isolate, and sucrose. Four oil-added diets were formulated by adding four levels of soybean oil (2%, 4%, 6%, and 8%) to the fat-free basal diet at the expense of cornstarch. All diets contained 26% sugar beet pulp and 0.40% chromic oxide. Results indicated that there were no differences between true ileal digestibility (TID) of fat and true total tract digestibility (TTTD) of fat when pigs were fed soybean oil. The TID of C18:0 and total saturated fatty acids (TSFA) was greater than TTTD (P < 0.05). The total ELF at the end of the ileum were not different from that over the entire intestinal tract. In addition, total endogenous losses of C18:0 and TSFA were greater for the entire intestinal tract than at the end of the ileum, whereas total endogenous losses of C18:2 and total unsaturated fatty acids were just the opposite. Similar results were observed for basal ELF and ELFA. As the inclusion level of soybean oil increased, apparent digestibility (AD) of fat and fatty acids increased linearly (P < 0.05) except for apparent ileal digestibility of C18:0. However, SD of fat and fatty acids was not influenced by the inclusion level of soybean oil. Estimation of ELF and ELFA observed by the regression and fat-free diet methods did not differ when measured at the end of the ileal or total tract. There were no differences between the estimations of TD and SD of fat and fatty acids for soybean oil. Collectively, the estimation of TD or SD of fat can be measured over the entire intestinal tract, whereas the ileal analysis method should be used to determine the ELF, ELFA, and TD or SD of fatty acids. Correcting AD for basal ELF and ELFA can accurately estimate SD values of fat and fatty acids.

Dietary Supplementation with Compound Probiotics and Berberine Alters Piglet Production Performance and Fecal Microbiota.

This study was conducted to investigate the effects of dietary supplementation with compound probiotics and berberine (CPB) on growth performance, nutrient digestibility and fecal microflora in weaned piglets. A total of 200 piglets 35 days old were randomly allocated to 5 groups, 4 replications in each group, and 10 piglets in each replication. Group A was the basal diet; group B was supplemented with antibiotics and zinc oxide; groups C, D and E were supplemented with 0.06%, 0.12% and 0.18% CPB, respectively. The experimental period was 42 d. The results indicated that there were no significant differences in average daily feed intake (ADFI), average daily gain (ADG) and feed conversion rate (FCR) among five groups (p > 0.05). However, mortality, diarrhea and rejection rates in the control group were higher than that in other groups. CPB could increase protein digestibility and serum IgG content (p < 0.05), while it could decrease serum urea nitrogen content and alkaline phosphatase activity (p < 0.05). Analysis of fecal microbiota showed that the relative abundances of Bacteroides and Firmicutes were increased, while the relative abundances of opportunistic pathogens such as Spirochaetae and Protebactreria were dramatically decreased in piglets fed with CPB or antibiotics, compared with the control group. Furthermore, CPB intervention increased the relative abundances of Prevotella_9, Megasphaera and Prevotella_2, while decreased the relative abundance of Prevotellaceae_NK3B31_group. Correlation analysis revealed that there was good correlation between serum indexes and fecal microbiota. It was suggested that CPB might be a promising antibiotic alternative for improving piglet health and immunity, decreasing mortality by positively altering gut microbiota.

Compound probiotics alleviating aflatoxin B1 and zearalenone toxic effects on broiler production performance and gut microbiota.

In order to alleviate toxic effects of aflatoxins B1 (AFB1) and zearalenone (ZEA) on broiler production performance and gut microbiota, three kinds of compound probiotics (CP) were selected. The optimal ratios of Bacillus subtilis, Lactobacillus casei and Candida utilis in broiler diets were 7, 5 and 6 log CFU/g for ZEA biodegradation (CP1); 6, 7 and 7 log CFU/g for AFB1 biodegradation (CP2); 7, 6 and 7 log CFU/g for ZEA + AFB1 biodegradation (CP3). A total of 350 1-day-old Ross broilers were randomly divided into 7 groups. Group A was the basal diet, group B-G contained ZEA, AFB1, ZEA + AFB1, ZEA + CP1, AFB1+CP2, ZEA + AFB1+CP3, respectively. The experiment showed that AFB1 or AFB1+ZEA significantly decreased broiler production performance, damaged liver and jejunum, increased mycotoxin residues in broiler body; however, three kinds of compound probiotics additions could alleviate mycotoxin negative effects on the above parameters (p < 0.05). The gut microbiota analysis indicated that AFB1+ZEA increased jejunal microbial richness, but which were decreased to almost the same level as the control group by CP3 addition. CP3 addition significantly increased jejunal Firmicutes and Lactobacillus aviarius abundances. The correlative analysis showed that gut Lactobacillus aviarius abundance was positively correlated with average daily gain (ADG) of broilers (p < 0.05), while AFB1+ZEA addition decreased its relative abundance, indicating that CP3 addition increased broiler growth by increasing Lactobacillus aviarius abundance. AFB1 and ZEA residues in broiler body were negatively correlated with the gut beneficial bacterial abundances (p < 0.01), but positively correlated with the potentially harmful bacterial abundances (p < 0.05), which inferred that CP3 addition could decrease mycotoxin residues through positively regulating gut relative bacterial abundances. In conclusion, compound probiotics could keep gut microbiota stable, degrade mycotoxins, alleviate histological lesions, increase production performance and reduce mycotoxin toxicity for broilers.

Zearalenone Biodegradation by the Combination of Probiotics with Cell-Free Extracts of Aspergillus oryzae and its Mycotoxin-Alleviating Effect on Pig Production Performance.

In order to remove zearalenone (ZEA) detriment-Bacillus subtilis, Candida utilis, and cell-free extracts from Aspergillus oryzae were used to degrade ZEA in this study. The orthogonal experiment in vitro showed that the ZEA degradation rate was 92.27% (p < 0.05) under the conditions that Candida utilis, Bacillus subtilis SP1, and Bacillus subtilis SP2 were mixed together at 0.5%, 1.0%, and 1.0%. When cell-free extracts from Aspergillus oryzae were combined with the above probiotics at a ratio of 2:1 to make mycotoxin-biodegradation preparation (MBP), the ZEA degradation rate reached 95.15% (p < 0.05). In order to further investigate the MBP effect on relieving the negative impact of ZEA for pig production performance, 120 young pigs were randomly divided into 5 groups, with 3 replicates in each group and 8 pigs for each replicate. Group A was given the basal diet with 86.19 µg/kg ZEA; group B contained 300 µg/kg ZEA without MBP addition; and groups C, D, and E contained 300 µg/kg ZEA added with 0.05%, 0.10%, and 0.15% MBP, respectively. The results showed that MBP addition was able to keep gut microbiota stable. ZEA concentrations in jejunal contents in groups A and D were 89.47% and 80.07% lower than that in group B (p < 0.05), indicating that MBP was effective in ZEA biodegradation. In addition, MBP had no significant effect on pig growth, nutrient digestibility, and the relative mRNA abundance of estrogen receptor alpha (ERα) genes in ovaries and the uterus (p > 0.05).

Effects of Saccharomyces cerevisiae on alleviating cytotoxicity of porcine jejunal epithelia cells induced by deoxynivalenol.

Deoxynivalenol (DON) is one of the mycotoxins most frequently encountering in cereal-based foods throughout the world. Saccharomyces cerevisiae was used to alleviate porcine jejunal epithelia cell (IPEC-J2) injury induced by DON in this study. The results indicated that cell viability and proliferation rates were significantly decreased when DON concentrations were increased from 0 to 64 µM after 24 h incubation (p < 0.05). The longer incubation time and higher DON concentrations would cause more serious effects on cell viability. S. cerevisiae could significantly degrade DON and decrease lactic dehydrogenase (LDH) release in the cells induced by DON (p < 0.05). DON (4 µM) could increase necrotic and apoptotic cell rates as well as decrease viable cell rates, compared with the control group (p < 0.05). However, S. cerevisiae addition in the DON group could decrease necrotic, late apoptotic and early apoptotic cell rates by 38.05%, 46.37% and 44.78% respectively, increase viable cell rates by 2.35%, compared with the single DON group (p < 0.05). In addition, S. cerevisiae addition could up-regulate mRNA abundances of IL-6, IL-8 and IL-10 in IPEC-J2 cells (p < 0.05), but down-regulate mRNA abundances of tight junction proteins (TJP-1) and occludin by 36.13% and 50.18% at 1 µM of DON (p < 0.05). It could be concluded that S. cerevisiae was able to alleviate IPEC-J2 cell damage exposed to DON.

Effect of Compound Probiotics and Mycotoxin Degradation Enzymes on Alleviating Cytotoxicity of Swine Jejunal Epithelial Cells Induced by Aflatoxin B1 and Zearalenone.

Zearalenone (ZEA) and aflatoxin B1 (AFB1) are two main kinds of mycotoxins widely existing in grain and animal feed that cause a lot of economic loss and health problems for animals and humans. In order to alleviate the cytotoxic effects of AFB1 and ZEA on swine jejunal epithelial cells (IPEC-J2), the combination of a cell-free supernatant of compound probiotics (CFSCP) with mycotoxin degradation enzymes (MDEs) from Aspergillus oryzae was tested. The results demonstrated that coexistence of AFB1 and ZEA had synergetic toxic effects on cell viability. The cell viability was decreased with mycotoxin concentrations increasing, but increased with incubation time extension. The necrotic cell rates were increased when 40 µg/L AFB1 and/or 500 µg/L ZEA were added, but the addition of CFSCP + MDE suppressed the necrotic effects of AFB1 + ZEA. The viable cell rates were decreased when AFB1 and/or ZEA were added: However, the addition of CFSCP + MDE recovered them. The relative mRNA abundances of Bcl-2, occludin, and ZO-1 genes were significantly upregulated, while Bax, caspase-3, GLUT2, ASCT2, PepT1, and IL6 genes were significantly downregulated by CFSCP + MDE addition, compared to the groups containing 40 µg/L AFB1 and 500 µg/L ZEA. This research provided an effective strategy in alleviating mycotoxin cytotoxicity and keeping normal intestinal cell structure and animal health.

Effect of the combined compound probiotics with mycotoxin-degradation enzyme on detoxifying aflatoxin B1 and zearalenone.

Aflatoxin B1 (AFB1) and zearalenone (ZEA) are the secondary toxic metabolites of fungi which contaminate a wide range of food and feedstuffs. Limiting exposure of humans and livestock to them is very essential. Among numerous methods of mycotoxin-degradation, biodegradation by microorganisms and enzymes is an effective and promising approach to eliminate their hazards. The present study aims to optimize the proportion of different species of beneficial microbes by means of response surface methodology (RSM) and its combination with mycotoxin-degradation enzymes. The results indicated that AFB1 and ZEA degradation rates were 38.38% and 42.18% by individual Bacillus subtilis (P < 0.05); however, AFB1 and ZEA degradation rates reached 45.49% and 44.90% (P < 0.05) when three probiotic species such as Bacillus subtilis, Lactobacillus casein and Candida utilis were at a ratio of 1:1:1, corresponding with the predictive value of the RSM model. The further experiment showed that AFB1 and ZEA degradation rates were 63.95% and 73.51% (P < 0.05) when the compound of three probiotic species was combined with mycotoxin-degradation enzymes from Aspergillus oryzae at a ratio of 3:2. This result indicated that the combination of probiotics with mycotoxin-degradation enzymes is a promising new approach for synchronous detoxification of AFB1 and ZEA.

Characterization of the complete mitochondrial genome of band-rumped Storm-petrel, Hydrobates castro.

In this study, the complete mitochondrial genome of band-rumped Storm-petrel, Hydrobates castro, was determined through sequencing of PCR fragments. The complete mitochondrial genome of H. castro was 17,065 bp in length and encoded 13 protein-coding genes, 22 transfer RNA (tRNA) genes, and two ribosomal RNA genes. The overall nucleotide composition is: 30.4% A, 24.5% T, 30.9% C, and 14.2% G, with a total G + C content of 45.1%. By phylogenetic analysis using Bayes method, H. castro showed the closest relationship with the white-faced storm petrel (Pelagodroma marina).

A Maternal Two-meal Feeding Sequence with Varying Crude Protein Affects Milk Lipid Profile in A Sow-Piglet Model.

The effects of a two-meal feeding sequence on production performance and milk lipid profile were investigated. Sixty pregnant sows (d 85 of gestation) were assigned to 3 groups: 2 C group (fed a control crude protein [CP] diet at 0600 and 1500 daily), LH group (fed a low CP diet and a high CP diet at 0600 and 1500), or HL group (fed a high CP diet and a low CP diet at 0600 and 1500). Reproductive performance of sows, and lipid profiles of plasma and milk were measured. Results showed that the HL feeding sequence dramatically increased average piglet weight/litter, average daily gain of piglet/litter, and milk production of sows. LH feeding sequence increased milk fat proportion, and HL feeding sequence significantly increased the proportion of milk MUFA on d 14 and 21 of lactation. Interestingly, the HL feeding sequence also reduced the ratio of C18:1 cis /C18:1 trans in milk, which may account for the greater milk production of sows and growth performance of piglets during lactation. These findings indicated that both the maternal two-meal feeding sequences with varying crude protein improved milk production and milk lipid profiles of sows, which might contribute to improving growth performance of piglets.

Effects of a two-meal daily feeding pattern with varied crude protein levels on growth performance and antioxidant indexes in pigs.

The present study aimed to evaluate the effects of daily feeding pattern on growth performance, blood biochemistry, and antioxidant indexes in pigs. One hundred and eighty female Duroc × Landrace × Yorkshire (DLY) pigs with similar body weight (11.00 ± 0.12 kg) were randomly assigned to 3 groups: the control group (fed 17.01% CP diet, twice daily); high-low group (H-L group, fed 18.33% CP diet in the morning, followed by 15.70% CP diet in the afternoon); and low-high group (L-H group, fed 15.70% CP diet in the morning, followed by 18.33% CP diet in the afternoon) (n = 6). Comparable amounts of their respective diets were given at 05:30 and 15:00 throughout the experimental periods to make all the treatments consumed the same type of food and the same amount of calories on a daily basis. On day 30, one pig was randomly selected per litter for blood samples. Compared with the control group, ADG in the H-L and L-H groups increased by 8.11% and 16.23%, but not significant (P > 0.05); and blood urea nitrogen (BUN) in the H-L and L-H groups decreased by 26.76% and 41.04% (P < 0.05), respectively. The H-L group feeding pattern could significantly improve levels of serum superoxide dismutase (SOD), when compared with the control group. These findings suggest that the two-meal daily feeding pattern with varied levels of CP affects serum levels of BUN and SOD. These changes could effectively silightly improve growth performance and antioxidant capacity in pigs without incurring increased feeding costs.