Small Intestinal Contents Research Articles

Identification of a natural recombinant transmissible gastroenteritis virus between Purdue and Miller clusters in China

Transmissible gastroenteritis virus (TGEV) is an infective coronavirus (CoV) that causes diarrhea-related morbidity and mortality in piglets. For the first time, a natural recombination strain of a TGEV Anhui Hefei (AHHF) virus between the Purdue and the Miller clusters was isolated from the small intestine content of piglets in China. A phylogenetic tree based on a complete genome sequence placed the TGEV AHHF strain between the Purdue and the Miller clusters. The results of a computational analysis of recombination showed that the TGEV AHHF strain is a natural recombinant strain between these clusters. Two breakpoints located in the open reading frame 1a (ORF1a) and spike (S) genes were identified. The pathogenicity of the TGEV AHHF strain was evaluated in piglets, and the results show that TGEV AHHF is an enteric pathogenic strain. These results provide valuable information about the recombination and evolution of CoVs and will facilitate future investigations of the molecular pathogenesis of TGEV.Emerging Microbes & Infections (2017) 6, e74; doi:10.1038/emi.2017.62; published online 23 August 2017

Synthesis and bio-evaluation of xylan-5-fluorouracil-1-acetic acid conjugates as prodrugs for colon cancer treatment

In the present study, xylan-5-fluorouracil-1-acetic acid (Xyl-5-FUAC) conjugates as colon specific prodrugs were synthesized and evaluated by in-vitro release study. The chemical stability of the conjugates was performed in acidic (pH 1.2) and basic buffers (pH 7.4), which showed their stability in upper gastrointestinal tract. The in-vitro drug release profiles of the conjugates were studied in the presence of rat’s gastrointestinal contents. The results showed that the low amounts of drug 3–4% and 5–7% were released in gastric and small intestine contents respectively, while 53–61% of the drug was released in cecum and colonic contents. The cytotoxicity studies of the conjugates were also evaluated on human colorectal cancer cell line (HTC-15 and HT-29), which showed that the conjugates are more cytotoxic than the free drug. Therefore the results reveal that Xyl-5-FUAC conjugates are potential candidates for colon specific drug delivery in the treatment of colonic cancer with minimal undesirable side effects.

Freeze dried extracts of Bidens biternata (Lour.) Merr. and Sheriff. show significant antidiarrheal activity in in-vivo models of diarrhea

Ethnopharmacological relevance of the studyDiarrhea remains one of the main killers of children aged below five years. Traditional antidiarrheal remedies form a potentially viable source of novel low cost efficacious treatments in low resource settings. There is therefore a pressing need to scientifically evaluate these remedies. Aim of the studyThis study aimed to investigate the in vivo and in vitro antidiarrheal activity of freeze dried Bidens biternata, a herb used in traditional Ayurvedic medicine in the management of diarrhea. Materials and methodsIn the castor oil test, twenty (20) adult Sprague-Dawley rats were randomized to a negative control (normal saline, n=5), a positive control (5mg/kg loperamide, n=5), and two test groups. The low dose test group received 200mg/kg Bidens biternata extract (n=5) while the high dose test group received 400mg/kg B. biternata extract (n=5). Castor oil (4ml/kg) was then administered to the animals one hour after administration of the respective treatments after which the total mass of fecal output excreted after four (4) hours was determined.In the charcoal meal test fifteen (15) Sprague Dawley rats were randomized to a control group (normal saline 5ml/kg orally, n=5), a positive control group (atropine sulfate 0.1mg/kg i.p., n=5) and a test group (400mg/kg B. biternata extract, n=5). Charcoal meal was then administered via oral gavage to each rat thirty (30) minutes after the administration of the various treatments. The distance covered by the charcoal meal from the pylorus was then determined after sacrifice of the animals thirty minutes after the meal.In the enteropooling test twenty (20) Sprague-Dawley rats were randomized to a control group (5% v/v ethanol in normal saline, n=5), a positive control group (5mg/kg loperamide, n=5) and a test group (400mg/kg B. biternata extract, n=5). For each group prostaglandin E2 (PGE2) (100μg/kg) was administered immediately after the treatments. The animals were then sacrificed half an hour later and the volume of the small intestine contents determined.The effects of different concentrations of B. biternata extract (0.5. 1.0, 2.0, 3.0 and 5.0mg/ml) on jejunal contraction were investigated and a dose-response curve constructed using the experimental data after which The ED50 dose was determined. The effect of tamsulosin (α1 adrenergic blocker), yohimbine (α2 adrenergic blocker), propranolol (β adrenergic blocker) and naloxone (μ opioid blocker) on the contractile activity of the extract were also investigated.The experimental data were expressed as mean±standard error of mean (SEM) and then analyzed using one-way ANOVA followed by Tukey's post hoc test in cases of significance (set at p<0.05). ResultsThe freeze dried extracts of B. biternata had significant antidiarrheal effects in the castor oil induced diarrhea model (p<0.01) with the highest activity being observed at the 400mg/kg dosage level (1.66±0.81g vs. 4.54±0.51g control, p=0.01). B. biternata extract had significant effects on intestinal motility in the charcoal meal test compared to the control group (43.61±4.42% vs. 60.54±3.33%: p<0.05). B. biternata extract had a significant effect on PGE2 induced enteropooling (3.06±0.07ml vs. 4.74±0.10ml; p<0.001).The freeze dried extracts of B. biternata had a significant negative effect on the contractility of the isolated rabbit jejunum (p<0.001). The effects of the extract were significantly attenuated by tamsulosin (53.94±4.20% vs. 80.57±4.09%; p<0.01) and naloxone (53.94±4.20% vs. 73.89±7.26%; p<0.05). Yohimbine (p>0.05) and propranolol (p>0.05) however did not have any significant effect on the contractile activity of the extract. ConclusionsThe freeze dried extract of B. biternata possess significant antidiarrheal activity in both in vitro and in vivo models which appears to be mediated by modulating both the intestinal motility as well as the secretory activity. The results of this study also validate its traditional use as an antidiarrheal remedy.

Characterisation of multidrug-resistant Shiga toxin-producing Escherichia coli cultured from pigs in China: co-occurrence of extended-spectrum β-lactamase- and mcr-1-encoding genes on plasmids

Identification of Enterobacteriaceae harbouring the plasmid-mediated transferable colistin resistance gene mcr-1 presents a new challenge to public health. The aim of this study was to characterise multidrug-resistant Shiga toxin-producing Escherichia coli (STEC) harbouring the mcr-1 gene on plasmids cultured from pigs in China. Using CHROMagar™ ECC plates combined with stx gene detection by PCR, 93 STEC were recovered from 326 faecal, 351 small intestine content and 326 colon content samples taken from healthy pigs in 2011 and 2012 in China. This study, in which ten colistin-resistant isolates with minimum inhibitory concentrations (MICs) of 8–12 mg/L were identified and found to be positive by PCR for the mcr-1 gene, is a follow-up to an earlier investigation. Plasmid profiling by S1-nuclease digestion followed by pulsed-field gel electrophoresis (PFGE) identified several high-molecular-weight plasmids and these were typed by PCR-based replicon typing (PBRT). Two of the ten isolates, namely STEC-CQ09 (O116:H11/CC23/ST88) and CQ10 (O2:H32/ST3628), were selected for further study as described in this report.

Sulfate-reducing bacteria slow intestinal transit in a bismuth-reversible fashion in mice.

Hydrogen sulfide (H2 S) serves as a mammalian cell-derived gaseous neurotransmitter. The intestines are exposed to a second source of this gas by sulfate-reducing bacteria (SRB). Bismuth subsalicylate binds H2 S rendering it insoluble. The aim of this study was to test the hypothesis that SRB may slow intestinal transit in a bismuth-reversible fashion. Eighty mice were randomized to five groups consisting of Live SRB, Killed SRB, SRB+Bismuth, Bismuth, and Saline. Desulfovibrio vulgaris, a common strain of SRB, was administered by gavage at the dose of 1.0×109 cells along with rhodamine, a fluorescent dye. Intestinal transit was measured 50minutes after gavage by euthanizing the animals, removing the small intestine between the pyloric sphincter and the ileocecal valve and visualizing the distribution of rhodamine across the intestine using an imaging system (IVIS, Perkin-Elmer). Intestinal transit (n=50) was compared using geometric center (1=minimal movement, 100=maximal movement). H2 S concentration (n=30) was also measured when small intestinal luminal content was allowed to generate this gas. The Live SRB group had slower intestinal transit as represented by a geometric center score of 40.2±5.7 when compared to Saline: 73.6±5.7, Killed SRB: 77.9±6.9, SRB+Bismuth: 81.0±2.0, and Bismuth: 73.3±4.2 (P<.0001). Correspondingly, the Live SRB group had the highest luminal H2 S concentration of 4181.0±968.0ppb compared to 0±0ppb for the SRB+Bismuth group (P<.0001). Live SRB slow intestinal transit in a bismuth-reversible fashion in mice. Our results demonstrate that intestinal transit is slowed by SRB and this effect could be abolished by H2 S-binding bismuth.

Ciprofloxacin blocked enterohepatic circulation of diclofenac and alleviated NSAID-induced enteropathy in rats partly by inhibiting intestinal β-glucuronidase activity.

Diclofenac is a non-steroidal anti-inflammatory drug (NSAID), which may cause serious intestinal adverse reactions (enteropathy). In this study we investigated whether co-administration of ciprofloxacin affected the pharmacokinetics of diclofenac and diclofenac-induced enteropathy in rats. The pharmacokinetics of diclofenac was assessed in rats after receiving diclofenac (10 mg/kg, ig, or 5 mg/kg, iv), with or without ciprofloxacin (20 mg/kg, ig) co-administered. After receiving 6 oral doses or 15 intravenous doses of diclofenac, the rats were sacrificed, and small intestine was removed to examine diclofenac-induced enteropathy. β-Glucuronidase activity in intestinal content, bovine liver and E coli was evaluated. Following oral or intravenous administration, the pharmacokinetic profile of diclofenac displayed typical enterohepatic circulation, and co-administration of ciprofloxacin abolished the enterohepatic circulation, resulted in significant reduction in the plasma content of diclofenac. In control rats, β-glucuronidase activity in small intestinal content was region-dependent: proximal intestine<distal intestine<ileal valve. Administration of ciprofloxac caused significant reduction of β-glucuronidase activity in distal small intestine, and particularly in ileal valve. Furthermore, ciprofloxacin (10-2000 μmol/L) dose-dependently inhibited β-glucuronidase activity in distal small intestine content or E coli incubated in vitro, but did not affect that in proximal small intestine content or bovine liver incubated in vitro. After receiving 6 oral doses or 15 intravenous doses of diclofenac, typical enteropathy was developed with severe enteropathy occurred in distal small intestine. Co-administration of ciprofloxacin significantly alleviated diclofenac-induced enteropathy. Co-administration of ciprofloxacin attenuated enterohepatic circulation of diclofenac and alleviated diclofenac-induced enteropathy in rats, partly via the inhibition of intestinal β-glucuronidase activity.

5-Aminosalicylic Acid Azo-Linked to Procainamide Acts as an Anticolitic Mutual Prodrug via Additive Inhibition of Nuclear Factor kappaB.

To improve the anticolitic efficacy of 5-aminosalicylic acid (5-ASA), a colon-specific mutual prodrug of 5-ASA was designed. 5-ASA was coupled to procainamide (PA), a local anesthetic, via an azo bond to prepare 5-(4-{[2-(diethylamino)ethyl]carbamoyl}phenylazo)salicylic acid (5-ASA-azo-PA). 5-ASA-azo-PA was cleaved to 5-ASA and PA up to about 76% at 10 h in the cecal contents while remaining stable in the small intestinal contents. Oral gavage of 5-ASA-azo-PA and sulfasalazine, a colon-specific prodrug currently used in clinic, to rats showed similar efficiency in delivery of 5-ASA to the large intestine, and PA was not detectable in the blood after 5-ASA-azo-PA administration. Oral gavage of 5-ASA-azo-PA alleviated 2,4,6-trinitrobenzenesulfonic acid-induced rat colitis. Moreover, combined intracolonic treatment with 5-ASA and PA elicited an additive ameliorative effect. Furthermore, combined treatment with 5-ASA and PA additively inhibited nuclear factor-kappaB (NFκB) activity in human colon carcinoma cells and inflamed colonic tissues. Finally, 5-ASA-azo-PA administered orally was able to reduce inflammatory mediators, NFκB target gene products, in the inflamed colon. 5-ASA-azo-PA may be a colon-specific mutual prodrug acting against colitis, and the mutual anticolitic effects occurred at least partly through the cooperative inhibition of NFκB activity.

Vomiting as a Symptom and Transmission Risk in Norovirus Illness: Evidence from Human Challenge Studies.

BackgroundIn the US, noroviruses are estimated to cause 21 million cases annually with economic losses reaching $2 billion. Outbreak investigations frequently implicate vomiting as a major transmission risk. However, little is known about the characteristics of vomiting as a symptom or the amount of virus present in emesis.Methodology and Principal FindingsEmesis samples and symptomology data were obtained from previous norovirus human challenge studies with GI.1 Norwalk virus, GII.2 Snow Mountain virus, and a pilot study with GII.1 Hawaii virus. Viral titers in emesis were determined using strain-specific quantitative RT-PCR. In all four studies, vomiting was common with 40–100% of infected subjects vomiting at least once. However, only 45% of subjects with vomiting also had diarrhea. Most of the emesis samples had detectable virus and the mean viral titers were 8.0 x 105 and 3.9 x 104 genomic equivalent copies (GEC)/ml for GI and GII viruses, respectively (p = 0.02). Sample pH was correlated with GII.2 Snow Mountain virus detection.Conclusions and SignificanceHalf of all subjects with symptomatic infection experienced vomiting and the average subject shed 1.7 x 108 GEC in emesis. Unlike shedding through stool, vomiting is more likely to result in significant environmental contamination, leading to transmission through fomites and airborne droplets. This quantitative data will be critical for risk assessment studies to further understand norovirus transmission and develop effective control measures. The correlation between sample pH and virus detection is consistent with a single site of virus replication in the small intestine and stomach contents becoming contaminated by intestinal reflux. Additionally, the frequency of vomiting without concurrent diarrhea suggests that epidemiology studies that enroll subjects based on the presence of diarrhea may be significantly underestimating the true burden of norovirus disease.

Ex vivo pharmacokinetic/pharmacodynamic relationship of valnemulin against Clostridium perfringens in plasma, the small intestinal and caecal contents of rabbits

The pharmacokinetic (PK) and ex vivo pharmacodynamic (PD) of valnemulin against Clostridium perfringens were investigated in plasma, the small intestinal and caecal contents of rabbits following intravenous (IV) or oral administration at 3 mg/kg bodyweight (BW). The postantibiotic effect (PAE) and postantibiotic sub-MIC effect (PA-SME) of valnemulin against C. perfringens ATCC13124 were also determined. The time-kill curves were established in vitro and ex vivo to evaluate the antibacterial activity of valnemulin against C. perfringens.The elimination half-lives (T1/2λz) of valnemulin in the jejunal fluids (7.82 h) or caecal contents (14.8 h) of rabbits was significantly longer than that in plasma (2.94 h). The MIC values of valnemulin against C. perfringens ATCC13124 were both 0.063 μg/mL in the artificial medium and jejunal fluids. The PAEs of valnemulin against C. perfringens were 2.9 h (1 × MIC) and 5.03 h (4 × MIC), and the PA-SMEs ranged from 7.9 h to 11.1 h. Valnemulin exhibited rapid, time-dependent killing feature, and the ex vivo dose-response profile was closely fitted to sigmoid Emax model (r2 = 0.9985). The surrogate index of AUC24 h/MIC ratios required to achieve the bactericidal and virtual bacterial elimination effects were 57.5 and 90.1 h, respectively. Accordingly, the calculated daily dosage regimens of valnemulin for the bactericidal activity (1.96 mg/kg) and bacterial elimination (3.08 mg/kg) would be therapeutically effective in rabbits against C. perfringens with MIC ≤0.5 μg/mL.

Formula diet driven microbial diversity and immunological consequence in small intestine of neonatal porcine model

Several studies have shown that the development and function of the neonatal immune system is enhanced by breast feeding compared to formula feeding. It is important to understand the interaction between infant feeding regimens and the developing GI tract, to identify the protective components of breast milk and mechanisms attributed in immune system development. We used the neonatal piglet model in which piglets were exclusively fed cow's milk formula (n=12) or sow fed (n = 12) from postnatal day (PND) 2 to PND 21 to study the microbiome and GI immune system development. Microbial richness and diversity were assessed from small intestinal contents (duodenum, jejunum and ileum) via 16sRNA amplicon sequencing using V4 variable primers. Data analyses was carried out by using QIIME 1.9.1. Principal component analyses (PCoA) indicated clear separation of the diet groups in all three regions of small intestine. The duodenum showed a significantly more diverse microbial population than jejunum and ileum (FDR corrected, P&lt;0.001) in both sow‐ and formula‐fed groups. However, formula feeding was associated with more microbial diversity in comparison to sow‐diet (FDR corrected, P&lt;0.01). In sow‐fed piglets, Lactobacillus spp was the most abundant and Clostridia spp was the next abundant (FDR corrected, P&lt;0.01) with 3 to 5‐fold higher amounts observed in comparison to formula fed piglets; whereas, Gamma proteobacteria was observed to be 3 to 5‐fold greater in formula fed piglets (FDR corrected, P&lt;0.01) in comparison to sow‐diet. We also assessed histomorphometric changes of Peyer's patches, surface area and inflammatory responses of ileum. In formula‐fed piglets, the lymphoid follicle size (P&lt;0.01) and germinal centers (P &lt;0.01) within the Peyer's patch were significantly decreased compared to to sow‐fed, suggesting less‐developed immune system. The surface area of the ileum mucosa significantly increased in formula group compared to sow‐fed pigs (P &lt;0.01) with no significant differences in growth rate or body weights between the diet groups. However, sow‐fed piglets had relatively longer small intestine than formula‐fed piglets (P&lt;0.05). The decrease in small intestine length in the formula group corresponds to 14%, suggesting compensation for surface area and possibly for nutrient absorption. Furthermore, in ileum, formula diet induced significant down‐regulation of anti‐inflammatory IL‐10 (P &lt;0.01) at protein level, indicating that formula diet impacts the immune response. We conclude that feeding a milk formula diet alters the microbiome, small intestine morphology and gut‐associated lymphoid tissue development and immune response.Funding: USDA‐ARS Project 6026‐51000‐010‐05S.Support or Funding InformationUSDA‐ARS Project 6026‐51000‐010‐05S.

Antibiotic-Induced Alterations of the Gut Microbiota Alter Secondary Bile Acid Production and Allow for Clostridium difficile Spore Germination and Outgrowth in the Large Intestine.

It is hypothesized that the depletion of microbial members responsible for converting primary bile acids into secondary bile acids reduces resistance to Clostridium difficile colonization. To date, inhibition of C.difficile growth by secondary bile acids has only been shown in vitro. Using targeted bile acid metabolomics, we sought to define the physiologically relevant concentrations of primary and secondary bile acids present in the murine small and large intestinal tracts and how these impact C.difficile dynamics. We treated mice with a variety of antibiotics to create distinct microbial and metabolic (bile acid) environments and directly tested their ability to support or inhibit C.difficile spore germination and outgrowth ex vivo. Susceptibility to C.difficile in the large intestine was observed only after specific broad-spectrum antibiotic treatment (cefoperazone, clindamycin, and vancomycin) and was accompanied by a significant loss of secondary bile acids (deoxycholate, lithocholate, ursodeoxycholate, hyodeoxycholate, and ω-muricholate). These changes were correlated to the loss of specific microbiota community members, the Lachnospiraceae and Ruminococcaceae families. Additionally, physiological concentrations of secondary bile acids present during C.difficile resistance were able to inhibit spore germination and outgrowth in vitro. Interestingly, we observed that C.difficile spore germination and outgrowth were supported constantly in murine small intestinal content regardless of antibiotic perturbation, suggesting that targeting growth of C.difficile will prove most important for future therapeutics and that antibiotic-related changes are organ specific. Understanding how the gut microbiota regulates bile acids throughout the intestine will aid the development of future therapies for C.difficile infection and other metabolically relevant disorders such as obesity and diabetes. IMPORTANCE Antibiotics alter the gastrointestinal microbiota, allowing for Clostridium difficile infection, which is a significant public health problem. Changes in the structure of the gut microbiota alter the metabolome, specifically the production of secondary bile acids. Specific bile acids are able to initiate C.difficile spore germination and also inhibit C.difficile growth in vitro, although no study to date has defined physiologically relevant bile acids in the gastrointestinal tract. In this study, we define the bile acids C.difficile spores encounter in the small and large intestines before and after various antibiotic treatments. Antibiotics that alter the gut microbiota and deplete secondary bile acid production allow C.difficile colonization, representing a mechanism of colonization resistance. Multiple secondary bile acids in the large intestine were able to inhibit C.difficile spore germination and growth at physiological concentrations and represent new targets to combat C.difficile in the large intestine.

The specific features of the distribution of 4-metoxyhydroxybenzene in the organism of the warm-blooded animals suffering lethal intoxication

This work was designed to study the distribution of 4-metoxyhydroxybenzene in the organism of the omnivorous warm-blooded animals (rats) after the intragastric administration of this poisonous compound at a dose three-fold greater than the LD50 value. The administered 4-metoxyhydroxybenzene was isolated from the organs and blood of the experimental animals by exposing the biological tissues to acetone with subsequent purification on a silica gel L 40/100 mcm using a hexane:dioxane:propanol-2 (20:5:1) as the mobile phase. The identification and quantitation of 4-metoxyhydroxybenzene were carried out with the use of TLC, GC-MS, and UF-spectrophotometry. It was shown that the administered 4-metoxyhydroxybenzene remained unmetabolized in the internal organs and blood of the poisoned experimental animals. The largest amounts of 4-metoxyhydroxybenzene were found in the stomach contents (2584,92±117,47), brain (59.49±6.05), contents of small intestines (28.21±3.77), and kidneys (26.13±1.64).

The specific features of the distribution of 2,6-di-tret-buthyl-4-methlhydroxybenzole in the organism of the warm-blooded animals

We have studied the specific features of the distribution of 2,6-di-tret-buthyl-4-methlhydroxybenzole in the organism of the omnivorous warm-blooded animals (rats) following the intragastric administration of the three-fold lethal dose of the poisonous substance. 2,6-di-tret-buthyl-4-methlhydroxybenzole was isolated from the blood and various organs of the animals by means of acetone extraction. It was further purified on the 40/100 mcm L silicagel column with the use of hexane/acetone for elution. TLC, GC-MS, and UV-spectrophotometry were employed to identify and quantify the material of interest. It was shown that 2,6-di-tret-buthyl-4-methlhydroxybenzole undergoes modification in the internal organs and blood of the poisoned animals. In was present in the highest amounts (100 mg/g) in the stomach and small intestine contents (787.78±52.31 and 18.31±3.47 respectively), spleen (12.46±1.02), and kidneys (8.48±0.61).

Nutrient restriction and realimentation in beef cows during early and mid-gestation and maternal and fetal hepatic and small intestinal in vitro oxygen consumption

Objectives were to determine the effects of advancing gestation, maternal nutrient restriction during early and mid-gestation, and realimentation on fetal liver and jejunal mass and energy use in both dams and fetuses. On day 30 of pregnancy, multiparous, non-lactating beef cows (initial BW=621±11.3 kg and body condition score=5.1±0.1) were assigned to one of the two dietary treatments: control (CON; 100% requirements; n=18) and restricted (R; 60% requirements; n=28). On day 85, cows were slaughtered (CON, n=6; R, n=6), and remaining cows continued on control (CC; n=12) and restricted (RR; n=12) diets, or were realimented to the control diet (RC; n=11). On day 140, cows were slaughtered (CC, n=6; RR, n=6; RC, n=5), remaining cows continued on the control diet (CCC, n=6; RCC, n=5), or were realimented to the control diet (RRC, n=6). On day 254, all remaining cows were slaughtered. Maternal liver O2 consumption linearly increased (P⩽0.04) and jejunal weight (g/kg) linearly decreased (P=0.04) as gestation advanced in CON groups. Fetal BW, and hepatic and small intestinal absolute mass, protein content and O2 consumption linearly increased (P⩽0.04) as pregnancy advanced in CON groups. However, mass and O2 consumption relative to BW linearly decreased (P⩽0.001) in the fetal liver in CON groups. When analyzing the effects of dietary treatment, at day 85, fetal jejunal O2 consumption (mol/min per kg BW) was lower (P=0.02) in the R group when compared with the CON group. At day 140, maternal hepatic weight (g) was lower (P=0.02) in RC and RR cows when compared with CC, and fetal jejunual O2 consumption (mmol/min per mg tissue and mmol/min per g protein) was greater (P⩽0.02) in RC when compared with RR. At day 254, maternal hepatic O2 consumption (absolute and relative to BW) was lower (P⩽0.04) in the RCC cows when compared with RRC. Fetal hepatic weight was lower (P=0.05) in the CCC group when compared with RCC and RRC. The changes in response to nutrient restriction and realimentation in both the dam and fetus may indicate an adaptation to a lower amount of available nutrients by altering tissue mass and metabolism.

Intestinal removal of free fatty acids from hosts by Lactobacilli for the treatment of obesity

Recent findings on the association of gut microbiota with various diseases, including obesity, prompted us to investigate the possibility of using a certain type of gut bacteria as a safe therapeutic for obesity. Lactobacillus mutants with enhanced capacity in absorption of free fatty acids (FFAs) were isolated to show reduced absorption of FFAs by the administered host, attributing to inhibition of body weight gain and body fat accumulation as well as amelioration of blood profiles. Consequently, high throughput screening of natural FFAs‐absorbing intestinal microbes led to the isolation of Lactobacillus reuteri JBD30 l. The administration of Lactobacillus JBD30l lowered the concentration of FFAs in the gut fluid content of small intestine, thus reducing intestinal absorption of FFAs whereas promoting fecal excretion of FFAs. Animal data also confirmed that the efficacy of Lactobacillus JBD30l on body weight similar to that of orlistat, an FDA‐approved pharmaceutical for long‐term use to treat obesity. In a subsequent random, double‐blind, placebo‐controlled clinical trial (KCT0000452 at Clinical Research Information Service of Korea), there was a statistically significant difference in the percentage change in body weight between the Lactobacillus JBD301 and the placebo group (P = 0.026) as well as in the BMI (P = 0.036) from the 0‐week assessment to the 12‐week assessment. Our results show that FFA‐absorbing Lactobacillus JBD301 effectively reduces dietary fat absorption, providing an ideal treatment for obesity with inherent safety.

Investigation of possible pumpkin seeds and ginger effects on gastrointestinal nematode infection indicators in meat goat kids and lambs

In four experiments, 77 naturally-infected Boer crossbred kids and 28 artificially-inoculated Katahdin lambs were used to evaluate the effect of pumpkin seeds (Exp 1; 21 kids), ginger or pumpkin seed drench (Exp 2; 30 kids) and pumpkin seed oil (Exp 3 and 4: 28 lambs and 26 kids, respectively) on gastrointestinal nematode (GIN) indicators. In all experiments, kids and lambs were placed in individual pens and received pre-weighed rations of a commercially pelleted meat goat or sheep diet daily. In Exp 1, kids were supplemented with ground pumpkin seeds (PUM; n=10) mixed into feed daily at a rate of 5g/kg body weight (BW) or were not supplemented (CON; n=11) for 21 days. In Exp 2, kids were orally drenched with water (CON; n=10), 5g pumpkin seed/kg BW (PUM; n=10) or 3g ginger/kg BW (GIR; n=10) every other day for 42 days. In Exp 3, lambs were orally drenched with 2ml/kg water (CON; n=7), 2.0ml/kg BW pumpkin seed oil once every 7 days (PUM1; n=10), or 2.0ml/kg BW pumpkin seed oil daily for 3 out of every 7 days (PUM2; n=11) for 28 days. In Exp 4, kids were orally drenched with 2ml/kg water (CON; n=13), or 2.0ml/kg BW pumpkin seed oil (PUM; n=13) every other day for 35 days. In all experiments, BW, daily feed intake and blood and fecal samples were collected every 7 days. All animals in Exp 2 were harvested at a USDA-inspected abattoir and abomasal and small intestinal contents were collected for total worm counts. The FEC were similar for treatments in all experiments. Treatment influenced PCV (P<0.05) only in Exp 1 and 4. In Exp 2, at harvest, there was a tendency (P=0.08) for CON animals to have a higher number of total GIN than GIR-treated animals, but PUM-treated animals were intermediate. BW were similar for treatments in Exp 1, 2 and 3 while CON animals in Exp 4 had a greater BW than PUM-treated animals on day 7 only and were similar thereafter (treatment by day interaction, P<0.05). In these studies, pumpkin and ginger treatments administered were not effective in reducing FEC in meat goat kids or lambs.

Regioselective synthesis, stability and release behaviors of the secondary 5-fluorouracil acetic acid/β-cyclodextrin conjugate for colon delivery

5-Fluorouracil acetic acid (5-FUAC) was regioselectively conjugated to the secondary C-2 hydroxyl group of β-cyclodextrin, and the chemical stability and in vitro drug release behaviors of the conjugate were investigated at 37 °C within 24 h. The conjugate in aqueous solutions was more stable at pH 1.2 and 4.4, and gradually hydrolyzed as a function of time at pH 6.8 and 7.4. According to the rate-pH profile, the half-lives of the conjugate at pH 1.2, 4.4, 6.8 and 7.4 were about 636, 576, 50 and 27 h respectively. In the contents of stomach, the conjugate did hardly release 5-FUAC, released 5-FUAC 19.3 % in the contents of small intestine, and released 5-FUAC significantly up to 62.7 % in the contents of colon. These results suggest that the secondary 5-FUAC/β-cyclodextrin conjugate may be of value as a colon-targeted and sustained-release prodrug.

Dextran-5-(4-ethoxycarbonylphenylazo)salicylic acid ester, a polymeric colon-specific prodrug releasing 5-aminosalicylic acid and benzocaine, ameliorates TNBS-induced rat colitis

Local anesthetics have beneficial effects on colitis. Dextran-5-(4-ethoxycarbonylphenylazo)salicylic acid ester (Dex-5-ESA), designed as a polymeric colon-specific prodrug liberating 5-ASA and benzocaine in the large intestine, was prepared and its therapeutic activity against colitis was evaluated using a TNBS-induced rat colitis model. Dex-5-ESA liberated 5-ASA and benzocaine in the cecal contents while (bio)chemically stable in the small intestinal contents and mucosa. Oral administration of Dex-5-ESA (equivalent to 10 mg 5-ASA/kg, twice a day) alleviated colonic injury and reduced MPO activity in the inflamed colon. In parallel, pro-inflammatory mediators, COX-2, iNOS and CINC-3, elevated by TNBS-induced colitis, were substantially diminished in the inflamed colon. Dex-5-ESA was much more effective for the treatment of colitis than 5-(4-ethoxycarbonylphenylazo)salicylic acid (5-ESA) that may not deliver benzocaine to the large intestine. Our data suggest that Dex-5-ESA is a polymeric colon-specific prodrug, liberating 5-ASA and benzocaine in the target site (large intestine), probably exerting anti-colitic effects by combined action of 5-ASA and benzocaine.

Study on the effects of microencapsulated Lactobacillus delbrueckii on the mouse intestinal flora

:Objective: To evaluate the protective effects of microencapsulation on Lactobacillus delbrueckii by random, parallel experimental design. Materials and methods: Lincomycin hydrochloride-induced intestinal malfunction mouse model was successfully established; then the L. delbrueckii microcapsule was given to the mouse. The clinical behaviour, number of intestinal flora, mucous IgA content in small intestine, IgG and IL-2 level in peripheral blood were monitored. The histological sections were also prepared. Results: The L. delbrueckii microcapsule could have more probiotic effects as indicated by higher bifidobacterium number in cecal contents. The sIgA content in microcapsule treated group was significantly higher than that in non-encapsulated L. delbrueckii treated group (p < 0.05). Intestine pathological damage of the L. delbrueckii microcapsule-treated group showed obvious restoration. Conclusion: The L. delbrueckii microcapsules could relieve the intestinal tissue pathological damage and play an important role in curing antibiotic-induced intestinal flora dysfunction.

Heat-treated colostrum feeding promotes beneficial bacteria colonization in the small intestine of neonatal calves

The present study investigated the effect of heat-treated colostrum feeding on the bacterial colonization in calf small intestine of neonatal calves within the first 12h of life. Newborn Holstein bull calves (n=32) were assigned to 3 treatment groups and fed with either fresh colostrum (FC, n=12) or heat-treated (60°C, 60min) colostrum (HC, n=12) soon after birth, whereas the control (NC, n=8) group did not receive colostrum or water. Small intestinal tissues and contents were collected from proximal jejunum, distal jejunum, and ileum at 6 and 12h after birth, following euthanasia. Quantitative real time-PCR was used to explore the colonization of total bacteria, Lactobacillus, Bifidobacterium, and Escherichia coli. The feeding of colostrum soon after birth increased the colonization of total bacteria in calf gut within the first 12h compared with NC. In contrast, the prevalence of Lactobacillus was lower in HC and FC compared to NC. Remarkable changes in the prevalence of small intestinal tissue-attached Bifidobacterium were observed with the feeding of HC, but not that in small intestinal contents. The prevalence of Bifidobacterium was 3.2 and 5.2 fold higher in HC than FC and NC, respectively, at 6h. Although the feeding of FC did not enhance the prevalence of tissue-attached Bifidobacterium at 6h compared with NC, it displayed a gradual increase over the time that was higher than NC, but similar to that of HC at 12h. Moreover, the colonization of E. coli was drastically reduced in HC calves compared with FC and NC. Thus, the present study suggests that the feeding of HC enhances the colonization of Bifidobacterium but lessens E. coli in the calf small intestine immediately postpartum compared with that of FC and NC. The increased colonization of beneficial bacteria along with the decreased colonization of potential pathogens in calf gut may also diminish the neonatal calf diarrhea when calves are fed heat-treated colostrum soon after birth.